WO2025252897A1 - Plant defense inducer - Google Patents

Plant defense inducer

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Publication number
WO2025252897A1
WO2025252897A1 PCT/EP2025/065691 EP2025065691W WO2025252897A1 WO 2025252897 A1 WO2025252897 A1 WO 2025252897A1 EP 2025065691 W EP2025065691 W EP 2025065691W WO 2025252897 A1 WO2025252897 A1 WO 2025252897A1
Authority
WO
WIPO (PCT)
Prior art keywords
aromatic
linkage
oligomer
aromatic monomer
plant
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/EP2025/065691
Other languages
French (fr)
Inventor
Bert Sels
Bruno Cammue
Karin Thevissen
Thomas NICOLAI
Bruno PANDALONE
Bert LAGRAIN
Bart Lievens
Joëlle MÜHLEMANN
Ludo CIALDELLA
Korneel VAN AELST
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Katholieke Universiteit Leuven
Original Assignee
Katholieke Universiteit Leuven
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US19/096,361 external-priority patent/US20250374921A1/en
Application filed by Katholieke Universiteit Leuven filed Critical Katholieke Universiteit Leuven
Publication of WO2025252897A1 publication Critical patent/WO2025252897A1/en
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P21/00—Plant growth regulators
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N31/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic oxygen or sulfur compounds
    • A01N31/08—Oxygen or sulfur directly attached to an aromatic ring system
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N31/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic oxygen or sulfur compounds
    • A01N31/08—Oxygen or sulfur directly attached to an aromatic ring system
    • A01N31/14—Ethers
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N31/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic oxygen or sulfur compounds
    • A01N31/08—Oxygen or sulfur directly attached to an aromatic ring system
    • A01N31/16—Oxygen or sulfur directly attached to an aromatic ring system with two or more oxygen or sulfur atoms directly attached to the same aromatic ring system
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
    • A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
    • A01N43/06—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom five-membered rings
    • A01N43/12—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom five-membered rings condensed with a carbocyclic ring

Definitions

  • the invention is broadly in the field of agrochemical or phytopharmaceutical c ompounds and compositions for plant treatment, and in particular relates to these 10 compounds or compositions as plant defense elicitors.
  • These are also commonly k nown as plant immune- system activator molecules, or ‘bio-pesticides’ in layman’s terms, a class of environmental- friendly molecules that induce or boost plant’s resistance against abiotic stressor or biotic stressors such as pests and for instance r etard the infection and propagation of microbial and viral pathogens on plants and 15 uses thereof.
  • Insects may be a vector of such pathogens.
  • B. Description of the Related Art There is a need for technology to efficiently produce high-quality agricultural products 20 in a limited amount of cultivated land. Therefore, it is of high interest in agriculture to control diseases caused by pests for instance parasites or pathogens such as fungi, oomycetes, bacteria, viruses, nematodes and insects.
  • Fungicides for instance, are such chemical compounds or biologic substances used to kill or inhibit fungi or oomycetes or their spores. Fungicides sometimes also have an effect on other plant 25 pathogens such as bacteria, viruses, nematodes or insects.
  • a drawback of using certain fungicides is that fungicide residues can be found, in the environment and on food for human consumption sometimes posing a danger to biodiversity, human or animal health.
  • traditional pesticides while effective, can harm the e nvironment in several ways. They can pollute soil and water sources, harm beneficial 30 insects and pollinators, and disrupt ecosystems. There is a need in the art to make them unnecessary or to decrease their use.
  • T hus in order to promote sustainable crop production, there is a need to use more safe and natural substances with biological activity that can decrease the amounts of35 pest control chemicals and in particular chemical fungicides needed. 1
  • Lignin is the second largest biopolymer on earth. Lignin, with its polyaromatic network is an aromatic polymer is a major constituent in e.g. wood, being the most abundant carbon source on Earth second only to cellulose. In recent years, with development and commercialization of technologies to extract lignin in a highly 5 purified, solid and particularized form from the pulp-making process, it has attracted significant attention as a possible renewable substitute to primarily aromatic chemical precursors currently sourced from the petrochemical industry. N ative or pristine lignin, a complex polymer consisting of aromatic building blocks, 10 results from a radical polymerization process of three 4-hydroxyphenylpropanoid building blocks or monolignols linking the latter via stable carbon-carbon bonds and more reactive ether bonds.
  • the complex native lignin polymer can be broken down by various catalytic or thermo-solvolytic methods of lignin depolymerization of the lignin source, such as reductive catalytic fractionation (RCF), non-catalytic thermo- 15 solvolytic depolymerization and fractioning or combinations thereof to cleave the i nterunit linkages within the lignin polymer (ether bonds) into a liquid (lignin oil) comprising low molecular weight oligomers (oligophenolics having 3 or more aromatic groups or a degree of polymerization (DP) of 3 and more) , dimers (diphenolics having 2 aromatic groups or a DP of 2) and monomers (monophenols with 1 aromatic 20 group, DP of 1).
  • RCF reductive catalytic fractionation
  • DP degree of polymerization
  • Such lignin oil can be further fractionated, for instance by ultrafiltration and solubility-based methods, into different fractions of aromatic c ompounds with a specific degree of polymerization (DP).
  • Plant cells have evolved a sophisticated immune system comprising two main layers 25 of defense known as pattern-triggered immunity (PTI) and effector-triggered immunity (ETI) (Song et al., New Phytologist (2022) 236:590–607), that constitute the so-called plant immune system.
  • PTI pattern-triggered immunity
  • ETI effector-triggered immunity
  • the plant immune system against insects involves a complex interplay of signaling pathways, defense mechanisms, and elicitors that work together to protect plants from herbivory.
  • ETI confers a narrow strain-specific resistance as it is initiated following the recognition of virulence effectors (A virulence-proteins) by cytoplasmic resistance genes (R-genes). This generally causes a strong site-specific accumulation of reactive o xygen species (ROS) leading to apoptosis.
  • R-genes cytoplasmic resistance genes
  • ROS reactive o xygen species
  • PAMPs pathogen-associated molecular- pattern
  • plant defense elicitor p lant pest defense elicitor(s)
  • plant immune system elicitor p lant pest defense elicitor(s)
  • plant defense elicitor p lant pest defense elicitor(s)
  • plant defense elicitor p lant pest defense elicitor(s)
  • plant immune system elicitor p lant pest defense elicitor(s)
  • plant defense elicitor(s) plant elicitor(s) or simply elicitor(s).
  • plant defense elicitor or plant defense elicitors is most used.
  • Natural compounds that induce or boost plant immunity are crucial for activating plant defense responses, thereby i nhibiting pathogen development and improving plant resilience against biotic and/or abiotic stress.
  • Compounds, which when perceived by a plant give rise to such defense 25 responses are commonly referred to as plant defense elicitor, plant pest defense elicitor(s), plant immune system elicitor(s), plant defense elicitor(s), plant elicitor(s) or simply elicitor(s).
  • plant defense elicitor or plant defense elicitors is most used.
  • Present invention solves this problem by a novel plant defense elicitor and demonstrates how this can be obtainable from a lignin depolymerisation process and fractioning in compositions comprising oligophenolics with a degree of polymerization ( DP) of 2 to 8 and preferable of 2 to 4 from depolymerized lignin or decomposed lignin or from structurally identical oligophenolics. They can cause the increase of 35 the resistance to adverse conditions (biotic or abiotic).
  • Present invention also demonstrates how these can be used in suitable compositions for plant defense. 3
  • the present invention solves the problems of the related art of plant protection against pathogens and stressors by more natural compounds or of natural sources 5 by a plant defense elicitor, that comprises as an active ingredient of the plant defense e licitor depolymerised lignin. Furthermore this depolymerized lignin comprises specific oligophenolics with a DP from 1 to 8.
  • present invention concerns new plant defense elicitors or new plant elicitor compositions, and the use of these in agricultural applications, more particularly to 10 protect plants against pests or pathogens or abiotic stressors.
  • an active ingredient(s) of the plant defense elicitor comprises lignin-derived oligophenoliocs or structural similar compounds with a DP 15 of 2 to 8, and yet more preferably of 2 to 4 (Fig. 2).
  • an active ingredient of the plant defense elicitor comprises depolymerized lignin containing as active ingredient lignin-derived oligophenoliocs w ith a DP of 2 to 8, and yet more preferably of 2 to 4 or selected structural similar 20 compounds (Fig. 2).
  • the invention also relate to uses of, and methods employing, depolymerized lignin o r decomposed lignin oligophenolics (Fig. 1A, 1B) with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4 or structural identical oligophenolics as plant defense elicitor.
  • compositions comprising depolymerized lignin or decomposed lignin oligophenolics w ith a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4 or structural identical oligophenolics, and applications thereof.
  • the compositions may further comprise other plant elicitors or may comprise antifungal, antimicrobials or antiviral compounds.
  • 30 the compositions may be produced by decomposition of lignin by reductive catalytic f ractionation (RCF) (Fig. 1A), non-catalytic thermo-solvolytic de-polymerization (Fig. 1A) and fractioning (Fig. 1C) of the lignin source. 4
  • RCF reductive catalytic f ractionation
  • An aspect of present invention concerns a plant elicitor (elicitor of natural plant defences), for instance plant defense elicitor, characterized in that, an active i ngredient of the plant defense elicitor comprises phenolics that are derived from depolymerized lignin, including lignin-derived dimeric (diphenolic), lignin-derived 5 trimeric compounds (triphenolic) and lignin derived tetrameric (tetraphenolic) compounds and engineered or synthesised structural similar compounds.
  • the present invention concerns a plant defense elicitor according to claim 1, characterized in that, an active ingredient of the plant defense elicitor comprises depolymerized lignin containing as active ingredient lignin-derived diphenolics. 10
  • an active ingredient of the plant defense elicitor comprises depolymerized lignin containing as active ingredient lignin-derived diphenolics. 10
  • the invention is broadly drawn to a method for controlling a plant disease c omprising treating a plant with a plant defense elicitor or elicitor containing phenolics that is derived from depolymerized lignin of the reductive catalytic fractioning (RCF) 15 or the non-catalytic thermos-solvolytic depolymerization process.
  • RCF reductive catalytic fractioning
  • This plant defense e licitor can be a lignin oil and fractions thereof (with an approach exemplified in figure 1C) applied, for instance at a concentration of 0.05 to 20 mg/ml , preferably 0.2 to 10 mg/ml, preferably 0.5 to 5 mg/ml, more preferably 0.8 to 1.2 mg/ml and most preferably 1 mg/mL and for instance comprising molecular mass (weight average 20 mass or Mw) of 180 g/mol to 1800 g/mol, preferably between 230 g/mol to 1000 g/mol, and yet more preferably between 230 g/mol to 650 g/mol of active ingredient compounds (Fig.
  • each R1, R3, and R4 is independently chosen from –H, -OH, O-CH3, a 5 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, 10 - wherein R2 is –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R5 is selected of –H, a ⁇ -O-4 link
  • each R12, R13, R15 and R16 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic 10 monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R11 and R14 is independently chosen from –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an ⁇ -
  • the invention is drawn to an engineered composition comprising aromatic compounds, whereby the molecular mass (weight average or Mw) of the aromatic c ompounds can be 180 g/mol between 230 g/mol to 1000 g/mol, and yet more 9
  • aromatic compounds comprise, consists essentially of or consists of lignin derived phenolics with a degree of polymerization (DP- of 2 to 8, preferably 2 to 4 or synthesized structurally similar compounds.
  • DP- degree of polymerization
  • Fig. 1A the reductive catalytic fractioning 5
  • Fig. 1B the non-catalytic thermos-solvolytic depolymerization
  • these defined phenolic structures are from lignin depolymerisation.
  • This term broadly encompasses any compounds or compositions that are 20 capable of eliciting natural plant defenses, of activating plant defense and resistance reactions against plant pathogen, of stimulating the production of plant defense molecules against plant pathogen and/or of preventing, controlling or treating a plant a gainst infection by a plant pathogen and/or against abiotic stressors, when administered to a plant, plant seed or an organ of a plant.
  • an elicitor when perceived by 25 a plant, can evoke molecular, biochemical and/or physiological defensive plant cell reactions, such as the synthesis, or increase of the synthesis, of plant d efense molecule(s), for example ethylene and/or salicylic and jasmonic acid, the production of reactive oxygen species (ROS), and/or expression of specific defense- related genes and proteins, for example polygalacturonase inhibitor proteins (PGIP).
  • ROS reactive oxygen species
  • PGIP polygalacturonase inhibitor proteins
  • biopesticides which mainly include bioinsecticides, biofimgicides, bionematicides, and others.
  • biopesticides colloquially known as biopesticides, which mainly include bioinsecticides, biofimgicides, bionematicides, and others.
  • 5 a plant infection by a plant pathogen may be prevented, controlled or treated.
  • preventing may in particular mean avoiding occurrence of at least one adverse effect or symptom, preferably all adverse effects or symptoms induced by a plant pathogen infection
  • controlling may in particular mean stopping the progression of a plant pathogen 10 infection, more precisely reducing or abolishing the plant pathogen spread across the healthy parts of a plant or of an organ of a plant, or from an infected plant to another p lant, typically to a neighboring plant
  • treating may in particular mean ameliorating the symptom(s) of an infection, or completely curing an infection, typically by reducing or completely eliminating a phytopathogen (typically a fungus 15 or bacterium), i.e., by eliminating any viable phytopathogen in the plant or in an organ or several or each organ(s) of the plant.
  • phytopathogen typically a fungus 15 or bacterium
  • a plant may be contacted with an effective amount of the present compounds or compositions, such as contacted via an organ of the plant, preferably an organ 20 selected from leaves, roots and/or fruits, or via seeds of the plant.
  • the contacting step may be performed once or several times (for example regularly or periodically, for example on the appropriate season or at the appropriate plant development stage).
  • an effective amount refers to an amount of the (active) compound or compounds as taught herein which induces or elicits plant natural defense, activate 25 plant defense and resistance reaction against abiotic stressors or against plant pathogen, and/or stimulates the production of plant defenses molecules against plant pathogen resulting in obtaining a plant that is resistant to pathogen(s).
  • the effective amount is understood to be variable, as it may be affected by many factors, including but not limited to the type of plant treated, treatment dosages and application rates, 30 method of contacting, weather and seasonal conditions experienced during the plant growing cycle, pathogen susceptibility, etc. Such variables are commonly encountered and understood by the skilled person, who may adjust the prophylactic or treatment regimen, e.g., application rate, application timings and/or frequencies, a nd application way. Particular suitable amounts or concentrations ranges are 35 discussed and exemplified elsewhere in this specification.
  • the terms “organ”, “organ of a plant” or “plant’s organ” interchangeably refer to a part of a plant or to a plant propagation material. Examples of plant’s organs include, but are not limited to, 11
  • the contacting step with the plant or organ can be performed in various ways, for example by spraying, drenching, soaking, dipping, injection, through soil feeding, and any combination thereof.
  • the compounds or compositions can be 5 applied on a plant or organ by supplying a volatile or vapor-based form of the compounds or compositions in the vicinity of the plant tissue and allowing the diffusion to the plant or organ through the atmosphere. The skilled person knows how to adapt the manner of administration to the particular use.
  • the term “plant” typically designates a plant i nfected by or presenting a susceptibility to infection by a plant pathogen or affected by an abiotic stressor.
  • the plant may belong to the clade of Angiosperm.
  • the plant may belong to the clade of dicots.
  • Examples of 15 plants from the dicots clade include, but are not limited to, the Solcmcicecie family, comprising Solcinum lycopersicum (tomato), Solcinum tuberosum (potatoes), Solcinum melongenci (eggplant), Capsicum genus (pepper) and Nicotiana tabacum (tobacco); the Vitaceae family comprising the Vitis genus (grapevines); the Brassicaceae family, comprising Brassica oleracea (cabbage), Brassica rapa (turnip 20 and Chinese cabbage), mustard species and A.
  • the Solcmcicecie family comprising Solcinum lycopersicum (tomato), Solcinum tuberosum (potatoes), Solcinum melongenci (eggplant), Capsicum genus (pepper) and Nicotiana tabacum (tobacco); the Vitaceae family comprising the Vitis genus (grapevines); the
  • the plant may belong to the clade of monocots.
  • An example o f plants from the monocot clade includes, but is not limited to, the Gramineae or Poaceae family, such as maize, rice, barley, or wheat.
  • the plant may be a dicot plant, preferably selected f rom Brassicaceae, Solanaceae or Rosacea families, such as A. thaliana, cabbage, tomato, grapevine, soybean, apple, pear, or strawberries, or wherein the plant is a m onocot plant, preferably selected from Gramineae family, such as maize, rice, barley, or wheat.
  • the plant pathogen may be a fungus or a bacterium.
  • the compositions as taught herein may also be conveniently 12
  • the pathogen may be a necrotrophic fungus or bacterium, 5 a hemibiotrophic fungus or bacterium, or a biotrophic fungus or bacterium.
  • biotrophy in which nutrients are obtained from living host cells
  • necrotrophy in which nutrients are obtained from host cells which have been 10 previously killed by the fungus.
  • a third mode of nutrition is hemibiotrophy, where the pathogen has an initial period of biotrophy followed by a period of necrotrophy.
  • Phytopathogenic pathogens in particular fungi, can thus be distinguished depending on their mode of nutrition: necrotrophic (e.g., Botrytis cinerea), biotrophic (e.g., U stilago maydis) or hemibiotrophic (e.g., Colletotrichum higginsianum).
  • the plant pathogen may be a fungus, typically a phytopathogenic fungus.
  • the expression “phytopathogen fungus” refers to fungi pathogens that infect plant organs.
  • phytopathogenic fungi include, but a re not limited to, fungi belonging to the Ascomycetes and Basidiomycetes classes, such as, for example, fungi of the order of Helotiales, such as, for example, family 20 Sclerotiniaceae, Botrytis/ Botryotinia, such as species Botrytis cinerea,' fungi of the order of Hypocreales, such as, for example, family Nectriaceae, genus Fusarium,' f ungi of the order of Uredinales, such as, for example, family Pucciniaceae, genus Puccinia,' fungi of the order of Ustilaginales, such as, for example, family U stilaginaceae, genus Ustilago),' fungi of the order of Sordariomycetes, such as, for 25 example, family Glomerellaceae, genus Colletotrichum.
  • the plant pathogen may be a bacterium, typically a phytopathogenic bacterium.
  • the expression “phytopathogen bacterium” refers to bacterial pathogens that infect plant organs.
  • phytopathogenic bacteria 30 include, but are not limited to, bacteria of the order of Pseudomonadales, such as, for example, family Pseudomonadaceae, genus Pseudomonas, such as species Pseudomonas syringae,' bacteria of the order of Burkholderiales, such as, for example, family Burkholderiaceae, genus Ralstonia, such as species Ralstonia solanacearum,' bacteria of the order of Enterobacterales, such as, for example, family 35 Erwiniaceae, genus Erwinia, such as species Erwinia amylovora, or family Pectobacteriaceae, genus 13
  • Pectobacterium such as species Pectobacterium carotovorum (formerly Erwinia carotovora, bacteria of the order of Xanthomonadales, such as, for example, family Xanthomonadaceae, genus Xylella, such as species Xylella fastidiosa, or genus Xanthomonas, such as species Xanthomonas campestris.
  • bacteria can also be classified into necrotrophic, biotrophic and hemibiotrophic sub-classes. Particularly preferred may be necrotrophic fungi, preferably as Botrytis cinerea.
  • the plant pathogen is a fungus or a bacterium, such 10 as a necrotrophic fungus or bacterium, such as Botrytis cinerea, a hemibiotrophic fungus or bacterium, or a biotrophic fungus or bacterium, such as Pseudomonas syringae.
  • the plant infection by a plant pathogen typically designates a 15 plant infection by at least one phytopathogen.
  • the infection can occur on any organ of the plant.
  • the plant infection may b e, e.g., a B. cinerea infection, for example a B. cinerea infection of A.
  • thaliana, tomato, strawberry, sunflower, grapevine, or apple a C. higginsianum infection, for example a C. higginsianum infection of turnip, Chinese cabbage, mustard, A. 20 thaliana, or apple
  • a U. maydis infection for example a U. maydis infection of maize
  • a R. solanacearum infection for example a R solanacearum infection of tomato, potatoes, eggplant,pepper, or tobacco
  • a Pseudomonas syringae infection for example a Pseudomonas syringae infection on apple or pear; or any combination thereof, such as a B. cinerea and/or C. higginsianum infection of A.
  • Statement 1 A phytopharmaceutical or agrochemical composition, wherein an effective dose of plant defense elicitor compounds that are lignin-derived phenolic oligomers with a degree of polymerization (DP) of 2 to 8, and yet m ore preferably of 2 to 4, or synthesized structural similar compounds. 5 2.
  • Statement 2 A phytopharmaceutical or agrochemical composition according to statement 1’, wherein the composition has a pH in the range of 4 to 10, preferably in the range of 5 to 8. 3 .
  • Statement 3 The phytopharmaceutical or agrochemical composition according to any one of the statements 1’ to 2’, wherein the lignin-derived 10 phenolic oligomers have a pH in the range of 4.0 to 6.0 in origin. 4.
  • Statement 4 The composition according to any one of the statements 1’ to 3’, whereby defense elicitor compounds are 0,5 to 30 wt% by dry weight of aromatic compounds, preferably 1 to 20 wt% by dry weight of aromatic compounds, more preferably 2 to 10 wt% of the composition in dry state. 15 5.
  • Statement 5 The composition according to any one of the statements 1’ to 4’, whereby defense elicitor compounds are of the group of lignin-derived dimerics (diphenolic) compounds, lignin-derived trimeric (triphenolic) compounds, lignin-derived tetrameric (tetraphenolic) compound or a c ombination thereof (Fig. 2). 20 6.
  • Statement 6 The composition according to any one of the statements 1’ to 5’, whereby lignin-derived phenolic oligomers are from depolymerized lignin or decomposed lignin. 7 .
  • Statement 7 The composition according to any one of the statements 1’ to 6’, whereby lignin-derived phenolic oligomers are from a lignin that is 25 depolymerized or decomposed by non-catalytic thermo-solvolytic d epolymerisation (Fig. 1B), reductive catalytic fractionation (RCF) (Fig. 1A) of lignocellulose. 8 .
  • Statement 8 The composition according to any one of the statements 1’ to 7’, whereby the lignin is from a lignocellulose biomass, for instance flax shives, 30 wood chips, pine, spruce or poplar sawdust.
  • a surfactant for instance flax shives, 30 wood chips, pine, spruce or poplar sawdust.
  • composition according to any one of the statements 1’ to 9’ further comprising a second plant defense elicitor, for instance of the group consisting of alginate, hexokinase, laminarin, sodium silicate, silicon, oligo- 15
  • a second plant defense elicitor for instance of the group consisting of alginate, hexokinase, laminarin, sodium silicate, silicon, oligo- 15
  • a plant defense e licitor selected from the group consisting of pectin fragment, oligogalacturonide, cellobiose, xyloglucan, non-branched P-l,3-glucan, chitin fragment, arabinose, arabinan, rhamnose, homogalacturonan, 5 rhamnogalacturonan I and II, xylogalacturonan, starch, and combinations thereof.
  • composition according to any one of the statements 1’ to 11’ further comprising a fungicide selected form the group consisting of phosphonates, benzamides, carbamates, dithiocarbamates , phtalimides, triazoles, quinolines, sulphur, and cyanoimidazoles or a fungicide selected from the group consisting of 2,6-dichloro-N-[3-chloro-5-(trifluoromethyl)-2- 15 pyridinylmethyl]benzamide; propyl 3-(dimethylamino)propylcarbamate hydrochloride; (2RS,3SR)-1-[3-(2-chlorophenyl)-2,3-epoxy-2-(4- fluorophenyl)propyl]-1H-1,2,4-triazole; 5,7-dichloro-4-quinolyl 4- fluorophenyl ether; sulphur; 4-chloro-2-cyano-N,N
  • the polymerization30 inhibitor is a compound of the group consisting of citric acid, salicylic acid, 2- Naphthol, phenolic acids (e.g., vanillic acid, syringic acid), ethylene glycol, glycerol, Mannitol (C 6 H 14 O 6 ), Sorbitol (C 6 H 14 O 6 ), Xylitol (C 5 H
  • composition according to any one of the statements 1’ to35 14’ further comprising a stabilizing agents of the group consisting of 1,4- butanediol, 2-hydroxy-1-naphthoic acid, 2-naphthol, 2-naphthol-7-sulfonat, 2-naththol, 3-hydroxy-2-naphthoic acid, 4-hydroxybenzoic acid, 6-hydroxy- 16
  • each R1, R3, and R4 is independently chosen from –H, -OH, O-CH3, a 15 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, 20 - wherein R2 is –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an aromatic compound selected from the - wherein each R1, R3, and R4 is independently chosen from –H, -OH, O-CH3, a 15 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic mono
  • R11 and R14 is independently chosen from –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic 5 monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or a romatic oligomer, and/or III) wherein at the aromatic compounds comprise at least one aromatic compound selected from the formula (viii) 10 , 19
  • each R22, R23, R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or an aromatic oligomer, or a 4 carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R21 is independently chosen from –H, a ⁇ -O-4 linkage to an aromatic 10 monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R24 is independently chosen from –H,
  • 21.Statement 21’ Use of the composition according any one of the statements 1’ to 18’, on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of an attack by a phytopathogenic of 15 the group consisting of fungi, bacteria, viruses, viroids, mycoplasma-like organisms, protozoa, insects, acari, and nematodes.
  • 22.Statement 22’ Use of the composition according any one of the statements 1’ to 18’, on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of abiotic stress.
  • 26.Statement 26’ A method for promoting induced systemic resistance, for 30 inducing latent host defenses or for priming the intrinsic resistance mechanisms in a plant, comprising applying to a plant or a plant part, the composition according to any one of the statements 1’ to 18’.
  • 27.Statement 27’ A method for promoting induced systemic resistance , for inducing latent host defenses or for priming the intrinsic resistance 35 mechanisms in a plant, comprising by spraying on said plant or contacting the roots of said plant with the composition according to any one of the statements 1’ to 21’.
  • a composition for protecting plants against plant pests 5 comprising a plant defense elicitor comprising, consisting of or essentially consisting of as active ingredient one or more aromatic compounds, with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4 or one or more aromatic compounds, with a molecular mass between 180 g /molbetween 230 g/mol to 1000 g/mol, and yet more preferably between 10 230 g/mol to 650 g/mol (Fig.
  • aromatic compounds comprise and/or I) at least one aromatic compound selected from the - wherein each R1, R3, and R4 is independently chosen from –H, -OH, O-CH3, a 15 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, 20 - wherein R2 is –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an 22
  • R11 and R14 is independently chosen from –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic 5 monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, a nd/or wherein at the aromatic compounds comprise at least one aromatic 10 , 24
  • a concentration of 0.05 to 20 mg/ml preferably 0.2 to 10 mg/ml, preferably 0.5 to 5 mg/ml, more preferably 0.8 to 1.2 mg/ml and most preferably 1 mg/mL or in case of a dry composition at 0,5 to 30 wt% by dry weight of aromatic compounds, preferably 1 to 20 wt% by dry weight 5 of aromatic compounds, more preferably 2 to 10 wt% by dry weight of aromatic compounds. 4 .
  • composition according to any one of the statements 1* to 3* further comprising a further plant defense elicitor such as alginate, hexokinase, laminarin, sodium silicate, silicon, oligo-galacturonan, 10 cellodextrin and/or chito-oligosaccharide or a plant defense elicitor selected from the group consisting of pectin fragment, oligogalacturonide, cellobiose, xyloglucan, non-branched P-l,3-glucan, chitin fragment, arabinose, arabinan, rhamnose, homogalacturonan, rhamnogalacturonan I and II, xylogalacturonan, starch, and combinations thereof.
  • a further plant defense elicitor such as alginate, hexokinase, laminarin, sodium silicate, silicon, oligo-galacturonan, 10 cellodextrin and/or chito-
  • composition according to any one of the statement 1* to 4* comprising a) the plant defense elicitor according to statement 1* and b) a fungicide, for instance a fungicide selected form the group consisting of selected from the group comprising: phosphonates, benzamides, carbamates, dithiocarbamates , phtalimides, triazoles, 20 quinolines, sulphur, and cyanoimidazoles or a fungicide selected from the group consisting of 2,6-dichloro-N-[3-chloro-5-(trifluoromethyl)-2- pyridinylmethyl]benzamide; propyl 3-(dimethylamino)propylcarbamate hydrochloride; (2RS,3SR)-1-[3-(2-chlorophenyl)-2,3-epoxy-2-(4- fluorophenyl)propyl]-1H-1,2,4-triazole; 5,7-dichloro-4
  • Statement 6* The composition according to any one of the statement 1* to 5*, comprising a) the plant defense elicitor according to statement 1* and b) a salt and/or sugar. 7 .
  • Statement 7* The composition according to any one of the statement 1* 35 to 6*, further comprising a co-formulant selected from the group comprising: detergents, emulsifiers, dispersing agents, anti-foaming agents, penetration enhancers, humectants, wetting agents of ionic or 26
  • non-ionic type anti-freeze agents, preservative agents, absorbent agents, thickeners, buffers, sticker agents, diluents or a mixture thereof, preferably a surfactant selected from the group comprising: detergents, emulsifiers, dispersing agents, anti-foaming agents, penetration 5 enhancers, humectants or wetting agents of ionic or non-ionic type, or a mixture thereof. 8 .
  • composition according to any one of the statement 1* to 7* further comprising a surfactant comprising one or more of the following components: castor oil ethoxylate, rapeseed methyl ester, alkyl 10 phosphates, tributyl phosphate, tripropyl phosphate, naphthalenesulphonic acid salts, a combination of organic sulfonate and 2-methylpentane-2,4-diol, alkylpolyglucoside, siloxanes derivates, alkylsulfonates, polycarboxylates, lignosulfonates, alkoxylated triglycerides, fatty amines polymers, dioctylsulfosuccinates or 15 polyoxyethylene (20) sorbitan monolaurate, preferably C18-castor-oil- ethoxylate, a combination of organic sulfonate and 2-methylpentane-2,4- diol,
  • composition is applied before harvest or post-harvest to the whole plant, the leaves, the flowers, fruits, seeds, seedlings or seedlings pricking out, propagation material such as tubers or rhizomes, plants 5 pricking out, and/or to the soil or inert substrate wherein the plant is growing or in which it is desired to grow, by spraying, drenching, soaking, dipping, injection or administration through fertilising or irrigation systems.
  • a plant defense elicitor characterized in that, the resistance inducing or defense eliciting active ingredient thereof comprises an effective dose of an aromatic compounds with a degree of polymerization (DP) of 2 to 8, 25 preferably of 2 to 4 or a molecular mass between 180 g/mol to 1800 g/mol, preferably between 230 g/mol to 1000 g/mol, and yet more preferably between 230 g/mol to 650 g/mol (Fig. 2), and is an aromatic compound and wherein and/or I) the aromatic compounds comprise at least one of the aromatic compounds from the formulae (i) 28
  • each R1, R3, and R4 is independently chosen from –H, -OH, O-CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to 5 an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R2 is –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic 10 oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R5 is selected of –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer,
  • the aromatic compounds comprise at least one aromatic compound - wherein each R12, R13, R15 and R16 is independently chosen from –H, -OH, O- 5 CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, 10 - wherein R11 and R14 is independently chosen from –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or a romatic oligomer, and/or III) wherein at the aromatic compound - wherein at
  • each R22, R23, R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic 5 monomer or an aromatic oligomer, or a 4 carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R21 is independently chosen from –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or aromatic 10 oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein the alky
  • Statement 2 The plant defense elicitor according to statement 1”, 25 characterized in that, the resistance inducing or defence eliciting active ingredient is extracted or derived from depolymerized lignin.
  • Statement 3 The plant elicitor according to statement 1” or 2”, characterized in that the resistance inducing or defense eliciting active i ngredients are diphenolics, triphenolics and or tetraphenolics comprising 30 compounds wherein the aromatic compounds have 2, 3 or 4 aromatic groups or a DP of 2, 3 or 4, respectively (Fig. 1). 4.
  • Statement 4 The plant defense elicitor according to any one of the statements 1” to 3”, whereby the resistance inducing or defence eliciting active ingredients are are diphenolics comprising compounds wherein the 35 aromatic compounds have 2 aromatic groups or a DP of 2 (Fig. 2). 5.
  • Statement 5 The plant defense elicitor, with in a dry composition a content of at least 0.5 % by dry weight of the resistance inducing or 32
  • Statement 8 The plant pest resistance inducer or plant defense elicitor 10 according to any one of the statement 1” to 6”, whereby the resistance inducing or defense eliciting active ingredient extracted from depolymerized lignin is from a lignocellulose or lignin source.
  • Statement 9 The plant defense elicitor according to any one of the statements 1” to 8”, comprising further a repolymerization inhibitor of the 15 group of a carbocation scavenger, aromatic scavengers and a polyhydric alcohol or a combination thereof.
  • the repolymerization inhibitor is a compound of the group consisting of citric 20 acid, salicylic acid, 2-Naphthol, phenolic acids (e.g., vanillic acid, syringic acid), ethylene glycol, glycerol, Mannitol (C6H14O6), Sorbitol (C6H14O6),
  • a polar aprotic solvent such as dimethyl sulfoxide, DMSO or a natural polar aprotic solvent of the group consisting of ⁇ - Valerolactone (DVL), Cyrene, (Dihydrolevoglucosenone); Ethyl Acetate, M ethyl Lactate, Acetone (Dimethyl Ketone), Limonene Oxide ) and a 5 surface-active agent such as surface active glycolipid or sophorolipid.
  • a polar aprotic solvent such as dimethyl sulfoxide, DMSO or a natural polar aprotic solvent of the group consisting of ⁇ - Valerolactone (DVL), Cyrene, (Dihydrolevoglucosenone); Ethyl Acetate, M ethyl Lactate, Acetone (Dimethyl Ketone), Limonene Oxide ) and a 5 surface-active agent such as surface active glycolipid or sophorolipid.
  • Statement 15 The plant defense elicitor according to any one of the 10 statement 1 to 14”, whereby the phenolics that are derived from depolymerized lignin are the reaction product of a reductive catalytic fractionation (RCF) of the lignin source.
  • RCF reductive catalytic fractionation
  • 6 The plant defense elicitor according to any one of the statement 1” to 15”, whereby the phenolics that are derived from 15 depolymerized lignin are the reaction product of a non-catalytic thermo- solvolytic de-polymerization and fractioning of the lignin source.
  • the plant defense elicitor according to any one of the statements 1” to 18” characterized in that it has a pH in the range of 4 to 10, preferably in the range of 5 to 8.
  • 2 0.Statement 20 The plant defense elicitor according to any one of the statements 1” to 18” , wherein the aromatic compounds are in origin an30 oil with a pH in the range of 4.0 to 6.0 in origin. 2 1.Statement 21” Use of the plant defense elicitor according any one of the statements 1” to 20”, on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of an attack by a p hytopathogenic pathogen or pest or by an abiotic stressor. 35 22.Statement 22” Use of the plant defense elicitor according any one of the statements 1” to 20”, on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of an attack by a 34
  • phytopathogenic pathogen or pest whereby the phytopathogenic pathogen or pest is selected from the group of a fungi, a bacteria, and insect.
  • 2 3.Statement 23” Use of the plant defense elicitor according any one of the statements 1” to 20”, on a plant or parts thereof whereby the latent host 5 defenses are activated preventive to or in the event of an attack by a phytopathogenic of the group consisting of fungi, bacteria, viruses, viroids, mycoplasma-like organisms, protozoa, insects, acari, and nematodes.
  • 29.Statement 29 A method for promoting induced systemic resistance of a plant, for inducing latent host defenses of a plant or for priming the 30 intrinsic resistance mechanisms in a plant, comprising by spraying on said plant or contacting the roots of said plant with the composition comprising the plant defense elicitoraccording to any one of the statements 1” to 20”.
  • 30.Statement 30 Any one of the statements according to 1” to 29”, whereby lignin-derived diphenolics are the reaction product of the lignin 35 depolymerization or 1) by a reductive catalytic fractionation (RCF) (Fig. 1A) of the lignin source with a heterogenous metal catalyst, including but not limited to Ru, Pd and Ni, on a support in an organic solvent or an 35
  • RCF reductive catalytic fractionation
  • organic solvent water mixture in a temperature range of 100°C to 300°C, preferably 150°C to 270°C and most preferably 200°C to 250°C and containing a hydrogen donor, including but not limited to H 2 or 2) by a non-catalytic thermo-solvolytic de-polymerization (Fig. 1B) of the lignin 5 source in an organic solvent or an organic solvent water mixture in a temperature range of 100°C to 300°C, preferably 150°C to 270°C and most preferably 200°C to 250°C and under an inert atmosphere.
  • a non-catalytic thermo-solvolytic de-polymerization Fig. 1B
  • Statement 31 A method for controlling a plant disease comprising treating a plant with a plant defense elicitor according to any one of the statements 10 1” to 29” comprising 0.05 to 20 mg/ml , preferably 0.2 to 10 mg/ml, preferably 0.5 to 5 mg/ml, more preferably 0.8 to 1.2 mg/ml and most preferably 1 mg/mL of active ingredient aromatic compounds or in case of a dry composition 0,5 to 30 wt% by dry weight of aromatic compounds, preferably 1 to 20 wt% by dry weight of aromatic compounds, more 15 preferably 2 to 10 wt% by dry weight of resistance inducing or defense eliciting active ingredient.
  • the present application also provides a spects and embodiments as set forth in the following statements (1° to 20 15°): 1 .
  • Statement 1° A plant defence elicitor comprising one or more lignin oligomers with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4. 25 2.
  • Statement 2° The elicitor according to statement 1°, characterized in that the lignin oligomers have a pH in the range of 5.0 to 8.0. 3 .
  • Statement 3° The elicitor according to any one of the statements 1° to 2°, 30 whereby the lignin-oligomers comprising, or essentially consisting of or consisting of I) at least one aromatic compound selected from the formulae 36
  • each R1, R3, and R4 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5 5-5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R2 is –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic 10 oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R5 is selected of –H, a ⁇ -O-4 linkage to an aromatic monomer 15 or aromatic oligomer
  • aromatic compounds comprise at least one aromatic 5 compound selected from the formulae (v) , (vi) - wherein each R12, R13, R15 and R16 is independently chosen from –H, - OH, O-CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, 10 a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein
  • aromatic compounds comprise at least one aromatic compound selected from the formula (viii) , (xi) (xviii) 39
  • each R12, R13, R15 and R16 is independently chosen from –H, - OH, O-CH3, 5 - wherein R11 and R14 is independently chosen from –H, and/or III) wherein at the aromatic compounds comprise at least one aromatic compound selected from the formula (viii) 42
  • R21 is independently chosen from –H, a ⁇ -O-4 linkage to an 5 aromatic monomer, a 4-O-5 linkage to an aromatic monomer, an ⁇ -O- 4 linkage to an aromatic monomer or a carbon-oxygen linkage to an aromatic monomer, - wherein each R22 and R23 is dependently chosen from -H, -OH, O-CH3 or where at least R22 or R23 is independently chosen from a 4-O-5 10 linkage to an aromatic monomer, a 5-5 linkage to an aromatic monomer, a ⁇ -5 linkage to an aromatic monomer, a carbon linkage to an aromatic monomer, or a carbon-oxygen linkage to an aromatic monomer, - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein 15 the alkyl group is derived from the alcohol solvent of the process, - wherein each R25 and
  • Statement 8° The elicitor according to any one of the statements 1° to 7°, wherein the one or more lignin oligomers is present in a concentration of 50% to 90% by weight of the elicitor. 15 9 .
  • Statement 9° The elicitor according to any one of the statements 1° to 8°, characterized in that it contains less than 0,1%, of each of acetic acid, methanol and ethanol.
  • a device 25 comprising means A and B
  • the scope of the expression “a device 25 comprising means A and B” should not be limited to the devices consisting only of components A and B. It means that with respect to the present invention, the only relevant components of the device are A and B.
  • the term “consisting of” refers to compositions, methods, and respective components thereof as described herein, which are exclusive of any element not30 recited in that description of the embodiment.
  • a s used herein the term “consisting essentially of” refers to those elements required for a given embodiment. The term permits the presence of additional elements that 50
  • a plant pest concerns anything that has a negative impact on a plant, including10 insects, fungi, bacteria, virus and parasites, Lignin is a complex organic substance that acts like a glue, binding the cells, fibers, and vessels that make up plants. It's the second most abundant biopolymer on earth, after cellulose, and plays a vital role in giving plants their strength and rigidity.
  • Lignin is a complex aromatic polymer made up of various phenolic subunits that are mainly 15 interlinked by ether and carbon-carbon bonds. Unlike cellulose, which is a carbohydrate, lignin is not easily broken down by microorganisms. Lignin depolymerization is a process that breaks down lignin, a complex molecule found in plants, into smaller, components. In this particular invention lignin depolymerization refers specifically to the breaking of ether bonds, and in particular 20 Beta-O-4 ether bonds, that occur in native lignin yielding a liquid lignin oil comprising of components like phenolic monomers, dimers and trimers or specific oligophenolics with a DP from 1 to 8.
  • Thermo-solvolytic depolymerization of lignin is a biorefinery method that combines lignocellulose biomass fractionation (solvent, heat and pressure disrupting 25 the structure of the biomass and dissolves lignin and some of the hemicellulose) with l ignin depolymerisation under an inert gas or atmosphere.
  • an inert gas or atmosphere selected from the group consisting of nitrogen, argon, helium and hydrogen.
  • the terms “induce” or “inducing” as used herein refers to cause, or causing and to enhance, or enhancing and to boost or boosting and to activate or activating.
  • the meaning of present application is non-catalytic thermo-solvolytic fractioning a 5 depolymerzation (Fig. 1B) process without the presence of a redox catalyst and under inert atmosphere.
  • Fig. 1B non-catalytic thermo-solvolytic fractioning a 5 depolymerzation
  • suppressors or stabilizing agents of the group consisting of 1,4-10 butanediol, 2-hydroxy-1-naphthoic acid, 2-naphthol, 2-naphthol-7-sulfonat, 2- naththol, 3-hydroxy-2-naphthoic acid, 4-hydroxybenzoic acid, 6-hydroxy-2- naphthoic acid, ascorbic acid, bovine serum albumin, citric acid, salicylic acid, cysteine, dimethylphloroglucinol, histidine, mannitol, o-dihydroxybenzene, p- benzenediol, phloroglucinol, resorcinol, soy protein isolate, syringic acid and vanillic 15 acid.
  • suppressors or stabilizing agents of the group consisting of 1,4-10 butanediol, 2-hydroxy-1-naphthoic acid, 2-naphthol, 2-naphthol-7-s
  • R eductive catalytic fractionation (RCF) (Fig. 1A) of lignocellulose is a biorefinery method that combines biomass fractionation (solvent, heat and pressure disrupting the structure of the biomass and dissolves lignin and some of the hemicellulose) with lignin depolymerisation enabled by heterogeneous catalysis, more particularly a 20 redox-active catalyst in a reductive environment (e.g. with the addition of hydrogen or H-donating agents).
  • the output is carbohydrate enriched pulp (mainly cellulose) and a lignin oil comprising fractions of low molecular weight oligomeric and m onomeric units of aromatic compounds which molecular weight fractions can be separated.
  • Reductive Catalytic Fractionation is a solvolytic process.
  • RCF 25 typically uses a solvent system (e.g., methanol, ethanol, or water) to facilitate the solvolytic depolymerisation of lignin from the lignocellulosic matrix.
  • a solvent system e.g., methanol, ethanol, or water
  • Catalysts operate within the solvent medium, enabling selective hydrogenolysis and 30 hydrogenation reactions to depolymerize and stabilize lignin-derived fragments.
  • the catalysts operate within the solvent medium, enabling selective hydrogenolysis and hydrogenation reactions to depolymerize and stabilize lignin-derived fragments.
  • Molecular hydrogen (H2) or hydrogen donors e.g., alcohol solvents like ethanol or formic acid
  • Solvolysis are 35 commonly used to create a reductive environment, further facilitating solvolytic 53
  • RCF in this application is also meant to comprise the reductive catalytic depolymerisation of 5 lignin solved in a suitable solvent (solvolytic catalytic depolymerisation).
  • pH of RCF liquids containing depolymerized lignin generally range from 4 to 6, reflecting the presence of phenolic and carboxylic acids and with hydrogenation stabilization this can shift to a neutral range (pH 6–7), as acidic groups are reduced or neutralized.
  • a suitable solvent for lignin depolymerization via RCF or non-catalytic thermo- solvolytic fractioning the process is an aliphatic alcohol such as methanol, ethanol, 1-propanol, 1-butanol, 1-pentanol, 2-pentanol, 2-butanol, 2-pentanol, 3-pentanol, 2-methyl-1-propanol, 2-methyl-1-butanol, or 3-methyl-1-butanol or binary mixture 20 with water thereof.
  • Micromixing means that the features of mixing are achieved at the molecular scale.
  • a polar aprotic solvent is one that has typically a dipole moment in thed range of 2– 25 5 D, a high dielectric constant of typically >15 and no hydrogen bond donors (i.e., t hey lack N-H or O-H groups).
  • Suitable polar aprotic solvents for the plant defene elicitor formulations of present invention are for instance ⁇ -Valerolactone (DVL), Cyrene, (Dihydrolevoglucosenone); Ethyl Acetate, Methyl Lactate, Acetone ( Dimethyl Ketone) and Limonene Oxide).
  • VL ⁇ -Valerolactone
  • Cyrene (Dihydrolevoglucosenone); Ethyl Acetate, Methyl Lactate, Acetone ( Dimethyl Ketone) and Limonene Oxide).
  • 30 PG is the abbreviation for the identified monomer, 4-propyl guaiacol.
  • PS is the abbreviation for the identified monomer, 4-propyl syringol. 54
  • DP is the abbreviation for degree of polymerization referring to the number of aromatic groups in the lignin oligomer
  • Pulping in terms of RCF or non-catalytic thermo-solvolytic fractioning is the separation cellulose fibers from the lignin or the other components, in generally via 5 the organic solvents and sometimes water. Fractionation in terms is the process of separating a mixture into its constituent parts based on their different properties. Fractionation allows processors to further separate the depolymerized lignin or lignin oils into fractions with different compositions, molecular weights or molecular sizes.
  • liquid-liquid extraction 10 is often used for separating lignin-derived phenolic monomers, dimers, and oligomers from the reaction mixture after depolymerization.
  • ISR is an abbreviation for induced systemic resistance. ISR is a defense mechanism in plants involving activation of the plant’s immune response systemically (throughout the entire plant) after exposure to certain beneficial elicitors. 15
  • a “disease index” is a numerical representations of disease severity or incidence and it can quantify the impact of a disease on a plant population.
  • the effect of ISR on disease suppression can be reflected indirectly and for evaluating the effectiveness of ISR-inducing treatments, researchers measure disease severity, pathogen growth, o r symptom development, measurements that contribute to disease indices.
  • a lower 20 disease index in ISR-treated plants indicates successful suppression of diseases due to induced resistance.
  • a "disease index" in the context of A. thaliana is a quantitative measure used to evaluate the resistance of A. thaliana accessions to specific pathogens. This index is particularly useful in studying the genetic basis of disease resistance in plants.
  • the 25 disease index is calculated based on the number of necrotic lesions (areas of plant t issue that have died) that develop on the leaves of A. thaliana after inoculation with a pathogen.
  • T he "disease index" for Hyaloperonospora arabidopsidis (H. arabidopsidis) in A. thaliana is a measure used to quantify the severity of infection by this oomycete 30 pathogen. This index is based on the number of sporangiophores (structures that bear spores) observed on the plants after inoculation. The disease severity is scored 55
  • biomass is used for the term “lignocellulosic material” and lignocellulosic material may be in the meaning of lignocellulose or 5 material comprising lignocellulose.
  • dry or “dried” referring to a compound, component or composition means that the water content has been significantly reduced from the original form thereof. This is typically achieved through processes like dehydration, which remove water 10 from the compound, component or composition by evaporation or other methods.
  • the amount of moisture left in a dry compound, component or composition powder can vary depending on the specific type of compound, component or composition and the drying method used. It has to be interpreted to have a moist content under 12%, preferably under 10 % and 7% and even having a moisture content of around15 5% or even having have a moisture content of around 3%.
  • the term “butter” as used herein is understood to be synonymous with the term “lipid” and may refer in general terms to a lipid or a composition comprising a lipid as a main constituent that retains solid, semi-solid, biphasic, or paste-like properties 20 at ordinary temperatures of use.
  • elicitor or “plant defense elicitor” as used herein refers to an exogenous defense-triggering molecule, for instance inducer of the plant immune system or elicitors of natural plant defences against pests (pathogens), abiotic and biotic 25 stressors.
  • pathogens pests
  • pathogens abiotic and biotic 25 stressors.
  • d efense mechanisms can be activated when receptors directly or indirectly come in contact with pathogens.
  • the ligands of these plant receptors are elicitors of the plant 30 immune system.
  • elicitors including so-called non-specific elicitors or PAMPs (pathogen associated molecular patterns) e.g. degradation products of cell wall components of pathogens or derived from a plant cell wall, and pathogen-specific elicitors or effectors e.g. avirulence gene products of pathogens such as AVR, such as AVR proteins, which play a crucial role in the gene-for-gene 35 interactions between plants and pathogens and which avirulence gene products are 56
  • pathogen associated molecular patterns e.g. degradation products of cell wall components of pathogens or derived from a plant cell wall
  • pathogen-specific elicitors or effectors e.g. avirulence gene products of pathogens such as AVR, such as AVR proteins, which play a crucial role in the gene-for-gene 35 interactions between plants and pathogens and which avirulence gene products are 56
  • Elicitors of the plant immune system comprise proteins, oligosaccharides, polysaccharides, lipids, glycolipids, glycoproteins, peptides of diverse origin, lipopeptides, algal extracts, extracts from the walls of plant material and/or fungal 5 material, fungi, bacterial material and viral material, or yeast material and/or extracts. Elicitors also comprise salicylic acid, jasmonic acid, lipid peroxidation products and/or one or more of their esters. Elicitors of the plant immune system against insects play a crucial role in activating defense mechanisms to protect plants from herbivory.
  • jasmonate pathway which is central in 10 promoting resistance to a broad spectrum of insects.
  • This pathway involves signal transduction pathways that include calcium ion fluxes, phosphorylation cascades, and the production of jasmonates, which are essential for plant defense against insect herbivores.
  • Such a plant defense elicitor triggers the activation of defense responses, such as the biosynthesis of jasmonic acid, to defend against insect 15 attacks.
  • S ome exogenous defense-triggering molecules Plant defense elicitor
  • Plant defense elicitor can induce a plant’s defense system associated with extensive transcriptional- and metabolic reprogramming of the genome.
  • Elicitation of plants with elicitor molecules can result 20 in the activation of a series of defense responses, including cell wall reinforcement by deposition of lignin and induction of an array of defense enzymes.
  • Diverse plant defense responses induced by elicitors involve de novo synthesis and accumulation of antimicrobial phytoalexins, induction of cell death (hypersensitive response), production of activated oxygen species (oxidative burst), and modification of plant 25 cell walls by deposition of callose (Bektas, 2022; Sudhamoy & al, 2010; Wang et al., 2004).
  • Elicitors can induce a range of responses in plants, including the production of antimicrobial compounds, cell death (hypersensitive response), generation of 30 reactive oxygen species, and modification of cell walls.
  • Elicitors might be specifically recognized by the plant and subsequently induce defense responses against pathogens or herbivores in the attacked host (Maffei et al., 2012). Elicitor recognition by the plant is assumed to be mediated by specific r eceptors in the plant cell, localized either on the cell surface for a number of fungal 5 elicitors or within the cell for certain bacterial elicitors, which initiate signaling processes that activate plant defenses (Angelova et al., 2006; Shinya et al., 2006). Some of such plant defense elicitors have been recently discovered and tested.
  • laminarin, a ⁇ -1,3 glucan oligosaccharide, oligo- galacturonan and sodium alginate are also plant defense elicitors and also found to be a plant defense elicitor (Priya Dey et al . PLoS ONE 14(9) (2019); Chalal et al. F ront. Plant Sci., 19 May 2015 Sec. Plant Pathogen Interactions Volume 6 - 2015 (2015) and Bektas (Horticulturae 2022, 8, 484).
  • Cellodextrins act as elicitors of plant 20 defense (Aziz et al., 2007 J. Exp. Bot.
  • C1-oxidized (aldonic) cellodextrins oligosaccharides composed of glucose units linked by ⁇ -1,4-glycosidic bonds
  • C4-oxidized cellodextrins with the same basic structure as cellodextrins (glucose chains linked together ß-1,4 glycosidic 25 bonds) but an additional oxygen atom (an oxygen-containing functional groups, such as hydroxyl (-OH) or carboxyl (-COOH) group) bonded to the fourth carbon (C4) of the glucose unit at the non-reducing end are both known improve solubility and reactivity.
  • oligo-galacturonan encompasses herein a chain of o(1-4)-linked D- galacturonic acids. Oligo-galacturonans are derived from pectin, which is a major 58
  • Pectin consists of a complex set of polysaccharides, including homogalacturonans, which are linear chains of a-(1-4)-linked D- galacturonic acids. Oligo-galacturonans are released from these galacturonans through the action of pectolytic enzymes. Oligo-galacturonans particularly suitable 5 as plant defense elicitor have a degree of polymerization higher than 8, preferably comprised between 9 and 20 or between 9 and 15.
  • chito-oligosaccharide and “chitosan oligosaccharide” are used interchangeably herein and refer to a linear oligosaccharide composed of randomly 10 distributed -(1-4)-linked D- glucosamine (deacetylated unit) and N-acetyl-D- glucosamine (acetylated unit).
  • Chitosan is naturally found in few organisms, but is mostly produced industrially by deacetylation of chitin, which is the structural element in the exoskeleton of crustaceans (crabs, shrimp, etc.), insects, and in the cell walls of some fungi and other organisms.
  • Chito-oligosaccharides used 15 particularly suitable as plant defense elicitor have a degree of acetylation lower than 50%, lower than 40%, or lower than 30%, preferably about 25% and a degree of polymerization higher than 5, preferably comprised between 5 and 10.
  • Salts can added to the composition according to the invention to ensure good ionic 20 conditions and sucrose can be added to the composition according to the invention.
  • sucrose is added in a concentration of from about 1 mM to 20 mM, most preferably about 5 to 10 mM sucrose.
  • Sucrose triggers signaling through hexokinase and is also a wetting agent. Sucrose can act as wetting agents improve the spreading and sticking of water-based sprays on plant surfaces.
  • fungicide encompasses chemical or biological substances or compositions 30 used to kill or inhibit fungi or oomycetes, e.g. by preventing sporulation, or their spores.
  • Fungicides can exert their biological effect by different modes of action, for example, but not limited to, by interference with nucleic acid synthesis, mitosis and c ell division, respiration, amino acids and protein synthesis, signal transduction, lipids and membrane synthesis, sterol biosynthesis, glucan synthesis in the pathogen or by35 inducing host plant defense.
  • Any fungicide can be included in the composition of the invention, such as, for example, a fungicide selected from: acylalanines (benalaxyl), anilinopyrimidines 59
  • the fungicide is chosen from the list comprising: phosphonates, benzamides, carbamates, dithiocarbamates, phtalimides, triazoles, quinolines,20 sulphur and cyanoimidazoles.
  • Phosphonates The mode of action of the phosphonates is largely unknown but could involve inhibition of mitochondrial ATP synthase. Suitable examples of phosphonates are phosphorous acid derivatives, including phosphorous acid itself and its alkali m etal or alkaline- earth metal salts.
  • the fungicides are 25 ethyl hydrogenphosphonates such as fosetyl-AI, fosetyl-K and fosetyl-Na.
  • Benzamides interfere with mitosis and cell division.
  • the benzamides used in the composition of the invention contain 2,6-dichloro-N-[3- c hloro-5- (trifluoromethyl)-2-pyridinyl]benzamide (fluopicolide) as active ingredient.
  • the carbamates used in the composition of the invention contain propamocarb, preferably propamocarb hydrochloride (propyl[3- (dimethylamino)propyl]carbamate hydrochloride) as the active ingredient.
  • propamocarb preferably propamocarb hydrochloride (propyl[3- (dimethylamino)propyl]carbamate hydrochloride) as the active ingredient.
  • dithiocarbamates show multi-site contact activity.
  • dithiocarbamates containing manganese ethylenebis(dithiocarbamate) (polymeric) complex with zinc salt (mancozeb) as active ingredient are used in the compostion of the invention.
  • the phtalimides used in t he plant defense elicitor composition of the invention comprise A/- (trichloromethylthio)phthalimide or 2-[(trichloromethyl)thio]-1 - -isoindole-1 ,3(2H)- 15 dione (folpet) as active ingredient.
  • A/- (trichloromethylthio)phthalimide or 2-[(trichloromethyl)thio]-1 - -isoindole-1 ,3(2H)- 15 dione (folpet) active ingredient.
  • the triazoles used in the composition of the invention contain (2RS,3RS)-3-(2-chloorfenyl)-2-(4-fluorfenyl)-[(1H-1,2,4-triazool-1- 20 yl)methyl]oxiraan (epoxyconazole) as active ingredient.
  • (2RS,3RS)-3-(2-chloorfenyl)-2-(4-fluorfenyl)-[(1H-1,2,4-triazool-1- 20 yl)methyl]oxiraan (epoxyconazole) as active ingredient.
  • Mention can be made of the triazole fungicide sold under the trade name Opus.
  • Cyanoimidazoles act by interfering with the electron transport chain at the level of complex III in the inner membrane of mitochondria, which blocks oxidative phosphorylation powered by electron transfer.
  • the 25 cyanoimidazoles used in the composition of the invention contain 4-chloro-2-cyano- /V,/ /-dimethyl-5-(4-methylphenyl)-1 - -imidazole-1- sulfonamide (cyazofamid) as active ingredient.
  • Quinolines act by interfering with, e.g. blocking, signal transduction.
  • the quinolines used in the composition of the invention contain 5,7- d ichloro-4- quinolyl 4-fluorophenyl ether (quinoxyfen) as active ingredient.
  • the compositions of the invention will typically contain additional components, known a s co- formulants or adjuvants, to obtain a product with good handling, efficacy and 61
  • co-formulant or “adjuvant” designate any substance other than the main oligosacharidic complex plant defense elicitor component defined herein, that is intentionally added to the plant defense elicitor composition of the invention.
  • the composition according to the invention further c omprises a co- formulant or adjuvant selected from the group comprising: surfactants, anti-freeze agents (including urea, ethylene glycol, propylene glycol or glycerol), preservative agents (including potassium sorbate, paraben and its derivates, 1 , 2-benzisothiazolin-3(2H)-one or essential oils), absorbent agents 10 (including raids of corn or sawdust), thickeners (including clays orxanthane gum), buffers, sticker agents (including latex, silicon or alkoxylated alkyl), diluents (including rapeseed methyl ester) or any standard inert ingredient conventionally used in agricultural compositions, or a mixture thereof.
  • surfactants including urea, ethylene glycol, propylene glycol or glycerol
  • preservative agents including potassium sorbate, paraben and its derivates, 1 , 2-benzisothiazolin-3
  • the composition according to the invention further comprises a 15 surfactant.
  • surfactant is meant herein a compound that lowers the surface tension of a liquid, allowing easier spreading.
  • the surfactant can be a detergent, an emulsifier (including alkyl polyglucosides glycerol ester or polyoxyethylene (20) sorbitan 20 monolaurate), or natural plant lecithin, or a biosurfactant, or a dispersing agent (including sodium chloride, potassium chloride, potassium nitrate, calcium chloride or starch of corn), a foaming agent (including derivates of tartric acid, malic acid or alcohols), a penetration enhancer, a humectant (including ammonium sulfate, g lycerin or urea) or a wetting agent of ionic or non-ionic type or a mixture of such 25 surfactants.
  • the surfactants used in the present invention are penetration enhancers, dispersing agents or emulsifiers.
  • penetration enhancer is understood herein as a compound that accelerates the uptake of active ingredient through the cuticle of a plant into the 30 plant, i.e. the rate of uptake, and/or increases the amount of active ingredient absorbed into the plant.
  • Classes of substances known as penetration enhancers include alkyl phosphates, such as tributyl phosphate and tripropyl phosphate, and naphthalenesulphonic acid salts.
  • Dehscofix® comprising castor oil and ethoxylated fatty 35 acids
  • Dehscofix CO 95 ® available from Huntsman, USA
  • dispersing agent is meant a substance added to a suspension, usually a colloid, to improve the separation of particles and to prevent settling or clumping. Mention can be made of the dispersing agent which is sold under the trade name Tensiofix Dp400 (available from Ajinomoto OmniChem), essentially comprising organic 5 sulfonate and 2-methylpentane-2,4- diol.
  • Tensiofix Dp400 available from Ajinomoto OmniChem
  • the term “abiotic” stress is used herein to refer to non-living chemical and/or physical factors in the environment that affect plant growth and/or development. Examples include extreme temperatures (heat or cold), water availability (e.g., drought), 10 salinity (e.g., salt), and the like.
  • the “dry” state in the present invention refers to a state that the water content is 15 about 20 mass% or less and the water activity value is 0.85 or less.
  • the water content is more preferably 15 mass% or less, more preferably 10 mass% or less, and more preferably 5 mass% or more.
  • the lower limit is not particularly limited and is usually 0.1 mass% or more.
  • the water activity value is preferably 0 .80 or less and more preferably 0.75 or less.
  • emulsifier refers to a substance that stabilizes an emulsion, i.e.
  • emulsifiers sold under the trade names Tween® 20, which essentially comprises polyoxyethylene (20) sorbitan monolaurate (polysorbate 20), and Radia®, which essentially 25 comprises alkyl polyglycosides.
  • said surfactant comprises one or more of the following components: castor oil ethoxylate, rapeseed methyl ester, alkyl phosphates, tributyl phosphate, tripropyl phosphate, n aphthalenesulphonic acid salts, organic sulfonate / 2-methylpentane-2,4- diol, alkylpolyglucoside, siloxanes derivates, alkylsulfonates, polycarboxylates 30 lignosulfonates, alkoxylated triglycerides, fatty amines polymers, dioctylsulfosuccinates or polyoxyethylene (20) sorbitan monolaurate (polysorbate 20), more preferably said surfactant is C18-castor-oil-ethoxylate (Dehscofix®), organic sulfonate / 2-methylpentane-2,4-diol (Tensiofix D
  • biosurfactant is understood a surface-active molecule produced by living organisms, typically microorganisms like bacteria or fungi. These special molecules can reduce surface tension and enhance the solubility and mobility of hydrophobic substances in aqueous environments and they have a unique structure with two key 5 parts: a hydrophilic head, often composed of things like sugars, amino acids, or phosphate groups and a hydrophobic tail usually made of fatty acids or long chains of hydrocarbons.
  • Biosurfactants are classified into different classes based on their chemical composition, including low molecular weight surface-active agents called biosurfactants and high molecular weight bioemulsifiers . 10 Any compound as intended herein may be a part of a composition.
  • composition generally refers to a thing composed of two or more components, and more specifically particularly denotes a mixture or a blend of two or more materials, such as elements, molecules, substances, biological molecules, or microbiological 15 materials, as well as reaction products and decomposition products formed from the materials of the composition.
  • a composition may comprise any compound as taught herein in combination with one or more other compounds or substances, be it one or more other compounds as taught herein or one or more other compounds or substances.
  • a composition may be obtained by 20 combining, such as admixing, a compound as taught herein with said one or more other compounds or substances.
  • a composition may be obtained by decomposing a starting material, such as cellulose, into a mixture of a plurality of decomposition products. 25
  • the present compositions may be configured as p hytopharmaceutical or agrochemical compositions for treatment of a plant or plant protection composition.
  • Phytopharmaceutical or agrochemical compositions typically comprise one or more active ingredients (chemically and/or biologically active materials having one or more beneficial effects on plant health) and one or more 30 phytopharmaceutically acceptable carriers.
  • the active ingredient o r one of the active ingredients is a plant defense elicitor.
  • compositions as typically used herein may be liquid, semisolid (e.g., gel), solid, or volatile or vapour-based, and may include solutions or dispersions, such as for example suspensions, emulsions, oil-in-water emulsions, water-in-oil emulsions, gelified aqueous solution 35 or dispersion, solutions comprising a volatile organic solvent, etc.
  • solutions or dispersions such as for example suspensions, emulsions, oil-in-water emulsions, water-in-oil emulsions, gelified aqueous solution 35 or dispersion, solutions comprising a volatile organic solvent, etc.
  • solid forms include, without limitation, powder, granules, pellets, water dispersible powder, water dispersible granules or water dispersible pellets.
  • the composition may 64
  • compositions can include active ingredients, carriers, adjuvants, and other additives that enhance their efficacy, stability, or application properties.
  • active ingredient or one of the active ingredients is a plant defense elicitor.
  • Such compositions are used to protect plants from biotic stress such as pests and diseases or from abiotic stress.
  • carrier broadly includes any and all solvents, diluents, bulking agents, buffers for pH control, dispersant, solubilisers, surfactants, wetting agents, emulsifiers, tackifiers, thickeners, binders, preservatives, antioxidants, cuticle solubilising molecules, natural orregenerated mineral substances, and the like, 15 and combinations thereof.
  • solvents diluents, bulking agents, buffers for pH control, dispersant, solubilisers, surfactants, wetting agents, emulsifiers, tackifiers, thickeners, binders, preservatives, antioxidants, cuticle solubilising molecules, natural orregenerated mineral substances, and the like, 15 and combinations thereof.
  • solvents diluents
  • bulking agents buffers for pH control
  • dispersant solubilisers
  • surfactants wetting agents
  • emulsifiers emulsifiers
  • tackifiers thickeners
  • solvolytic lignin depolymerisation means any one of the following processes reductive catalytic fractionation (RCF) of lignin or lignocellulose, non- 20 catalytic thermo-solvolytic depolymerisation of lignin or lignocellulose or oxidative catalytic fractioning (OCF).
  • RCF reductive catalytic fractionation
  • OCF oxidative catalytic fractioning
  • Typical salicylic acid pathway activators are salicylic acid (SA), benzothiadiazole (BTH), acibenzolar-S-methyl (ASM), chitosan, oligogalacturonides (OGs) and typical jasmonic acid pathway activator are jasmonic acid (JA), methyl jasmonate (MeJA),25 coronatine, hexanoic acid and volicitin.
  • SA salicylic acid
  • BTH benzothiadiazole
  • ASM acibenzolar-S-methyl
  • chitosan oligogalacturonides
  • OGs oligogalacturonides
  • typical jasmonic acid pathway activator are jasmonic acid (JA), methyl jasmonate (MeJA),25 coronatine, hexanoic acid and volicitin.
  • a preferred example of a plant treating phytopharmaceutically or agrochemically acceptable solvent is water, hence, compositions as taught herein may comprise water, i.e.,
  • suitable 30 solvents include, but are not limited to, aromatic hydrocarbons, such as, for example, xylene mixtures or substituted naphthalenes; phthalates, such as, for example, dibutyl phthalate or dioctyl phthalate; aliphatic hydrocarbons, such as, for example, cyclohexane or paraffins; alcohols and glycols and their ethers and esters, such as, for example, ethanol, ethylene glycol, ethylene glycol mono methyl or monoethyl 35 ether; ketones, such as, for example, cyclohexanone; strongly polar solvents, such a s, for example, N-methyl- 2-pyrrolidone, dimethyl sulfoxide or dimethylformamide; 65
  • aromatic hydrocarbons such as, for example, xylene mixtures or substituted naphthalenes
  • phthalates such as, for example, dibutyl phthalate or dioctyl phthalate
  • Non-limiting examples of solid carriers include, but are not limited to, natural mineral fillers, such as, for example, calcite, talcum, kaolin, montmorillonite or attapulgite; highly dispersed silicic acid or highly dispersed absorbent polymers; pumice, broken brick, sepiolite or bentonite; calcite or sand; dolomite or pulverized plant residues.
  • natural mineral fillers such as, for example, calcite, talcum, kaolin, montmorillonite or attapulgite; highly dispersed silicic acid or highly dispersed absorbent polymers; pumice, broken brick, sepiolite or bentonite; calcite or sand; dolomite or pulverized plant residues.
  • the compositions may comprise one or more surfactant, such as an anionic, non-ionic, amphoteric, or cationic surfactant, or a combination thereof, such as without limitation Triton X-100, non-ionic surfactant that has a hydrophilic polyethylene oxide chain (such as on average 9.5 ethylene oxide units) and an aromatic hydrocarbon hydrophobic group, 4-(l,l,3,3-tetramethylbutyl)- 15 phenyl); a polysorbate-type non-ionic surfactant such as polyoxyethylene (20) sorbitan monolaurate (Tween-20), polyoxyethylene (20) sorbitan monopalmitate (Tween-40); and/ or a non-ionic organosilicone surfactant such as Silwet® L-77 (3- (2 -methoxyethoxy )propyl-methyl-bis(trimethylsilyloxy)silane).
  • a surfactant such as an anionic, non-ionic, amphoteric, or
  • compositions may comprise one or more compounds miscible in organic solvents as well as water and having a weak acidity, such as without limitation dimethyl sulfoxide (DMSO) that enable the emulsification of lignin oil in water.
  • DMSO dimethyl sulfoxide
  • An embodiment of present invention is a) a plant defense elicitor characterized in that it comprises one or more a plant defense elicitor comprising, consisting of or essentially consisting of as active ingredient one or more aromatic compounds, with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4 or one or more aromatic compounds, with a molecular mass 180 g/molbetween 230 g/mol to 1000 g/mol, 30 and yet more preferably between 230 g/mol to 650 g/mol (Fig. 2), and wherein and/or I) the aromatic compounds comprise at least one aromatic compound selected 66
  • each R1, R3, and R4 is independently chosen from –H, -OH, O-CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to 5 an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or a romatic oligomer, and - wherein R2 is –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic 10 oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, and - wherein R5 is selected of –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer,
  • each R12, R13, R15 and R16 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic 10 monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R11 and R14 is independently chosen from –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic
  • each R22, R23, R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic 5 monomer or an aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R21 is independently chosen from –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or aromatic 10 oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein the alkyl
  • said surfactant comprises one or more of the following components: castor oil ethoxylate, rapeseed methyl ester, alkyl phosphates, tributyl phosphate, tripropyl phosphate, naphthalenesulphonic acid salts, organic sulfonate / 2-methylpentane-2,4-diol, alkylpolyglucoside, 5 siloxanes derivates, alkylsulfonates, polycarboxylates, lignosulfonates, alkoxylated triglycerides, fatty amines polymers, dioctylsulfosuccinates or polyoxyethylene (20) sorbitan monolaurate, most preferably said surfactant is C 18- castor-oil-ethoxylate (Dehscofix®), organic sulfonate / 2-methylpentane- 2,4-diol (Tensio
  • compositions according to the invention can further comprises a further plant immune system elicitor chosen among silica, copper, sulfur, aluminium, vanadium, cobalt, nickel, iron, silver, salicylic acid and its derivates (including acetyl-salicylic acid, isonicotinic acid, acibenzolar-S-methyl), jasmonic acid and its derivates (including methyl 20 jasmonate), ethylene and its derivates, polysaccharides (including glucans, xyloglucans, cellodextrins in particular with modified structure containing oxygen- containing functional groups at the C1 and C4 positions, chitin, chitosans, fucans, galactofucans, xylans, galactans, alginates, galacturonans, apiogalacturonans, fructans including inulin, mannans, xylomannans, galactomannans, 25 glucomannans and
  • Puccinia Colletotrichum, Verticillium, Magna porthe
  • bacterial extracts including extracts from Escherichia, Rhyzobia, Pseudomonas
  • BABA probenazole
  • isothianil phosphorous acid and its derivates (including aluminium, sodium and potassium fosetyl)
  • horsetail extracts potassium iodide and potassium 5 thiocyanate
  • Citrus extracts Yucca extracts Salix extracts and plant decoctions (including nettle decoction).
  • said further plant immune system elicitor c ontains laminarin (a linear ⁇ (1 ⁇ 3)- glucan with (1 ⁇ 6)-linkages) such as, for example Vacciplant Fruit®.
  • Yet another embodiment of present invention is a) a plant defense elicitor c haracterized in that it comprises one or more a plant defense elicitor comprising, consisting of or essentially consisting of as active ingredient one or more aromatic compounds, with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4 or one or more aromatic compounds, with a molecular mass 180 g/molbetween 230 15 g/mol to 1000 g/mol, and yet more preferably between 230 g/mol to 650 g/mol, and w herein and/or I) the aromatic compounds comprise at least one aromatic compound - wherein each R1, R3, and R4 is independently chosen from –H, -OH, O-CH3, a 20 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic 72
  • R2 is –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic 5 oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer
  • R5 is selected of –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic 10 oligomer, a ⁇ - ⁇ linkage to an aromatic monomer or aromatic oligomer, a ⁇ -1 linkage to an aromatic monomer or aromatic oligomer, an end-unit selected of CH3, -CH2CH3, -(CH2)2CH
  • each R12, R13, R15 and R16 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic 5 monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R11 and R14 is independently chosen from –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or 10 aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or a romatic oligomer, and/or III) wherein at the aromatic compounds comprise at least one aromatic compound selected from the formula (viii) 15
  • each R22, R23, R25 and R26 is independently chosen from –H, -OH, O- 5 CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or an aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, 10 - wherein R21 is independently chosen from –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, 15 - wherein R24 is independently chosen from –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer
  • a s econd plant defense elicitor preferably a second plant defense elicitor chosen among silica, copper, sulfur, aluminium, vanadium, cobalt, nickel, iron, silver, 5 salicylic acid and its derivates (including acetyl-salicylic acid, isonicotinic acid, acibenzolar-S-methyl), jasmonic acid and its derivates (including methyl jasmonate), ethylene and its derivates, polysaccharides (including glucans, xyloglucans, chitin, chitosans, fucans, galactofucans, xylans, galactans, alginates, galacturonans,
  • compositions according to the invention further comprise a further plant immune system elicitor that contains silicon or silicium (Si), such as, for example, silica (Si02) or silicates, including sodium silicate (Na2Si03).
  • a further plant immune system elicitor that contains silicon or silicium (Si), such as, for example, silica (Si02) or silicates, including sodium silicate (Na2Si03).
  • said further plant immune system elicitor is a silicate, more preferably sodium silicate.
  • the present invention also discloses a composition
  • a composition comprising: a ) a plant defense elicitor characterized in that it comprises one or more a plant defense elicitor comprising, consisting of or essentially consisting of as active 5 ingredient one or more aromatic compounds, with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4 or one or more aromatic compounds, with a molecular m ass 180 g/mol between 230 g/mol to 1000 g/mol, and yet more preferably between 230 g/mol to 650 g/mol (Fig.
  • DP degree of polymerization
  • the aromatic compounds comprise at least one aromatic compound selected from the formulae (i) 10 and - wherein each R1, R3, and R4 is independently chosen from –H, -OH, O-CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic 15 monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, and - wherein R2 is –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an20 ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, and 77
  • R11 and R14 is independently chosen from –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or 5 aromatic oligomer, and/or III) wherein at the aromatic compounds comprise at least one aromatic compound selected from the formula (viii) 10 , 79
  • each R22, R23, R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or an aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R21 is independently chosen from –H, a ⁇ -O-4 linkage to an aromatic 10 monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R24 is independently chosen from –H, OH, or
  • compositions of the invention comprising a plant defense elicitor and a second plant defense 5 elicitor further comprise an adjuvant, preferably a surfactant, more preferably a surfactant comprising polyoxyethylene (20) sorbitan monolaurate such as Tween® 20, a biosurfactant or a plant lecithin.
  • an adjuvant preferably a surfactant, more preferably a surfactant comprising polyoxyethylene (20) sorbitan monolaurate such as Tween® 20, a biosurfactant or a plant lecithin.
  • compositions according to the present invention encompass not only 10 compositions which are ready to be applied to the plant by means of a suitable device, such as a spraying device, but also the commercial concentrated compositions which have to be diluted before application to the plant.
  • the compositions according to the invention are themselves in quite diverse, solid or liquid forms.
  • solid composition forms mention may be made of powders for 15 dusting and granules, in particular those obtained by extrusion, by compacting, by impregnation of a granulated support or by granulation from a powder, tablets or effervescent lozenges.
  • liquid composition forms or forms intended to constitute liquid compositions when applied mention may be made of solutions, in particular water-soluble concentrates, emulsions, concentrated suspensions, dispersions, 20 aerosols and wettable granules and powders (or powders for spraying), pastes, gels and water soluble packaging.
  • the present invention relates to the use of the compositions of the invention in agricultural applications, more particularly for protecting plants against (infection by) plant pathogens.
  • the present invention not only provides in the simultaneous use of the different 25 components of the compositions, i.e. the use of the compositions, but also provides in the sequential use of the different components of the compositions.
  • the sequential use of the plant defense elicitor from a lignin depolymerisation process and fractioning in compositions comprising oligophenolics with a degree of polymerization (DP) of 2 to 5, preferably of 2 to 3 30 from depolymerized lignin or decomposed lignin (Fig. 2) or from structural identical oligophenolic and a fungicide also results in enhanced efficacy of the fungicide.
  • the present invention not only provides in the simultaneous use of the different components of the compositions, i.e. the use of the composition, but also provides in the sequential use of the different components of the compositions.
  • the35 inventors have found that the sequential use of the plant defense elicitor characterized in that it comprises one or more plant defense elicitors comprising, 81
  • each R1, R3, and R4 is independently chosen from –H, -OH, O-CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to 10 an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, and - wherein R2 is –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic 15 oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, and - wherein R5 is selected of –H, a
  • each R22, R23, R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or an aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R21 is independently chosen from –H, a ⁇ -O-4 linkage to an aromatic 10 monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R24 is independently chosen from –H,
  • organisms that cause infectious diseases in plants include fungi, oomycetes, bacteria, viruses, viroids, virus-like organisms, phytoplasmas, protozoa, nematodes and parasitic plants.
  • the plant pathogens are fungi, oomycetes, bacteria, viruses, nematodes and insects. 5
  • the majority of phytopathogenic fungi belong to the Ascomycetes and the Basidiomycetes, reproducing both sexually and asexually via the production of spores that can be spread through air (wind) or water, or can be soil borne such as zoospores that are capable of living saprotrophically, carrying out the first part of their lifecycle in the soil.
  • Deuteromycetes are fungi from which only the asexual 10 form of reproduction is known, meaning that this group of fungus produces their spores asexually.
  • the Oomycetes are not true fungi but are fungal-like organisms that use the same mechanisms as fungi to infect plants.
  • Fungal and fungal-like organisms are heterotrophic, i.e. they need an external source of nutrients for growth, development and reproduction.
  • An understanding of other 15 key features of these organisms can assist in their identification: - Hyphae: thread-like strands with a filamentous growth habit are a common feature in most fungi. The hyphae colonize (grow through) substrates so that the organism can obtain nutrients.
  • Plant pathogenic species colonize plants through the h ost surface, sometimes through direct penetration of intact plant surfaces.
  • 20 Saprophytic fungi tend to penetrate and colonize diseased plant tissue, senescing (dying) plants and plant residues. These fungi are major decomposers of organic matter in soil.
  • Hyphal cell walls true fungi have cell walls composed mainly of glucans and chitin, whereas fungal-like organisms have cell walls composed of cellulose and glycans.
  • 25 - Septate hyphae true fungi have cross walls within the hyphae, whereas fungal-like organisms do not. This can aid in the differentiation of these two groups under microscopic examination.
  • Motile spores true fungi do not have motile spores, with the exception of Chytrids. Motile zoospores (asexually produced spores) are common in many species 30 in the Oomycota (e.g. Pythium and Phytophthora) and some downy mildews. Zoospores enable dispersal through water in soil and on plant surfaces.
  • Wind dispersed spores many species of true fungi produce asexual or sexual spores for dispersal in the wind. This is a common feature of foliar fungal pathogens (e.g. Erysiphe). However some spores are adapted to splash dispersal.
  • 35 - Survival structures thick walled spores (e.g. oospores and chlamydospores), sclerotia and multicellular reproductive structures (e.g. pycnidia and perithecia) are important in the diseasecycle. During unfavourable environmental conditions or in 86
  • Non-limiting examples of phytopathogenic fungi and fungal-like organisms include 5 Pyricularia oryzae (Magnaporthe grisea) on rice and wheat and other Pyricularia spp. on other hosts; Puccinia spp. e.g. Puccinia sorghi, Puccinia graminis f.sp.
  • Drechslera spp. e.g. Helminthosporium turcicum, Helminthosporium carbonum, Helminthosporium mavdis or Helminthosporium sigmoideum
  • Drechslera s pp. Polyrenophora spp. e.g.
  • Pyrenophora tritici- repentens or Pyrenophora teres 20 Rhynchosporium spp., Mycosphaerella gramninicola (Septoria tritici) and P haeosphaeria nodorum (Stagonospora nodoruni or Septoria nodorum), Pseudocercosporella herpotrichoides and Gaeumannomyces graminis on cereals (for example wheat, barley, rye), turf and other hosts (e.g.
  • Septoria lycopersici Septoria g lycines, Septoria
  • Cercospora arachidicola and Cercosporidium personatum on 25 peanuts and other Cercospora spp. e.g. Cercospora kikuchii or Cercospora zaea- maydis
  • B otrytis spp. e.g. Botrytis cinerea or Botryotinia fuckeliana), Botrytis cinerea (grey mould) on tomatoes, strawberries, vegetables, vines and other hosts and other B otrytis spp. on other hosts
  • Alternaria spp. e.g.
  • Ascochyta spp. e.g. Ascochyta pisi
  • Stemphylium spp. Pleospora s pp.
  • summer diseases for example bitter rot (Glomerella cingulata), black rot or frogeye leaf spot (Botryosphaeria obtusa), 5 Brooks fruit spot (Mycosphaerellapomi), Cedar apple rust (Gymnosporangiumjuniperi-virginianae), sooty blotch (Gloeodespomigena), flyspeck (Schizothyrium pomi) and white rot (Botryosphaeria dothidea)) on apples and pears; P lasmopara viticola on vines; other downy mildews, such as Bremia lactucae on
  • Peronospora manshurica or Peronospora tabacina on 10 soybeans, tobacco, onions and other hosts, Pseudoperonospora humuli on hops and Pseudoperonospora cubensis on cucurbits; Pythium spp. (including Pythium ultimum) on turf and other hosts (e.g. Pythium aphanidermatum); Phytophthora infestans on potatoes and tomatoes and other Phytophthora spp. on vegetables, strawberries, avocado, pepper, ornamentals, tobacco, cocoa and other hosts (e.g.
  • Phytophthora 15 cinnamomi, Phytophthora cactorum, Phytophthora phaseoli, Phytophthora parasitica, Phytophthora porri, Phytophthora citrophthora, Phytophthora megasperma f.sp. s oiae or Phytophthora infestans); Thanatephorus cucumeris on rice and turf and other Rhizoctonia spp. on various hosts such as wheat and barley, peanuts, vegetables, cotton and turf; Sclerotinia spp. on turf, peanuts, potatoes, oil-seed rape 20 and other hosts (e.g.
  • Sclerotinia sclerotiorum Sclerotium spp. on turf, peanuts and other hosts; Gibberellafujikuroi on rice; Colletotrichum spp. (e.g. Colletotrichum lindemuthianum) on a range of hosts including turf, coffee and vegetables; Laetisaria f uciformis on turf; Mycosphaerella spp. on bananas, peanuts, citrus, pecans, papaya and other hosts; Diaporthe spp. on citrus, soybean, melon, pears, lupin and other 25 hosts; Elsinoe spp on citrus, vines, olives, pecans, roses and other hosts; Verticillium s pp.
  • Colletotrichum spp. e.g. Colletotrichum lindemuthianum
  • Verticillium dahliae or Verticillium albo-atrum on a range of hosts including hops, potatoes and tomatoes; Pyrenopeziza spp. on oil-seed rape and other hosts; Oncobasidium theobromae on cocoa causing vascular streak dieback; Fusarium spp. (e.g.
  • Fusarium nivale Fusarium sporotrichioides, Fusarium oxysporum, Fusarium 30 graminearum, Fusarium germinearum, Fusarium culmorum, Fusarium solani, F usarium moniliforme or Fusarium roseum), Typhula spp., Microdochium nivale, Ustilago spp. e.g. Ustilago maydis (e.g. corn smut), Urocystis spp., Tilletia spp. and Clavicepspurpurea on a variety of hosts but particularly wheat, barley, turf and maize; Ramularia spp.
  • post-harvest diseases 35 particularly of fruit for example Penicillium expansum, Penicilliumn digitatum, Penicillium italicum and Trichoderma viride on oranges, Colletotrichum musae and Gloeosporium musarum on bananas and Botrytis cinerea on grapes; other 88
  • pathogens on vines notably Eutypa lata, Guignardia bidwellii, Phellinus igniarus, Phomopsis viticola, Pseudopeziza tracheiphila and Stereum hirsutum; other pathogens on trees (for example Lophodermiunm seditiosum) or lumber, notably Cephaloascusfragrans, Ceratocystis spp., Ophiostoma piceae, Penicillium spp., 5 Trichoderma pseudokoningii, Trichoderma viride, Trichoderma harzianum, A spergillus niger, Leptographium liindbergi and Aureobasidium pullulans; and fungal vectors of viral diseases (for example Polymyxa graminis on cereals as the vector of b arley yellow mosaic virus (BYMV) and Polymyxa betae on sugar beet as the vector of
  • Emericella ssp. Encephalitozoon spp., Eremothecium spp.
  • Gaeumanomyces spp. e.g. Gaeumanomyces graminis f.sp. 15 tritici
  • Geomyces spp. Gibberella spp. (e.g. Gibberella zeae), Gloeophyllum spp., Glomus spp., Hypocrea spp., Kluyveromyces spp., Lentinula spp., Leptosphaeria s alvinii, Leucosporidium spp., Macrophomina spp.
  • Rhizocystis spp. Pyronema spp., Rhincosporium secalis, Rhizoctonia spp. (e.g. Rhizoctonia solani, Rhizoctonia oryzae or Rhizoctonia cerealis), Rhizopus spp. (e.g. Rhizopus chinensid), Saccharomyces spp., Scerotium spp. (e.g. Scerotium rolfsii), 25 Spizellomyces spp., Thermomyces spp., Thielaviopsis spp. (e.g.
  • Plant diseases caused by fungi including yeasts, rusts, smuts, mildews, molds, mushrooms and toadstools that can be treated using the plant defense elicitor30 compositions according to the present invention are for example: "Rust” is a fungal diseases in plants, which produces reddish-brown discoloration of the stems and leaves. "Black Rot” is characterized by the darkening and decaying of leaves of fruit and vegetable plants. 35 "Black Spot” is one of the many fungal diseases in plants. It is named “black spot” because it produces small black spots on plants. 89
  • Bottom Rot is a fungal disease found on lettuce plants. The characteristic of this fungus is that it first affects the leaves on the lower part of the plant and then moves upward to affect the upper part.
  • Canker affects the roots and bark, is found on woody trees and is notorious for 5 causing localized damage to the bark of trees.
  • Cotton Ball is notorious for attacking cranberry plants.
  • Crown Wart like canker attacks on the barks of woody trees, this fungus attacks the stem of the alfalfa plants. It forms white protrusions at the base of the stem of the plant.
  • “Potato Wart” is a fungal disease that causes dark, warty, spongy excrescences in the eyes of potato tubers, similar to the crown wart in alfalfa plants. "Damping Off” causes excessive moisture conditions of the seedlings. "Dry Rot” causes the drying and crumbling of timber, bulbs, potatoes or fruits.”Rhizoctinia Disease” is caused by fungi called Pellicularia and Corticium. It is 15 often seen to affect small potatoes. "Root Rot” infects the roots causing root decay, eventually causing the plant to die. “Sclerotium Rot” is caused by Fungus of the genus Sclerotium causing the formation of sclerotia on plants.
  • Dutch Elm Disease is a fungal disease affecting Elms. It spreads from one plant to20 another through root grafts or by the elm beetles that feed on small twigs. "Pinkroot” attacks onion plants and makes them unsuitable for consumption. "Soft Rot” is a slimy, mushy decay caused by fungi. "Yellow Spot” is characterized by a yellow spotting on the leaves of plants. "Powdery Mildews” is often specific to the host that it invades. It is normally seen on25 roses, lilac, English oak, zinnias, etc. "Plant Wilting” gets it name because it causes the plant it infects to wilt.
  • the fungus invasion starts in the roots and then slowly makes its way into the stem and plugs the vascular system of the plant.
  • "Decay” is decomposition of wood that is caused by fungi. When it attacks living plant30 tissue, it kills the plants.
  • the pine family are conifers or shrubs including the commercial important cedars, firs, hemlocks, pinons, larches, pines and spruces. In practice they are referred to as "softwood lumber” a broad industry term that refers to all commercial timber derived 35 from gymnosperms, specifically members of the pine family (Pinaceae) and a few other coniferous families.
  • the lignin composition among members of the Pine family (Pinaceae) is highly similar, as they are all gymnosperms (softwoods) that produce 90
  • G-type lignin predominantly guaiacyl (G-type) lignin. While guaiacyl (G-type) lignin is the dominant lignin in gymnosperms (softwoods) like wood of the pine family, Syringyl- Guaiacyl (SG-type) lignin is typical in hardwoods like wood from birch family, beech family and willow family. Present invention demonstrates that such lignin are useful 5 to produce the plant defence elicitor.
  • Non-limiting examples of phytopathogenic bacteria include the genii Erwinia (including Erwinia amylovora, causing fire blight on pears), Pseudomonas (including P seudomonas syringae), Xanthomonas (including Xanthomonas orizae, 10 Xanthomonas citri, Xanthomonas fuscans (citrus cancer) and Xanthomonas fragariae) and Ralstonia.
  • genii Erwinia including Erwinia amylovora, causing fire blight on pears
  • Pseudomonas including P seudomonas syringae
  • Xanthomonas including Xanthomonas orizae, 10 Xanthomonas citri, Xanthomonas fuscans (citrus cancer) and Xanthomonas fragariae
  • Non-limiting examples of phytopathogenic viruses include Cucumber Mosaic Virus, Barley Yellow Mosaic Virus, Strawberry Mild Yellow Edge Virus, Strawberry Latent Ringspot Virus, Beet Necrotic Yellow Vein Virus and Potato Virus Y.
  • Phytopathogenic insects that can be targeted by application of the compositions according to the invention include aphids, beetles, bugs, hoppers, locusts, mites, ants, ticks, trips, whiteflies, rootworms, maggots, weevils, (stem)borers, caterpillars, b utterflies, leaf-rolers, leaf- miners, etc.
  • Plant protection refers to the activation of mechanisms aimed at 20 controlling or reducing the pathogens and/or to minimize their effects on the plant. Plant protection can be achieved by killing the pathogens, by delaying their growth a nd/or reproduction, by reducing sporulation, etc.. According to another aspect of the present invention, there is provided a method for protecting plants against (infection by) plant pathogens, characterized in that an effective and substantially 25 non-phytotoxic amount of a composition according to the invention is applied to the plants.
  • the expression "effective and non-phytotoxic amount” means an amount of p lant defense elicitor composition according to the invention that is sufficient to induce control or destruction of the plant pathogens present or liable to appear on the plants, and that does not entail any appreciable symptom of phytotoxicity for 30 said plants. Such an amount can vary within a wide range depending on the plant pathogen to be controlled, the type of plant, the climatic conditions and the compounds included in the composition according to the invention. This amount can be determined by systematic field trials that are within the capabilities of a person s killed in the art. 35
  • the fungicide in the composition of the invention is applied at a reduced rate.
  • the rate of the fungicide is reduced by at least a factor 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 25, 40, 50, 60, 70, 80, 90, or 100 91
  • the rate of the fungicide is reduced by 50% to 90%, 6 0% to 90%, 70% to 90%, 80% to 90%, 60% to 80%, or 60% to 70% of the 5 recommended rate for said plant and/or conditions.
  • Application of the composition according to the invention can be carried out in accordance with techniques well known to persons skilled in the art.
  • the composition according to the invention can be applied to the whole plant, or to leaves, flowers, fruits, seeds and/or roots of the plant, as well as to the soil or inert substrate wherein 10 the plant is growing or in which it is desired to grow (e.g.
  • inorganic substrates like sand, rockwool, glasswool; expanded minerals like perlite, vermiculite, zeolite or expanded clay), pumice, pyroclastic materials or stuff, synthetic organic substrates (e.g. polyurethane), organic substrates (e.g. peat, composts, tree waste products like coir, wood fibre or chips, tree bark) or to a liquid substrate (e.g. floating 15 hydroponic systems, Nutrient Film Technique, Aeroponics).
  • the application can be done by spraying, drenching, soaking, dipping, injection, etc., or via fertigation systems.
  • compositions according to the invention can also be useful to apply the compositions according to the invention to propagation material such as tubers or rhizomes, but also seeds, seedlings or 20 seedlings pricking out and plants or plants pricking out.
  • the compositions according t o the invention can also be applied post-harvest to control decay.
  • the plants that can be protected by the method according to the invention mention can be made of cotton; flax; vine; fruit or vegetable crops such as Rosaceae sp.
  • Solanaceae sp. for instance tomatoes
  • Liliaceae sp. for 30 instance lettuces
  • Umbelliferae sp. for instance Cruciferae sp.
  • Chenopodiaceae sp. C ucurbitaceae sp.
  • Papilionaceae sp. for instance peas
  • Rosaceae sp. for instance strawberries
  • major crops such as Graminae sp. (for instance maize, lawn or cereals such as wheat, rice, barley and triticale), Asteraceae sp. (for instance sunflower), Brassicaceae sp.
  • Fabacae sp. for instance 35 peanuts
  • Papilionaceae sp. for instance soybean
  • Solanaceae sp. for instance tomatoes and potatoes
  • Chenopodiaceae sp. for instance beetroots
  • horticultural and forest crops as well as genetically modified homologues of these crops.
  • the term “monophenolic compounds” means molecules with o ne phenolic group. If the molecules result from the chemical modification of lignin, they are referred to as “lignin-derived monophenolics”, “lignin-derived monomers”, 5 “lignin monomers”, or “phenolic monomers”. These terms are used interchangeably. Chemical modification herein means depolymerisation and/or partial reduction. T he lignin-derived monophenolics comprise compounds having the formulae: 10 93
  • diphenolic compounds means molecules with two phenolic centers chemically linked to each other. Thus having a degree of p olymerization (DP) of 2 and two phenol molecules. If the molecules result from the chemical modification of lignin, they are referred to as “lignin-derived diphenolics”, 5 “lignin-derived dimers”, or “phenolic dimers”. These terms are used interchangeably. Chemical modification herein means depolymerisation and/or partial reduction In the present invention, the term “triphenolic compounds” or trimers means molecules with 3 phenolic centers chemically linked to each other. Thus having a 10 degree of polymerization (DP) of 3 and tree phenol molecules.
  • DP degree of polymerization
  • lignin-derived triphenolics molecules with three phenolic centers chemically linked to 15 each other resulting from the chemical modification of lignin are referred to as “lignin- derived triphenolics”, “lignin-derived trimers”, or “phenolic trimers”. These terms are used interchangeably.
  • tetraphenolic compounds or tetramers means 20 molecules with 4 phenolic centers chemically linked to each other. Thus having a degree of polymerization (DP) of 4 and four phenol molecules.
  • molecules with four phenolic centers chemically linked to e ach other resulting from the chemical modification of lignin are referred to as “lignin- derived tetraphenolics”, “lignin-derived tetramers”, or “phenolic tetramers”. These terms are used interchangeably.
  • polyphenolic compounds means molecules r esulting from the chemical modification of lignin.
  • lignin-derived polyphenolics lignin-derived oligomers
  • phenolic oligomers phenolic oligomers
  • the term “phenolic compounds” and “phenolic products mixture” are used interchangeably to indicate the mixture comprising monophenolic compounds, diphenolic compounds, triphenolic compounds, tetraphenolic c ompounds and other phenolic oligomers.
  • the term “hemicellulose-derived polyols” or simply “polyols” means aliphatic alcohols comprising at least two hydroxyl groups.
  • the term “hemicellulose-derived polyols” or “polyols” does not include monosaccharides or oligosaccharides.
  • Hemicellulose-derived polyols include sugar 10 alcohols derived from hydrogenation of monosaccharides.
  • the polyols result from the chemical modification of hemicellulose. Chemical modification herein means hydrolysis and hydrogenation. Hemicellulose-derived polyols primarily include xylitol, arabitol, dulcitol, mannitol, sorbitol, ethylene glycol, glycerol.
  • the term “C5 polyols” is used to indicate the group of polyols comprising 5 carbon atoms, such as xylitol 15 and arabitol.
  • C6 polyols is used to indicate the group of polyols comprising 6 carbon atoms, such as dulcitol, mannitol, and sorbitol.
  • C5 sugars is used to indicate the group of sugars comprising 5 carbon atoms, such as xylose and arabinose.
  • C6 sugars is used to indicate the group of sugars comprising 6 carbon atoms, such as glucose, mannose, and galactose.
  • hemicellulose-derived oligosaccharides is used to indicate molecules comprising two or more saccharide monomers or saccharide-derived monomers, linked to each other by a glycosidic bond.
  • oligosaccharides is used to denote saccharide oligomers with a non- reduced terminal saccharide group as well as molecules with a reduced terminal saccharide group. 30 Examples of such oligosaccharides include, but are not limited to, 97
  • unstable compounds is used to refer to compounds that are unstable 5 under the reaction conditions of the lignin depolymerisation process or in the formulation of the ISR product, and that cause unwanted side-reactions, such as recondensation. Unstable compounds typically bear a C ⁇ O or C ⁇ C functional group. Examples of such unstable compounds derived from carbohydrates include, but are not limited to, xylose, glucose, furfural, and hydroxymethylfurfural. Examples of 10 unstable compounds derived from lignin include, but are not limited to, coniferyl alcohol, sinapyl alcohol, phenolic compounds with C2-aldehyde substituents, and so- called Hibbert's ketones.
  • stable compounds can be transformed to “stable compounds” by transforming the C ⁇ O and/or C ⁇ C functional groups, for instance t hrough hydrogenation.
  • stable compounds is used to refer to compounds 15 that are stable under the reaction conditions or in the formulation of the ISR product, and that do not cause unwanted side-reactions, such as recondensation.
  • stable compounds derived from carbohydrates include, but are not limited to, xylitol, arabitol, dulcitol, mannitol, sorbitol, ethylene glycol, glycerol.
  • Examples of s table compounds derived from lignin include, but are not limited to, 4-n- 20 propanolsyringol, 4-n-propanolguaiacol, 4-n-propylsyringol, 4-n-propylguaiacol.
  • “n-Butanol” or simply “butanol” is abbreviated as “BuOH”.
  • MeOH is abbreviated as “MeOH”.
  • 25 “Ethanol” is abbreviated as “EtOH”. 98
  • Non limiting examples of “lignin” sources are from woody plants (vascular plants (tracheophytes) which includes most trees, shrubs) of the group of softwoods (softwood lumber) and hardwoods (oak, maple, birch), non-woody plants such as 5 grasses (wheat straw, rice straw, switchgrass), herbaceous plants (bamboo, sugar cane), seed coats (nuts, legume pulses, beans), flax stems and hemp stem, jute and bagasse (sugarcane residue).
  • C rops subjected to the plant defense elicitor of the present invention are not 10 particularly limited and any general cultivated plants can be subjected.
  • Examples t hereof include the Poaceae plants (such as rice, barley, wheat, corn, oat or lawn grass), the Solanaceae plants (such as tomato, eggplant or potato), the Cucurbitaceae plants (such as cucumber, melon or pumpkin), the Leguminosae plants (such as pea, soybean, kidney bean, alfalfa, peanut, fava bean), the 15 Brassicaceae plants (such as daikon radish, Chinese cabbage, cabbage, komatsuna, rape blossoms, bok choy or A.
  • the Poaceae plants such as rice, barley, wheat, corn, oat or lawn grass
  • the Solanaceae plants such as tomato, eggplant or potato
  • the Cucurbitaceae plants such as cucumber, melon or pumpkin
  • the Leguminosae plants such as pea, soybean, kidney bean, alfalfa, peanut, fava bean
  • the 15 Brassicaceae plants such as daikon radish, Chinese cabbage, cabbage
  • the Rosaceae plants such as strawberry, apple or pear
  • the Moraceae such as mulberry
  • the Malvaceae such as cotton
  • the Umbelliferae such as carrot, parsley or celery
  • the Liliaceae such as green o nion, onion or asparagus
  • the Compositae such as burdock, sunflower, 20 chrysanthemum, crown daisy, safflower, lettuce
  • the Vitaceae such as grape. Since the reaction which gives rise to plant disease resistance is generally nonspecific to pathogens, all the plant diseases caused by fungus, bacteria and viruses are included as subject diseases.
  • Examples thereof include diseases caused by 25 Magnaporthe grisea, Cochliobolus miyabeanus, Pseudomonas syringae pv. maculicola, Spongospora subterranea, Phytophthora infestans, Peronospora manshurica, Eryshiphe graminis f. sp. hordei, Eryshiphe graminis f. sp.
  • T he plant defense elicitor of the present invention can be used on plants in any forms 5 such as solution, powder, granule, emulsion, wettable powder, oil, aerosol, flowable by mixing the lignin derived dimers with appropriate additives such as zinc and/or copper, bicarbonates, carbocation scavenger or polyhydric alcohols.
  • the pH thereof can be adjusted by adding buffer, and properties such as penetration properties to plants or spreading properties can be modified by adding a 10 spreading agent, surfactant such a plant lecithin’s, such as lyso-lecithin, or the like and amino acids such as branched-chain amino acids, proline, glutamic acid, aspartic acid, and histidine.
  • a 10 spreading agent such as a plant lecithin’s, such as lyso-lecithin, or the like and amino acids such as branched-chain amino acids, proline, glutamic acid, aspartic acid, and histidine.
  • RCF reductive catalytic fractionation
  • the RCF experiment was performed in a 2 L stainless steel batch reactor (Parr Instruments & Co.).
  • 150 g l ignocellulose biomass e.g., pine or spruce, further referred to as ‘Pine’), or poplar
  • 150 g l ignocellulose biomass e.g., pine or spruce, further referred to as ‘Pine’
  • Pine pine or spruce
  • poplar mL methanol
  • the reactor was sealed, flushed three times with N 2 (10 bar) 25 and then pressurized with H2 (30 bar at room temperature).
  • the reaction mixture was stirred (750 rpm) and simultaneously heated to 235 °C ( ⁇ 30 min. heating time). After the reaction time of 3h, the reactor was cooled and depressurized at room temperature.
  • the reactor contents were quantitatively collected by washing the reactor with ethanol. 30
  • the solid pulp was separated by filtration and washed thoroughly with ethanol.
  • the resulting filtrate was evaporated and a brown oil was obtained, which was subjected to a threefold liquid-liquid extraction using ethyl acetate and water in order to remove extracted sugars.
  • the ethyl acetate-extracted phase was dried to o btain the lignin oil.
  • the resulting oils are named with reference to the biomass and 35 catalyst used during the RCF (e.g., pineRuOil). 100
  • Example 2 Preparation of depolymerized lignin via non-catalytic thermo-solvolytic fractioning.
  • F igure 1 B shows typical input and output streams of a non-catalytic thermos- solvolytic fractioning where no catalyst is used.
  • the non-catalytic thermo-solvolytic 5 fractioning experiment was performed in an identical set-up to the RCF and biomass feedstock with the difference of excluding the redox catalyst and under inert atmosphere, pressurizing with N2 (to 30 bar at room temperature). All other procedures were kept identical.
  • the resulting lignin oils are named with reference to t he biomass used during the non-catalytic thermo-solvolytic fractioning (e.g., pine 10 non-catalytic thermo-solvolytic fractioning oils or PineOil).
  • Example 3 Fractionation by sequential liquid-liquid solvent extraction.
  • F igure 1 C shows fractionation of lignin oil through liquid-liquid extraction.
  • An initial fractionation step involved the lignin oil from RCF or non-catalytic thermo-solvolytic fractioning and heptane solvent at a 1:5 ratio (g/mL) with threefold extraction at 15 80°C for 0.5 h.
  • Figure 1 C shows a subsequent fractionation steps on the heptane residue, if performed, using a mixture of heptane 20 and ethyl acetate in the v:v ratio of 80% heptane/20% ethyl acetate.
  • a lignin sample was solubilized in THF (5 mg mL -1 ) and subsequently filtered with a 0.2 ⁇ m PTFE membrane to remove any particulate matter to prevent plugging of the column.
  • GPC- SEC analyses were performed at 40 °C on a Waters E2695 equipped with a PL-Gel 3 101
  • gDNA plant genomic DNA
  • plant samples were grinded using the Precellys 24 tissue homogenizer at 6000 rpm for 10 seconds. After homogenization, 30 400 ⁇ l of Edwards buffer (200 mM Tris-HCl pH 7.5, 250 mM NaCl, 25 mM EDTA, and 0.5% (v/v) SDS) was added to each sample. The samples were vortexed and incubated at 55°C for 15 min. After incubation, samples were centrifuged for 2 min at 13000 rpm.
  • Edwards buffer 200 mM Tris-HCl pH 7.5, 250 mM NaCl, 25 mM EDTA, and 0.5% (v/v) SDS
  • the protocol for pathogen growth quantification via qPCR was as follows.
  • the hydroponics tanks were filled with 1.6 l plant nutrient solution containing the macronutrients 30 MgSO4.7H2O (500 mg/l), KH2PO4 (270 mg/l), KNO3 (200 mg/l), K2SO4 (100mg/l), Ca(NO3)2.4H2O (500 mg/l), and FeEDTA sodium salt (25 mg/l); and the micronutrients H 3 BO 3 (4.1 mg/), MnSO 4 .H 2 O (3.7 mg/l), CuCl 2 .2H 2 O (0.2 mg/l), (NH 4 )6Mo7O24.4H 2 O (0.0825 mg/l with 81.2% MoO3), and ZnSO 4 .7H 2 O (0.649 mg/l).
  • Tomato cultivar seeds were sown in the hydroponic systems in seed holders 35 (18 seeds/system) containing a solidified 0.65% (w/v) agar in water solution.
  • the hydroponic tanks were covered with plastic lids and placed in the plant growth chamber. One week later, the seeds had germinated so the lids were removed and 103
  • aeration pumps were installed to aerate the root compartments. After 24 days, 8 plants were selected per system and were treated with plant defense elicitor compounds by spraying the leaves with compound solution until run-off. Treatment with the solvent 1% v/v DMSO was included as mock treatment. Three, twelve and 5 seventeen days after treatment, five leaflets per plant were inoculated with 5 ⁇ l d roplets of a B. cinerea R16 strain spore suspension of 5 x 105 spores/ml in 1 ⁇ 2 potato dextrose broth. The hydroponics tanks were placed inside an infection box, containing a moist mat to obtain high humidity, in the growth chamber.
  • plants Post germination, seeds were seeded in soil. Thirty-eight days post seeding, plants were 20 treated with mock (1% v/v DMSO) and PineRuH100R (1 mg/ml) by spraying leaves till run-off. Afterwards, plants were placed in air inflated plant cages (60 cm ⁇ 40 cm ⁇ 40 cm with mesh size 0.25 mm ⁇ 0.25 mm, Entomologie-Speciaalzaak Vermandel V.O.F., the Netherlands), which could be accessed by opening the zipper. Three days after initial treatment, the plants received a second treatment 3 hours prior to insect 25 infestation. Plants were infested by introducing one Nesidiocoris tenuis female per plant cage.
  • Figure 8 depicts decreased transpiration rate of tomato plants (Solanum lycopersicum) by RCF lignin treatment.
  • Solanum lycopersicum (cultivar: Alisa Craig) were sown in seed holders containing 0.65% (w/v) agar in a water solution placed in a hydroponics tank (Araponics Surrey, Belgium) filled with 1.6 L of nutrient solution 5 containing the macronutrients MgSO4.7H2O (500 mg/l), KH2PO4 (270 mg/l), KNO3 (200 mg/l), K2SO4 (100 mg/l), Ca(NO3)2.4H2O (500 mg/l), and FeEDTA sodium salt (25 mg/l); and the micronutrients H3BO3 (4.1 mg/), MnSO4.H2O (3.7 mg/l), CuCl2.2H2O (0.2 mg/l), (NH4)6Mo7O24.4H2O (0.0825 mg/l with 81.2 % MoO3), and Z
  • Example 10 Improved survival of Arabidopsis thaliana under heat stress by RCF lignin treatment.
  • F igure 9 depicts improved survival rate of Arabidopsis thaliana under heat stress by RCF lignin treatment.
  • thaliana – heat stress protocol was adapted from Silva- 20 Correi et al., 2014. Briefly, Arabidopsis thaliana seeds were surface sterilized by 10 minute exposure to 30% bleach. Subsequently, 50 seeds were sown on 1X Murashige and Skoog (MS) plates. Four days post seeding, plants were treated with either mock (1% v/v DMSO) or PineRuH100R (1 mg/ml in 1% v/v DMSO) by submerging seedlings in 5 ⁇ l droplets. Three days after, heat stress was imposed by submersion 25 of parafilm-sealed plates into a water bath (45 °C) for 18 minutes.
  • the obtained liquid was acidified by HCl until pH 10 2 ⁇ 3.
  • the acidified liquid was extracted with chloroform or ethyl acetate (EtOAc) until the organic phase was colorless.
  • a small amount of NaHCO3 was then added into the organic phase to neutralize the residual acid.
  • Anhydrous Na2SO4 was used to remove water.
  • the water 15 phase was centrifuged to separate acid-insoluble oligomers and the acid (and water)- soluble portion.
  • the acid-insoluble oligomers were washed with deionized water until the pH of the eluent was 7.
  • the acid (and water)-soluble portion was vacuum distilled at 60 °C to remove H2O and HCl, and solid salts(including NaCl) were obtained. The salts were freeze-dried.
  • the resulting depolymerized lignin mixtures are named OCF20 4.1 (OCF monomers) and OCF 4.2 (OCF oligomers)
  • OCF20 4.1 OCF monomers
  • OCF 4.2 OCF oligomers
  • FIG.12 shows the relative pathogen proliferation of Hyaloperonospora arabidopsidis in A.
  • a phytopharmaceutical or agrochemical composition wherein an effective dose of plant defense elicitor compounds that are lignin-derived oligomer aromatics with a degree of p olymerization (DP) of 2 to 8, preferably of 2 to 4, or synthesised structurally similar 5 compounds.
  • DP degree of p olymerization
  • the present invention a further embodiment also provides that the plant defense elicitor compounds are from a lignin that is depolymerized or decomposed by reductive catalytic fractionation (RCF) of lignin or lignocellulose or that the plant defense elicitor compounds are from a lignin that is depolymerized or decomposed 10 by non-catalytic thermo-solvolytic depolymerisation of lignin or lignocellulose.
  • the present invention provides that the plant defense elicitor compounds are from a lignin that is depolymerized or decomposed or 1) by a reductive catalytic fractionation (RCF) (Fig.
  • a heterogenous metal catalyst including but not limited to Ru, Pd and Ni
  • a support 15 in an organic solvent or an organic solvent water mixture in a temperature range of 100°C to 300°C, preferably 150°C to 270°C and most preferably 200°C to 250°C and containing a hydrogen donor, including but not limited to H2 or 2) by a non-catalytic thermo-solvolytic de-polymerization (Fig. 1B) of the lignin source in an organic solvent or an organic solvent water mixture in a temperature range of 100°C to 20 300°C, preferably 150°C to 270°C and most preferably 200°C to 250°C and under an inert atmosphere.
  • a heterogenous metal catalyst including but not limited to Ru, Pd and Ni
  • plant defense elicitor compounds can thus be the reaction product of these processes of lignin depolymerization.
  • This embodiment of the invention advantageously comprises that the plant defense elicitor compounds are obtained from lignin depolymerization or decomposing by reductive catalytic25 fractionation (RCF), as this resulted to the most stable compositions when no re- polymerization where add.
  • the present invention provides that the phytopharmaceutical or agrochemical composition has a pH in the range of 4 to 10, preferably in the range of 5 to 8 or that the plant defense elicitor compounds in origin have a pH in the range30 of 4.0 to 6.0.
  • compositions described above may be embodied as substantially free of acetic acid, methanol and ethanol, meaning it contains less than 0,1%, of each acetic acid, methanol and ethanol.
  • the compositions described above is characterised in that the plant defense elicitor compounds comprise, or essentially 35 consist of or consist of I) at least one aromatic compound selected from the formulae 107
  • each R1, R3, and R4 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5 5-5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R2 is –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic 10 oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R5 is selected of –H, a ⁇ -O-4 linkage to an aromatic monomer 15 or aromatic oligomer, a ⁇ -5
  • aromatic compounds comprise at least one aromatic 5 compound selected from the formulae (v) , (vi) - wherein each R12, R13, R15 and R16 is independently chosen from –H, - OH, O-CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, 10 a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein
  • aromatic compounds comprise at least one aromatic compound selected from the formula (viii) , (xi) (xviii) 110
  • each R12, R13, R15 and R16 is independently chosen from –H, - OH, O-CH3, 5 - wherein R11 and R14 is independently chosen from –H, and/or III) wherein at the aromatic compounds comprise at least one aromatic compound selected from the formula (viii) 113
  • R21 is independently chosen from –H, a ⁇ -O-4 linkage to an 5 aromatic monomer, a 4-O-5 linkage to an aromatic monomer, an ⁇ -O- 4 linkage to an aromatic monomer or a carbon-oxygen linkage to an aromatic monomer, - wherein each R22 and R23 is dependently chosen from -H, -OH, O-CH3 or where at least R22 or R23 is independently chosen from a 4-O-5 10 linkage to an aromatic monomer, a 5-5 linkage to an aromatic monomer, a ⁇ -5 linkage to an aromatic monomer, a carbon linkage to an aromatic monomer, or a carbon-oxygen linkage to an aromatic monomer, - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein 15 the alkyl group is derived from the alcohol solvent of the process, - wherein each R25 and
  • compositions described above is characterised in that the plant defense elicitor compounds comprise, or essentially consist of or consist of at least one aromatic compound selected from the formulae 116
  • compositions described above is characterised in that the plant defense elicitor compounds are phenolic oligomers 5 comprising two benzene rings directly bridged or bridged with a common bridging 118
  • the compositions described above is characterised in that the plant defense elicitor compounds are 0,5 to 30 wt% by dry weight of aromatic compounds, preferably 1 to 20 wt% by dry weight of aromatic 10 compounds, more preferably 2 to 10 wt% of the composition in dry state.
  • the phytopharmaceutical or agrochemical composition according to the present invention further comprises an ingredient of the group consisting of a surfactant, a biosurfactant, a penetration enhancer, a dispersing agent, an emulsifier and a carrier or a combination thereof.
  • the phytopharmaceutical or agrochemical composition according to the present invention further comprises a polymerization inhibitor of the group of a carbocation scavenger, aromatic scavengers and a polyhydric alcohol or a combination thereof.
  • the phytopharmaceutical or agrochemical composition 20 further comprises a polymerization inhibitor whereby the repolymerization inhibitor is a compound of the group consisting of citric acid, salicylic acid, 2-naphthol, phenolic acids (e.g., vanillic acid, syringic acid), ethylene glycol, glycerol, mannitol (C 6 H1 4 O 6 ), sorbitol (C 6 H 14 O 6 ), xylitol (C 5 H 12 O 5 ), erythritol, maltitol (C 12 H 24 O 11 ).
  • the repolymerization inhibitor is a compound of the group consisting of citric acid, salicylic acid, 2-naphthol, phenolic acids (e.g., vanillic acid, syringic acid), ethylene glycol, glycerol, mannitol (C 6 H1 4 O 6 ), sorbitol (C 6 H 14 O 6 ),
  • the phytopharmaceutical or agrochemical composition according to the present invention further comprises a stabilizing agents of the group consisting of 1,4-butanediol, 2-hydroxy-1-naphthoic acid, 2-naphthol, 2- naphthol-7-sulfonat, 2-naththol, 3-hydroxy-2-naphthoic acid, 4-hydroxybenzoic acid, 6-hydroxy-2-naphthoic acid, ascorbic acid, bovine serum albumin, citric acid,30 salicylic acid, cysteine, dimethylphloroglucinol, histidine, mannitol, o- dihydroxybenzene, p-benzenediol, phloroglucinol, resorcinol, soy protein isolate, syringic acid and vanillic acid so to prevent repolymerisation.
  • the phytopharmaceutical or agrochemical composition according to the present invention further comprises a second plant
  • the phytopharmaceutical or agrochemical composition according to the present invention further comprises a fungicide, an antimicrobial, 10 an insecticidal, and/or an antiviral for instance.
  • the phytopharmaceutical or agrochemical composition according to the present invention and described above comprises the lignin-derived oligomer aromatics with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 15 4 from lignin that is from coconut husk, softwood trees, hardwood trees, a grass (such as bamboo, corn stalks & stover; wheat straw, rice straw, barley straw, miscanthus), flax shives or hemp stalk or a combination thereof.
  • DP degree of polymerization
  • the phytopharmaceutical or agrochemical composition 20 comprises an ingredient of the group consisting of a surfactant, a biosurfactant, a penetration enhancer, a dispersing agent, an emulsifier and a carrier or a combination thereof.
  • the phytopharmaceutical or agrochemical composition 25 according to the present invention and described above further comprises a second plant defense elicitor, for instance of the group consisting of alginate, hexokinase, laminarin, sodium silicate, silicon, oligo-galacturonan, cellodextrin and/or chito- oligosaccharide or a plant defense elicitor selected from the group consisting of pectin fragment, oligogalacturonide, cellobiose, xyloglucan, non-branched P-l,3-glucan, 30 chitin fragment, arabinose, arabinan, rhamnose, homogalacturonan, rhamnogalacturonan I and II, xylogalacturonan, starch, and combinations thereof.
  • the phytopharmaceutical or agrochemical composition according to the present invention and described above further comprises a 35 fungicide, antimicrobial,
  • the phytopharmaceutical or agrochemical composition according to the present invention and described above further comprises a fungicide selected form the group consisting of phosphonates, benzamides, carbamates, dithiocarbamates , phtalimides, triazoles, quinolines, sulphur, and 5 cyanoimidazoles or a fungicide selected from the group consisting of 2,6-dichloro-N- [3-chloro-5-(trifluoromethyl)-2-pyridinylmethyl]benzamide; propyl 3- (dimethylamino)propylcarbamate hydrochloride; (2RS,3SR)-1-[3-(2-chlorophenyl)- 2,3-epoxy-2-(4-fluorophenyl)propyl]-1H-1,2,4-triazole; 5,7-dichloro-4-quinolyl 4- fluorophenyl ether; sulphur; 4-chloro-2-cyano-
  • the present invention provides the phytopharmaceutical or agrochemical composition according to the present invention and described above further comprises stabilizing agents of the group consisting of 1,4-butanediol, 2- hydroxy-1-naphthoic acid, 2-naphthol, 2-naphthol-7-sulfonat, 2-naththol, 3- hydroxy-2-naphthoic acid, 4-hydroxybenzoic acid, 6-hydroxy-2-naphthoic acid, 20 ascorbic acid, bovine serum albumin, citric acid, salicylic acid, cysteine, dimethylphloroglucinol, histidine, mannitol, o-dihydroxybenzene, p-benzenediol, phloroglucinol, resorcinol, soy protein isolate, syringic acid and vanillic acid so to prevent repolymerization.
  • the present invention provides the phytopharmaceutical or agrochemical composition according to the present invention comprises a) the plant defense elicitor of present invention and as described hereabove and b) a fungicide, for instance a fungicide selected form the group consisting of selected from the group comprising: phosphonates, benzamides, carbamates, dithiocarbamates , 30 phtalimides, triazoles, quinolines, sulphur, and cyanoimidazoles or a fungicide selected from the group consisting of 2,6-dichloro-N-[3-chloro-5-(trifluoromethyl)-2- pyridinylmethyl]benzamide; propyl 3-(dimethylamino)propylcarbamate hydrochloride; (2RS,3SR)-1-[3-(2-chlorophenyl)-2,3-epoxy-2-(4- fluorophenyl)propyl]-1H-1,2,4-triazole
  • the present invention provides the phytopharmaceutical or 5 agrochemical composition according to the present invention and described above comprises also another type of plant defense elicitor such as alginate, hexokinase, laminarin, sodium silicate, silicon, oligo-galacturonan, cellodextrin and/or chito- oligosaccharide or a plant defense elicitor selected from the group consisting of pectin fragment, oligogalacturonide, cellobiose, xyloglucan, non-branched P-l,3-glucan, 10 chitin fragment, arabinose, arabinan, rhamnose, homogalacturonan, rhamnogalacturonan I and II, x
  • plant defense elicitor such as alginate, hexokinase, laminarin, sodium silicate, silicon, oligo-galacturonan, cellodextrin and/or chito- oligosaccharide
  • the present invention provides the phytopharmaceutical or agrochemical composition according to the present invention and described above 15 further comprises a co-formulant selected from the group comprising: detergents, emulsifiers, dispersing agents, anti-foaming agents, penetration enhancers, humectants, wetting agents of ionic or non-ionic type, anti-freeze agents, preservative agents, absorbent agents, thickeners, buffers, sticker agents, diluents or a mixture thereof, preferably a surfactant selected from the group comprising: 20 detergents, emulsifiers, dispersing agents, anti-foaming agents, penetration enhancers, humectants or wetting agents of ionic or non-ionic type, or a mixture thereof.
  • a co-formulant selected from the group comprising: detergents, emulsifiers, dispersing agents, anti-foaming agents, penetration enhancers, humectants or wetting agents of i
  • the present invention provides the phytopharmaceutical or 25 agrochemical composition according to the present invention and described above further comprises a surfactant comprising one or more of the following components: castor oil ethoxylate, rapeseed methyl ester, alkyl phosphates, tributyl phosphate, tripropyl phosphate, naphthalenesulphonic acid salts, a combination of organic sulfonate and 2-methylpentane-2,4-diol, alkylpolyglucoside, siloxanes derivates, 30 alkylsulfonates, polycarboxylates, lignosulfonates, alkoxylated triglycerides, fatty amines polymers, dioctylsulfosuccinates or polyoxyethylene (20) sorbitan monolaurate, preferably C18-castor-oil-ethoxylate, a combination of organic sulfonate and 2-methylpentane
  • the present invention provides that the lignin-derived aromatic oligomers are obtained from a lignin that is depolymerized or decomposed by reductive catalytic fractionation (RCF) of lignin or lignocellulose.
  • RCF reductive catalytic fractionation
  • the lignin-derived aromatic oligomers comprise, or essentially consist of or consist of I) at least one - wherein each R1, R3, and R4 is independently chosen from –H, -OH, O- 20 CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 123
  • R2 is –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic 5 oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R5 is selected of –H, a ⁇ -O-4 linkage to an aromatic monomer 10 or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ - ⁇ linkage to an aromatic monomer or aromatic oligomer, a ⁇ -1 linkage to an aromatic monomer or aromatic oligomer, an end-unit selected of CH3, -CH2CH3, -(CH)
  • aromatic compounds comprise at least one aromatic compound selected from the formulae (v) - wherein each R12, R13, R15 and R16 is independently chosen from –H, - 5 OH, O-CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a ⁇ -5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, 10 - wherein R11 and R14 is independently chosen from –H, a ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an ⁇ -O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, 125
  • aromatic compounds comprise at least one aromatic compound selected from the formula (viii) , (xi) (xviii) 126
  • each R12, R13, R15 and R16 is independently chosen from –H, - OH, O-CH3, 5 - wherein R11 and R14 is independently chosen from –H, and/or III) wherein at the aromatic compounds comprise at least one aromatic compound selected from the formula (viii) 129
  • R21 is independently chosen from –H, a ⁇ -O-4 linkage to an 5 aromatic monomer, a 4-O-5 linkage to an aromatic monomer, an ⁇ -O- 4 linkage to an aromatic monomer or a carbon-oxygen linkage to an aromatic monomer, - wherein each R22 and R23 is dependently chosen from -H, -OH, O-CH3 or where at least R22 or R23 is independently chosen from a 4-O-5 10 linkage to an aromatic monomer, a 5-5 linkage to an aromatic monomer, a ⁇ -5 linkage to an aromatic monomer, a carbon linkage to an aromatic monomer, or a carbon-oxygen linkage to an aromatic monomer, - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein 15 the alkyl group is derived from the alcohol solvent of the process, - wherein each R25 and
  • the lignin-derived 5 aromatic oligomers comprise, or essentially consist of or consist of at least one aromatic compound selected from the formulae (i) , (v) 132
  • lignin-derived 5 aromatic oligomers are phenolics comprising two benzene rings directly bridged or bridged with a common bridging group of the group consisting of aliphatic chains, 134
  • these lignin-derived aromatic oligomers are substantially free of acetic acid, methanol and ethanol, meaning it contains less than 0,1%, of each acetic acid, 10 methanol and ethanol.
  • these lignin-derived aromatic oligomers are comprised in a composition with a pH in the range of 4 to 10, preferably in the range of 5 to 8 or in origin have a pH in the range of 4.0 to 6.0.
  • the present invention provides that the lignin-derived aromatic 15 oligomers are comprised in a composition, further comprising an ingredient of the group consisting of a surfactant, a biosurfactant, a penetration enhancer, a dispersing agent, an emulsifier and a carrier or a combination thereof.
  • the present invention provides that these lignin-derived aromatic 20 oligomers are comprised in a composition, further comprising a repolymerization inhibitor of the group of a carbocation scavenger, aromatic scavengers and a polyhydric alcohol or a combination thereof or a repolymerization inhibitor of the group consisting of citric acid, salicylic acid, 2-naphthol, phenolic acids (e.g., vanillic acid, syringic acid), ethylene glycol, glycerol, mannitol (C 6 H1 4 O 6 ), sorbitol (C 6 H 14 O 6 ),25 xylitol (C5H12O5), erythritol, maltitol (C12H24O11) or a combination thereof.
  • a repolymerization inhibitor of the group of a carbocation scavenger, aromatic scavengers and a polyhydric alcohol or a combination thereof or a
  • the present invention provides that these lignin-derived aromatic oligomers are 0,5 to 30 wt% by dry weight of aromatic compounds, preferably 1 to 20 wt% by dry weight of aromatic compounds, more preferably 2 to 10 wt% of the 30 composition in dry state.
  • S ome of the methods described above may be embodied as promoting induced systemic resistance, for inducing latent host defenses or for priming the intrinsic resistance mechanisms in a plant, comprising applying to a plant or a plant part, the 35 composition described in these methods above. 135
  • Some of the methods described above may be embodied as promoting induced systemic resistance , for inducing latent host defenses or for priming the intrinsic resistance mechanisms in a plant, comprising by spraying on said plant or contacting the roots of said plant with the lignin-derived aromatic oligomers or a composition 5 therewith.
  • Some of the methods described above may be embodied as these method of present invention for protecting plants against plant pests, comprising applying an effective a nd substantially non-phytotoxic amount of the lignin-derived aromatic oligomers to 10 said plants, for instance wherein said plant pests are selected from the group comprising: fungi, oomycetes, bacteria, viruses, nematodes and insects.
  • compositions are 15 applied before harvest or post-harvest to the whole plant, the leaves, the flowers, fruits, seeds, seedlings or seedlings pricking out, propagation material such as tubers or rhizomes, plants pricking out, and/or to the soil or inert substrate wherein the plant is growing or in which it is desired to grow, by spraying, drenching, soaking, dipping, injection or administration through fertilising or irrigation systems.
  • inventive methods of plant treatment comprising applying to a plant or a plant part, a composition comprising the plant defense elicitor of present invention are suitable for promoting induced systemic resistance in a plant, for inducing latent host defenses of a plant or for priming the intrinsic resistance mechanisms in the plant.
  • inventive methods of plant treatment by spraying on said plant or contacting the roots of said plant with the plant defense elicitor according to any one of above stated methods are suitable for promoting induced systemic resistance of a plant, for inducing latent host defenses of a plant or for priming the intrinsic resistance 5 mechanisms in a plant.
  • a further embodiment of the invention concerns use of a lignin-derived aromatic oligomers of present invention and described here above as a plant defense elicitor. With this use the induced systemic resistance can be promoted in a plant or the latent host defenses in a plant can be induced by priming of the intrinsic disease10 resistance mechanisms of a plant.
  • Present invention also concerns the use of the plant defense elicitor of present invention on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of an attack by a phytopathogenic pathogen or pest.
  • Present invention also concerns the use of the plant defense elicitor of present 15 invention on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of an attack by a phytopathogenic pathogen or pest whereby the phytopathogenic pathogen or pest is selected from the group of a fungi, a bacteria, and insect.
  • Present invention also concerns the use of the plant defense elicitor of present 20 invention on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of an attack by a phytopathogenic of the group consisting of fungi, bacteria, viruses, viroids, mycoplasma-like organisms, protozoa, insects, acari, and nematodes.
  • Present invention also concerns the use of the plant defense elicitor of present 25 invention on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of abiotic stress.
  • Present invention also concerns the use of the plant defense elicitor of present i nvention on a plant or parts thereof to prime the intrinsic resistance mechanisms of a plant for stronger or faster induced plant defense preventive to or in the event of 30 an attack by a phytopathogenic pathogen or pest or of abiotic stress, as compared to control or other plant defense inducers.
  • H ere by the plant defense elicitor can be used in foliar spray agent, in a root drench. 137
  • Present invention also concerns the use of the plant defense elicitor of present invention in agricultural applications or to protect plants against plant pests.
  • plant pests can be selected from the group comprising: phytopestic fungi, oomycetes, bacteria, viruses, nematodes and insects.
  • a further embodiment of the invention concerns use of a lignin-derived aromatic o ligomers of present invention a as a plant defense elicitor to enhance the efficacy of said fungicide in said composition, or to stimulate the plant immune system.
  • Such use thereof can be on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of an attack by a phytopathogenic pathogen 10 or pest or by an abiotic stressor, or such use can be on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of an attack by a phytopathogenic pathogen or pest whereby the phytopathogenic pathogen or pest is selected from the group of a fungi, a bacteria, and insect.
  • the latent host defenses are activated 15 preventive to or in the event of an attack by a phytopathogenic of the group consisting of fungi, bacteria, viruses, viroids, mycoplasma-like organisms, protozoa, insects, acari, and nematodes, on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of abiotic stress or on a plant or parts thereof to prime the intrinsic resistance mechanisms of a plant for stronger or 20 faster induced plant defense preventive to or in the event of an attack by a phytopathogenic pathogen or pest or of abiotic stress, as compared to control or other plant defense inducers.
  • a further embodiment is a method for producing phytopharmaceutical or agrochemical compositionof present invention, the method comprising a. Subjecting 25 lignin or lignocellulose in a liquid phase to solvolytic lignin depolymerisation, b. Fractionating the extract to obtain a purified fraction enriched in lignin oligomers, c. Formulating the purified fraction into a phytopharmaceutically or agrochemically acceptable dosage form.
  • Y et a further embodiment is a method for producing phytopharmaceutical or 30 agrochemical composition of present invention, whereby the liquid phase described here above contains one or more solvents selected from the group comprising water, methanol, ethanol, n-propanol and isopropylalcohol and mixtures of two or more thereof. 138
  • solvolytic lignin depolymerisation and fractioning is of the group consisting of reductive catalytic fractionation (RCF), non-catalytic thermo-solvolytic depolymerisation and oxidative 5 catalytic fractioning (OCF).
  • RCF reductive catalytic fractionation
  • OCF oxidative 5 catalytic fractioning
  • FIG. 1 T ables Description Number Primer name Sequence 1 PFIN760 5’ – GTG TCG CAC ACT GTA CCC ATT TAT – 3’ 2 PFIN761 5’ – ATC TTC ATC ATG TAG TCG GTC AAG T – 3’ 3 PFIN762 5’ – AAT CAC AGC ACT TGC ACC A – 3’ 4 PFIN763 5’- GAG GGA AGC AAG AAT GGA AC – 3’ Table 1 15 Drawing Description BRIEF DESCRIPTION OF THE DRAWINGS A figure can contain different panels for instance figure 1 contains panel A, B, C.. w hich can be marked Fig. 1A, Fig. 1B and Fig 1C.
  • F IG. 1 is a schematic diagram showing depolymerized lignin extraction from biomass 25 through reductive catalytic fractionation (RCF) (Fig 1A), with use of heterogenous 139
  • lignin oil fractions
  • the RCF 5 and non-catalytic lignin oil (fractions) are named after i) the biomass used (e.g., p oplar, Fig. 2A and 2B; or pine, Fig. 2C, 2D, 2E) ii) the presence of a catalyst used during the lignin extraction process or not as indicated by the acronym of the catalyst (e.g.
  • the final solvent composition 10 used during subsequent liquid-liquid extraction being a mixture of heptane and ethyl acetate in a certain ratio (% v/v, e.g., 80 % heptane and 20 % ethyl acetate, H 80E20) and iv) whether the obtained fraction is either the dissolved liquid (‘L’) or the residue (‘R’) when using the respective solvent composition.
  • FIG. 2 are graphics showing the gel permeation chromatography (GPC) of depolymerized lignin oil and fractions thereof obtained through liquid-liquid extraction from pine and poplar biomass obtained through RCF in panel A, B, C and 20 D or in a reductive environment or a non-catalytic thermo-solvolytic fractionation of p ine biomass without catalyst and under inert atmosphere in panel E.
  • FIG. 3 are graphics that show in panel A the relative pathogen proliferation of Hyaloperonospora arabidopsidis in A.
  • RCF reductive catalytic fractionation
  • Capabilities of PineRuH100R compared to the PimeRu80E20L fraction obtained through liquid-liquid extraction of PineRuH100R (Fig. 1C).
  • A. thaliana thaliana after the treatment with the lignin derivative H. arabidopsidis pathogen spores were used as shown in panel A) to identify the most active lignin fraction; B) to optimize the treatment concentration; and in C) identify the active IR 140
  • FIG. 4 shows the IR activity of the heptane insoluble fraction from a depolymerized lignin obtained from the catalytic RCF process (PineRuH100R) in comparison to the heptane insoluble fraction and a non-catalytic thermo-solvolytic fractioning process (PineH100R) both emulsified in 1 % v/v DMSO at a concentration of 1 mg/ml prior 5 to immediate testing (Panel A) or after an accelerated stability test at higher temperatures for a shorter period of time (Panel B, 30 days 54 °C) and stability test at lower temperature for a prolonged period of time (Panel C, 58 days, 4 °C).
  • PineRuH100R catalytic RCF process
  • Panel A shows induced plant disease resistance activity of the depolymerized lignin fraction of the non-catalytic thermo-solvolytic fractioning process (PineH100R) and of the RCF p rocess (PineRuH100R);
  • Panel B shows the difference in stability of the PineRuH100R and PineH100R with the lignin-derived di/triphenolics after storage for 30 days at 54 15 °C and panel C 58 days at 4 °C.
  • PineRuH100R shows higher s tability than the non-catalytic thermo-solvolytic fractioning equivalent PineH100R, 25 when kept at similar conditions at 54°C for 30 days and 4°C for 58 days.
  • F IG. 5 is a graphic display that shows infection of A. thaliana by Hyaloperonospora arabidopsidis pathogen to determine optimal time between treatment by the plant disease resistance inducer comprises lignin-derived di/triphenolics (PineRuH100R 30 from RCF of pine biomass) and infection. Based on this data we concluded that the biggest relative reduction in pathogen proliferation is observed when plants are t reated 24h before exposure to H.
  • FIG.6 is a graphic display that in panel A shows the plant disease resistance activity 35 of various tomato (Solanum lycopersicum) cultivars after treatment with PineRuH100R (from RCF of pine biomass), which comprises lignin-derived 142
  • Solid,darker bars are the infected tomato cultivars that were not treated with the P ineRuH100R which comprises lignin-derived diphenolics, triphenolics and tetraphenolics.
  • Panel B shows that IR induction is at least maintained for 12 days 5 post PineRuH100R treatment (dpt) in S. lycopersicum when infected with B. cinerea. Briefly, 24 days post seeding, S. lycopersicum plants were treated with PineRuH100R by spraying the leaves with compound solution until run-off.
  • Treatment with the s olvent 1% v/v DMSO was included as mock treatment (indicated with .
  • the hydroponics tanks were placed inside an infection box, containing a moist mat to obtain high humidity, in the growth chamber.
  • the disease symptoms were quantified by measuring the diameter parallel to the midrib of the developing necrotic lesions at 2 dpi.
  • FIG. 7 concerns induction by RCF lignin of increased resistance in tomato (Solanum lycopersicum) against insects and it a graphic display that quantifies that number of necrotic rings caused by Nesidiocoris tenuis (also known as the tomato bug, tobacco leaf bug, tomato mirid, or green tobacco capsid) feeding post mock (1% v/v DMSO) a nd PineRuH100R (1 mg/ml, emulsified in 1% v/v DMSO) treatment in accordance 25 with Example 9.
  • Nesidiocoris tenuis also known as the tomato bug, tobacco leaf bug, tomato mirid, or green tobacco capsid
  • F IG. 8 is a graphic display that demonstrates the difference in tomato transpiration rate between mock and PineRuH100R sprayed plants under heat stress conditions30 (38 °C), in a treatment in accordance with Example 10.
  • F IG. 9 is a display that shows the seedling survival post mock (1% v/v DMSO) and PineRuH100R (1 mg/ml in 1% v/v DMSO) six days after heat stress exposure. The PineRuH100R renders that the seedling are more protected to heat stress had a35 higher survival rate. 143
  • FIG. 10 is a schematic diagram showing depolymerized lignin extraction from biomass through oxidative catalytic fractionation (OCF), with use of heterogenous catalysts with use of an alkaline aqueous solution in an oxidative environment.
  • OCF oxidative catalytic fractionation
  • heterogenous catalysts CuO
  • F IG. 11 shows the A.
  • thaliana ecotype: Columbia-0 plants seedlings in little pots (20 well-developed, freestanding seedlings) treated after eight days after sowing treated with mock (1% v/v DMSO) (Fig 11A.), and depolymerized lignin monomers 10 from OCF (OCF 4.1, 1 mg/ml) (Fig 11 B.) fraction by spraying the leaves with compound solution until run-off.
  • OCF OCF 4.1
  • a depolymerized lignin obtained from the catalytic RCF process of pine wood (PineRuH100R) or OCF oligomers (OCF 4.2) obtained from birch wood using a catalytic OCF process demonstrating equal elicitor activity of depolymerized lignin oligomers (DP ⁇ 2) from RCF and OCF.
  • H 80E20L the dissolved liquid (‘L’) subfraction (of the H100R) obtained in the fractioning of lignin oil through liquid-liquid extraction from mixture of heptane and 25 ethyl acetate in a certain ratio (% v/v, e.g., 80 % heptane and 20 % ethyl acetate), see figure 1C.
  • H 80E20R the residue (‘R’) subfraction (of the H100R) obtained in the fractioning of lignin oil through liquid-liquid extraction from mixture of heptane and ethyl acetate in a certain ratio (% v/v, e.g., 80 % heptane and 20 % ethyl acetate), see30 figure 1C.
  • PineH100R the depolymerized lignin (lignin oil) residue (‘R’) fraction from the 100% heptane extraction of the non-catalytic thermo-solvolytic fractioning process using pine biomass, see figure 1C.
  • PineH100L the depolymerized lignin (lignin oil) dissolved liquid (‘L’) fraction from 35 the 100% heptane extraction of the non-catalytic thermo-solvolytic fractioning process using pine biomass.
  • PineH100R the depolymerized lignin (lignin oil) dissolved liquid (‘L’) fraction from 35
  • PineRuH100R the depolymerized lignin (lignin oil) residue (‘R’) fraction from the 1 00% heptane extraction of the reductive catalytic fractionation (RCF) process using pine biomass with ruthenium catalyst, see figure 1C.
  • PineCdH100R the depolymerized lignin (lignin oil) residue (‘R’) fraction from the 5 100% heptane extraction of the reductive catalytic fractionation (RCF) process using pine biomass with cadmium catalyst, see figure 1C.
  • P ineRuH80E20L the dissolved liquid (‘L’) subfraction (of the PineRuH100R) obtained in the fractioning of lignin oil through liquid-liquid extraction from mixture of heptane and ethyl acetate in a certain ratio (% v/v, e.g., 80 % heptane and 20 % 10 ethyl acetate), see figure 1C.
  • PineCdH80E20L the dissolved liquid (‘L’) subfraction (of the PineCdH100R) obtained in the fractioning of lignin oil through liquid-liquid extraction from mixture of heptane and ethyl acetate in a certain ratio (% v/v, e.g., 80 % heptane and 20 % ethyl acetate), see figure 1C.
  • PopH100R the depolymerized lignin (lignin oil) residue (‘R’) fraction from the 100% heptane extraction of the non-catalytic thermo-solvolytic fractioning process using poplar biomass, see figure 1C.
  • PopH100L the depolymerized lignin (lignin oil) dissolved liquid (‘L’) fraction from the 100% heptane extraction of the non-catalytic thermo-solvolytic fractioning20 process using poplar biomass, see figure 1C.
  • PopRuH100R the depolymerized lignin (lignin oil) residue (‘R’) fraction from the 1 00% heptane extraction of the reductive catalytic fractionation (RCF) process using poplar biomass with ruthenium catalyst, see figure 1C.
  • PopCdH100R the depolymerized lignin (lignin oil) residue (‘R’) fraction from the 25 100% heptane extraction of the reductive catalytic fractionation (RCF) process using poplar biomass with cadmium catalyst, see figure 1C.
  • P opRuH80E20L the dissolved liquid (‘L’) subfraction (of the PopRuH100R) obtained in the fractioning of lignin oil through liquid-liquid extraction from mixture of heptane and ethyl acetate in a certain ratio (% v/v, e.g., 80 % heptane and 20 % 30 ethyl acetate), see figure 1C.
  • PopRuOil the lignin oil obtained by lignin depolymerized lignin by reductive c atalytic fractionation (RCF) process using poplar biomass with ruthenium catalyst.
  • PopRuOil the lignin oil obtained by lignin depolymerized lignin by reductive catalytic fractionation (RCF) process using poplar biomass with ruthenium catalyst.
  • 35 PineRuOil the lignin oil obtained by lignin depolymerized lignin by reductive catalytic fractionation (RCF) process using pine biomass with ruthenium catalyst.
  • PineRuOil the lignin oil obtained by lignin depolymerized lignin by reductive catalytic fractionation (RCF) process using pine biomass with ruthenium catalyst.
  • Fytosol a promising plant defense elicitor, controls early blight (alternaria solani) disease in the tomato by inducing host resistance- associated gene expression.
  • Sesquiterpene volatile organic compounds (vocs) are markers of elicitation by sulfated laminarine in grapevine. Frontiers in Plant Science, 6. - Hatami, M., Badi, H., & Ghorbanpour, M.

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Abstract

The present invention concerns the use of plant defense elicitor compositions, more particularly to protect plants against pests or pathogens. This includes the protection of plants and crops by application of these plant elicitor compositions. The active ingredient(s) of the plant defense elicitor compositions comprise lignin-derived phenolic oligomers with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4 (such as lignin-derived dimeric (diphenolic), lignin-derived trimeric compounds (triphenolic) and lignin-derived tetrameric compounds) and the and engineered or synthesized structural similar compounds.

Description

PLANT DEFENSE INDUCER Background and Summary BACKGROUND OF THE INVENTION 5 A. Field of the Invention The invention is broadly in the field of agrochemical or phytopharmaceutical compounds and compositions for plant treatment, and in particular relates to these 10 compounds or compositions as plant defense elicitors. These are also commonly known as plant immune- system activator molecules, or ‘bio-pesticides’ in layman’s terms, a class of environmental- friendly molecules that induce or boost plant’s resistance against abiotic stressor or biotic stressors such as pests and for instance retard the infection and propagation of microbial and viral pathogens on plants and 15 uses thereof. Insects may be a vector of such pathogens. B. Description of the Related Art There is a need for technology to efficiently produce high-quality agricultural products 20 in a limited amount of cultivated land. Therefore, it is of high interest in agriculture to control diseases caused by pests for instance parasites or pathogens such as fungi, oomycetes, bacteria, viruses, nematodes and insects. Fungicides, for instance, are such chemical compounds or biologic substances used to kill or inhibit fungi or oomycetes or their spores. Fungicides sometimes also have an effect on other plant 25 pathogens such as bacteria, viruses, nematodes or insects. A drawback of using certain fungicides is that fungicide residues can be found, in the environment and on food for human consumption sometimes posing a danger to biodiversity, human or animal health. In general, traditional pesticides, while effective, can harm the environment in several ways. They can pollute soil and water sources, harm beneficial 30 insects and pollinators, and disrupt ecosystems. There is a need in the art to make them unnecessary or to decrease their use. Thus, in order to promote sustainable crop production, there is a need to use more safe and natural substances with biological activity that can decrease the amounts of35 pest control chemicals and in particular chemical fungicides needed. 1
Lignin is the second largest biopolymer on earth. Lignin, with its polyaromatic network is an aromatic polymer is a major constituent in e.g. wood, being the most abundant carbon source on Earth second only to cellulose. In recent years, with development and commercialization of technologies to extract lignin in a highly 5 purified, solid and particularized form from the pulp-making process, it has attracted significant attention as a possible renewable substitute to primarily aromatic chemical precursors currently sourced from the petrochemical industry. Native or pristine lignin, a complex polymer consisting of aromatic building blocks, 10 results from a radical polymerization process of three 4-hydroxyphenylpropanoid building blocks or monolignols linking the latter via stable carbon-carbon bonds and more reactive ether bonds. The complex native lignin polymer can be broken down by various catalytic or thermo-solvolytic methods of lignin depolymerization of the lignin source, such as reductive catalytic fractionation (RCF), non-catalytic thermo- 15 solvolytic depolymerization and fractioning or combinations thereof to cleave the interunit linkages within the lignin polymer (ether bonds) into a liquid (lignin oil) comprising low molecular weight oligomers (oligophenolics having 3 or more aromatic groups or a degree of polymerization (DP) of 3 and more) , dimers (diphenolics having 2 aromatic groups or a DP of 2) and monomers (monophenols with 1 aromatic 20 group, DP of 1). Such lignin oil can be further fractionated, for instance by ultrafiltration and solubility-based methods, into different fractions of aromatic compounds with a specific degree of polymerization (DP). Plant cells have evolved a sophisticated immune system comprising two main layers 25 of defense known as pattern-triggered immunity (PTI) and effector-triggered immunity (ETI) (Song et al., New Phytologist (2022) 236:590–607), that constitute the so-called plant immune system. The plant immune system against insects involves a complex interplay of signaling pathways, defense mechanisms, and elicitors that work together to protect plants from herbivory. 30 ETI confers a narrow strain-specific resistance as it is initiated following the recognition of virulence effectors (A virulence-proteins) by cytoplasmic resistance genes (R-genes). This generally causes a strong site-specific accumulation of reactive oxygen species (ROS) leading to apoptosis. In contrast, PTI provide a broad-35 spectrum protection. Evolutionarily-conserved pathogen-associated molecular- pattern (PAMPs) are sensed by plants through a plethora of plasma membrane- 2
anchored pattern-recognition receptors (PRRs) (Albert et al. Surface Sensor Systems in Plant Immunity. Plant Physiol. 2020, vol. 182(4), 1582-1596). Compounds, which when perceived by a plant give rise to such defense responses to 5 abiotic and/or biotic stressors, are commonly referred to as plant defense elicitor, plant pest defense elicitor(s), plant immune system elicitor(s), plant defense elicitor(s), plant elicitor(s) or simply elicitor(s). Through the description the term plant defense elicitor or plant defense elicitors is most used. 10 The agricultural industry is engaged in a relentless fight against plant pathogens, exacerbated by an ever-changing environment due to inter alia climate changes, striving to avoid major economically early losses and uncertainty in the food supply chain. While only a small variety of chemicals are in common use as pesticides or fungicides, even this reservoir is diminishing, due to emerging biological resistance 15 in plant pathogens, and because of the side effects of some of these chemicals compounds on human health. Therefore, there is an urgent need for new plant protection compounds and compositions. The importance of natural plant defense elicitors lies in their ability to enhance plant 20 defense mechanisms against various pest (pathogens and stressors) such as fungi, oomycetes, bacteria, viruses, nematodes and insects. Natural compounds that induce or boost plant immunity are crucial for activating plant defense responses, thereby inhibiting pathogen development and improving plant resilience against biotic and/or abiotic stress. Compounds, which when perceived by a plant give rise to such defense 25 responses, are commonly referred to as plant defense elicitor, plant pest defense elicitor(s), plant immune system elicitor(s), plant defense elicitor(s), plant elicitor(s) or simply elicitor(s). Through the description the term plant defense elicitor or plant defense elicitors is most used. 30 Present invention solves this problem by a novel plant defense elicitor and demonstrates how this can be obtainable from a lignin depolymerisation process and fractioning in compositions comprising oligophenolics with a degree of polymerization (DP) of 2 to 8 and preferable of 2 to 4 from depolymerized lignin or decomposed lignin or from structurally identical oligophenolics. They can cause the increase of 35 the resistance to adverse conditions (biotic or abiotic). Present invention also demonstrates how these can be used in suitable compositions for plant defense. 3
SUMMARY OF THE INVENTION The present invention solves the problems of the related art of plant protection against pathogens and stressors by more natural compounds or of natural sources 5 by a plant defense elicitor, that comprises as an active ingredient of the plant defense elicitor depolymerised lignin. Furthermore this depolymerized lignin comprises specific oligophenolics with a DP from 1 to 8. In general present invention concerns new plant defense elicitors or new plant elicitor compositions, and the use of these in agricultural applications, more particularly to 10 protect plants against pests or pathogens or abiotic stressors. This includes the corresponding methods of and uses in the protection of plants and crops by application of these new plant defense elicitors or new plant elicitor compositions. This is characterized in that, an active ingredient(s) of the plant defense elicitor comprises lignin-derived oligophenoliocs or structural similar compounds with a DP 15 of 2 to 8, and yet more preferably of 2 to 4 (Fig. 2). More particularly the present invention concerns a plant defense elicitor or plant elicitor according to claim 1, characterized in that, an active ingredient of the plant defense elicitor comprises depolymerized lignin containing as active ingredient lignin-derived oligophenoliocs with a DP of 2 to 8, and yet more preferably of 2 to 4 or selected structural similar 20 compounds (Fig. 2). The invention also relate to uses of, and methods employing, depolymerized lignin or decomposed lignin oligophenolics (Fig. 1A, 1B) with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4 or structural identical oligophenolics as plant defense elicitor. Also provided are phytopharmaceutical or agrochemical 25 compositions comprising depolymerized lignin or decomposed lignin oligophenolics with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4 or structural identical oligophenolics, and applications thereof. In certain preferred embodiments, the compositions may further comprise other plant elicitors or may comprise antifungal, antimicrobials or antiviral compounds. In certain preferred embodiments, 30 the compositions may be produced by decomposition of lignin by reductive catalytic fractionation (RCF) (Fig. 1A), non-catalytic thermo-solvolytic de-polymerization (Fig. 1A) and fractioning (Fig. 1C) of the lignin source. 4
An aspect of present invention concerns a plant elicitor (elicitor of natural plant defences), for instance plant defense elicitor, characterized in that, an active ingredient of the plant defense elicitor comprises phenolics that are derived from depolymerized lignin, including lignin-derived dimeric (diphenolic), lignin-derived 5 trimeric compounds (triphenolic) and lignin derived tetrameric (tetraphenolic) compounds and engineered or synthesised structural similar compounds. More particularly the present invention concerns a plant defense elicitor according to claim 1, characterized in that, an active ingredient of the plant defense elicitor comprises depolymerized lignin containing as active ingredient lignin-derived diphenolics. 10 In accordance with the purpose of the invention, as embodied and broadly described herein, the invention is broadly drawn to a method for controlling a plant disease comprising treating a plant with a plant defense elicitor or elicitor containing phenolics that is derived from depolymerized lignin of the reductive catalytic fractioning (RCF) 15 or the non-catalytic thermos-solvolytic depolymerization process. This plant defense elicitor can be a lignin oil and fractions thereof (with an approach exemplified in figure 1C) applied, for instance at a concentration of 0.05 to 20 mg/ml , preferably 0.2 to 10 mg/ml, preferably 0.5 to 5 mg/ml, more preferably 0.8 to 1.2 mg/ml and most preferably 1 mg/mL and for instance comprising molecular mass (weight average20 mass or Mw) of 180 g/mol to 1800 g/mol, preferably between 230 g/mol to 1000 g/mol, and yet more preferably between 230 g/mol to 650 g/mol of active ingredient compounds (Fig. 2) or in case of a dry composition 0.5 to 30 wt% by dry weight of aromatic compounds, preferably 1 to 20 wt% by dry weight of aromatic compounds, more preferably 2 to 10 wt% by dry weight of aromatic compounds, wherein the 25 aromatic compound comprise at least one aromatic compound from the selected formula having the formula of: 5
and/or I) at least one aromatic compound selected from the formulae (i) and - wherein each R1, R3, and R4 is independently chosen from –H, -OH, O-CH3, a 5 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, 10 - wherein R2 is –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R5 is selected of –H, a β-O-4 linkage to an aromatic monomer or 15 aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, an end-unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, -(CH2)2CH2OH, - (CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, -CH=CHCH2OCH3, -20 (CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, - CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, -CH=CHCH2OCH(CH3)2, - 6
(CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, a carbon linkage to an aromatic monomer or aromatic oligomer; and/or II) at least one aromatic compound selected from the formulae (v) 5 and - wherein each R12, R13, R15 and R16 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic 10 monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R11 and R14 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or 15 aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, 7
and/or III) wherein at the aromatic compounds comprise at least one aromatic 8
each with at least one linkage to an aromatic monomer or aromatic oligomer and - wherein each R22, R23, R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or an aromatic oligomer, or a 4 carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R21 is independently chosen from –H, a β-O-4 linkage to an aromatic 10 monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein the 15 alkyl group is derived from the alcohol solvent of the process, wherein R27 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, end-unit selected of 20 CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, -(CH2)2CH2OH, - (CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, -CH=CHCH2OCH3, - (CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, - CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, -CH=CHCH2OCH(CH3)2, - (CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, a carbon linkage to an25 aromatic monomer or an aromatic oligomer. In accordance with the purpose of the invention, as embodied and broadly described herein, the invention is drawn to an engineered composition comprising aromatic compounds, whereby the molecular mass (weight average or Mw) of the aromatic compounds can be 180 g/mol between 230 g/mol to 1000 g/mol, and yet more 9
preferably between 230 g/mol to 650 g/mol or wherein the aromatic compounds comprise, consists essentially of or consists of lignin derived phenolics with a degree of polymerization (DP- of 2 to 8, preferably 2 to 4 or synthesized structurally similar compounds. These compounds were obtained -the reductive catalytic fractioning 5 (RCF) (Fig. 1A) or the non-catalytic thermos-solvolytic depolymerization (Fig. 1B) process of lignin, lignocellulose or biomass comprising lignocellulose or lignin. In one aspect of the invention, these defined phenolic structures are from lignin depolymerisation. Further scope of applicability of the present invention will become apparent from the 10 detailed description given hereinafter. However, it should be understood that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description. It is to be understood that both 15 the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed. The compounds, and combinations of or composition comprising the compounds as taught herein, are according to the present disclosure put to use as plant defense elicitor. This term broadly encompasses any compounds or compositions that are 20 capable of eliciting natural plant defenses, of activating plant defense and resistance reactions against plant pathogen, of stimulating the production of plant defense molecules against plant pathogen and/or of preventing, controlling or treating a plant against infection by a plant pathogen and/or against abiotic stressors, when administered to a plant, plant seed or an organ of a plant. Hence, when perceived by 25 a plant, an elicitor can evoke molecular, biochemical and/or physiological defensive plant cell reactions, such as the synthesis, or increase of the synthesis, of plant defense molecule(s), for example ethylene and/or salicylic and jasmonic acid, the production of reactive oxygen species (ROS), and/or expression of specific defense- related genes and proteins, for example polygalacturonase inhibitor proteins (PGIP). 30 The activation of signal transduction pathway(s) can subsequently lead to long- lasting defence gene expression and secondary metabolites production. As a result, a plant defense elicitor can enhance the ability of the plant to resist or battle a given pathogen. Plant defense elicitors are often subsumed by the group of agents 10
colloquially known as biopesticides, which mainly include bioinsecticides, biofimgicides, bionematicides, and others. Hence, in certain embodiments, by means of the present compounds or compositions, 5 a plant infection by a plant pathogen may be prevented, controlled or treated. While these terms are well-known as such, by means of further guidance, “preventing” may in particular mean avoiding occurrence of at least one adverse effect or symptom, preferably all adverse effects or symptoms induced by a plant pathogen infection; “controlling” may in particular mean stopping the progression of a plant pathogen 10 infection, more precisely reducing or abolishing the plant pathogen spread across the healthy parts of a plant or of an organ of a plant, or from an infected plant to another plant, typically to a neighboring plant; and “treating” may in particular mean ameliorating the symptom(s) of an infection, or completely curing an infection, typically by reducing or completely eliminating a phytopathogen (typically a fungus 15 or bacterium), i.e., by eliminating any viable phytopathogen in the plant or in an organ or several or each organ(s) of the plant. A plant may be contacted with an effective amount of the present compounds or compositions, such as contacted via an organ of the plant, preferably an organ 20 selected from leaves, roots and/or fruits, or via seeds of the plant. The contacting step may be performed once or several times (for example regularly or periodically, for example on the appropriate season or at the appropriate plant development stage). The term “an effective amount” refers to an amount of the (active) compound or compounds as taught herein which induces or elicits plant natural defense, activate 25 plant defense and resistance reaction against abiotic stressors or against plant pathogen, and/or stimulates the production of plant defenses molecules against plant pathogen resulting in obtaining a plant that is resistant to pathogen(s). The effective amount is understood to be variable, as it may be affected by many factors, including but not limited to the type of plant treated, treatment dosages and application rates, 30 method of contacting, weather and seasonal conditions experienced during the plant growing cycle, pathogen susceptibility, etc. Such variables are commonly encountered and understood by the skilled person, who may adjust the prophylactic or treatment regimen, e.g., application rate, application timings and/or frequencies, and application way. Particular suitable amounts or concentrations ranges are 35 discussed and exemplified elsewhere in this specification. The terms “organ”, “organ of a plant” or “plant’s organ” interchangeably refer to a part of a plant or to a plant propagation material. Examples of plant’s organs include, but are not limited to, 11
leaves, stems, fruits, seeds, cuttings, tubers, roots, bulbs, rhizomes, and the like. The contacting step with the plant or organ can be performed in various ways, for example by spraying, drenching, soaking, dipping, injection, through soil feeding, and any combination thereof. Alternatively, the compounds or compositions can be 5 applied on a plant or organ by supplying a volatile or vapor-based form of the compounds or compositions in the vicinity of the plant tissue and allowing the diffusion to the plant or organ through the atmosphere. The skilled person knows how to adapt the manner of administration to the particular use. 10 In the context of the present invention, the term “plant” typically designates a plant infected by or presenting a susceptibility to infection by a plant pathogen or affected by an abiotic stressor. For example, the plant may belong to the clade of Angiosperm. In certain embodiments, the plant may belong to the clade of dicots. Examples of 15 plants from the dicots clade include, but are not limited to, the Solcmcicecie family, comprising Solcinum lycopersicum (tomato), Solcinum tuberosum (potatoes), Solcinum melongenci (eggplant), Capsicum genus (pepper) and Nicotiana tabacum (tobacco); the Vitaceae family comprising the Vitis genus (grapevines); the Brassicaceae family, comprising Brassica oleracea (cabbage), Brassica rapa (turnip 20 and Chinese cabbage), mustard species and A. thaliana,' the Rosaceae family, comprising Malus pumila (apple), Pyrus species (pear), and Fragaria ananassa (strawberry); the Fabaceae family comprising legumes such as pea, bean and soybean; the Asteraceae family, comprising sunflower; Amaranthaceae family, comprising sugar beet. 25 In certain embodiments, the plant may belong to the clade of monocots. An example of plants from the monocot clade includes, but is not limited to, the Gramineae or Poaceae family, such as maize, rice, barley, or wheat. 30 Hence, in certain embodiments, the plant may be a dicot plant, preferably selected from Brassicaceae, Solanaceae or Rosacea families, such as A. thaliana, cabbage, tomato, grapevine, soybean, apple, pear, or strawberries, or wherein the plant is a monocot plant, preferably selected from Gramineae family, such as maize, rice, barley, or wheat. 35 In preferred embodiments, the plant pathogen may be a fungus or a bacterium. Hence, in such case the compositions as taught herein may also be conveniently 12
denoted as antifungal and/or an antibacterial adjuvants, i.e., products that assist in the prevention or treatment of a plant disease typically caused by fungi or bacteria. In certain embodiments, the pathogen may be a necrotrophic fungus or bacterium, 5 a hemibiotrophic fungus or bacterium, or a biotrophic fungus or bacterium. During the colonization of plant hosts, most fungal pathogens exhibit one of two modes of nutrition: biotrophy, in which nutrients are obtained from living host cells, and necrotrophy, in which nutrients are obtained from host cells which have been 10 previously killed by the fungus. A third mode of nutrition is hemibiotrophy, where the pathogen has an initial period of biotrophy followed by a period of necrotrophy. Phytopathogenic pathogens, in particular fungi, can thus be distinguished depending on their mode of nutrition: necrotrophic (e.g., Botrytis cinerea), biotrophic (e.g., Ustilago maydis) or hemibiotrophic (e.g., Colletotrichum higginsianum). In 15 particularly embodiments, the plant pathogen may be a fungus, typically a phytopathogenic fungus. The expression “phytopathogen fungus” refers to fungi pathogens that infect plant organs. Examples of phytopathogenic fungi include, but are not limited to, fungi belonging to the Ascomycetes and Basidiomycetes classes, such as, for example, fungi of the order of Helotiales, such as, for example, family 20 Sclerotiniaceae, Botrytis/ Botryotinia, such as species Botrytis cinerea,' fungi of the order of Hypocreales, such as, for example, family Nectriaceae, genus Fusarium,' fungi of the order of Uredinales, such as, for example, family Pucciniaceae, genus Puccinia,' fungi of the order of Ustilaginales, such as, for example, family Ustilaginaceae, genus Ustilago),' fungi of the order of Sordariomycetes, such as, for 25 example, family Glomerellaceae, genus Colletotrichum. In further embodiments, the plant pathogen may be a bacterium, typically a phytopathogenic bacterium. The expression “phytopathogen bacterium” refers to bacterial pathogens that infect plant organs. Examples of phytopathogenic bacteria 30 include, but are not limited to, bacteria of the order of Pseudomonadales, such as, for example, family Pseudomonadaceae, genus Pseudomonas, such as species Pseudomonas syringae,' bacteria of the order of Burkholderiales, such as, for example, family Burkholderiaceae, genus Ralstonia, such as species Ralstonia solanacearum,' bacteria of the order of Enterobacterales, such as, for example, family 35 Erwiniaceae, genus Erwinia, such as species Erwinia amylovora, or family Pectobacteriaceae, genus 13
Pectobacterium, such as species Pectobacterium carotovorum (formerly Erwinia carotovora, bacteria of the order of Xanthomonadales, such as, for example, family Xanthomonadaceae, genus Xylella, such as species Xylella fastidiosa, or genus Xanthomonas, such as species Xanthomonas campestris. Similarly to the fungi and 5 based on the type of plant colonisation, bacteria can also be classified into necrotrophic, biotrophic and hemibiotrophic sub-classes. Particularly preferred may be necrotrophic fungi, preferably as Botrytis cinerea. Hence, in certain embodiments, the plant pathogen is a fungus or a bacterium, such 10 as a necrotrophic fungus or bacterium, such as Botrytis cinerea, a hemibiotrophic fungus or bacterium, or a biotrophic fungus or bacterium, such as Pseudomonas syringae. In the present context, the plant infection by a plant pathogen typically designates a 15 plant infection by at least one phytopathogen. The infection can occur on any organ of the plant. By means of examples and without limitation, the plant infection may be, e.g., a B. cinerea infection, for example a B. cinerea infection of A. thaliana, tomato, strawberry, sunflower, grapevine, or apple; a C. higginsianum infection, for example a C. higginsianum infection of turnip, Chinese cabbage, mustard, A. 20 thaliana, or apple; a U. maydis infection, for example a U. maydis infection of maize; a R. solanacearum infection, for example a R solanacearum infection of tomato, potatoes, eggplant,pepper, or tobacco; a Pseudomonas syringae infection, for example a Pseudomonas syringae infection on apple or pear; or any combination thereof, such as a B. cinerea and/or C. higginsianum infection of A. thaliana, an apple 25 infection by B. cinerea and/or C. higginsianum or a tomato infection by B. cinerea and/ or R. solanacearum. In certain embodiments, the concentration of one or more oligophenolics with a 30 degree of polymerization (DP) of 2 to 8, preferably of 2 to 4 (Fig. 2) from depolymerized lignin or decomposed lignin or from structural identical oligophenolics (e.g., the structures and the structures in a (phytopharmaceutical or agrochemical) composition as taught herein) is 180 g/mol between 230 g/mol to 1000 g/mol, and yet more preferably between 230 g/mol to 650 g/mol 35 In view of the foregoing discussion, the present application also provides aspects and embodiments as set forth in the following statements (1’ to 27’): 14
1. Statement 1’ A phytopharmaceutical or agrochemical composition, wherein an effective dose of plant defense elicitor compounds that are lignin-derived phenolic oligomers with a degree of polymerization (DP) of 2 to 8, and yet more preferably of 2 to 4, or synthesized structural similar compounds. 5 2. Statement 2’ A phytopharmaceutical or agrochemical composition according to statement 1’, wherein the composition has a pH in the range of 4 to 10, preferably in the range of 5 to 8. 3. Statement 3’ The phytopharmaceutical or agrochemical composition according to any one of the statements 1’ to 2’, wherein the lignin-derived 10 phenolic oligomers have a pH in the range of 4.0 to 6.0 in origin. 4. Statement 4’ The composition according to any one of the statements 1’ to 3’, whereby defense elicitor compounds are 0,5 to 30 wt% by dry weight of aromatic compounds, preferably 1 to 20 wt% by dry weight of aromatic compounds, more preferably 2 to 10 wt% of the composition in dry state. 15 5. Statement 5’ The composition according to any one of the statements 1’ to 4’, whereby defense elicitor compounds are of the group of lignin-derived dimerics (diphenolic) compounds, lignin-derived trimeric (triphenolic) compounds, lignin-derived tetrameric (tetraphenolic) compound or a combination thereof (Fig. 2). 20 6. Statement 6’ The composition according to any one of the statements 1’ to 5’, whereby lignin-derived phenolic oligomers are from depolymerized lignin or decomposed lignin. 7. Statement 7’ The composition according to any one of the statements 1’ to 6’, whereby lignin-derived phenolic oligomers are from a lignin that is 25 depolymerized or decomposed by non-catalytic thermo-solvolytic depolymerisation (Fig. 1B), reductive catalytic fractionation (RCF) (Fig. 1A) of lignocellulose. 8. Statement 8’ The composition according to any one of the statements 1’ to 7’, whereby the lignin is from a lignocellulose biomass, for instance flax shives,30 wood chips, pine, spruce or poplar sawdust. 9. Statement 9’ The composition according to any one of the statements 1’ to 8’, further comprising an ingredient of the group consisting of a surfactant, a biosurfactant, a penetration enhancer, a dispersing agent, an emulsifier and a carrier or a combination thereof. 35 10.Statement 10’ The composition according to any one of the statements 1’ to 9’, further comprising a second plant defense elicitor, for instance of the group consisting of alginate, hexokinase, laminarin, sodium silicate, silicon, oligo- 15
galacturonan, cellodextrin and/or chito-oligosaccharide or a plant defense elicitor selected from the group consisting of pectin fragment, oligogalacturonide, cellobiose, xyloglucan, non-branched P-l,3-glucan, chitin fragment, arabinose, arabinan, rhamnose, homogalacturonan, 5 rhamnogalacturonan I and II, xylogalacturonan, starch, and combinations thereof. 11.Statement 11’ The composition according to any one of the statements 1’ to 10’, further comprising an fungicide, antimicrobial, an insecticidal, or an antiviral. 10 12.Statement 12’ The composition according to any one of the statements 1’ to 11’, further comprising a fungicide selected form the group consisting of phosphonates, benzamides, carbamates, dithiocarbamates , phtalimides, triazoles, quinolines, sulphur, and cyanoimidazoles or a fungicide selected from the group consisting of 2,6-dichloro-N-[3-chloro-5-(trifluoromethyl)-2- 15 pyridinylmethyl]benzamide; propyl 3-(dimethylamino)propylcarbamate hydrochloride; (2RS,3SR)-1-[3-(2-chlorophenyl)-2,3-epoxy-2-(4- fluorophenyl)propyl]-1H-1,2,4-triazole; 5,7-dichloro-4-quinolyl 4- fluorophenyl ether; sulphur; 4-chloro-2-cyano-N,N-dimethyl-5-(4- methylphenyl)-1H-imidazole-1-sulfonamide; N- 20 (trichloromethylthio)phthalimide; manganese ethylenebis(dithiocarbamate) (polymeric) complex with zinc salt and methyl (E)-methoxyimino-{(E)-α-[1- (α,α,α-trifluoro-m-tolyl)ethylideneaminooxy]-o-tolyl}acetate; or a combination thereof. 13.Statement 13’ The composition according to any one of the statements 1’ to 25 12’, further comprising a polymerization inhibitor of the group of a carbocation scavenger, aromatic scavengers and a polyhydric alcohol or a combination thereof. 14.Statement 14’ The composition according to any one of the statements 1 to 12, further comprising a polymerization inhibitor whereby the polymerization30 inhibitor is a compound of the group consisting of citric acid, salicylic acid, 2- Naphthol, phenolic acids (e.g., vanillic acid, syringic acid), ethylene glycol, glycerol, Mannitol (C6H14O6), Sorbitol (C6H14O6), Xylitol (C5H12O5), Erythritol, Maltitol (C12H24O11). 15.Statement 15’ The composition according to any one of the statements 1’ to35 14’, further comprising a stabilizing agents of the group consisting of 1,4- butanediol, 2-hydroxy-1-naphthoic acid, 2-naphthol, 2-naphthol-7-sulfonat, 2-naththol, 3-hydroxy-2-naphthoic acid, 4-hydroxybenzoic acid, 6-hydroxy- 16
2-naphthoic acid, ascorbic acid, bovine serum albumin, citric acid, salicylic acid, cysteine, dimethylphloroglucinol, histidine, mannitol, o- dihydroxybenzene, p-benzenediol, phloroglucinol, resorcinol, soy protein isolate, syringic acid and vanillic acid so to prevent repolymerization. 5 16.Statement 16’ The composition according to any one of the statements 1 to 15, wherein the phenolic oligomers comprise one or more aromatic compounds, with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4 (Fig. 1) or one or more aromatic compounds, with a molecular mass 180 g/molbetween 230 g/mol to 1000 g/mol, and yet more preferably between 10 230 g/mol to 650 g/mol (Fig. 1), and wherein the aromatic compounds comprise and/or I) at least one aromatic compound selected from the - wherein each R1, R3, and R4 is independently chosen from –H, -OH, O-CH3, a 15 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, 20 - wherein R2 is –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an 17
α-O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R5 is selected of –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, an end-unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, -(CH2)2CH2OH, - (CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, -CH=CHCH2OCH3, - (CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, - CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, -CH=CHCH2OCH(CH3)2, - (CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, a carbon linkage to an aromatic monomer or aromatic oligomer; - and/or II) at least one aromatic compound selected from the formulae (v) - wherein each R12, R13, R15 and R16 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic 18
monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R11 and R14 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic 5 monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, and/or III) wherein at the aromatic compounds comprise at least one aromatic compound selected from the formula (viii) 10 , 19
linkage to an aromatic monomer or aromatic oligomer and - wherein each R22, R23, R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or an aromatic oligomer, or a 4 carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R21 is independently chosen from –H, a β-O-4 linkage to an aromatic 10 monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein the 15 alkyl group is derived from the alcohol solvent of the process, wherein R27 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, end-unit selected of20 CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, -(CH2)2CH2OH, - (CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, -CH=CHCH2OCH3, - (CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, - CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, -CH=CHCH2OCH(CH3)2, - (CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, a carbon linkage to an25 aromatic monomer or an aromatic oligomer. 17. Statement 17’ The composition according to any one of the statements 1 to 16, wherein the phenolic oligomers are diphenolics comprising compounds wherein the aromatic compounds have 2 aromatic groups or a DP of 2 (Fig. 2). 18.Statement 18’ The composition according to any one of the statements 1’ to 30 17’, for promoting induced systemic resistance in a plant or for inducing 20
latent host defenses in a plant by priming of the intrinsic disease resistance mechanisms of a plant. 19.Statement 19’ Use of the composition according any one of the statements 1’ to 18’, on a plant or parts thereof whereby the latent host defenses are 5 activated preventive to or in the event of an attack by a phytopathogenic pathogen or pest. 20.Statement 20’ Use of the composition according any one of the statements 1’ to 18’, on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of an attack by a phytopathogenic 10 pathogen or pest whereby the phytopathogenic pathogen or pest is selected from the group of a fungi, a bacteria, and insect. 21.Statement 21’ Use of the composition according any one of the statements 1’ to 18’, on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of an attack by a phytopathogenic of 15 the group consisting of fungi, bacteria, viruses, viroids, mycoplasma-like organisms, protozoa, insects, acari, and nematodes. 22.Statement 22’ Use of the composition according any one of the statements 1’ to 18’, on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of abiotic stress. 20 23.Statement 23’ Use of the composition according any one of the statements 1’ to 18’, on a plant or parts thereof to prime the intrinsic resistance mechanisms of a plant for stronger or faster induced plant defense preventive to or in the event of an attack by a phytopathogenic pathogen or pest or of abiotic stress, as compared to control or other plant defense inducers. 25 24.Statement 24’ Use of the composition according any one of the statements 1’ to 15’, whereby the composition is a foliar spray agent. 25. Statement 25’ Use of the composition according any one of the statements 1’ to 21’, whereby the composition is a root drench. 26.Statement 26’ A method for promoting induced systemic resistance, for 30 inducing latent host defenses or for priming the intrinsic resistance mechanisms in a plant, comprising applying to a plant or a plant part, the composition according to any one of the statements 1’ to 18’. 27.Statement 27’ A method for promoting induced systemic resistance , for inducing latent host defenses or for priming the intrinsic resistance 35 mechanisms in a plant, comprising by spraying on said plant or contacting the roots of said plant with the composition according to any one of the statements 1’ to 21’. 21
In view of the foregoing discussion, the present application also provides aspects and embodiments as set forth in the following statements (1* to 17*): 1. Statement 1* A composition for protecting plants against plant pests 5 comprising a plant defense elicitor comprising, consisting of or essentially consisting of as active ingredient one or more aromatic compounds, with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4 or one or more aromatic compounds, with a molecular mass between 180 g/molbetween 230 g/mol to 1000 g/mol, and yet more preferably between 10 230 g/mol to 650 g/mol (Fig. 2) , and wherein the aromatic compounds comprise and/or I) at least one aromatic compound selected from the - wherein each R1, R3, and R4 is independently chosen from –H, -OH, O-CH3, a 15 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, 20 - wherein R2 is –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an 22
α-O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R5 is selected of –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic 5 oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, an end-unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, -(CH2)2CH2OH, - (CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, -CH=CHCH2OCH3, - (CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, -10 CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, -CH=CHCH2OCH(CH3)2, - (CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, and/or II) the aromatic compounds comprise at least one aromatic compound selected from the formulae (v) , (vi) 15 - wherein each R12, R13, R15 and R16 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic 23
monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R11 and R14 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic 5 monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, and/or wherein at the aromatic compounds comprise at least one aromatic 10 , 24
each with at least one linkage to an aromatic monomer or aromatic oligomer and - wherein each R22, R23, R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or an aromatic oligomer, or a 4 carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R21 is independently chosen from –H, a β-O-4 linkage to an aromatic 10 monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein the 15 alkyl group is derived from the alcohol solvent of the process, - wherein R27 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, end-20 unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, - (CH2)2CH2OH, -(CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, - CH=CHCH2OCH3, -(CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, - (CH2)2CH2O(CH2)2CH3, -CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, - CH=CHCH2OCH(CH3)2, -(CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, a25 carbon linkage to an aromatic monomer or an aromatic oligomer. 2. Statement 2* The composition according to statement 1*, whereby the one or more aromatic compound active ingredient are dimers, trimers and/or tetramers. 3. Statement 3* The composition according to any one of the statements 1* 30 to 2*, whereby the one or more aromatic compound active ingredient are 25
at a concentration of 0.05 to 20 mg/ml , preferably 0.2 to 10 mg/ml, preferably 0.5 to 5 mg/ml, more preferably 0.8 to 1.2 mg/ml and most preferably 1 mg/mL or in case of a dry composition at 0,5 to 30 wt% by dry weight of aromatic compounds, preferably 1 to 20 wt% by dry weight 5 of aromatic compounds, more preferably 2 to 10 wt% by dry weight of aromatic compounds. 4. Statement 4* The composition according to any one of the statements 1* to 3*, further comprising a further plant defense elicitor such as alginate, hexokinase, laminarin, sodium silicate, silicon, oligo-galacturonan, 10 cellodextrin and/or chito-oligosaccharide or a plant defense elicitor selected from the group consisting of pectin fragment, oligogalacturonide, cellobiose, xyloglucan, non-branched P-l,3-glucan, chitin fragment, arabinose, arabinan, rhamnose, homogalacturonan, rhamnogalacturonan I and II, xylogalacturonan, starch, and combinations thereof. 15 5. Statement 5* The composition according to any one of the statement 1* to 4*, comprising a) the plant defense elicitor according to statement 1* and b) a fungicide, for instance a fungicide selected form the group consisting of selected from the group comprising: phosphonates, benzamides, carbamates, dithiocarbamates , phtalimides, triazoles, 20 quinolines, sulphur, and cyanoimidazoles or a fungicide selected from the group consisting of 2,6-dichloro-N-[3-chloro-5-(trifluoromethyl)-2- pyridinylmethyl]benzamide; propyl 3-(dimethylamino)propylcarbamate hydrochloride; (2RS,3SR)-1-[3-(2-chlorophenyl)-2,3-epoxy-2-(4- fluorophenyl)propyl]-1H-1,2,4-triazole; 5,7-dichloro-4-quinolyl 4-25 fluorophenyl ether; sulphur; 4-chloro-2-cyano-N,N-dimethyl-5-(4- methylphenyl)-1H-imidazole-1-sulfonamide; N- (trichloromethylthio)phthalimide; manganese ethylenebis(dithiocarbamate) (polymeric) complex with zinc salt and methyl (E)-methoxyimino-{(E)-α-[1-(α,α,α-trifluoro-m-30 tolyl)ethylideneaminooxy]-o-tolyl}acetate; or a combination thereof 6. Statement 6* The composition according to any one of the statement 1* to 5*, comprising a) the plant defense elicitor according to statement 1* and b) a salt and/or sugar. 7. Statement 7* The composition according to any one of the statement 1* 35 to 6*, further comprising a co-formulant selected from the group comprising: detergents, emulsifiers, dispersing agents, anti-foaming agents, penetration enhancers, humectants, wetting agents of ionic or 26
non-ionic type, anti-freeze agents, preservative agents, absorbent agents, thickeners, buffers, sticker agents, diluents or a mixture thereof, preferably a surfactant selected from the group comprising: detergents, emulsifiers, dispersing agents, anti-foaming agents, penetration 5 enhancers, humectants or wetting agents of ionic or non-ionic type, or a mixture thereof. 8. Statement 8* The composition according to any one of the statement 1* to 7*, further comprising a surfactant comprising one or more of the following components: castor oil ethoxylate, rapeseed methyl ester, alkyl 10 phosphates, tributyl phosphate, tripropyl phosphate, naphthalenesulphonic acid salts, a combination of organic sulfonate and 2-methylpentane-2,4-diol, alkylpolyglucoside, siloxanes derivates, alkylsulfonates, polycarboxylates, lignosulfonates, alkoxylated triglycerides, fatty amines polymers, dioctylsulfosuccinates or15 polyoxyethylene (20) sorbitan monolaurate, preferably C18-castor-oil- ethoxylate, a combination of organic sulfonate and 2-methylpentane-2,4- diol, or polyoxyethylene (20) sorbitan monolaurate. 9. Statement 9* The composition according to any one of the statement 1* to 8*, whereby the composition has a pH in the range of 4 to 10, preferably in 20 the range of 5 to 8. 10.Statement 10* The composition according to any one of the statement 1* to 9*, whereby the ingredients have in origin a pH in the range of 4.0 to 6.0. 11.Statement 11* Use of the composition according to any of statements 1* to 10* in agricultural applications or to protect plants against plant pests. 25 12.Statement 12* Use according to statement 11*, wherein said plant pests are selected from the group comprising: phytopestic fungi, oomycetes, bacteria, viruses, nematodes and insects. 13.Statement 13* Use of the composition according to any of statements 1* to 10*, to enhance the efficacy of said fungicide in said composition, or to30 stimulate the plant immune system. 14.Statement 14* A method for protecting plants against plant pests, comprising applying an effective and substantially non-phytotoxic amount of the composition according to any of statements 1* to 10* to said plants. 15.Statement 15* The method according to statement 12*, wherein said 35 plant pests are selected from the group comprising: fungi, oomycetes, bacteria, viruses, nematodes and insects, 27
16.Statement 16* The method according to any of statements 14* to 114*, wherein the composition is applied before harvest or post-harvest to the whole plant, the leaves, the flowers, fruits, seeds, seedlings or seedlings pricking out, propagation material such as tubers or rhizomes, plants 5 pricking out, and/or to the soil or inert substrate wherein the plant is growing or in which it is desired to grow, by spraying, drenching, soaking, dipping, injection or administration through fertilising or irrigation systems. 17.Statement 17* The method according to anyone of statements 14* to 16*, 10 wherein said plant is selected from the group comprising: cotton, flax, vine, fruit, vegetable, major horticultural and forest crops such as: Rosaceae sp., Ribesioidae sp., Juglandaceae sp., Betulaceae sp., Anacardiaceae sp., Fagaceae sp., Moraceae sp., Oleaceae sp., Actinidaceae sp., Lauraceae sp., Musaceae sp., Rubiaceae sp., Theaceae 15 sp., Sterculiceae sp., Rutaceae sp., Solanaceae sp., Vitaceae sp., Liliaceae sp., Asteraceae sp., Umbelliferae sp., Cruciferae sp., Chenopodiaceae sp., Cucurbitaceae sp., Papilionaceae sp., such as Graminae sp., Fabacae sp.. The present application also provides aspects and embodiments as set forth in 20 the following statements (1” – 31”) : 1. A plant defense elicitor, characterized in that, the resistance inducing or defense eliciting active ingredient thereof comprises an effective dose of an aromatic compounds with a degree of polymerization (DP) of 2 to 8, 25 preferably of 2 to 4 or a molecular mass between 180 g/mol to 1800 g/mol, preferably between 230 g/mol to 1000 g/mol, and yet more preferably between 230 g/mol to 650 g/mol (Fig. 2), and is an aromatic compound and wherein and/or I) the aromatic compounds comprise at least one of the aromatic compounds from the formulae (i) 28
and (iv) and - wherein each R1, R3, and R4 is independently chosen from –H, -OH, O-CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to 5 an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R2 is –H, a β-O-4 linkage to an aromatic monomer or aromatic 10 oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R5 is selected of –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic 15 oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, an end-unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, -(CH2)2CH2OH, - (CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, -CH=CHCH2OCH3, - (CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, -20 CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, -CH=CHCH2OCH(CH3)2, - (CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, 29
- and/or II) the aromatic compounds comprise at least one aromatic compound - wherein each R12, R13, R15 and R16 is independently chosen from –H, -OH, O- 5 CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, 10 - wherein R11 and R14 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, and/or III) wherein at the aromatic compounds comprise 15 at least one aromatic compound selected from the formula (viii) 30
linkage to an aromatic monomer or aromatic oligomer and 31
- wherein each R22, R23, R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic 5 monomer or an aromatic oligomer, or a 4 carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R21 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic 10 oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein the alkyl group is derived from the alcohol solvent of the process, - wherein R27 is independently chosen from –H, a β-O-4 linkage to an aromatic 15 monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, end- unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, - (CH2)2CH2OH, -(CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, -20 CH=CHCH2OCH3, -(CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, - (CH2)2CH2O(CH2)2CH3, -CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, - CH=CHCH2OCH(CH3)2, -(CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, a carbon linkage to an aromatic monomer or an aromatic oligomer. 2. Statement 2”: The plant defense elicitor according to statement 1”, 25 characterized in that, the resistance inducing or defence eliciting active ingredient is extracted or derived from depolymerized lignin. 3. Statement 3”: The plant elicitor according to statement 1” or 2”, characterized in that the resistance inducing or defense eliciting active ingredients are diphenolics, triphenolics and or tetraphenolics comprising 30 compounds wherein the aromatic compounds have 2, 3 or 4 aromatic groups or a DP of 2, 3 or 4, respectively (Fig. 1). 4. Statement 4”: The plant defense elicitor according to any one of the statements 1” to 3”, whereby the resistance inducing or defence eliciting active ingredients are are diphenolics comprising compounds wherein the 35 aromatic compounds have 2 aromatic groups or a DP of 2 (Fig. 2). 5. Statement 5” The plant defense elicitor, with in a dry composition a content of at least 0.5 % by dry weight of the resistance inducing or 32
defense eliciting active ingredient according to any one of the statement 1” to 4”. 6. Statement 6” The plant defense elicitor, with in a dry composition a content of at least 2% by dry weight of the resistance inducing or defense 5 eliciting active ingredient according to any one of the statement 1” to 4”. 7. Statement 7” The plant defense elicitor, with in a dry composition a content of at least 5% the resistance inducing or defense eliciting active ingredient according to any one of the statement 1” to 4”. 8. Statement 8” The plant pest resistance inducer or plant defense elicitor 10 according to any one of the statement 1” to 6”, whereby the resistance inducing or defense eliciting active ingredient extracted from depolymerized lignin is from a lignocellulose or lignin source. 9. Statement 9” The plant defense elicitor according to any one of the statements 1” to 8”, comprising further a repolymerization inhibitor of the 15 group of a carbocation scavenger, aromatic scavengers and a polyhydric alcohol or a combination thereof. 10.Statement 10” The plant defense elicitor according to any one of the statement 1” to 9”, comprising a repolymerization inhibitor whereby the repolymerization inhibitor is a compound of the group consisting of citric 20 acid, salicylic acid, 2-Naphthol, phenolic acids (e.g., vanillic acid, syringic acid), ethylene glycol, glycerol, Mannitol (C6H14O6), Sorbitol (C6H14O6), Xylitol (C5H12O5), Erythritol, Maltitol (C12H24O11). 11.Statement 11” The plant defense elicitor according to any one of the statement 1” to 9”, with a stabilizing agents of the group consisting of25 1,4-butanediol, 2-hydroxy-1-naphthoic acid, 2-naphthol, 2-naphthol-7- sulfonat, 2-naththol, 3-hydroxy-2-naphthoic acid, 4-hydroxybenzoic acid, 6-hydroxy-2-naphthoic acid, ascorbic acid, bovine serum albumin, citric acid, salicylic acid, cysteine, dimethylphloroglucinol, histidine, mannitol, o-dihydroxybenzene, p-benzenediol, phloroglucinol, resorcinol, soy 30 protein isolate, syringic acid and vanillic acid so to prevent repolymerisation. 12.Statement 12” The plant defense elicitor according to any one of the statements 1” to 11”, further comprising a surfactant. 13.Statement 13” The plant defense elicitor according to statement 12”,35 whereby the surfactant is of the group of a lecithin (such as lyso- lecithin), a cyclic lipopeptides biosurfactant (such as surfactin or 33
pseudofactin), a polar aprotic solvent (such as dimethyl sulfoxide, DMSO or a natural polar aprotic solvent of the group consisting of δ- Valerolactone (DVL), Cyrene, (Dihydrolevoglucosenone); Ethyl Acetate, Methyl Lactate, Acetone (Dimethyl Ketone), Limonene Oxide ) and a 5 surface-active agent such as surface active glycolipid or sophorolipid. 14.Statement 14” The plant defense elicitor according to any one of the statements 1” to 13”, further comprising a metal such as zinc and/or copper. 15.Statement 15” The plant defense elicitor according to any one of the 10 statement 1 to 14”, whereby the phenolics that are derived from depolymerized lignin are the reaction product of a reductive catalytic fractionation (RCF) of the lignin source. 16.Statement 16” The plant defense elicitor according to any one of the statement 1” to 15”, whereby the phenolics that are derived from15 depolymerized lignin are the reaction product of a non-catalytic thermo- solvolytic de-polymerization and fractioning of the lignin source. 17.Statement 17” The plant defense elicitor according to any one of the statement 1” to 15”, whereby the phenolics that are derived from depolymerized lignin and whereby the depolymerized lignin is the reaction20 product of a solvolytic lignin depolymerization of the lignin source. 18.Statement 18” The plant defense elicitor according to any one of the statements 1” to 17”, for promoting induced systemic resistance in a plant or for inducing latent host defenses in a plant by priming of the intrinsic disease resistance mechanisms of a plant. 25 19.Statement 19” The plant defense elicitor according to any one of the statements 1” to 18” , characterized in that it has a pH in the range of 4 to 10, preferably in the range of 5 to 8. 20.Statement 20” The plant defense elicitor according to any one of the statements 1” to 18” , wherein the aromatic compounds are in origin an30 oil with a pH in the range of 4.0 to 6.0 in origin. 21.Statement 21” Use of the plant defense elicitor according any one of the statements 1” to 20”, on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of an attack by a phytopathogenic pathogen or pest or by an abiotic stressor. 35 22.Statement 22” Use of the plant defense elicitor according any one of the statements 1” to 20”, on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of an attack by a 34
phytopathogenic pathogen or pest whereby the phytopathogenic pathogen or pest is selected from the group of a fungi, a bacteria, and insect. 23.Statement 23” Use of the plant defense elicitor according any one of the statements 1” to 20”, on a plant or parts thereof whereby the latent host 5 defenses are activated preventive to or in the event of an attack by a phytopathogenic of the group consisting of fungi, bacteria, viruses, viroids, mycoplasma-like organisms, protozoa, insects, acari, and nematodes. 24.Statement 24” Use of the plant defense elicitor according any one of the statements 1” to 20”, on a plant or parts thereof whereby the latent host10 defenses are activated preventive to or in the event of abiotic stress. 25.Statement 25” Use of the plant defense elicitor according any one of the statements 1” to 20”, on a plant or parts thereof to prime the intrinsic resistance mechanisms of a plant for stronger or faster induced plant defense preventive to or in the event of an attack by a phytopathogenic 15 pathogen or pest or of abiotic stress, as compared to control or other plant defense inducers. 26.Statement 26” Use of the plant defense elicitor according any one of the statements 1” to 20”, whereby the plant resistance inducer is a foliar spray agent. 20 27.Statement 25” Use of the plant defense elicitor according any one of the statements 1” to 20”, whereby the plant resistance inducer is a root drench. 28.Statement 28” A method for promoting induced systemic resistance in a plant, for inducing latent host defenses of a plant or for priming the 25 intrinsic resistance mechanisms in a plant, comprising applying to a plant or a plant part, a composition comprising the plant defense elicitor according to any one of the statements 1 to 20. 29.Statement 29” A method for promoting induced systemic resistance of a plant, for inducing latent host defenses of a plant or for priming the 30 intrinsic resistance mechanisms in a plant, comprising by spraying on said plant or contacting the roots of said plant with the composition comprising the plant defense elicitoraccording to any one of the statements 1” to 20”. 30.Statement 30” Any one of the statements according to 1” to 29”, whereby lignin-derived diphenolics are the reaction product of the lignin 35 depolymerization or 1) by a reductive catalytic fractionation (RCF) (Fig. 1A) of the lignin source with a heterogenous metal catalyst, including but not limited to Ru, Pd and Ni, on a support in an organic solvent or an 35
organic solvent water mixture in a temperature range of 100°C to 300°C, preferably 150°C to 270°C and most preferably 200°C to 250°C and containing a hydrogen donor, including but not limited to H2 or 2) by a non-catalytic thermo-solvolytic de-polymerization (Fig. 1B) of the lignin 5 source in an organic solvent or an organic solvent water mixture in a temperature range of 100°C to 300°C, preferably 150°C to 270°C and most preferably 200°C to 250°C and under an inert atmosphere. 31.Statement 31”A method for controlling a plant disease comprising treating a plant with a plant defense elicitor according to any one of the statements 10 1” to 29” comprising 0.05 to 20 mg/ml , preferably 0.2 to 10 mg/ml, preferably 0.5 to 5 mg/ml, more preferably 0.8 to 1.2 mg/ml and most preferably 1 mg/mL of active ingredient aromatic compounds or in case of a dry composition 0,5 to 30 wt% by dry weight of aromatic compounds, preferably 1 to 20 wt% by dry weight of aromatic compounds, more 15 preferably 2 to 10 wt% by dry weight of resistance inducing or defense eliciting active ingredient. In view of the foregoing discussion, the present application also provides aspects and embodiments as set forth in the following statements (1° to 20 15°): 1. Statement 1° A plant defence elicitor comprising one or more lignin oligomers with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4. 25 2. Statement 2° The elicitor according to statement 1°, characterized in that the lignin oligomers have a pH in the range of 5.0 to 8.0. 3. Statement 3° The elicitor according to any one of the statements 1° to 2°, 30 whereby the lignin-oligomers comprising, or essentially consisting of or consisting of I) at least one aromatic compound selected from the formulae 36
and (iv) and - wherein each R1, R3, and R4 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R2 is –H, a β-O-4 linkage to an aromatic monomer or aromatic 10 oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R5 is selected of –H, a β-O-4 linkage to an aromatic monomer 15 or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, an end-unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, - CH=CHCH3, -(CH2)2CH2OH, -(CH2)2CHO, -CH=CHCH2OH, -20 (CH2)2CH2OCH3, -CH=CHCH2OCH3, -(CH2)2CH2OCH2CH3, - CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, -CH=CHCH2O(CH2)2CH3, 37
-(CH2)2CH2OCH(CH3)2, -CH=CHCH2OCH(CH3)2, - (CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, a carbon linkage to an aromatic monomer or aromatic oligomer; and/or II) wherein the aromatic compounds comprise at least one aromatic 5 compound selected from the formulae (v) , (vi) - wherein each R12, R13, R15 and R16 is independently chosen from –H, - OH, O-CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, 10 a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R11 and R14 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an 15 aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, 38
and/or III) wherein at the aromatic compounds comprise at least one aromatic compound selected from the formula (viii) , (xi) (xviii) 39
each with at least one linkage to an aromatic monomer or aromatic oligomer and - wherein R21 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an 5 aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein each R22 and R23 is dependently chosen from -H, -OH, O-CH3 or from a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 10 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or an aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein the alkyl group is derived from the alcohol solvent of the process,15 - wherein each R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or an aromatic oligomer, or a carbon- 20 oxygen linkage to an aromatic monomer or aromatic oligomer, wherein R27 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, end-25 unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, - (CH2)2CH2OH, -(CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, - CH=CHCH2OCH3, -(CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, - (CH2)2CH2O(CH2)2CH3, -CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, - CH=CHCH2OCH(CH3)2, -(CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, a 30 carbon linkage to an aromatic monomer or an aromatic oligomer. 4. Statement 4° The elicitor according to any one of the statements 1° to 3°, whereby the lignin-oligomers comprising, or essentially consisting of or consisting of I) at least one aromatic compound selected from the formulae 40
and (iv) and - wherein each R1, R3, and R4 is independently chosen from –H, -OH, O- CH3, 5 - wherein R2 is –H, - wherein R5 is selected of –H, an end-unit selected of CH3, -CH2CH3, - (CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, -(CH2)2CH2OH, -(CH2)2CHO, - CH=CHCH2OH, -(CH2)2CH2OCH3, -CH=CHCH2OCH3, - (CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, -10 CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, - CH=CHCH2OCH(CH3)2, -(CH2)2CH2O(CH2)3CH3, - CH=CHCH2O(CH2)3CH3, - and/or II) wherein the aromatic compounds comprise at least one aromatic compound selected from the formulae (v) 41
- wherein each R12, R13, R15 and R16 is independently chosen from –H, - OH, O-CH3, 5 - wherein R11 and R14 is independently chosen from –H, and/or III) wherein at the aromatic compounds comprise at least one aromatic compound selected from the formula (viii) 42
and (xiv) each with at least one linkage to an aromatic monomer or aromatic oligomer and - wherein R21 is independently chosen from –H, a β-O-4 linkage to an 5 aromatic monomer, a 4-O-5 linkage to an aromatic monomer, an α-O- 4 linkage to an aromatic monomer or a carbon-oxygen linkage to an aromatic monomer, - wherein each R22 and R23 is dependently chosen from -H, -OH, O-CH3 or where at least R22 or R23 is independently chosen from a 4-O-5 10 linkage to an aromatic monomer, a 5-5 linkage to an aromatic monomer, a β-5 linkage to an aromatic monomer, a carbon linkage to an aromatic monomer, or a carbon-oxygen linkage to an aromatic monomer, - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein 15 the alkyl group is derived from the alcohol solvent of the process, - wherein each R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer, a 5-5 linkage to an aromatic monomer, a β-5 linkage to an aromatic monomer, a carbon linkage to an aromatic monomer, or a carbon-oxygen linkage to an 20 aromatic monomer, - wherein R27 is independently chosen from –H, end-unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, -(CH2)2CH2OH, - (CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, -CH=CHCH2OCH3, - 44
(CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, - CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, - CH=CHCH2OCH(CH3)2, -(CH2)2CH2O(CH2)3CH3, - CH=CHCH2O(CH2)3CH3. 5. Statement 5° The elicitor according to any one of the statements 1° to 4°, whereby the lignin-oligomers comprising, or essentially consisting of or consisting of at least one aromatic compound selected from the formulae (i) 45
or a combination thereof. 6. Statement 6° The elicitor according to any one of the statements 1° to 5°, 5 wherein the lignin-oligomers are phenolic oligomers comprising two benzene rings directly bridged or bridged with a common bridging group of the group 47
consisting of aliphatic chains, alkene groups, carbonyl groups and ether linkages or wherein the phenolic oligomers have two aromatic groups. 5 7. Statement 7° The elicitor according to any one of the statements 1° to 6°, wherein the lignin-oligomers are phenolics oligomers comprising two benzene rings directly bridged or bridged with a common bridging group of the group consisting of -CH₂- groups, -CH=CH-, -C(=O)-and -O-. 10 8. Statement 8° The elicitor according to any one of the statements 1° to 7°, wherein the one or more lignin oligomers is present in a concentration of 50% to 90% by weight of the elicitor. 15 9. Statement 9° The elicitor according to any one of the statements 1° to 8°, characterized in that it contains less than 0,1%, of each of acetic acid, methanol and ethanol. 20 10.Statement 10° The elicitor according to any one of the statements 1° to 9°, comprised in phytopharmaceutical or agrochemical composition, whereby the dry weight of the composition contains between 0,5 to 30 wt% by dry weight of lignin-oligomers, preferably 1 to 20 wt% by dry weight of lignin-oligomers, more preferably 2 to 10 wt% by dry weight of lignin-oligomers. 25 11.Statement 11° The elicitor according to any one of the statements 1° to 10°, comprised in phytopharmaceutical or agrochemical composition, further comprising one or more one or more anionic, non-ionic, amphoteric, or cationic surfactant, or a combination thereof. 30 12.Statement 12° The elicitor according to any one of the statements 1° to 11°, comprised in phytopharmaceutical or agrochemical composition, further comprising a lignin oligomer solubilizing agent, preferably one or more polar aprotic solvent. 35 13. Statement 13° A method for producing phytopharmaceutical or agrochemical composition according to any one of the statements 10° to 12°, the method comprising a. Subjecting lignin or lignocellulose in a liquid phase to solvolytic 48
lignin depolymerisation, b. Fractionating the extract to obtain a purified fraction enriched in lignin oligomers, c. Formulating the purified fraction into a phytopharmaceutically or agrochemically acceptable dosage form. 5 14.Statement 14° The method for producing phytopharmaceutical or agrochemical composition according to statement 13°, whereby the liquid phase contains one or more solvents selected from the group comprising water, methanol, ethanol, n-propanol and isopropylalcohol and mixtures of two or more thereof. 10 15.Statement 15° The method for producing phytopharmaceutical or agrochemical composition according to any one of the statements 13° to 14°, whereby the solvolytic lignin depolymerisation and fractioning is of the group consisting of reductive catalytic fractionation (RCF), non-catalytic thermo-15 solvolytic depolymerisation and oxidative catalytic fractioning (OCF). Detailed Description DETAILED DESCRIPTION OF EMBODIMENTS OF THE INVENTION The following detailed description of the invention refers to the accompanying drawings. The same reference numbers in different drawings identify the same or 20 similar elements. Also, the following detailed description does not limit the invention. Instead, the scope of the invention is defined by the appended claims and equivalents thereof. The following detailed description of the invention refers to the accompanying drawings. The same reference numbers in different drawings identify the same or 25 similar elements. Also, the following detailed description does not limit the invention. Instead, the scope of the invention is defined by the appended claims and equivalents thereof. Several documents are cited throughout the text of this specification. Each of the documents herein (including any manufacturer’s specifications, instructions etc.) are 30 hereby incorporated by reference; however, there is no admission that any document cited is indeed prior art of the present invention. 49
The present invention will be described with respect to particular embodiments and with reference to certain drawings but the invention is not limited thereto but only by the claims. The drawings described are only schematic and are non-limiting. In the drawings, the size of some of the elements may be exaggerated and not drawn 5 to scale for illustrative purposes. The dimensions and the relative dimensions do not correspond to actual reductions to practice of the invention. Furthermore, the terms first, second, third and the like in the description and in the claims, are used for distinguishing between similar elements and not necessarily for describing a sequential or chronological order. It is to be understood that the terms 10 so used are interchangeable under appropriate circumstances and that the embodiments of the invention described herein are capable of operation in other sequences than described or illustrated herein. Moreover, the terms upper, top, bottom, over, under and the like in the description and the claims are used for descriptive purposes and not necessarily for describing 15 relative positions. It is to be understood that the terms so used are interchangeable under appropriate circumstances and that the embodiments of the invention described herein are capable of operation in other orientations than described or illustrated herein. It is to be noticed that the term “comprising”, used in the claims, should not be 20 interpreted as being restricted to the means listed thereafter; it does not exclude other elements or steps. It is thus to be interpreted as specifying the presence of the stated features, integers, steps or components as referred to, but does not preclude the presence or addition of one or more other features, integers, steps or components, or groups thereof. Thus, the scope of the expression “a device 25 comprising means A and B” should not be limited to the devices consisting only of components A and B. It means that with respect to the present invention, the only relevant components of the device are A and B. The term "consisting of" refers to compositions, methods, and respective components thereof as described herein, which are exclusive of any element not30 recited in that description of the embodiment. As used herein the term "consisting essentially of" refers to those elements required for a given embodiment. The term permits the presence of additional elements that 50
do not materially affect the basic and novel or functional characteristic(s) of that embodiment of the technology. Reference throughout this specification to “one embodiment” or “an embodiment” means that a particular feature, structure or characteristic described in connection 5 with the embodiment is included in at least one embodiment of the present invention. Thus, appearances of the phrases “in one embodiment” or “in an embodiment” in various places throughout this specification are not necessarily all referring to the same embodiment, but may. Furthermore, the particular features, structures or characteristics may be combined in any suitable manner, as would be apparent to10 one of ordinary skill in the art from this disclosure, in one or more embodiments. Similarly it should be appreciated that in the description of exemplary embodiments of the invention, various features of the invention are sometimes grouped together in a single embodiment, figure, or description thereof for the purpose of streamlining the disclosure and aiding the understanding of one or more of the various inventive 15 aspects. This method of disclosure, however, is not to be interpreted as reflecting an intention that the claimed invention requires more features than are expressly recited in each claim. Rather, as the following claims reflect, inventive aspects lie in less than all features of a single foregoing disclosed embodiment. Thus, the claims following the detailed description are hereby expressly incorporated into this detailed 20 description, with each claim standing on its own as a separate embodiment of this invention. Furthermore, while some embodiments described herein include some but not other features included in other embodiments, combinations of features of different embodiments are meant to be within the scope of the invention, and form different 25 embodiments, as would be understood by those in the art. For example, in the following claims, any of the claimed embodiments can be used in any combination. In the description provided herein, numerous specific details are set forth. However, it is understood that embodiments of the invention may be practiced without these specific details. In other instances, well-known methods, structures and techniques 30 have not been shown in detail in order not to obscure an understanding of this description. Other embodiments of the invention will be apparent to those skilled in the art from consideration of the specification and practice of the invention disclosed herein. 51
It is intended that the specification and examples be considered as exemplary only. Each and every claim is incorporated into the specification as an embodiment of the present invention. Thus, the claims are part of the description and are a further description and are in addition to the preferred embodiments of the present 5 invention. Each of the claims set out a particular embodiment of the invention. The following terms are provided solely to aid in the understanding of the invention. Definitions A plant pest concerns anything that has a negative impact on a plant, including10 insects, fungi, bacteria, virus and parasites, Lignin is a complex organic substance that acts like a glue, binding the cells, fibers, and vessels that make up plants. It's the second most abundant biopolymer on earth, after cellulose, and plays a vital role in giving plants their strength and rigidity. Lignin is a complex aromatic polymer made up of various phenolic subunits that are mainly15 interlinked by ether and carbon-carbon bonds. Unlike cellulose, which is a carbohydrate, lignin is not easily broken down by microorganisms. Lignin depolymerization is a process that breaks down lignin, a complex molecule found in plants, into smaller, components. In this particular invention lignin depolymerization refers specifically to the breaking of ether bonds, and in particular 20 Beta-O-4 ether bonds, that occur in native lignin yielding a liquid lignin oil comprising of components like phenolic monomers, dimers and trimers or specific oligophenolics with a DP from 1 to 8. This can be fractionated in fractions with selected DP. Thermo-solvolytic depolymerization of lignin (Fig. 1B) is a biorefinery method that combines lignocellulose biomass fractionation (solvent, heat and pressure disrupting 25 the structure of the biomass and dissolves lignin and some of the hemicellulose) with lignin depolymerisation under an inert gas or atmosphere. For instance an inert gas or atmosphere selected from the group consisting of nitrogen, argon, helium and hydrogen. 52
In the meaning of the present application the terms “induce” or “inducing” as used herein refers to cause, or causing and to enhance, or enhancing and to boost or boosting and to activate or activating. In the meaning of present application is non-catalytic thermo-solvolytic fractioning a 5 depolymerzation (Fig. 1B) process without the presence of a redox catalyst and under inert atmosphere. As we experienced the resulting lignin oils from such process, when not stored refrigerated, are prone to re-polymerisation in an aqueous solution containing DMSO. In that case that process of re-polymerisation has to be suppressed or prevented by suppressors or stabilizing agents of the group consisting of 1,4-10 butanediol, 2-hydroxy-1-naphthoic acid, 2-naphthol, 2-naphthol-7-sulfonat, 2- naththol, 3-hydroxy-2-naphthoic acid, 4-hydroxybenzoic acid, 6-hydroxy-2- naphthoic acid, ascorbic acid, bovine serum albumin, citric acid, salicylic acid, cysteine, dimethylphloroglucinol, histidine, mannitol, o-dihydroxybenzene, p- benzenediol, phloroglucinol, resorcinol, soy protein isolate, syringic acid and vanillic15 acid. Reductive catalytic fractionation (RCF) (Fig. 1A) of lignocellulose is a biorefinery method that combines biomass fractionation (solvent, heat and pressure disrupting the structure of the biomass and dissolves lignin and some of the hemicellulose) with lignin depolymerisation enabled by heterogeneous catalysis, more particularly a 20 redox-active catalyst in a reductive environment (e.g. with the addition of hydrogen or H-donating agents). The output is carbohydrate enriched pulp (mainly cellulose) and a lignin oil comprising fractions of low molecular weight oligomeric and monomeric units of aromatic compounds which molecular weight fractions can be separated. Reductive Catalytic Fractionation (RCF) is a solvolytic process. As RCF 25 typically uses a solvent system (e.g., methanol, ethanol, or water) to facilitate the solvolytic depolymerisation of lignin from the lignocellulosic matrix. In addition, as the solvent plays a dual role by dissolving lignin fragments and stabilizing them against recondensation. Catalysts (often heterogeneous, like supported Ru, Ni, or Pd) operate within the solvent medium, enabling selective hydrogenolysis and 30 hydrogenation reactions to depolymerize and stabilize lignin-derived fragments. The catalysts (often heterogeneous, like supported Ru, Ni, or Pd) operate within the solvent medium, enabling selective hydrogenolysis and hydrogenation reactions to depolymerize and stabilize lignin-derived fragments. Molecular hydrogen (H₂) or hydrogen donors (e.g., alcohol solvents like ethanol or formic acid) in Solvolysis are 35 commonly used to create a reductive environment, further facilitating solvolytic 53
depolymerisation. The solvent helps target lignin selectively, leaving cellulose and much of the hemicellulose largely unaffected in the solid phase (selective depolymerisation), consistent with solvolytic principles. However, RCF in this application is also meant to comprise the reductive catalytic depolymerisation of 5 lignin solved in a suitable solvent (solvolytic catalytic depolymerisation). Without post-treatment the pH of RCF liquids containing depolymerized lignin generally range from 4 to 6, reflecting the presence of phenolic and carboxylic acids and with hydrogenation stabilization this can shift to a neutral range (pH 6–7), as acidic groups are reduced or neutralized. When using ethanol/water systems with Ru/C catalysts, 10 the pH after RCF typically stabilizes around 5–6 due to phenolic compounds. While employing stronger hydrogen donors or additional base (e.g., NaOH) to assist depolymerisation can push the pH closer to neutral. We found that catalysis overcomes the tendency of lignin oils or lignin oil fractions to repolymerize. This can occur for lignin oils obtained from thermo-solvolytic methods in absence of15 hydrogenation catalysis. A suitable solvent for lignin depolymerization via RCF or non-catalytic thermo- solvolytic fractioning the process is an aliphatic alcohol such as methanol, ethanol, 1-propanol, 1-butanol, 1-pentanol, 2-pentanol, 2-butanol, 2-pentanol, 3-pentanol, 2-methyl-1-propanol, 2-methyl-1-butanol, or 3-methyl-1-butanol or binary mixture 20 with water thereof. Preferably the solvent of the group consisting of ethanol, methanol, butanol, acetone, ethyl acetate, toluene, tetrahydrofuran (THF), tetrahydrofuran (THF), gamma-valerolactone (GVL) and ionic liquid. Micromixing means that the features of mixing are achieved at the molecular scale. A polar aprotic solvent is one that has typically a dipole moment in thed range of 2– 25 5 D, a high dielectric constant of typically >15 and no hydrogen bond donors (i.e., they lack N-H or O-H groups). Suitable polar aprotic solvents for the plant defene elicitor formulations of present invention are for instance δ-Valerolactone (DVL), Cyrene, (Dihydrolevoglucosenone); Ethyl Acetate, Methyl Lactate, Acetone (Dimethyl Ketone) and Limonene Oxide). 30 PG is the abbreviation for the identified monomer, 4-propyl guaiacol. PS is the abbreviation for the identified monomer, 4-propyl syringol. 54
DP is the abbreviation for degree of polymerization referring to the number of aromatic groups in the lignin oligomer Pulping in terms of RCF or non-catalytic thermo-solvolytic fractioning is the separation cellulose fibers from the lignin or the other components, in generally via 5 the organic solvents and sometimes water. Fractionation in terms is the process of separating a mixture into its constituent parts based on their different properties. Fractionation allows processors to further separate the depolymerized lignin or lignin oils into fractions with different compositions, molecular weights or molecular sizes. In RCF, liquid-liquid extraction 10 (LLE) is often used for separating lignin-derived phenolic monomers, dimers, and oligomers from the reaction mixture after depolymerization. ISR is an abbreviation for induced systemic resistance. ISR is a defense mechanism in plants involving activation of the plant’s immune response systemically (throughout the entire plant) after exposure to certain beneficial elicitors. 15 A “disease index” is a numerical representations of disease severity or incidence and it can quantify the impact of a disease on a plant population. The effect of ISR on disease suppression can be reflected indirectly and for evaluating the effectiveness of ISR-inducing treatments, researchers measure disease severity, pathogen growth, or symptom development, measurements that contribute to disease indices. A lower 20 disease index in ISR-treated plants indicates successful suppression of diseases due to induced resistance. A "disease index" in the context of A. thaliana is a quantitative measure used to evaluate the resistance of A. thaliana accessions to specific pathogens. This index is particularly useful in studying the genetic basis of disease resistance in plants. The 25 disease index is calculated based on the number of necrotic lesions (areas of plant tissue that have died) that develop on the leaves of A. thaliana after inoculation with a pathogen. The "disease index" for Hyaloperonospora arabidopsidis (H. arabidopsidis) in A. thaliana is a measure used to quantify the severity of infection by this oomycete 30 pathogen. This index is based on the number of sporangiophores (structures that bear spores) observed on the plants after inoculation. The disease severity is scored 55
by determining the number of sporangiophores per plant, with more sporangiophores indicating a higher level of infection or disease severity. In the present invention, the term “biomass” is used for the term “lignocellulosic material” and lignocellulosic material may be in the meaning of lignocellulose or 5 material comprising lignocellulose. The term “dry” or “dried” referring to a compound, component or composition means that the water content has been significantly reduced from the original form thereof. This is typically achieved through processes like dehydration, which remove water 10 from the compound, component or composition by evaporation or other methods. The amount of moisture left in a dry compound, component or composition powder can vary depending on the specific type of compound, component or composition and the drying method used. It has to be interpreted to have a moist content under 12%, preferably under 10 % and 7% and even having a moisture content of around15 5% or even having have a moisture content of around 3%. The term “butter” as used herein is understood to be synonymous with the term “lipid” and may refer in general terms to a lipid or a composition comprising a lipid as a main constituent that retains solid, semi-solid, biphasic, or paste-like properties20 at ordinary temperatures of use. The term "elicitor" or “plant defense elicitor” as used herein refers to an exogenous defense-triggering molecule, for instance inducer of the plant immune system or elicitors of natural plant defences against pests (pathogens), abiotic and biotic 25 stressors. When plants are attacked by pest such as pathogens, they defend themselves with an arsenal of mechanisms directed to fight infection or make the plant less attractive to that pest. As in most cellular responses to the environment, defense mechanisms can be activated when receptors directly or indirectly come in contact with pathogens. The ligands of these plant receptors are elicitors of the plant 30 immune system. There is a wide variety of elicitors, including so-called non-specific elicitors or PAMPs (pathogen associated molecular patterns) e.g. degradation products of cell wall components of pathogens or derived from a plant cell wall, and pathogen-specific elicitors or effectors e.g. avirulence gene products of pathogens such as AVR, such as AVR proteins, which play a crucial role in the gene-for-gene 35 interactions between plants and pathogens and which avirulence gene products are 56
recognized by plant resistance (R) genes, triggering defense responses in the host plant. Elicitors of the plant immune system comprise proteins, oligosaccharides, polysaccharides, lipids, glycolipids, glycoproteins, peptides of diverse origin, lipopeptides, algal extracts, extracts from the walls of plant material and/or fungal 5 material, fungi, bacterial material and viral material, or yeast material and/or extracts. Elicitors also comprise salicylic acid, jasmonic acid, lipid peroxidation products and/or one or more of their esters. Elicitors of the plant immune system against insects play a crucial role in activating defense mechanisms to protect plants from herbivory. One such key elicitor is the jasmonate pathway, which is central in 10 promoting resistance to a broad spectrum of insects. This pathway involves signal transduction pathways that include calcium ion fluxes, phosphorylation cascades, and the production of jasmonates, which are essential for plant defense against insect herbivores. Such a plant defense elicitor triggers the activation of defense responses, such as the biosynthesis of jasmonic acid, to defend against insect15 attacks. Some exogenous defense-triggering molecules (Plant defense elicitor) can induce a plant’s defense system associated with extensive transcriptional- and metabolic reprogramming of the genome. Elicitation of plants with elicitor molecules can result 20 in the activation of a series of defense responses, including cell wall reinforcement by deposition of lignin and induction of an array of defense enzymes. Diverse plant defense responses induced by elicitors involve de novo synthesis and accumulation of antimicrobial phytoalexins, induction of cell death (hypersensitive response), production of activated oxygen species (oxidative burst), and modification of plant 25 cell walls by deposition of callose (Bektas, 2022; Sudhamoy & al, 2010; Wang et al., 2004). Elicitors can induce a range of responses in plants, including the production of antimicrobial compounds, cell death (hypersensitive response), generation of 30 reactive oxygen species, and modification of cell walls. These responses contribute to enhancing plant resistance to pathogens and herbivores (Wang et al., 2004). The effects of elicitors on plants can vary depending on factors such as plant genotype, developmental stage, and environmental conditions (Chalal et al., 2015). Additionally, the application of elicitors can lead to the enhanced production of 35 secondary metabolites in plants, which are crucial for defense against biotic stresses (Hatami et al., 2019). 57
Elicitors might be specifically recognized by the plant and subsequently induce defense responses against pathogens or herbivores in the attacked host (Maffei et al., 2012). Elicitor recognition by the plant is assumed to be mediated by specific receptors in the plant cell, localized either on the cell surface for a number of fungal 5 elicitors or within the cell for certain bacterial elicitors, which initiate signaling processes that activate plant defenses (Angelova et al., 2006; Shinya et al., 2006). Some of such plant defense elicitors have been recently discovered and tested. For instance silicon has also been identified as a key element that can upregulate plant 10 defense pathways against insects, providing physical and biochemical defense mechanisms (Alhousari & Greger, Plants 2018, 7(2), 33). Plant cells also recognize chitin fragments for defense signaling through a plasma membrane receptor. Chitin oligosaccharides are a representative general plant defense elicitor inducing defense responses in a wide range of plant cells ( Kaku et al. (2006) 11086 –11091 PNAS15 July 18, 2006 vol. 103 no. 29). laminarin, a β-1,3 glucan oligosaccharide, oligo- galacturonan and sodium alginate are also plant defense elicitors and also found to be a plant defense elicitor (Priya Dey et al . PLoS ONE 14(9) (2019); Chalal et al. Front. Plant Sci., 19 May 2015 Sec. Plant Pathogen Interactions Volume 6 - 2015 (2015) and Bektas (Horticulturae 2022, 8, 484). Cellodextrins act as elicitors of plant 20 defense (Aziz et al., 2007 J. Exp. Bot. 58, 1463–1472; Souza et al., 2017 Plant Physiol. 173, 2383–2398; Johnson et al., 2018 Plant Physiol. 176, 2496–2514). In particular C1-oxidized (aldonic) cellodextrins, oligosaccharides composed of glucose units linked by β-1,4-glycosidic bonds, and the C4-oxidized cellodextrins, with the same basic structure as cellodextrins (glucose chains linked together ß-1,4 glycosidic 25 bonds) but an additional oxygen atom (an oxygen-containing functional groups, such as hydroxyl (-OH) or carboxyl (-COOH) group) bonded to the fourth carbon (C4) of the glucose unit at the non-reducing end are both known improve solubility and reactivity. With their modified structure containing oxygen-containing functional groups at the C1 and C4 positions, can act as molecular signals that plants recognize 30 as indicators of potential danger. When plants perceive these signals, they may initiate defense responses, such as the production of defense-related proteins, activation of signaling pathways, or synthesis of secondary metabolites with antimicrobial properties (Josman Velasco et al. ACS SustainableChem.Eng.2022,10, 16969−16984). 35 The term "oligo-galacturonan" encompasses herein a chain of o(1-4)-linked D- galacturonic acids. Oligo-galacturonans are derived from pectin, which is a major 58
constituent of plant cell walls. Pectin consists of a complex set of polysaccharides, including homogalacturonans, which are linear chains of a-(1-4)-linked D- galacturonic acids. Oligo-galacturonans are released from these galacturonans through the action of pectolytic enzymes. Oligo-galacturonans particularly suitable 5 as plant defense elicitor have a degree of polymerization higher than 8, preferably comprised between 9 and 20 or between 9 and 15. The terms "chito-oligosaccharide" and "chitosan oligosaccharide" are used interchangeably herein and refer to a linear oligosaccharide composed of randomly10 distributed -(1-4)-linked D- glucosamine (deacetylated unit) and N-acetyl-D- glucosamine (acetylated unit). Chitosan is naturally found in few organisms, but is mostly produced industrially by deacetylation of chitin, which is the structural element in the exoskeleton of crustaceans (crabs, shrimp, etc.), insects, and in the cell walls of some fungi and other organisms. Chito-oligosaccharides used 15 particularly suitable as plant defense elicitor have a degree of acetylation lower than 50%, lower than 40%, or lower than 30%, preferably about 25% and a degree of polymerization higher than 5, preferably comprised between 5 and 10. Salts can added to the composition according to the invention to ensure good ionic 20 conditions and sucrose can be added to the composition according to the invention. Preferably, sucrose is added in a concentration of from about 1 mM to 20 mM, most preferably about 5 to 10 mM sucrose. Sucrose triggers signaling through hexokinase and is also a wetting agent. Sucrose can act as wetting agents improve the spreading and sticking of water-based sprays on plant surfaces. This is crucial for applying 25 pesticides, herbicides, and fungicides effectively. And hexokinase, the enzyme sucrose binds to, acts as a sensor, a binding that can activate genes involved in defense against pathogens or insects. The term "fungicide" encompasses chemical or biological substances or compositions 30 used to kill or inhibit fungi or oomycetes, e.g. by preventing sporulation, or their spores. Fungicides can exert their biological effect by different modes of action, for example, but not limited to, by interference with nucleic acid synthesis, mitosis and cell division, respiration, amino acids and protein synthesis, signal transduction, lipids and membrane synthesis, sterol biosynthesis, glucan synthesis in the pathogen or by35 inducing host plant defense. Any fungicide can be included in the composition of the invention, such as, for example, a fungicide selected from: acylalanines (benalaxyl), anilinopyrimidines 59
(cyprodinil or pyrimethanil), benzamides (fluopicolide or zoxamide), benzimidazoles (fuberidazole, thiabendazole or metrafenone), benzothiadiazoles (acibenzolar-S- methyl), carbamates (benthiavalicarb, iprovalicarb or propamocarb), carboxamides (boscalid), chloronitriles (chlorothalonil), chlorophenyls (tolclophos-methyl), 5 cyanoacetamide oximes (cymoxanil), cyanoimidazoles (cyazofamid), dicarboximides (iprodione), dithiocarbamates (thiram, metiram, mancozeb, manebe or propineb), guanidines (dodine), hydroxyanilides (fenhexamid), imidazoles (fenamidone, imazalil or triflumizole), morpholines (dimethomorph, fenpropimorph, spiroxamine or dodemorph), phosphonates (fosetyl), oxathiins (flutolanil), oxazoles (famoxadone or 10 hymexazol), phenylamides (metalaxyl or metalaxy-M), phenylpyridinamides (fluazinam), phenylpyrroles (fludioxonil), phtalimides (captan or folpet), quinazolinones (proquinazide), quinolins (quinoxyfen), strobilurins (dimoxystrobin, fluoxastrobin, kresomin- methyl, pyraclostrobin, trifloxystrobin or picoxystrobin), thiophenes (silthiofam), triazoles (difenoconazole, epoxyconazole, fenbuconazole, 15 flusilazole, metconazole, myclobutanil, penconazole, propiconazole, tebuconazole, tetraconazole, triadimenol, triticonazole orprothioconazole), copper derivates (copper oxychloride, copper hydrochloride, copper oxide or copper sulphate) and sulphur. Preferably, the fungicide is chosen from the list comprising: phosphonates, benzamides, carbamates, dithiocarbamates, phtalimides, triazoles, quinolines,20 sulphur and cyanoimidazoles. Phosphonates: The mode of action of the phosphonates is largely unknown but could involve inhibition of mitochondrial ATP synthase. Suitable examples of phosphonates are phosphorous acid derivatives, including phosphorous acid itself and its alkali metal or alkaline- earth metal salts. In a preferred embodiment the fungicides are 25 ethyl hydrogenphosphonates such as fosetyl-AI, fosetyl-K and fosetyl-Na. Mention can be made of the phosphonates sold under the trade names Aliette, Autograph, Avalon, Flanker, Legion, Linebacker, Novasource, Prodigy Signature and Quali-Pro, which all comprise fosetyl-AI as active ingredient, and Magellan and Phostrol, which comprise phosphorous acid as active ingredient. 30 Benzamides interfere with mitosis and cell division. In a preferred embodiment, the benzamides used in the composition of the invention contain 2,6-dichloro-N-[3- chloro-5- (trifluoromethyl)-2-pyridinyl]benzamide (fluopicolide) as active ingredient. Mention can be made of Infinito. Carbamates act by interfering with lipids and membrane synthesis. In a preferred 35 embodiment, the carbamates used in the composition of the invention contain propamocarb, preferably propamocarb hydrochloride (propyl[3- (dimethylamino)propyl]carbamate hydrochloride) as the active ingredient. Mention 60
can be made of the carbamates sold under the trade names Infinito and Stellar (comprising fluopicolide and propamocarb hydrochloride), Banol, Previcur, Proplant (comprising propamocarb hydrochloride) and Previcur Energy (comprising propamocarb and fosetyl-AI). - Dithiocarbamates show multi-site contact activity. In 5 a preferred embodiment, dithiocarbamates containing manganese ethylenebis(dithiocarbamate) (polymeric) complex with zinc salt (mancozeb) as active ingredient, are used in the compostion of the invention. Mention can be made of the dithiocarbamates sold under the trade names Acrobat MZ, Clevis, Cuprofix MZ, Dithane, Evolve, Fore, Gaucho, Gavel, Junction, Mancozide, Manhandle, Manzate, 10 Maxim, Moncoat, Nubark, Penncozeb, Pentathlon, Potato Seed Treater, Protect, Ridomil Gold MZ, SA-50, Stature, Tops MZ, Wingman and Zyban.- Phtalimides also show multi-site contact activity. In a preferred embodiment, the phtalimides used in the plant defense elicitor composition of the invention comprise A/- (trichloromethylthio)phthalimide or 2-[(trichloromethyl)thio]-1 - -isoindole-1 ,3(2H)- 15 dione (folpet) as active ingredient. Mention can be made of the phtalimides sold under the trade names Folpet and Fungitrol. Triazoles act by interfering with sterol biosynthesis in membranes. In a preferred embodiment, the triazoles used in the composition of the invention contain (2RS,3RS)-3-(2-chloorfenyl)-2-(4-fluorfenyl)-[(1H-1,2,4-triazool-1- 20 yl)methyl]oxiraan (epoxyconazole) as active ingredient. Mention can be made of the triazole fungicide sold under the trade name Opus. Cyanoimidazoles act by interfering with the electron transport chain at the level of complex III in the inner membrane of mitochondria, which blocks oxidative phosphorylation powered by electron transfer. In a preferred embodiment, the25 cyanoimidazoles used in the composition of the invention contain 4-chloro-2-cyano- /V,/ /-dimethyl-5-(4-methylphenyl)-1 - -imidazole-1- sulfonamide (cyazofamid) as active ingredient. Mention can be made of the cyanoimidazole sold under the trade name Ranman. Quinolines act by interfering with, e.g. blocking, signal transduction. In a preferred30 embodiment, the quinolines used in the composition of the invention contain 5,7- dichloro-4- quinolyl 4-fluorophenyl ether (quinoxyfen) as active ingredient. Mention can be made of the quinoline fungicide sold under the trade names Legend or Quintec. Sulphur-containing fungicides show multi-site contact activity and contain sulphur as the active ingredient. Mention can be made of the sulphur-containing fungicide sold35 under the trade name Thiovit®. The compositions of the invention will typically contain additional components, known as co- formulants or adjuvants, to obtain a product with good handling, efficacy and 61
stability properties. As used herein, the terms "co-formulant" or "adjuvant" designate any substance other than the main oligosacharidic complex plant defense elicitor component defined herein, that is intentionally added to the plant defense elicitor composition of the invention. 5 In a certain embodiment, the composition according to the invention further comprises a co- formulant or adjuvant selected from the group comprising: surfactants, anti-freeze agents (including urea, ethylene glycol, propylene glycol or glycerol), preservative agents (including potassium sorbate, paraben and its derivates, 1 , 2-benzisothiazolin-3(2H)-one or essential oils), absorbent agents 10 (including raids of corn or sawdust), thickeners (including clays orxanthane gum), buffers, sticker agents (including latex, silicon or alkoxylated alkyl), diluents (including rapeseed methyl ester) or any standard inert ingredient conventionally used in agricultural compositions, or a mixture thereof. In a particularly preferred embodiment, the composition according to the invention further comprises a15 surfactant. With "surfactant" is meant herein a compound that lowers the surface tension of a liquid, allowing easier spreading. The surfactant can be a detergent, an emulsifier (including alkyl polyglucosides glycerol ester or polyoxyethylene (20) sorbitan 20 monolaurate), or natural plant lecithin, or a biosurfactant, or a dispersing agent (including sodium chloride, potassium chloride, potassium nitrate, calcium chloride or starch of corn), a foaming agent (including derivates of tartric acid, malic acid or alcohols), a penetration enhancer, a humectant (including ammonium sulfate, glycerin or urea) or a wetting agent of ionic or non-ionic type or a mixture of such 25 surfactants. The surfactants used in the present invention are penetration enhancers, dispersing agents or emulsifiers. The term "penetration enhancer" is understood herein as a compound that accelerates the uptake of active ingredient through the cuticle of a plant into the 30 plant, i.e. the rate of uptake, and/or increases the amount of active ingredient absorbed into the plant. Classes of substances known as penetration enhancers, include alkyl phosphates, such as tributyl phosphate and tripropyl phosphate, and naphthalenesulphonic acid salts. Mention may be made, for example, of surfactants sold under the trade name Dehscofix®, comprising castor oil and ethoxylated fatty 35 acids, such as Dehscofix CO 95 ® (available from Huntsman, USA), comprising C18 ethoxylated fatty acids from castor oil. 62
With "dispersing agent" is meant a substance added to a suspension, usually a colloid, to improve the separation of particles and to prevent settling or clumping. Mention can be made of the dispersing agent which is sold under the trade name Tensiofix Dp400 (available from Ajinomoto OmniChem), essentially comprising organic 5 sulfonate and 2-methylpentane-2,4- diol. The term “abiotic” stress is used herein to refer to non-living chemical and/or physical factors in the environment that affect plant growth and/or development. Examples include extreme temperatures (heat or cold), water availability (e.g., drought), 10 salinity (e.g., salt), and the like. Such abiotic factors are considered “stressors” when they influence the environment beyond its normal range of variation to adversely affect plant growth and/or development. The "dry" state in the present invention refers to a state that the water content is 15 about 20 mass% or less and the water activity value is 0.85 or less. In addition, the water content is more preferably 15 mass% or less, more preferably 10 mass% or less, and more preferably 5 mass% or more. The lower limit is not particularly limited and is usually 0.1 mass% or more. Furthermore, the water activity value is preferably 0.80 or less and more preferably 0.75 or less. 20 The term "emulsifier" as used herein refers to a substance that stabilizes an emulsion, i.e. a mixture of two or more liquids. Mention can be made of the emulsifiers sold under the trade names Tween® 20, which essentially comprises polyoxyethylene (20) sorbitan monolaurate (polysorbate 20), and Radia®, which essentially 25 comprises alkyl polyglycosides. In a preferred embodiment, said surfactant comprises one or more of the following components: castor oil ethoxylate, rapeseed methyl ester, alkyl phosphates, tributyl phosphate, tripropyl phosphate, naphthalenesulphonic acid salts, organic sulfonate / 2-methylpentane-2,4- diol, alkylpolyglucoside, siloxanes derivates, alkylsulfonates, polycarboxylates 30 lignosulfonates, alkoxylated triglycerides, fatty amines polymers, dioctylsulfosuccinates or polyoxyethylene (20) sorbitan monolaurate (polysorbate 20), more preferably said surfactant is C18-castor-oil-ethoxylate (Dehscofix®), organic sulfonate / 2-methylpentane-2,4-diol (Tensiofix Dp40) or polyoxyethylene (20) sorbitan monolaurate (Tween®20). The present invention also discloses a 35 composition comprising polar aprotic compounds with weak acidity, like for example dimethyl sulfoxide (DMSO), enabling emulsification of the depolymerized lignin in water. 63
The term “biosurfactant” is understood a surface-active molecule produced by living organisms, typically microorganisms like bacteria or fungi. These special molecules can reduce surface tension and enhance the solubility and mobility of hydrophobic substances in aqueous environments and they have a unique structure with two key 5 parts: a hydrophilic head, often composed of things like sugars, amino acids, or phosphate groups and a hydrophobic tail usually made of fatty acids or long chains of hydrocarbons. Biosurfactants are classified into different classes based on their chemical composition, including low molecular weight surface-active agents called biosurfactants and high molecular weight bioemulsifiers . 10 Any compound as intended herein may be a part of a composition. The term “composition” generally refers to a thing composed of two or more components, and more specifically particularly denotes a mixture or a blend of two or more materials, such as elements, molecules, substances, biological molecules, or microbiological 15 materials, as well as reaction products and decomposition products formed from the materials of the composition. By means of an example, a composition may comprise any compound as taught herein in combination with one or more other compounds or substances, be it one or more other compounds as taught herein or one or more other compounds or substances. For example, a composition may be obtained by 20 combining, such as admixing, a compound as taught herein with said one or more other compounds or substances. For example, a composition may be obtained by decomposing a starting material, such as cellulose, into a mixture of a plurality of decomposition products. 25 In certain embodiments, the present compositions may be configured as phytopharmaceutical or agrochemical compositions for treatment of a plant or plant protection composition. Phytopharmaceutical or agrochemical compositions typically comprise one or more active ingredients (chemically and/or biologically active materials having one or more beneficial effects on plant health) and one or more 30 phytopharmaceutically acceptable carriers. In present invention the active ingredient or one of the active ingredients is a plant defense elicitor. Compositions as typically used herein may be liquid, semisolid (e.g., gel), solid, or volatile or vapour-based, and may include solutions or dispersions, such as for example suspensions, emulsions, oil-in-water emulsions, water-in-oil emulsions, gelified aqueous solution 35 or dispersion, solutions comprising a volatile organic solvent, etc. Examples of solid forms include, without limitation, powder, granules, pellets, water dispersible powder, water dispersible granules or water dispersible pellets. The composition may 64
be formulated as a concentrate to be diluted before use, such as, for example, a soluble concentrate, an emulsifiable concentrate, a liquid concentrate and the like. Such composition may also be described as agrochemical composition. This a formulated mixture of chemical or biological substances designed for agricultural use. 5 These compositions can include active ingredients, carriers, adjuvants, and other additives that enhance their efficacy, stability, or application properties. In present invention the active ingredient or one of the active ingredients is a plant defense elicitor. Such compositions are used to protect plants from biotic stress such as pests and diseases or from abiotic stress. 10 As used herein, the term “carrier” broadly includes any and all solvents, diluents, bulking agents, buffers for pH control, dispersant, solubilisers, surfactants, wetting agents, emulsifiers, tackifiers, thickeners, binders, preservatives, antioxidants, cuticle solubilising molecules, natural orregenerated mineral substances, and the like, 15 and combinations thereof. Such materials should not be non-toxic to the plants and should not interfere with the activity of the actives. As used herein “solvolytic lignin depolymerisation” means any one of the following processes reductive catalytic fractionation (RCF) of lignin or lignocellulose, non- 20 catalytic thermo-solvolytic depolymerisation of lignin or lignocellulose or oxidative catalytic fractioning (OCF). Typical salicylic acid pathway activators are salicylic acid (SA), benzothiadiazole (BTH), acibenzolar-S-methyl (ASM), chitosan, oligogalacturonides (OGs) and typical jasmonic acid pathway activator are jasmonic acid (JA), methyl jasmonate (MeJA),25 coronatine, hexanoic acid and volicitin. A preferred example of a plant treating phytopharmaceutically or agrochemically acceptable solvent is water, hence, compositions as taught herein may comprise water, i.e., may be aqueous solutions or dispersions. Further examples of suitable 30 solvents include, but are not limited to, aromatic hydrocarbons, such as, for example, xylene mixtures or substituted naphthalenes; phthalates, such as, for example, dibutyl phthalate or dioctyl phthalate; aliphatic hydrocarbons, such as, for example, cyclohexane or paraffins; alcohols and glycols and their ethers and esters, such as, for example, ethanol, ethylene glycol, ethylene glycol mono methyl or monoethyl 35 ether; ketones, such as, for example, cyclohexanone; strongly polar solvents, such as, for example, N-methyl- 2-pyrrolidone, dimethyl sulfoxide or dimethylformamide; 65
vegetable oils or epoxidised vegetable oils, such as, for example, epoxidised coconut oil or soybean oil; and water. In a particular aspect, the solvent is a volatile solvent, such as methanol and ethanol. 5 Non-limiting examples of solid carriers include, but are not limited to, natural mineral fillers, such as, for example, calcite, talcum, kaolin, montmorillonite or attapulgite; highly dispersed silicic acid or highly dispersed absorbent polymers; pumice, broken brick, sepiolite or bentonite; calcite or sand; dolomite or pulverized plant residues. 10 In certain embodiments, the compositions may comprise one or more surfactant, such as an anionic, non-ionic, amphoteric, or cationic surfactant, or a combination thereof, such as without limitation Triton X-100, non-ionic surfactant that has a hydrophilic polyethylene oxide chain (such as on average 9.5 ethylene oxide units) and an aromatic hydrocarbon hydrophobic group, 4-(l,l,3,3-tetramethylbutyl)- 15 phenyl); a polysorbate-type non-ionic surfactant such as polyoxyethylene (20) sorbitan monolaurate (Tween-20), polyoxyethylene (20) sorbitan monopalmitate (Tween-40); and/ or a non-ionic organosilicone surfactant such as Silwet® L-77 (3- (2 -methoxyethoxy )propyl-methyl-bis(trimethylsilyloxy)silane). 20 In certain embodiments, the compositions may comprise one or more compounds miscible in organic solvents as well as water and having a weak acidity, such as without limitation dimethyl sulfoxide (DMSO) that enable the emulsification of lignin oil in water. 25 An embodiment of present invention is a) a plant defense elicitor characterized in that it comprises one or more a plant defense elicitor comprising, consisting of or essentially consisting of as active ingredient one or more aromatic compounds, with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4 or one or more aromatic compounds, with a molecular mass 180 g/molbetween 230 g/mol to 1000 g/mol, 30 and yet more preferably between 230 g/mol to 650 g/mol (Fig. 2), and wherein and/or I) the aromatic compounds comprise at least one aromatic compound selected 66
- wherein each R1, R3, and R4 is independently chosen from –H, -OH, O-CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to 5 an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, and - wherein R2 is –H, a β-O-4 linkage to an aromatic monomer or aromatic 10 oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, and - wherein R5 is selected of –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic 15 oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, an end-unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, -(CH2)2CH2OH, - (CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, -CH=CHCH2OCH3, - (CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, -20 CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, -CH=CHCH2OCH(CH3)2, - 67
(CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, or a carbon linkage to an aromatic monomer or aromatic oligomer; - and/or the aromatic compounds comprise at least one aromatic compound selected from the formulae (v) 5 - wherein each R12, R13, R15 and R16 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic 10 monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R11 and R14 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or 15 aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, and/or III) wherein at the aromatic compounds comprise at least one aromatic compound selected from the formula (viii) 68
linkage to an aromatic monomer or aromatic oligomer and 69
- wherein each R22, R23, R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic 5 monomer or an aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R21 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic 10 oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein the alkyl group is derived from the alcohol solvent of the process, - wherein R27 is independently chosen from –H, a β-O-4 linkage to an aromatic 15 monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, end- unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, (CH2)2CH2OH, -(CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, -20 CH=CHCH2OCH3, -(CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, - (CH2)2CH2O(CH2)2CH3, -CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, - CH=CHCH2OCH(CH3)2, -(CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, a carbon linkage to an aromatic monomer or an aromatic oligomer and b) a co-formulant selected from the group comprising: surfactants, anti- 25 freeze agents (including urea, ethylene glycol, propylene glycol or glycerol), preservative agents (including potassium sorbate, paraben and its derivates, 1 , 2-benzisothiazolin-3(2H)- one or essential oils), absorbent agents (including raids of corn or sawdust), thickeners (including clays or xanthane gum), buffers, sticker agents (including latex, silicon or alkoxylated alkyl), diluents (including rapeseed 30 methyl ester) or any standard inert ingredient conventionally used in agricultural compositions, or a mixture thereof, preferably said co-formulant is a surfactant selected among a detergent, an emulsifier (including alkyl polyglucosides, glycerol ester or polyoxyethylene (20) sorbitan monolaurate (polysorbate 20)), a dispersing agent (including sodium chloride, potassium chloride, potassium 35 nitrate, calcium chloride or starch of corn), a foaming agent (including derivates of tartric acid, malic acid or alcohols), a penetration enhancer, a humectant (including ammonium sulfate, glycerin or urea) or a wetting agent of ionic or non- 70
ionic type or a mixture thereof, more preferably said surfactant comprises one or more of the following components: castor oil ethoxylate, rapeseed methyl ester, alkyl phosphates, tributyl phosphate, tripropyl phosphate, naphthalenesulphonic acid salts, organic sulfonate / 2-methylpentane-2,4-diol, alkylpolyglucoside, 5 siloxanes derivates, alkylsulfonates, polycarboxylates, lignosulfonates, alkoxylated triglycerides, fatty amines polymers, dioctylsulfosuccinates or polyoxyethylene (20) sorbitan monolaurate, most preferably said surfactant is C18- castor-oil-ethoxylate (Dehscofix®), organic sulfonate / 2-methylpentane- 2,4-diol (Tensiofix Dp40) or polyoxyethylene (20) sorbitan monolaurate 10 (Tween®20).In another embodiment, the compositions according to the invention also comprise one or more other active compounds selected from the group comprising: herbicides, insecticides, plant growth regulators or other plant immune system elicitors. 15 In a further embodiment of present invention this compositions according to the invention can further comprises a further plant immune system elicitor chosen among silica, copper, sulfur, aluminium, vanadium, cobalt, nickel, iron, silver, salicylic acid and its derivates (including acetyl-salicylic acid, isonicotinic acid, acibenzolar-S-methyl), jasmonic acid and its derivates (including methyl 20 jasmonate), ethylene and its derivates, polysaccharides (including glucans, xyloglucans, cellodextrins in particular with modified structure containing oxygen- containing functional groups at the C1 and C4 positions, chitin, chitosans, fucans, galactofucans, xylans, galactans, alginates, galacturonans, apiogalacturonans, fructans including inulin, mannans, xylomannans, galactomannans, 25 glucomannans and galactomannans), algae extracts (green algae extracts including ulvans, brown algae extracts including laminarin, and red algae extracts including carragenans), oligosaccharides (including trehalose), peptides (including systemin, 13-pep, flg-22, glutathion), amino acids, proteins (including harpin and flagellin), peptone, beef extract, essential oils (including cumin, anise, 30 mint, cinnamon, thyme, basil, cardamom, coriander, oregano, manzanilla, clove, jojoba and tea tree oils), lipids (including ergosterol, amphotericin, sphingolipids, cerebrosides), glycolipids (including syringolids), glycoproteins (including cryptogeins), lipopeptides, lipoproteins (including volicitin), yeast extracts (including extracts from Saccharomyces, Candida, Pichia, Aureobasidium and 35 more particularly Saccharomyces cerevisiae, Candida famata, Candida oleophila, Pichia guilliermondii, Aureobasidium pullulans), fungal extracts (including extracts from Trichoderma, Megasperma, Pyricularia, Alternaria, Pythium, 71
Puccinia, Colletotrichum, Verticillium, Magna porthe), bacterial extracts (including extracts from Escherichia, Rhyzobia, Pseudomonas), BABA, probenazole, isothianil, phosphorous acid and its derivates (including aluminium, sodium and potassium fosetyl), horsetail extracts, potassium iodide and potassium 5 thiocyanate, Citrus extracts, Yucca extracts Salix extracts and plant decoctions (including nettle decoction). Preferably, said further plant immune system elicitor contains laminarin (a linear β(1→3)- glucan with (1→6)-linkages) such as, for example Vacciplant Fruit®. 10 Yet another embodiment of present invention is a) a plant defense elicitor characterized in that it comprises one or more a plant defense elicitor comprising, consisting of or essentially consisting of as active ingredient one or more aromatic compounds, with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4 or one or more aromatic compounds, with a molecular mass 180 g/molbetween 230 15 g/mol to 1000 g/mol, and yet more preferably between 230 g/mol to 650 g/mol, and wherein and/or I) the aromatic compounds comprise at least one aromatic compound - wherein each R1, R3, and R4 is independently chosen from –H, -OH, O-CH3, a 20 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic 72
monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, and - wherein R2 is –H, a β-O-4 linkage to an aromatic monomer or aromatic 5 oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, and - wherein R5 is selected of –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic 10 oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, an end-unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, -(CH2)2CH2OH, - (CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, -CH=CHCH2OCH3, - (CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, -15 CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, -CH=CHCH2OCH(CH3)2, - (CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, or a carbon linkage to an aromatic monomer or aromatic oligomer; - and/or the aromatic compounds comprise at least one aromatic compound 20 73
- wherein each R12, R13, R15 and R16 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic 5 monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R11 and R14 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or 10 aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, and/or III) wherein at the aromatic compounds comprise at least one aromatic compound selected from the formula (viii) 15 74
, linkage to an aromatic monomer or aromatic oligomer and - wherein each R22, R23, R25 and R26 is independently chosen from –H, -OH, O- 5 CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or an aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, 10 - wherein R21 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, 15 - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein the alkyl group is derived from the alcohol solvent of the process, - wherein R27 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a β-β linkage to an aromatic monomer or aromatic20 oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, end- unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, (CH2)2CH2OH, -(CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, - CH=CHCH2OCH3, -(CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, - (CH2)2CH2O(CH2)2CH3, -CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, - 75
CH=CHCH2OCH(CH3)2, -(CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, a carbon linkage to an aromatic monomer or an aromatic oligomer, and b) a second plant defense elicitor, preferably a second plant defense elicitor chosen among silica, copper, sulfur, aluminium, vanadium, cobalt, nickel, iron, silver, 5 salicylic acid and its derivates (including acetyl-salicylic acid, isonicotinic acid, acibenzolar-S-methyl), jasmonic acid and its derivates (including methyl jasmonate), ethylene and its derivates, polysaccharides (including glucans, xyloglucans, chitin, chitosans, fucans, galactofucans, xylans, galactans, alginates, galacturonans, apiogalacturonans, fructans including inulin, 10 mannans, xylomannans, galactomannans, glucomannans and galactomannans), polyols, algae extracts (green algae extracts including ulvans, brown algae extracts including laminarin, and red algae extracts including carragenans), oligosaccharides (including trehalose), peptides (including systemin, 13-pep, flg-22, glutathion), amino acids, proteins 15 (including harpin and flagellin), peptone, beef extract, essential oils (including cumin, anise, mint, cinnamon, thyme, basil, cardamom, coriander, oregano, manzanilla, clove, jojoba and tea tree oils), lipids (including ergosterol, amphotericin, sphingolipids, cerebrosides), glycolipids (including syringolids), glycoproteins (including cryptogeins), lipopeptides, lipoproteins (including 20 volicitin), yeast extracts (including extracts from Saccharomyces, Candida, Pichia, Aureobasidium and more particularly Saccharomyces cerevisiae, Candida famata, Candida oleophila, Pichia guilliermondii, Aureobasidium pullulans), fungal extracts (including extracts from Trichoderma, Megasperma, Pyricularia, Alternaria, Pythium, Puccinia, Colletotrichum, 25 Verticillium, Magnaporthe), bacterial extracts (including extracts from Escherichia, Rhyzobia, Pseudomonas), BABA, probenazole, isothianil, phosphorous acid and its derivates (including aluminium, sodium and potassium fosetyl), horsetail extracts, potassium iodide and potassium thiocyanate, Citrus extracts, Yucca extracts Salix extracts and plant 30 decoctions (including nettle decoction), more preferably said second plant defense elicitor contains i-3(1-6)glucane (laminarin). In another preferred embodiment, said compositions according to the invention further comprise a further plant immune system elicitor that contains silicon or silicium (Si), such as, for example, silica (Si02) or silicates, including sodium silicate (Na2Si03). 35 Preferably, said further plant immune system elicitor is a silicate, more preferably sodium silicate. 76
The present invention also discloses a composition comprising: a) a plant defense elicitor characterized in that it comprises one or more a plant defense elicitor comprising, consisting of or essentially consisting of as active 5 ingredient one or more aromatic compounds, with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4 or one or more aromatic compounds, with a molecular mass 180 g/mol between 230 g/mol to 1000 g/mol, and yet more preferably between 230 g/mol to 650 g/mol (Fig. 2) , and wherein and/or I) the aromatic compounds comprise at least one aromatic compound selected from the formulae (i) 10 and - wherein each R1, R3, and R4 is independently chosen from –H, -OH, O-CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic 15 monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, and - wherein R2 is –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an20 α-O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, and 77
- wherein R5 is selected of –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, an end-unit selected 5 of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, -(CH2)2CH2OH, - (CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, -CH=CHCH2OCH3, - (CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, - CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, -CH=CHCH2OCH(CH3)2, - (CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, or a carbon linkage to an 10 aromatic monomer or aromatic oligomer; - and/or the aromatic compounds comprise at least one aromatic compound - wherein each R12, R13, R15 and R16 is independently chosen from –H, -OH, O- 15 CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, 78
- wherein R11 and R14 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or 5 aromatic oligomer, and/or III) wherein at the aromatic compounds comprise at least one aromatic compound selected from the formula (viii) 10 , 79
linkage to an aromatic monomer or aromatic oligomer and - wherein each R22, R23, R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or an aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R21 is independently chosen from –H, a β-O-4 linkage to an aromatic 10 monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein the 15 alkyl group is derived from the alcohol solvent of the process, - wherein R27 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, end- 20 unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, (CH2)2CH2OH, -(CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, - CH=CHCH2OCH3, -(CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, - (CH2)2CH2O(CH2)2CH3, -CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, - CH=CHCH2OCH(CH3)2, -(CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, a 25 carbon linkage to an aromatic monomer or an aromatic oligomer, said first plant elicitor in proportions ranging from 1 :50 to 50:1 , preferably from 1 :40 to 40:1 , more preferably from 1 :30 to 30:1 , even more preferably from 1 :20 to 20:1 , most preferably from 1 :10 to 10:1 and for instance is 1 :9, 1 :8, 1 :7, 1 :6, 1 :5, 1 :4, 1 :3, 1 :2, 1 :1 , 2: 1 , 3: 1 , 4:1 , 5:1 , 6:1 , 7:1 , 30 8:1 or 9:1 , and b) a second plant defense elicitor, preferably a second plant 80
defense elicitor that contains silicon, more preferably said second plant defense elicitor is a silicate, even more preferably said second plant defense elicitor is sodium silicate.In a preferred embodiment, said compositions of the invention comprising a plant defense elicitor and a second plant defense 5 elicitor further comprise an adjuvant, preferably a surfactant, more preferably a surfactant comprising polyoxyethylene (20) sorbitan monolaurate such as Tween® 20, a biosurfactant or a plant lecithin. The compositions according to the present invention encompass not only 10 compositions which are ready to be applied to the plant by means of a suitable device, such as a spraying device, but also the commercial concentrated compositions which have to be diluted before application to the plant. The compositions according to the invention are themselves in quite diverse, solid or liquid forms. As solid composition forms, mention may be made of powders for 15 dusting and granules, in particular those obtained by extrusion, by compacting, by impregnation of a granulated support or by granulation from a powder, tablets or effervescent lozenges. As liquid composition forms or forms intended to constitute liquid compositions when applied, mention may be made of solutions, in particular water-soluble concentrates, emulsions, concentrated suspensions, dispersions, 20 aerosols and wettable granules and powders (or powders for spraying), pastes, gels and water soluble packaging. In another aspect, the present invention relates to the use of the compositions of the invention in agricultural applications, more particularly for protecting plants against (infection by) plant pathogens. The present invention not only provides in the simultaneous use of the different 25 components of the compositions, i.e. the use of the compositions, but also provides in the sequential use of the different components of the compositions. For instance, the inventors have found that the sequential use of the plant defense elicitor from a lignin depolymerisation process and fractioning in compositions comprising oligophenolics with a degree of polymerization (DP) of 2 to 5, preferably of 2 to 3 30 from depolymerized lignin or decomposed lignin (Fig. 2) or from structural identical oligophenolic and a fungicide also results in enhanced efficacy of the fungicide. The present invention not only provides in the simultaneous use of the different components of the compositions, i.e. the use of the composition, but also provides in the sequential use of the different components of the compositions. For instance, the35 inventors have found that the sequential use of the plant defense elicitor characterized in that it comprises one or more plant defense elicitors comprising, 81
consisting of or essentially consisting of as active ingredient one or more aromatic compounds, with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4 or one or more aromatic compounds, with a molecular mass 180 g/molbetween 230 g/mol to 1000 g/mol, and yet more preferably between 230 g/mol to 650 g/mol (Fig. 5 2), and wherein and/or I) the aromatic Icompounds comprise at least one aromatic - wherein each R1, R3, and R4 is independently chosen from –H, -OH, O-CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to 10 an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, and - wherein R2 is –H, a β-O-4 linkage to an aromatic monomer or aromatic 15 oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, and - wherein R5 is selected of –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic 20 oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, an end-unit selected 82
of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, -(CH2)2CH2OH, - (CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, -CH=CHCH2OCH3, - (CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, - CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, -CH=CHCH2OCH(CH3)2, - 5 (CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, or a carbon linkage to an aromatic monomer or aromatic oligomer; - and/or the aromatic compounds comprise at least one aromatic compound 10 - wherein each R12, R13, R15 and R16 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic 15 monomer or aromatic oligomer, - wherein R11 and R14 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or 20 aromatic oligomer, and/or III) wherein at the aromatic compounds comprise 83
at least one aromatic compound selected from the formula (viii) 84
linkage to an aromatic monomer or aromatic oligomer and - wherein each R22, R23, R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or an aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R21 is independently chosen from –H, a β-O-4 linkage to an aromatic 10 monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein the 15 alkyl group is derived from the alcohol solvent of the process, - wherein R27 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, end- 20 unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, (CH2)2CH2OH, -(CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, - CH=CHCH2OCH3, -(CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, - (CH2)2CH2O(CH2)2CH3, -CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, - CH=CHCH2OCH(CH3)2, -(CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, a 25 carbon linkage to an aromatic monomer or an aromatic oligomer, and a fungicide also results in enhanced efficacy of the fungicide. By "sequential use" is meant herein that first the oligosaccharide plant defense elicitor is added and subsequently the fungicide, adjuvant, surfactant or other plant defense elicitor is applied to the plant, or vice versa. "Plant pathogens" refer to 85
organisms that cause infectious diseases in plants and include fungi, oomycetes, bacteria, viruses, viroids, virus-like organisms, phytoplasmas, protozoa, nematodes and parasitic plants. In a preferred embodiment, the plant pathogens are fungi, oomycetes, bacteria, viruses, nematodes and insects. 5 The majority of phytopathogenic fungi belong to the Ascomycetes and the Basidiomycetes, reproducing both sexually and asexually via the production of spores that can be spread through air (wind) or water, or can be soil borne such as zoospores that are capable of living saprotrophically, carrying out the first part of their lifecycle in the soil. Deuteromycetes (Fungi imperfecti) are fungi from which only the asexual 10 form of reproduction is known, meaning that this group of fungus produces their spores asexually. The Oomycetes are not true fungi but are fungal-like organisms that use the same mechanisms as fungi to infect plants. Fungal and fungal-like organisms are heterotrophic, i.e. they need an external source of nutrients for growth, development and reproduction. An understanding of other15 key features of these organisms can assist in their identification: - Hyphae: thread-like strands with a filamentous growth habit are a common feature in most fungi. The hyphae colonize (grow through) substrates so that the organism can obtain nutrients. Plant pathogenic species colonize plants through the host surface, sometimes through direct penetration of intact plant surfaces. 20 Saprophytic fungi tend to penetrate and colonize diseased plant tissue, senescing (dying) plants and plant residues. These fungi are major decomposers of organic matter in soil. Hyphal cell walls: true fungi have cell walls composed mainly of glucans and chitin, whereas fungal-like organisms have cell walls composed of cellulose and glycans. 25 - Septate hyphae: true fungi have cross walls within the hyphae, whereas fungal-like organisms do not. This can aid in the differentiation of these two groups under microscopic examination. - Motile spores: true fungi do not have motile spores, with the exception of Chytrids. Motile zoospores (asexually produced spores) are common in many species 30 in the Oomycota (e.g. Pythium and Phytophthora) and some downy mildews. Zoospores enable dispersal through water in soil and on plant surfaces. - Wind dispersed spores: many species of true fungi produce asexual or sexual spores for dispersal in the wind. This is a common feature of foliar fungal pathogens (e.g. Erysiphe). However some spores are adapted to splash dispersal. 35 - Survival structures: thick walled spores (e.g. oospores and chlamydospores), sclerotia and multicellular reproductive structures (e.g. pycnidia and perithecia) are important in the diseasecycle. During unfavourable environmental conditions or in 86
the absence of a suitable plant host or other substrate, these organisms persist in such specialised survival structures. Non-limiting examples of phytopathogenic fungi and fungal-like organisms include 5 Pyricularia oryzae (Magnaporthe grisea) on rice and wheat and other Pyricularia spp. on other hosts; Puccinia spp. e.g. Puccinia sorghi, Puccinia graminis f.sp. tritici, Puccinia asparagi, Puccinia recondita or Puccinia arachidis, Puccinia triticina (or recondita), Puccinia striiformis and other rusts on wheat, Puccinia hordei, Puccinia striiformis and other rusts on barley, and rusts on other hosts (for example turf, rye, 10 coffee, pears, apples, peanuts, sugar beet, vegetables and ornamental plants); Erysiphe cichoracearum on cucurbits (for example melon); Erysiphe necator (Uncinula necator) on grape, Blumeria (or Erysiphe) graminis (powdery mildew) on barley, wheat, rye and turf and other powdery mildews on various hosts, such as Sphaerotheca macularis on hops, Sphaerotheca fusca (Sphaerotheca fuliginea) on 15 cucurbits (for example cucumber), Leveillula taurica on tomatoes, aubergine and green pepper, Podosphaera leucotricha on apples; Cochliobolus spp., Helminthosporium spp. (e.g. Helminthosporium turcicum, Helminthosporium carbonum, Helminthosporium mavdis or Helminthosporium sigmoideum), Drechslera spp. (Pyrenophora spp. e.g. Pyrenophora tritici- repentens or Pyrenophora teres), 20 Rhynchosporium spp., Mycosphaerella gramninicola (Septoria tritici) and Phaeosphaeria nodorum (Stagonospora nodoruni or Septoria nodorum), Pseudocercosporella herpotrichoides and Gaeumannomyces graminis on cereals (for example wheat, barley, rye), turf and other hosts (e.g. Septoria lycopersici, Septoria glycines, Septoria); Cercospora arachidicola and Cercosporidium personatum on 25 peanuts and other Cercospora spp. (e.g. Cercospora kikuchii or Cercospora zaea- maydis) on other hosts, for example sugar beet, bananas, soya beans and rice; Botrytis spp. (e.g. Botrytis cinerea or Botryotinia fuckeliana), Botrytis cinerea (grey mould) on tomatoes, strawberries, vegetables, vines and other hosts and other Botrytis spp. on other hosts; Alternaria spp. (e.g. Alternaria brassicola or Alternaria 30 solani) on vegetables (for example carrots), oil-seed rape, apples, tomatoes, potatoes, cereals (for example wheat) and other hosts; Venturia spp. (including Venturia inaequalis (scab) or Venturia pirina) on apples, pears, stone fruit, tree nuts and other hosts; Cladosporium spp. (e.g. Cladosporium fulvum) on a range of hosts including cereals (for example wheat) and tomatoes; Monilinia spp. on stone fruit, 35 tree nuts and other hosts; Didymella spp. on tomatoes, turf, wheat, cucurbits and other hosts; Phoma spp. (e.g. Phoma betae on sugar beet and Phoma lingam on oil- seed rape), on turf, rice, potatoes, wheat and other hosts; Aspergillus spp. and 87
Aureobasidium spp. on wheat, lumber and other hosts; Ascochyta spp. (e.g. Ascochyta pisi) on peas, wheat, barley and other hosts; Stemphylium spp. (Pleospora spp.) on apples, pears, onions and other hosts; summer diseases (for example bitter rot (Glomerella cingulata), black rot or frogeye leaf spot (Botryosphaeria obtusa), 5 Brooks fruit spot (Mycosphaerellapomi), Cedar apple rust (Gymnosporangiumjuniperi-virginianae), sooty blotch (Gloeodespomigena), flyspeck (Schizothyrium pomi) and white rot (Botryosphaeria dothidea)) on apples and pears; Plasmopara viticola on vines; other downy mildews, such as Bremia lactucae on lettuce, Peronospora spp. (e.g. Peronospora manshurica or Peronospora tabacina) on 10 soybeans, tobacco, onions and other hosts, Pseudoperonospora humuli on hops and Pseudoperonospora cubensis on cucurbits; Pythium spp. (including Pythium ultimum) on turf and other hosts (e.g. Pythium aphanidermatum); Phytophthora infestans on potatoes and tomatoes and other Phytophthora spp. on vegetables, strawberries, avocado, pepper, ornamentals, tobacco, cocoa and other hosts (e.g. Phytophthora 15 cinnamomi, Phytophthora cactorum, Phytophthora phaseoli, Phytophthora parasitica, Phytophthora porri, Phytophthora citrophthora, Phytophthora megasperma f.sp. soiae or Phytophthora infestans); Thanatephorus cucumeris on rice and turf and other Rhizoctonia spp. on various hosts such as wheat and barley, peanuts, vegetables, cotton and turf; Sclerotinia spp. on turf, peanuts, potatoes, oil-seed rape 20 and other hosts (e.g. Sclerotinia sclerotiorum); Sclerotium spp. on turf, peanuts and other hosts; Gibberellafujikuroi on rice; Colletotrichum spp. (e.g. Colletotrichum lindemuthianum) on a range of hosts including turf, coffee and vegetables; Laetisaria fuciformis on turf; Mycosphaerella spp. on bananas, peanuts, citrus, pecans, papaya and other hosts; Diaporthe spp. on citrus, soybean, melon, pears, lupin and other 25 hosts; Elsinoe spp on citrus, vines, olives, pecans, roses and other hosts; Verticillium spp. (e.g. Verticillium dahliae or Verticillium albo-atrum) on a range of hosts including hops, potatoes and tomatoes; Pyrenopeziza spp. on oil-seed rape and other hosts; Oncobasidium theobromae on cocoa causing vascular streak dieback; Fusarium spp. (e.g. Fusarium nivale, Fusarium sporotrichioides, Fusarium oxysporum, Fusarium 30 graminearum, Fusarium germinearum, Fusarium culmorum, Fusarium solani, Fusarium moniliforme or Fusarium roseum), Typhula spp., Microdochium nivale, Ustilago spp. e.g. Ustilago maydis (e.g. corn smut), Urocystis spp., Tilletia spp. and Clavicepspurpurea on a variety of hosts but particularly wheat, barley, turf and maize; Ramularia spp. on sugar beet, barley and other hosts; post-harvest diseases 35 particularly of fruit (for example Penicillium expansum, Penicilliumn digitatum, Penicillium italicum and Trichoderma viride on oranges, Colletotrichum musae and Gloeosporium musarum on bananas and Botrytis cinerea on grapes); other 88
pathogens on vines, notably Eutypa lata, Guignardia bidwellii, Phellinus igniarus, Phomopsis viticola, Pseudopeziza tracheiphila and Stereum hirsutum; other pathogens on trees (for example Lophodermiunm seditiosum) or lumber, notably Cephaloascusfragrans, Ceratocystis spp., Ophiostoma piceae, Penicillium spp., 5 Trichoderma pseudokoningii, Trichoderma viride, Trichoderma harzianum, Aspergillus niger, Leptographium liindbergi and Aureobasidium pullulans; and fungal vectors of viral diseases (for example Polymyxa graminis on cereals as the vector of barley yellow mosaic virus (BYMV) and Polymyxa betae on sugar beet as the vector of rhizomania), Acremoniella spp., Allomyces spp., Amorphothec spp., 10 Aspergilliusspp., Blastocladiella spp., Candida spp., Chaetomium spp., Coccidioides spp., Conidiobolus spp., Coprinopsis spp., Corynascus spp., Cryphonectria spp., Cryptococcus spp., Cunninghamella spp., Curvularia spp., Debarymyces spp., Diplodia spp. (e.g. Diplodia maydis), Emericella ssp., Encephalitozoon spp., Eremothecium spp., Gaeumanomyces spp. (e.g. Gaeumanomyces graminis f.sp. 15 tritici), Geomyces spp., Gibberella spp. (e.g. Gibberella zeae), Gloeophyllum spp., Glomus spp., Hypocrea spp., Kluyveromyces spp., Lentinula spp., Leptosphaeria salvinii, Leucosporidium spp., Macrophomina spp. (e.g. Macrophomina phaseolina), Magnaportha spp. (e.g. Magnaporthe oryzae), Metharhizium spp., Mucor spp., Neurospora spp., Nectria spp. (e.g. Nectria heamatococca), Paracocidioides spp., 20 Phaeopsheria spp., Phanerochaete spp., Phakopsora spp. (e.g. Phakopsora pachyrhizi), Phymatotrichum spp. (e.g. Phymatotrichum omnivorum), Pneumocystis spp., Pyronema spp., Rhincosporium secalis, Rhizoctonia spp. (e.g. Rhizoctonia solani, Rhizoctonia oryzae or Rhizoctonia cerealis), Rhizopus spp. (e.g. Rhizopus chinensid), Saccharomyces spp., Scerotium spp. (e.g. Scerotium rolfsii), 25 Spizellomyces spp., Thermomyces spp., Thielaviopsis spp. (e.g. Thielaviopsis basicola), Tra metes spp., Trichophyton spp., or Yarrwia spp. Plant diseases caused by fungi including yeasts, rusts, smuts, mildews, molds, mushrooms and toadstools that can be treated using the plant defense elicitor30 compositions according to the present invention are for example: "Rust" is a fungal diseases in plants, which produces reddish-brown discoloration of the stems and leaves. "Black Rot" is characterized by the darkening and decaying of leaves of fruit and vegetable plants. 35 "Black Spot" is one of the many fungal diseases in plants. It is named "black spot" because it produces small black spots on plants. 89
"Bottom Rot" is a fungal disease found on lettuce plants. The characteristic of this fungus is that it first affects the leaves on the lower part of the plant and then moves upward to affect the upper part. "Canker" affects the roots and bark, is found on woody trees and is notorious for 5 causing localized damage to the bark of trees. "Cotton Ball" is notorious for attacking cranberry plants. "Crown Wart" like canker attacks on the barks of woody trees, this fungus attacks the stem of the alfalfa plants. It forms white protrusions at the base of the stem of the plant. 10 "Potato Wart" is a fungal disease that causes dark, warty, spongy excrescences in the eyes of potato tubers, similar to the crown wart in alfalfa plants. "Damping Off" causes excessive moisture conditions of the seedlings. "Dry Rot" causes the drying and crumbling of timber, bulbs, potatoes or fruits."Rhizoctinia Disease" is caused by fungi called Pellicularia and Corticium. It is15 often seen to affect small potatoes. "Root Rot" infects the roots causing root decay, eventually causing the plant to die. "Sclerotium Rot" is caused by Fungus of the genus Sclerotium causing the formation of sclerotia on plants. "Dutch Elm Disease" is a fungal disease affecting Elms. It spreads from one plant to20 another through root grafts or by the elm beetles that feed on small twigs. "Pinkroot" attacks onion plants and makes them unsuitable for consumption. "Soft Rot" is a slimy, mushy decay caused by fungi. "Yellow Spot" is characterized by a yellow spotting on the leaves of plants. "Powdery Mildews" is often specific to the host that it invades. It is normally seen on25 roses, lilac, English oak, zinnias, etc. "Plant Wilting" gets it name because it causes the plant it infects to wilt. The fungus invasion starts in the roots and then slowly makes its way into the stem and plugs the vascular system of the plant. "Decay" is decomposition of wood that is caused by fungi. When it attacks living plant30 tissue, it kills the plants. The pine family are conifers or shrubs including the commercial important cedars, firs, hemlocks, pinons, larches, pines and spruces. In practice they are referred to as "softwood lumber" a broad industry term that refers to all commercial timber derived 35 from gymnosperms, specifically members of the pine family (Pinaceae) and a few other coniferous families. The lignin composition among members of the Pine family (Pinaceae) is highly similar, as they are all gymnosperms (softwoods) that produce 90
predominantly guaiacyl (G-type) lignin. While guaiacyl (G-type) lignin is the dominant lignin in gymnosperms (softwoods) like wood of the pine family, Syringyl- Guaiacyl (SG-type) lignin is typical in hardwoods like wood from birch family, beech family and willow family. Present invention demonstrates that such lignin are useful 5 to produce the plant defence elicitor. Non-limiting examples of phytopathogenic bacteria include the genii Erwinia (including Erwinia amylovora, causing fire blight on pears), Pseudomonas (including Pseudomonas syringae), Xanthomonas (including Xanthomonas orizae, 10 Xanthomonas citri, Xanthomonas fuscans (citrus cancer) and Xanthomonas fragariae) and Ralstonia. Non-limiting examples of phytopathogenic viruses include Cucumber Mosaic Virus, Barley Yellow Mosaic Virus, Strawberry Mild Yellow Edge Virus, Strawberry Latent Ringspot Virus, Beet Necrotic Yellow Vein Virus and Potato Virus Y. 15 Phytopathogenic insects that can be targeted by application of the compositions according to the invention include aphids, beetles, bugs, hoppers, locusts, mites, ants, ticks, trips, whiteflies, rootworms, maggots, weevils, (stem)borers, caterpillars, butterflies, leaf-rolers, leaf- miners, etc.. "Plant protection" as used herein refers to the activation of mechanisms aimed at 20 controlling or reducing the pathogens and/or to minimize their effects on the plant. Plant protection can be achieved by killing the pathogens, by delaying their growth and/or reproduction, by reducing sporulation, etc.. According to another aspect of the present invention, there is provided a method for protecting plants against (infection by) plant pathogens, characterized in that an effective and substantially 25 non-phytotoxic amount of a composition according to the invention is applied to the plants. The expression "effective and non-phytotoxic amount" means an amount of plant defense elicitor composition according to the invention that is sufficient to induce control or destruction of the plant pathogens present or liable to appear on the plants, and that does not entail any appreciable symptom of phytotoxicity for 30 said plants. Such an amount can vary within a wide range depending on the plant pathogen to be controlled, the type of plant, the climatic conditions and the compounds included in the composition according to the invention. This amount can be determined by systematic field trials that are within the capabilities of a person skilled in the art. 35 In a particularly preferred embodiment, the fungicide in the composition of the invention is applied at a reduced rate. Preferably, the rate of the fungicide is reduced by at least a factor 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 25, 40, 50, 60, 70, 80, 90, or 100 91
when compared to the recommended rate, or is reduced by 10, 20, 30, 40, 50, 60, 70, 80, 90, or even 95% or more of the recommended dosage for said plant and/or conditions. More preferably, the rate of the fungicide is reduced by 50% to 90%, 60% to 90%, 70% to 90%, 80% to 90%, 60% to 80%, or 60% to 70% of the 5 recommended rate for said plant and/or conditions. Application of the composition according to the invention can be carried out in accordance with techniques well known to persons skilled in the art. The composition according to the invention can be applied to the whole plant, or to leaves, flowers, fruits, seeds and/or roots of the plant, as well as to the soil or inert substrate wherein 10 the plant is growing or in which it is desired to grow (e.g. inorganic substrates like sand, rockwool, glasswool; expanded minerals like perlite, vermiculite, zeolite or expanded clay), pumice, pyroclastic materials or stuff, synthetic organic substrates (e.g. polyurethane), organic substrates (e.g. peat, composts, tree waste products like coir, wood fibre or chips, tree bark) or to a liquid substrate (e.g. floating 15 hydroponic systems, Nutrient Film Technique, Aeroponics). The application can be done by spraying, drenching, soaking, dipping, injection, etc., or via fertigation systems. It can also be useful to apply the compositions according to the invention to propagation material such as tubers or rhizomes, but also seeds, seedlings or 20 seedlings pricking out and plants or plants pricking out. The compositions according to the invention can also be applied post-harvest to control decay. Among the plants that can be protected by the method according to the invention, mention can be made of cotton; flax; vine; fruit or vegetable crops such as Rosaceae sp. (for instance pip fruit such as apples and pears, but also stone fruit such as apricots, almonds and 25 peaches), Ribesioidae sp., Juglandaceae sp., Betulaceae sp., Anacardiaceae sp., Fagaceae sp., Moraceae sp., Oleaceae sp., Actinidaceae sp., Lauraceae sp., Musaceae sp. (for instance banana trees and plantins), Rubiaceae sp., Theaceae sp., Sterculiceae sp., Vitaceae sp., Rutaceae sp. (for instance lemons, oranges and grapefruit); Solanaceae sp. (for instance tomatoes), Liliaceae sp., Asteraceae sp. (for 30 instance lettuces), Umbelliferae sp., Cruciferae sp., Chenopodiaceae sp., Cucurbitaceae sp., Papilionaceae sp. (for instance peas), Rosaceae sp. (for instance strawberries); major crops such as Graminae sp. (for instance maize, lawn or cereals such as wheat, rice, barley and triticale), Asteraceae sp. (for instance sunflower), Brassicaceae sp. (for instance rapeseed and colza), Fabacae sp. (for instance 35 peanuts), Papilionaceae sp. (for instance soybean), Solanaceae sp. (for instance tomatoes and potatoes), Chenopodiaceae sp. (for instance beetroots); horticultural and forest crops; as well as genetically modified homologues of these crops. 92
In the present invention, the term “monophenolic compounds” means molecules with one phenolic group. If the molecules result from the chemical modification of lignin, they are referred to as “lignin-derived monophenolics”, “lignin-derived monomers”, 5 “lignin monomers”, or “phenolic monomers”. These terms are used interchangeably. Chemical modification herein means depolymerisation and/or partial reduction. The lignin-derived monophenolics comprise compounds having the formulae: 10 93
(ix) each with at least one linkage to an aromatic monomer or aromatic oligomer and - wherein each R22, R23, R25 and R26 is independently chosen from –H, -OH or O-CH3, 5 - wherein R21 is –H - wherein R24 is independently chosen from –H or OH, - wherein R27 is independently chosen from –H or an end-unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, (CH2)2CH2OH, -(CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, -CH=CHCH2OCH3, -(CH2)2CH2OCH2CH3, -10 CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, -CH=CHCH2O(CH2)2CH3, - (CH2)2CH2OCH(CH3)2, -CH=CHCH2OCH(CH3)2, -(CH2)2CH2O(CH2)3CH3, - CH=CHCH2O(CH2)3CH3 Specific examples of said phenolic monomers are: 94
(1) is referred to as “4-(3-hydroxy-propyl)-guaiacol”, “propanol-guaiacol”, or simply “propanol-G” (2) is referred to as “4-(3-hydroxy-propyl)-syringol”, “propanol-syringol”, or simply5 “propanol-S” (3) is referred to as “4-n-propyl-guaiacol”, “propyl-guaiacol”, or simply “propyl-G” (4) is referred to as “4-n-propyl-syringol”, “propyl-syringol”, or simply “propyl-S” 95
In the present invention, the term “diphenolic compounds” means molecules with two phenolic centers chemically linked to each other. Thus having a degree of polymerization (DP) of 2 and two phenol molecules. If the molecules result from the chemical modification of lignin, they are referred to as “lignin-derived diphenolics”, 5 “lignin-derived dimers”, or “phenolic dimers”. These terms are used interchangeably. Chemical modification herein means depolymerisation and/or partial reduction In the present invention, the term “triphenolic compounds” or trimers means molecules with 3 phenolic centers chemically linked to each other. Thus having a 10 degree of polymerization (DP) of 3 and tree phenol molecules. If the molecules result from the chemical modification of lignin, they are referred to as “lignin-derived triphenolics”, “lignin-derived trimers”, or “phenolic trimers”. In the present invention, molecules with three phenolic centers chemically linked to15 each other resulting from the chemical modification of lignin are referred to as “lignin- derived triphenolics”, “lignin-derived trimers”, or “phenolic trimers”. These terms are used interchangeably. In the present invention, the term “tetraphenolic compounds” or tetramers means 20 molecules with 4 phenolic centers chemically linked to each other. Thus having a degree of polymerization (DP) of 4 and four phenol molecules. If the molecules result from the chemical modification of lignin, they are referred to as “lignin-derived tetraphenolics”, “lignin-derived tetramers”, or “phenolic tetramers”. 25 In the present invention, molecules with four phenolic centers chemically linked to each other resulting from the chemical modification of lignin are referred to as “lignin- derived tetraphenolics”, “lignin-derived tetramers”, or “phenolic tetramers”. These terms are used interchangeably. 30 In the present invention, the term “polyphenolic compounds” means molecules resulting from the chemical modification of lignin. Hence they are referred to as “lignin-derived polyphenolics”, “lignin-derived oligomers”, or “phenolic oligomers”. These terms are used interchangeably. Chemical modification herein means depolymerisation and/or partial reduction. 35 96
In the present invention, the term “phenolic compounds” and “phenolic products mixture” are used interchangeably to indicate the mixture comprising monophenolic compounds, diphenolic compounds, triphenolic compounds, tetraphenolic compounds and other phenolic oligomers. 5 In the present invention, the term “hemicellulose-derived polyols” or simply “polyols” means aliphatic alcohols comprising at least two hydroxyl groups. In the present invention, the term “hemicellulose-derived polyols” or “polyols” does not include monosaccharides or oligosaccharides. Hemicellulose-derived polyols include sugar 10 alcohols derived from hydrogenation of monosaccharides. The polyols result from the chemical modification of hemicellulose. Chemical modification herein means hydrolysis and hydrogenation. Hemicellulose-derived polyols primarily include xylitol, arabitol, dulcitol, mannitol, sorbitol, ethylene glycol, glycerol. The term “C5 polyols” is used to indicate the group of polyols comprising 5 carbon atoms, such as xylitol 15 and arabitol. The term “C6 polyols” is used to indicate the group of polyols comprising 6 carbon atoms, such as dulcitol, mannitol, and sorbitol. Likewise, the term “C5 sugars” is used to indicate the group of sugars comprising 5 carbon atoms, such as xylose and arabinose. The term “C6 sugars” is used to indicate the group of sugars comprising 6 carbon atoms, such as glucose, mannose, and galactose. 20 The term “hemicellulose-derived oligosaccharides”, “hemicellulose oligomers” or simply “oligosaccharides” is used to indicate molecules comprising two or more saccharide monomers or saccharide-derived monomers, linked to each other by a glycosidic bond. 25 The term “oligosaccharides” is used to denote saccharide oligomers with a non- reduced terminal saccharide group as well as molecules with a reduced terminal saccharide group. 30 Examples of such oligosaccharides include, but are not limited to, 97
with n between 0-7, for example 0-3. The term “unstable compounds” is used to refer to compounds that are unstable 5 under the reaction conditions of the lignin depolymerisation process or in the formulation of the ISR product, and that cause unwanted side-reactions, such as recondensation. Unstable compounds typically bear a C═O or C═C functional group. Examples of such unstable compounds derived from carbohydrates include, but are not limited to, xylose, glucose, furfural, and hydroxymethylfurfural. Examples of 10 unstable compounds derived from lignin include, but are not limited to, coniferyl alcohol, sinapyl alcohol, phenolic compounds with C2-aldehyde substituents, and so- called Hibbert's ketones. These unstable compounds can be transformed to “stable compounds” by transforming the C═O and/or C═C functional groups, for instance through hydrogenation. The term “stable compounds” is used to refer to compounds 15 that are stable under the reaction conditions or in the formulation of the ISR product, and that do not cause unwanted side-reactions, such as recondensation. Examples of stable compounds derived from carbohydrates include, but are not limited to, xylitol, arabitol, dulcitol, mannitol, sorbitol, ethylene glycol, glycerol. Examples of stable compounds derived from lignin include, but are not limited to, 4-n-20 propanolsyringol, 4-n-propanolguaiacol, 4-n-propylsyringol, 4-n-propylguaiacol. “n-Butanol” or simply “butanol” is abbreviated as “BuOH”. “Methanol” is abbreviated as “MeOH”. 25 “Ethanol” is abbreviated as “EtOH”. 98
Non limiting examples of “lignin” sources are from woody plants (vascular plants (tracheophytes) which includes most trees, shrubs) of the group of softwoods (softwood lumber) and hardwoods (oak, maple, birch), non-woody plants such as 5 grasses (wheat straw, rice straw, switchgrass), herbaceous plants (bamboo, sugar cane), seed coats (nuts, legume pulses, beans), flax stems and hemp stem, jute and bagasse (sugarcane residue). Crops subjected to the plant defense elicitor of the present invention are not 10 particularly limited and any general cultivated plants can be subjected. Examples thereof include the Poaceae plants (such as rice, barley, wheat, corn, oat or lawn grass), the Solanaceae plants (such as tomato, eggplant or potato), the Cucurbitaceae plants (such as cucumber, melon or pumpkin), the Leguminosae plants (such as pea, soybean, kidney bean, alfalfa, peanut, fava bean), the 15 Brassicaceae plants (such as daikon radish, Chinese cabbage, cabbage, komatsuna, rape blossoms, bok choy or A. thaliana), the Rosaceae plants (such as strawberry, apple or pear), the Moraceae (such as mulberry), the Malvaceae (such as cotton), the Umbelliferae (such as carrot, parsley or celery), the Liliaceae (such as green onion, onion or asparagus), the Compositae (such as burdock, sunflower, 20 chrysanthemum, crown daisy, safflower, lettuce) and the Vitaceae (such as grape). Since the reaction which gives rise to plant disease resistance is generally nonspecific to pathogens, all the plant diseases caused by fungus, bacteria and viruses are included as subject diseases. Examples thereof include diseases caused by 25 Magnaporthe grisea, Cochliobolus miyabeanus, Pseudomonas syringae pv. maculicola, Spongospora subterranea, Phytophthora infestans, Peronospora manshurica, Eryshiphe graminis f. sp. hordei, Eryshiphe graminis f. sp. tritici, Gibberella zeae, Mycosphaerella pinodes, Sclerotinia borealis, Puccinia recondita, Ustilago maydis, Ceratobasidium gramineum, Rhizoctonia solani, Rhizoctonia solani, 30 Alternaria solani, Cercospora kikuchii, Fusarium oxysporum f. sp. batatas, Fusarium oxysporum f sp. melonis, Fusarium oxysporum f. sp. lactucae, Fusarium oxysporum f. sp. lycopersici, Fusarium oxysporum f. sp. spinaciae, Verticillium dahliae, Plasmodiophora brassicae, Pythium debaryanum, Botrytis cinerea, Colletotrichum phomoides, Hordeum vulgare, Pseudomas syringae pv. syringae, Erwiniasubsp. 35 atroseptica, Xanthomonas campestris pv. oryzae, Streptomyces scabies, Soil-borne 99
wheat mosaic virus, Soybean mosaic virus, Alfalfa mosaic virus and Potato leafroll virus. The plant defense elicitor of the present invention can be used on plants in any forms 5 such as solution, powder, granule, emulsion, wettable powder, oil, aerosol, flowable by mixing the lignin derived dimers with appropriate additives such as zinc and/or copper, bicarbonates, carbocation scavenger or polyhydric alcohols. Further, optionally, the pH thereof can be adjusted by adding buffer, and properties such as penetration properties to plants or spreading properties can be modified by adding a 10 spreading agent, surfactant such a plant lecithin’s, such as lyso-lecithin, or the like and amino acids such as branched-chain amino acids, proline, glutamic acid, aspartic acid, and histidine. EXAMPLES 15 Example 1: Preparation of depolymerized lignin via reductive catalytic fractionation (RCF). Figure 1 A shows typical input and output of an RCF reaction that can be performed under varying conditions with use of a heterogenous catalyst and under a reductive 20 environment or containing a hydrogen donor as detailed below. The RCF experiment was performed in a 2 L stainless steel batch reactor (Parr Instruments & Co.). 150 g lignocellulose biomass [e.g., pine or spruce, further referred to as ‘Pine’), or poplar] was loaded into the reactor, together with 15.0 g Pd/C or 15.0 g Ru/C and 800 mL methanol. Subsequently, the reactor was sealed, flushed three times with N2 (10 bar) 25 and then pressurized with H2 (30 bar at room temperature). Next, the reaction mixture was stirred (750 rpm) and simultaneously heated to 235 °C (~30 min. heating time). After the reaction time of 3h, the reactor was cooled and depressurized at room temperature. The reactor contents were quantitatively collected by washing the reactor with ethanol. 30 The solid pulp was separated by filtration and washed thoroughly with ethanol. Next, the resulting filtrate was evaporated and a brown oil was obtained, which was subjected to a threefold liquid-liquid extraction using ethyl acetate and water in order to remove extracted sugars. Lastly, the ethyl acetate-extracted phase was dried to obtain the lignin oil.The resulting oils are named with reference to the biomass and 35 catalyst used during the RCF (e.g., pineRuOil). 100
Example 2: Preparation of depolymerized lignin via non-catalytic thermo-solvolytic fractioning. Figure 1 B shows typical input and output streams of a non-catalytic thermos- solvolytic fractioning where no catalyst is used. The non-catalytic thermo-solvolytic 5 fractioning experiment was performed in an identical set-up to the RCF and biomass feedstock with the difference of excluding the redox catalyst and under inert atmosphere, pressurizing with N2 (to 30 bar at room temperature). All other procedures were kept identical. The resulting lignin oils are named with reference to the biomass used during the non-catalytic thermo-solvolytic fractioning (e.g., pine 10 non-catalytic thermo-solvolytic fractioning oils or PineOil). Example 3: Fractionation by sequential liquid-liquid solvent extraction. Figure 1 C shows fractionation of lignin oil through liquid-liquid extraction. An initial fractionation step involved the lignin oil from RCF or non-catalytic thermo-solvolytic fractioning and heptane solvent at a 1:5 ratio (g/mL) with threefold extraction at 15 80°C for 0.5 h. After each extraction, the soluble fraction (H100 liquid, HeptFra, H100L) was separated from the residual fraction (H100 residue, HeptRes, H100R), as shown in figure 1 C, and all solvents of both fractions were removed by rotary evaporator and dried at 80°C in an oven. Figure 1 C also shows a subsequent fractionation steps on the heptane residue, if performed, using a mixture of heptane 20 and ethyl acetate in the v:v ratio of 80% heptane/20% ethyl acetate. Specific fractions are denoted based on the origin of the lignin oil and the solvent composition that was used to obtain the fraction: for example PineRu H80E20 residue (PineRuH80E20R). 25 Example 4: GPC analysis. Figure 2 shows the distribution of the molar mass of the refined lignin oils and fractions obtained from various CF reactions with different feedstocks and catalyst (poplar and Ru/C in Fig.2A, poplar and Pd/C in Fig 2B, pine and Ru/C in Fig.2C, pine 30 and Pd/C in Fig. 2D and pine without catalyst in Fig. 2E), as investigated using gel permeation chromatography – size exclusion (GPC-SEC). Therefore, a lignin sample was solubilized in THF (5 mg mL-1) and subsequently filtered with a 0.2 µm PTFE membrane to remove any particulate matter to prevent plugging of the column. GPC- SEC analyses were performed at 40 °C on a Waters E2695 equipped with a PL-Gel 3 101
µm Mixed-E column with at length of 300 mm, using THF as a solvent with a flow of 1 mL min-1. The detection was UV based at a wavelength of 280 nm. Calibration was based on calibration with commercial polystyrene standards of Agilent. 5 Example 5: Elicitor treatment on A. thaliana– Hyaloperonospora arabidopsidis pathogen system. Figure 3A depicts the A. thaliana – H. arabidopsidis disease assay A. thaliana 10 (ecotype: Columbia-0) plants were seeded in little pots (approximately 40 seeds/pot). Five to seven days after sowing, seedlings were selected so that 20 well- developed, freestanding seedlings were retained in each pot. Eight days after sowing, the plants were treated with mock (1% v/v DMSO), any of the depolymerized lignin fractions (varying concentrations) (1 mg/ml) fraction by spraying the leaves with 15 compound solution until run-off (~20 ml was used per 8 biological replicates). Twenty-four hours after treatment, the leaves of the plants were inoculated by spraying them with a H. arabidopsidis (per 23 pots, 15 ml of spore solution) Noks1 spore suspension of 4.875-8.125 x 104 spores/ml in cold dH2O until run-off. Plants were placed in a closed infection box with high humidity and the infection box was 20 placed in a growth chamber under controlled conditions (16°C, 70% humidity, 12 h day-night cycle, light intensity of approximately 100 μmol/m2s). Three days post infection, leaves of 15 plants per pot were taken, transferred to an Eppendorf tube containing 250 µl dH2O and stored at -80 °C and used for genomic DNA extraction with qPCR to determine relative pathogen proliferation. 25 Example 6: Genomic DNA extraction and qPCR. For isolation of plant genomic DNA (gDNA), plant samples were grinded using the Precellys 24 tissue homogenizer at 6000 rpm for 10 seconds. After homogenization, 30 400 μl of Edwards buffer (200 mM Tris-HCl pH 7.5, 250 mM NaCl, 25 mM EDTA, and 0.5% (v/v) SDS) was added to each sample. The samples were vortexed and incubated at 55°C for 15 min. After incubation, samples were centrifuged for 2 min at 13000 rpm. Subsequently, 20 μl of the supernatant was transferred to a new, sterile 1.5 ml Eppendorf tube and an equal amount of isopropanol was added to 35 precipitate the gDNA. The mixture was incubated for 10 min at RT, after which it was centrifuged for 10 minutes at 13000 rpm. The supernatant was discarded and the pellet was washed with 70% ethanol. After another centrifugation round of 5 min at 102
13000 rpm, the ethanol was removed and the pellet was dried for one hour at 37°C. The pellet, containing the isolated gDNA, was resuspended in 40 μl sterile demi water. The concentration of gDNA was determined using the NanoDropTM One Microvolume UV-Vis Spectrophotometer (Thermo Fisher Scientific, US) by measuring the 5 absorbance at 260 nm. The gDNA samples were diluted to 10 ng/μl and stored at - 20°C. The protocol for pathogen growth quantification via qPCR was as follows. Samples for qPCR were prepared by mixing 2.5 μl gDNA (= 25 ng) with 6.125 μl 2X SYBR® Green Mastermix (Thermo Fisher Scientfic, US) and 500 nM of the reverse10 and forward primer each. The final volume was adjusted to 12.5 μl using sterile Milli- Q® water. The samples were loaded in an MicroAmpTM Fast Optical 96-Well Reaction Plate (Thermo Fischer Scientific, US) which was kept on ice. Post loading, the plate was shortly centrifuged. The qPCR was performed in triplicate for each biological sample on the StepOnePlusTM Real-Time PCR system (Thermo Fisher Scientific, US). 15 After 10 min at 95°C, samples were run for 40 cycles of 15 s at 95°C, 15 s at 57°C, and 15 s at 72°C. After each run, melting curves were acquired to check for amplification specificity by heating the samples from 60°C to 95°C. The Ct values were determined by the included StepOneTM software. For each biological replicate, the relative amount of pathogen gDNA over plant gDNA was calculated using the 20 formula below (adapted from Livak and Schmittgen (2001)). Relative amount gDNA pathogen/gDNA plant = 2−∆Ct = 2−(Ct pathogen−Ct plant) Primers used are as in Table 1 Example 7: Elicitor treatment on Solanum lycopersicum – Botrytis cinerea pathogen25 system Figure 6 depicts the hydroponics tomato – B. cinerea assay, with tomato plants grown in a lab-scale hydroponics setup (Araponics Liège, Belgium). The hydroponics tanks were filled with 1.6 l plant nutrient solution containing the macronutrients 30 MgSO4.7H2O (500 mg/l), KH2PO4 (270 mg/l), KNO3 (200 mg/l), K2SO4 (100mg/l), Ca(NO3)2.4H2O (500 mg/l), and FeEDTA sodium salt (25 mg/l); and the micronutrients H3BO3 (4.1 mg/), MnSO4.H2O (3.7 mg/l), CuCl2.2H2O (0.2 mg/l), (NH4)6Mo7O24.4H2O (0.0825 mg/l with 81.2% MoO3), and ZnSO4.7H2O (0.649 mg/l). Tomato cultivar seeds were sown in the hydroponic systems in seed holders 35 (18 seeds/system) containing a solidified 0.65% (w/v) agar in water solution. The hydroponic tanks were covered with plastic lids and placed in the plant growth chamber. One week later, the seeds had germinated so the lids were removed and 103
aeration pumps were installed to aerate the root compartments. After 24 days, 8 plants were selected per system and were treated with plant defense elicitor compounds by spraying the leaves with compound solution until run-off. Treatment with the solvent 1% v/v DMSO was included as mock treatment. Three, twelve and 5 seventeen days after treatment, five leaflets per plant were inoculated with 5 μl droplets of a B. cinerea R16 strain spore suspension of 5 x 105 spores/ml in ½ potato dextrose broth. The hydroponics tanks were placed inside an infection box, containing a moist mat to obtain high humidity, in the growth chamber. The disease symptoms were quantified by measuring the diameter parallel to the midrib of the developing10 necrotic lesions at 2 dpi. The lesion area was calculated using the formula below Lesion area = (Lesion diameter / 2)^2 * π Example 8: Protecting Solanum lycopersicum against insect feeding - Induction by RCF lignin of increased resistance in tomato against insects 15 Figure 7 shows protection of tomato plants (Solanum lycopersicum) against insect feeding (Nesidiocoris tenuis) by RCF lignin treatment. Solanum lycopersicum (cultivar: Moneymaker) seeds were disinfected by 5 min exposure to 20% bleach and subsequently germinated in petri dish covered with moisturized paper. Post germination, seeds were seeded in soil. Thirty-eight days post seeding, plants were 20 treated with mock (1% v/v DMSO) and PineRuH100R (1 mg/ml) by spraying leaves till run-off. Afterwards, plants were placed in air inflated plant cages (60 cm × 40 cm × 40 cm with mesh size 0.25 mm × 0.25 mm, Entomologie-Speciaalzaak Vermandel V.O.F., the Netherlands), which could be accessed by opening the zipper. Three days after initial treatment, the plants received a second treatment 3 hours prior to insect 25 infestation. Plants were infested by introducing one Nesidiocoris tenuis female per plant cage. To force the insects to feed on the plant no additional food or prey were present in the cage. The total number of necrotic rings on leaves and shoots were assessed 7 days after infestation as a measure of feeding damage. Statistical differences were determined by using a student’s t test (p< 0.05). Experiments were 30 conducted in a fully randomized block design in a climate controlled greenhouse compartments (T = 20°C +/- 4°C, RH = 70%, and a 18L:6D photoperiod). Example 9: Induction by RCF lignin of increased resistance in tomato against heat stress 104
Figure 8 depicts decreased transpiration rate of tomato plants (Solanum lycopersicum) by RCF lignin treatment. Solanum lycopersicum (cultivar: Alisa Craig) were sown in seed holders containing 0.65% (w/v) agar in a water solution placed in a hydroponics tank (Araponics Liege, Belgium) filled with 1.6 L of nutrient solution 5 containing the macronutrients MgSO4.7H2O (500 mg/l), KH2PO4 (270 mg/l), KNO3 (200 mg/l), K2SO4 (100 mg/l), Ca(NO3)2.4H2O (500 mg/l), and FeEDTA sodium salt (25 mg/l); and the micronutrients H3BO3 (4.1 mg/), MnSO4.H2O (3.7 mg/l), CuCl2.2H2O (0.2 mg/l), (NH4)6Mo7O24.4H2O (0.0825 mg/l with 81.2 % MoO3), and ZnSO4.7H2O (0.649 mg/l). After 24 days S. lycopersicum leaves were sprayed with 10 mock (1% v/v DMSO) or PineRuH100R (1 mg/ml) till run off. Three-days post spraying plants were placed in a heat stress cabinet (38 °C). Determination of transpiration was carried out by clipping the apical side of the youngest fully developed leaf into the LI-COR 600 device at 0h, 2h, 4h, 6h and 6h + 2h recovery time post heat-stress initiation. 15 Example 10: Improved survival of Arabidopsis thaliana under heat stress by RCF lignin treatment. Figure 9 depicts improved survival rate of Arabidopsis thaliana under heat stress by RCF lignin treatment. The A. thaliana – heat stress protocol was adapted from Silva- 20 Correi et al., 2014. Briefly, Arabidopsis thaliana seeds were surface sterilized by 10 minute exposure to 30% bleach. Subsequently, 50 seeds were sown on 1X Murashige and Skoog (MS) plates. Four days post seeding, plants were treated with either mock (1% v/v DMSO) or PineRuH100R (1 mg/ml in 1% v/v DMSO) by submerging seedlings in 5 µl droplets. Three days after, heat stress was imposed by submersion 25 of parafilm-sealed plates into a water bath (45 °C) for 18 minutes. Following heat treatment, plates were returned to the growth chamber and allowed to recover for six days. Next, seedling survival was assessed by counting seedlings that remained green. Statistical differences were determined by using a student’s t test (p< 0.05). Plants were grown vertically in climate controlled growth chambers (T = 22°C, RH = 30 70%, and a 12L:12D photoperiod). Fig 10. demonstrates that the seedling survival is improved in seedlings that were treated with PineRuH100R. Example 11: Preparation of depolymerized lignin via oxidative catalytic fractionation (OCF). 35 Figure 10 shows typical input and output of an OCF reaction that can be performed under varying conditions with use of a heterogenous catalyst and under oxygen gas (O2) or air. The OCF experiment was performed in a 50 mL stainless steel batch 105
reactor with an alkali-resisting polyphenylene liner. 0.5 g lignocellulose biomass (birch wood) was loaded into the reactor, together with 1.26 mmol of catalyst (CuO) and 25 mL of NaOH aqueous solution . Subsequently, the reactor was sealed, flushed and then pressurized with O2 (1MPa at room temperature). Next, the 5 reaction mixture was stirred (150 to 1100 rpm) and simultaneously heated up to 160 °C (~30 min. heating time). After the reaction time of max 2h, the reactor was cooled and depressurized at room temperature. After reaction, the slurry was centrifuged to separate the solid residue (including the pulp and spent catalyst) and liquid. The obtained liquid was acidified by HCl until pH 10 2−3. The acidified liquid was extracted with chloroform or ethyl acetate (EtOAc) until the organic phase was colorless. A small amount of NaHCO3 was then added into the organic phase to neutralize the residual acid. Anhydrous Na2SO4 was used to remove water. By evaporation of the organic phase, crude monophenolics and oligomers were obtained, and chloroform/EtOAc can be reused in the extraction step. The water15 phase was centrifuged to separate acid-insoluble oligomers and the acid (and water)- soluble portion. The acid-insoluble oligomers were washed with deionized water until the pH of the eluent was 7. The acid (and water)-soluble portion was vacuum distilled at 60 °C to remove H2O and HCl, and solid salts(including NaCl) were obtained. The salts were freeze-dried. The resulting depolymerized lignin mixtures are named OCF20 4.1 (OCF monomers) and OCF 4.2 (OCF oligomers) Example 12: Elicitor treatment on A. thaliana– Hyaloperonospora arabidopsidis pathogen system with OCF lignin Figure 11 shows the A. thaliana (ecotype: Columbia-0) plants seedlings in little pots (20 well-developed, freestanding seedlings) treated after eight days after sowing 25 treated with mock (1% v/v DMSO) (Fig 11A), and depolymerized lignin monomers from OCF (OCF 4.1, 1 mg/ml) (Fig 11B) fraction by spraying the leaves with compound solution until run-off. The Figures clearly show the toxic impact of the monomers from OCF (OCF 4.1) on the plants that resulted in limited growth as compared to the plant seedlings without the treatment. 30 FIG.12 shows the relative pathogen proliferation of Hyaloperonospora arabidopsidis in A. thaliana plants treated with the heptane insoluble fraction from a depolymerized lignin obtained from the catalytic RCF process of pine wood (PineRuH100R) or OCF oligomers (OCF 4.2) obtained from birch wood using a catalytic OCF process demonstrating equal elicitor activity of depolymerized lignin oligomers (DP ≥ 2) from35 RCF and OCF 106
According to the present invention there is provided a phytopharmaceutical or agrochemical composition, wherein an effective dose of plant defense elicitor compounds that are lignin-derived oligomer aromatics with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4, or synthesised structurally similar 5 compounds. The present invention a further embodiment also provides that the plant defense elicitor compounds are from a lignin that is depolymerized or decomposed by reductive catalytic fractionation (RCF) of lignin or lignocellulose or that the plant defense elicitor compounds are from a lignin that is depolymerized or decomposed 10 by non-catalytic thermo-solvolytic depolymerisation of lignin or lignocellulose. In another aspect, the present invention provides that the plant defense elicitor compounds are from a lignin that is depolymerized or decomposed or 1) by a reductive catalytic fractionation (RCF) (Fig. 1A) of the lignin source with a heterogenous metal catalyst, including but not limited to Ru, Pd and Ni, on a support 15 in an organic solvent or an organic solvent water mixture in a temperature range of 100°C to 300°C, preferably 150°C to 270°C and most preferably 200°C to 250°C and containing a hydrogen donor, including but not limited to H2 or 2) by a non-catalytic thermo-solvolytic de-polymerization (Fig. 1B) of the lignin source in an organic solvent or an organic solvent water mixture in a temperature range of 100°C to 20 300°C, preferably 150°C to 270°C and most preferably 200°C to 250°C and under an inert atmosphere. These plant defense elicitor compounds can thus be the reaction product of these processes of lignin depolymerization. This embodiment of the invention advantageously comprises that the plant defense elicitor compounds are obtained from lignin depolymerization or decomposing by reductive catalytic25 fractionation (RCF), as this resulted to the most stable compositions when no re- polymerization where add. In another aspect, the present invention provides that the phytopharmaceutical or agrochemical composition has a pH in the range of 4 to 10, preferably in the range of 5 to 8 or that the plant defense elicitor compounds in origin have a pH in the range30 of 4.0 to 6.0. Some of the compositions described above may be embodied as substantially free of acetic acid, methanol and ethanol, meaning it contains less than 0,1%, of each acetic acid, methanol and ethanol. In a further embodiment of the invention, the compositions described above is characterised in that the plant defense elicitor compounds comprise, or essentially 35 consist of or consist of I) at least one aromatic compound selected from the formulae 107
- wherein each R1, R3, and R4 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R2 is –H, a β-O-4 linkage to an aromatic monomer or aromatic 10 oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R5 is selected of –H, a β-O-4 linkage to an aromatic monomer 15 or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, an end-unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, - CH=CHCH3, -(CH2)2CH2OH, -(CH2)2CHO, -CH=CHCH2OH, -20 (CH2)2CH2OCH3, -CH=CHCH2OCH3, -(CH2)2CH2OCH2CH3, - CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, -CH=CHCH2O(CH2)2CH3, 108
-(CH2)2CH2OCH(CH3)2, -CH=CHCH2OCH(CH3)2, - (CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, a carbon linkage to an aromatic monomer or aromatic oligomer; and/or II) wherein the aromatic compounds comprise at least one aromatic 5 compound selected from the formulae (v) , (vi) - wherein each R12, R13, R15 and R16 is independently chosen from –H, - OH, O-CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, 10 a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R11 and R14 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an 15 aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, 109
and/or III) wherein at the aromatic compounds comprise at least one aromatic compound selected from the formula (viii) , (xi) (xviii) 110
each with at least one linkage to an aromatic monomer or aromatic oligomer and - wherein R21 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an 5 aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein each R22 and R23 is dependently chosen from -H, -OH, O-CH3 or from a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 10 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or an aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein the alkyl group is derived from the alcohol solvent of the process,15 - wherein each R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or an aromatic oligomer, or a carbon- 20 oxygen linkage to an aromatic monomer or aromatic oligomer, wherein R27 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or arom- atic oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, end-25 unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, - (CH2)2CH2OH, -(CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, - CH=CHCH2OCH3, -(CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, - (CH2)2CH2O(CH2)2CH3, -CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, - CH=CHCH2OCH(CH3)2, -(CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, a 30 carbon linkage to an aromatic monomer or an aromatic oligomer. In a further embodiment of the invention, the compositions described above is characterised in that the plant defense elicitor compounds comprise, or essentially consist of or consist of I) at least one aromatic compound selected from the 111
- wherein each R1, R3, and R4 is independently chosen from –H, -OH, O- CH3, 5 - wherein R2 is –H, - wherein R5 is selected of –H, an end-unit selected of CH3, -CH2CH3, - (CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, -(CH2)2CH2OH, -(CH2)2CHO, - CH=CHCH2OH, -(CH2)2CH2OCH3, -CH=CHCH2OCH3, - (CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, -10 CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, - CH=CHCH2OCH(CH3)2, -(CH2)2CH2O(CH2)3CH3, - CH=CHCH2O(CH2)3CH3, - and/or II) wherein the aromatic compounds comprise at least one aromatic compound selected from the formulae (v) 112
- wherein each R12, R13, R15 and R16 is independently chosen from –H, - OH, O-CH3, 5 - wherein R11 and R14 is independently chosen from –H, and/or III) wherein at the aromatic compounds comprise at least one aromatic compound selected from the formula (viii) 113
and (xiv) each with at least one linkage to an aromatic monomer or aromatic oligomer and - wherein R21 is independently chosen from –H, a β-O-4 linkage to an 5 aromatic monomer, a 4-O-5 linkage to an aromatic monomer, an α-O- 4 linkage to an aromatic monomer or a carbon-oxygen linkage to an aromatic monomer, - wherein each R22 and R23 is dependently chosen from -H, -OH, O-CH3 or where at least R22 or R23 is independently chosen from a 4-O-5 10 linkage to an aromatic monomer, a 5-5 linkage to an aromatic monomer, a β-5 linkage to an aromatic monomer, a carbon linkage to an aromatic monomer, or a carbon-oxygen linkage to an aromatic monomer, - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein 15 the alkyl group is derived from the alcohol solvent of the process, - wherein each R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer, a 5-5 linkage to an aromatic monomer, a β-5 linkage to an aromatic monomer, a carbon linkage to an aromatic monomer, or a carbon-oxygen linkage to an 20 aromatic monomer, - wherein R27 is independently chosen from –H, end-unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, -(CH2)2CH2OH, - (CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, -CH=CHCH2OCH3, - 115
(CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, - CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, - CH=CHCH2OCH(CH3)2, -(CH2)2CH2O(CH2)3CH3, - CH=CHCH2O(CH2)3CH3. - In a further embodiment of the invention, the compositions described above is characterised in that the plant defense elicitor compounds comprise, or essentially consist of or consist of at least one aromatic compound selected from the formulae 116
or a combination thereof. In a further embodiment of the invention, the compositions described above is characterised in that the plant defense elicitor compounds are phenolic oligomers 5 comprising two benzene rings directly bridged or bridged with a common bridging 118
group of the group consisting of aliphatic chains, alkene groups, carbonyl groups and ether linkages or wherein the phenolic oligomers have two aromatic groups. In a further embodiment of the invention, the compositions described above is characterised in that the plant defense elicitor compounds are phenolics oligomers 5 comprising two benzene rings directly bridged or bridged with a common bridging group of the group consisting of -CH₂- groups, -CH=CH-, -C(=O)-and -O-. In a further embodiment of the invention, the compositions described above is characterised in that the plant defense elicitor compounds are 0,5 to 30 wt% by dry weight of aromatic compounds, preferably 1 to 20 wt% by dry weight of aromatic 10 compounds, more preferably 2 to 10 wt% of the composition in dry state. In another aspect, the phytopharmaceutical or agrochemical composition according to the present invention, further comprises an ingredient of the group consisting of a surfactant, a biosurfactant, a penetration enhancer, a dispersing agent, an emulsifier and a carrier or a combination thereof. 15 In yet another aspect, the phytopharmaceutical or agrochemical composition according to the present invention, further comprises a polymerization inhibitor of the group of a carbocation scavenger, aromatic scavengers and a polyhydric alcohol or a combination thereof. In yet another aspect, the phytopharmaceutical or agrochemical composition 20 according to the present invention, further comprises a polymerization inhibitor whereby the repolymerization inhibitor is a compound of the group consisting of citric acid, salicylic acid, 2-naphthol, phenolic acids (e.g., vanillic acid, syringic acid), ethylene glycol, glycerol, mannitol (C6H14O6), sorbitol (C6H14O6), xylitol (C5H12O5), erythritol, maltitol (C12H24O11). 25 In yet another aspect, the phytopharmaceutical or agrochemical composition according to the present invention, further comprises a stabilizing agents of the group consisting of 1,4-butanediol, 2-hydroxy-1-naphthoic acid, 2-naphthol, 2- naphthol-7-sulfonat, 2-naththol, 3-hydroxy-2-naphthoic acid, 4-hydroxybenzoic acid, 6-hydroxy-2-naphthoic acid, ascorbic acid, bovine serum albumin, citric acid,30 salicylic acid, cysteine, dimethylphloroglucinol, histidine, mannitol, o- dihydroxybenzene, p-benzenediol, phloroglucinol, resorcinol, soy protein isolate, syringic acid and vanillic acid so to prevent repolymerisation. In yet another aspect, the phytopharmaceutical or agrochemical composition according to the present invention, further comprises a second plant defense elicitor 119
of the group consisting of alginate, hexokinase, laminarin, sodium silicate, silicon, oligo-galacturonan, cellodextrin and/or chito-oligosaccharide or a plant defense elicitor selected from the group consisting of pectin fragment, oligogalacturonide, cellobiose, xyloglucan, non-branched P-l,3-glucan, chitin fragment, arabinose, 5 arabinan, rhamnose, homogalacturonan, rhamnogalacturonan I and II, xylogalacturonan and starch or combinations thereof In yet another aspect, the phytopharmaceutical or agrochemical composition according to the present invention, further comprises a fungicide, an antimicrobial, 10 an insecticidal, and/or an antiviral for instance. In a practical embodiment, the phytopharmaceutical or agrochemical composition according to the present invention and described above comprises the lignin-derived oligomer aromatics with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 15 4 from lignin that is from coconut husk, softwood trees, hardwood trees, a grass (such as bamboo, corn stalks & stover; wheat straw, rice straw, barley straw, miscanthus), flax shives or hemp stalk or a combination thereof. In yet a practical embodiment, the phytopharmaceutical or agrochemical composition 20 according to the present invention and described above comprises an ingredient of the group consisting of a surfactant, a biosurfactant, a penetration enhancer, a dispersing agent, an emulsifier and a carrier or a combination thereof. In yet a practical embodiment, the phytopharmaceutical or agrochemical composition 25 according to the present invention and described above further comprises a second plant defense elicitor, for instance of the group consisting of alginate, hexokinase, laminarin, sodium silicate, silicon, oligo-galacturonan, cellodextrin and/or chito- oligosaccharide or a plant defense elicitor selected from the group consisting of pectin fragment, oligogalacturonide, cellobiose, xyloglucan, non-branched P-l,3-glucan, 30 chitin fragment, arabinose, arabinan, rhamnose, homogalacturonan, rhamnogalacturonan I and II, xylogalacturonan, starch, and combinations thereof. In yet a practical embodiment, the phytopharmaceutical or agrochemical composition according to the present invention and described above further comprises a 35 fungicide, antimicrobial, an insecticidal, or an antiviral. 120
In yet a practical embodiment, the phytopharmaceutical or agrochemical composition according to the present invention and described above further comprises a fungicide selected form the group consisting of phosphonates, benzamides, carbamates, dithiocarbamates , phtalimides, triazoles, quinolines, sulphur, and 5 cyanoimidazoles or a fungicide selected from the group consisting of 2,6-dichloro-N- [3-chloro-5-(trifluoromethyl)-2-pyridinylmethyl]benzamide; propyl 3- (dimethylamino)propylcarbamate hydrochloride; (2RS,3SR)-1-[3-(2-chlorophenyl)- 2,3-epoxy-2-(4-fluorophenyl)propyl]-1H-1,2,4-triazole; 5,7-dichloro-4-quinolyl 4- fluorophenyl ether; sulphur; 4-chloro-2-cyano-N,N-dimethyl-5-(4-methylphenyl)- 10 1H-imidazole-1-sulfonamide; N-(trichloromethylthio)phthalimide; manganese ethylenebis(dithiocarbamate) (polymeric) complex with zinc salt and methyl (E)- methoxyimino-{(E)-α-[1-(α,α,α-trifluoro-m-tolyl)ethylideneaminooxy]-o- tolyl}acetate; or a combination thereof. 15 In another aspect, the present invention provides the phytopharmaceutical or agrochemical composition according to the present invention and described above further comprises stabilizing agents of the group consisting of 1,4-butanediol, 2- hydroxy-1-naphthoic acid, 2-naphthol, 2-naphthol-7-sulfonat, 2-naththol, 3- hydroxy-2-naphthoic acid, 4-hydroxybenzoic acid, 6-hydroxy-2-naphthoic acid, 20 ascorbic acid, bovine serum albumin, citric acid, salicylic acid, cysteine, dimethylphloroglucinol, histidine, mannitol, o-dihydroxybenzene, p-benzenediol, phloroglucinol, resorcinol, soy protein isolate, syringic acid and vanillic acid so to prevent repolymerization. 25 In another aspect, the present invention provides the phytopharmaceutical or agrochemical composition according to the present invention comprises a) the plant defense elicitor of present invention and as described hereabove and b) a fungicide, for instance a fungicide selected form the group consisting of selected from the group comprising: phosphonates, benzamides, carbamates, dithiocarbamates , 30 phtalimides, triazoles, quinolines, sulphur, and cyanoimidazoles or a fungicide selected from the group consisting of 2,6-dichloro-N-[3-chloro-5-(trifluoromethyl)-2- pyridinylmethyl]benzamide; propyl 3-(dimethylamino)propylcarbamate hydrochloride; (2RS,3SR)-1-[3-(2-chlorophenyl)-2,3-epoxy-2-(4- fluorophenyl)propyl]-1H-1,2,4-triazole; 5,7-dichloro-4-quinolyl 4-fluorophenyl35 ether; sulphur; 4-chloro-2-cyano-N,N-dimethyl-5-(4-methylphenyl)-1H-imidazole- 1-sulfonamide; N-(trichloromethylthio)phthalimide; manganese ethylenebis(dithiocarbamate) (polymeric) complex with zinc salt and methyl (E)- 121
methoxyimino-{(E)-α-[1-(α,α,α-trifluoro-m-tolyl)ethylideneaminooxy]-o- tolyl}acetate; or a combination thereof In another aspect, the present invention provides the phytopharmaceutical or 5 agrochemical composition according to the present invention and described above comprises also another type of plant defense elicitor such as alginate, hexokinase, laminarin, sodium silicate, silicon, oligo-galacturonan, cellodextrin and/or chito- oligosaccharide or a plant defense elicitor selected from the group consisting of pectin fragment, oligogalacturonide, cellobiose, xyloglucan, non-branched P-l,3-glucan, 10 chitin fragment, arabinose, arabinan, rhamnose, homogalacturonan, rhamnogalacturonan I and II, xylogalacturonan, starch, and combinations thereof. In another aspect, the present invention provides the phytopharmaceutical or agrochemical composition according to the present invention and described above 15 further comprises a co-formulant selected from the group comprising: detergents, emulsifiers, dispersing agents, anti-foaming agents, penetration enhancers, humectants, wetting agents of ionic or non-ionic type, anti-freeze agents, preservative agents, absorbent agents, thickeners, buffers, sticker agents, diluents or a mixture thereof, preferably a surfactant selected from the group comprising: 20 detergents, emulsifiers, dispersing agents, anti-foaming agents, penetration enhancers, humectants or wetting agents of ionic or non-ionic type, or a mixture thereof. In another aspect, the present invention provides the phytopharmaceutical or 25 agrochemical composition according to the present invention and described above further comprises a surfactant comprising one or more of the following components: castor oil ethoxylate, rapeseed methyl ester, alkyl phosphates, tributyl phosphate, tripropyl phosphate, naphthalenesulphonic acid salts, a combination of organic sulfonate and 2-methylpentane-2,4-diol, alkylpolyglucoside, siloxanes derivates, 30 alkylsulfonates, polycarboxylates, lignosulfonates, alkoxylated triglycerides, fatty amines polymers, dioctylsulfosuccinates or polyoxyethylene (20) sorbitan monolaurate, preferably C18-castor-oil-ethoxylate, a combination of organic sulfonate and 2-methylpentane-2,4-diol, or polyoxyethylene (20) sorbitan monolaurate. 35 The invention provides a method of activating plant defense against a pathogen stressor and/or abiotic stressor, comprising contacting said plant with an effective 122
amount of a lignin-derived aromatic oligomers with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4, or a synthesised structurally similar compound. This embodiment of the invention advantageously comprises activating plant defense preventive or activating plant defense in case of a stressor plant attack. In an aspect, 5 the present invention provides that the lignin-derived aromatic oligomers are obtained from a lignin that is depolymerized or decomposed by reductive catalytic fractionation (RCF) of lignin or lignocellulose. This embodiment of the invention advantageously provide very stable lignin-derived aromatic oligomers oils, even at room temperature and when no re-polymerization inhibitors are added. In another 10 application, the method according to the present invention activating plant defense against a pathogen stressor and/or abiotic stressor provides that the lignin-derived aromatic oligomers are from a lignin that is depolymerized or decomposed by non- catalytic thermo-solvolytic depolymerisation of lignin or lignocellulose. 15 Some of the methods described above may be embodied as that the lignin-derived aromatic oligomers comprise, or essentially consist of or consist of I) at least one - wherein each R1, R3, and R4 is independently chosen from –H, -OH, O- 20 CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 123
linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R2 is –H, a β-O-4 linkage to an aromatic monomer or aromatic 5 oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R5 is selected of –H, a β-O-4 linkage to an aromatic monomer 10 or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, an end-unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, - CH=CHCH3, -(CH2)2CH2OH, -(CH2)2CHO, -CH=CHCH2OH, -15 (CH2)2CH2OCH3, -CH=CHCH2OCH3, -(CH2)2CH2OCH2CH3, - CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, -CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, -CH=CHCH2OCH(CH3)2, - (CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, a carbon linkage to an aromatic monomer or aromatic oligomer; 124
and/or II) wherein the aromatic compounds comprise at least one aromatic compound selected from the formulae (v) - wherein each R12, R13, R15 and R16 is independently chosen from –H, - 5 OH, O-CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, 10 - wherein R11 and R14 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, 125
and/or III) wherein at the aromatic compounds comprise at least one aromatic compound selected from the formula (viii) , (xi) (xviii) 126
each with at least one linkage to an aromatic monomer or aromatic oligomer and - wherein R21 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an 5 aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein each R22 and R23 is dependently chosen from -H, -OH, O-CH3 or from a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 10 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or an aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein the alkyl group is derived from the alcohol solvent of the process,15 - wherein each R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or an aromatic oligomer, or a carbon- 20 oxygen linkage to an aromatic monomer or aromatic oligomer, wherein R27 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or arom- atic oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, end-25 unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, - (CH2)2CH2OH, -(CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, - CH=CHCH2OCH3, -(CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, - (CH2)2CH2O(CH2)2CH3, -CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, - CH=CHCH2OCH(CH3)2, -(CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, a 30 carbon linkage to an aromatic monomer or an aromatic oligomer. Some of the methods described above may be embodied as that the lignin- derived aromatic oligomers comprise, or essentially consist of or consist of I) at least one aromatic compound selected from the formulae (i) 127
(iv) and - wherein each R1, R3, and R4 is independently chosen from –H, -OH, O- CH3, 5 - wherein R2 is –H, - wherein R5 is selected of –H, an end-unit selected of CH3, -CH2CH3, - (CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, -(CH2)2CH2OH, -(CH2)2CHO, - CH=CHCH2OH, -(CH2)2CH2OCH3, -CH=CHCH2OCH3, - (CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, -10 CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, - CH=CHCH2OCH(CH3)2, -(CH2)2CH2O(CH2)3CH3, - CH=CHCH2O(CH2)3CH3, - and/or II) wherein the aromatic compounds comprise at least one aromatic compound selected from the formulae (v) 128
and - wherein each R12, R13, R15 and R16 is independently chosen from –H, - OH, O-CH3, 5 - wherein R11 and R14 is independently chosen from –H, and/or III) wherein at the aromatic compounds comprise at least one aromatic compound selected from the formula (viii) 129
and (xiv) each with at least one linkage to an aromatic monomer or aromatic oligomer and - wherein R21 is independently chosen from –H, a β-O-4 linkage to an 5 aromatic monomer, a 4-O-5 linkage to an aromatic monomer, an α-O- 4 linkage to an aromatic monomer or a carbon-oxygen linkage to an aromatic monomer, - wherein each R22 and R23 is dependently chosen from -H, -OH, O-CH3 or where at least R22 or R23 is independently chosen from a 4-O-5 10 linkage to an aromatic monomer, a 5-5 linkage to an aromatic monomer, a β-5 linkage to an aromatic monomer, a carbon linkage to an aromatic monomer, or a carbon-oxygen linkage to an aromatic monomer, - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein 15 the alkyl group is derived from the alcohol solvent of the process, - wherein each R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer, a 5-5 linkage to an aromatic monomer, a β-5 linkage to an aromatic monomer, a carbon linkage to an aromatic monomer, or a carbon-oxygen linkage to an 20 aromatic monomer, - wherein R27 is independently chosen from –H, end-unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, -(CH2)2CH2OH, - (CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, -CH=CHCH2OCH3, - (CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, - 131
CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, - CH=CHCH2OCH(CH3)2, -(CH2)2CH2O(CH2)3CH3, - CH=CHCH2O(CH2)3CH3. Some of the methods described above may be embodied as that the lignin-derived 5 aromatic oligomers comprise, or essentially consist of or consist of at least one aromatic compound selected from the formulae (i) , (v) 132
or a combination thereof. Some of the methods described above may be embodied as that the lignin-derived 5 aromatic oligomers are phenolics comprising two benzene rings directly bridged or bridged with a common bridging group of the group consisting of aliphatic chains, 134
alkene groups, carbonyl groups and ether linkages or wherein the phenolic oligomers have two aromatic groups. Yet some of the methods described above may be embodied as that the lignin-derived 5 aromatic oligomers are phenolics comprising two benzene rings directly bridged or bridged with a common bridging group of the group consisting of -CH₂- groups, - CH=CH-, -C(=O)-and -O-. Preferably these lignin-derived aromatic oligomers are substantially free of acetic acid, methanol and ethanol, meaning it contains less than 0,1%, of each acetic acid, 10 methanol and ethanol. Furthermore preferably these lignin-derived aromatic oligomers are comprised in a composition with a pH in the range of 4 to 10, preferably in the range of 5 to 8 or in origin have a pH in the range of 4.0 to 6.0. In another aspect, the present invention provides that the lignin-derived aromatic 15 oligomers are comprised in a composition, further comprising an ingredient of the group consisting of a surfactant, a biosurfactant, a penetration enhancer, a dispersing agent, an emulsifier and a carrier or a combination thereof. In another aspect, the present invention provides that these lignin-derived aromatic 20 oligomers are comprised in a composition, further comprising a repolymerization inhibitor of the group of a carbocation scavenger, aromatic scavengers and a polyhydric alcohol or a combination thereof or a repolymerization inhibitor of the group consisting of citric acid, salicylic acid, 2-naphthol, phenolic acids (e.g., vanillic acid, syringic acid), ethylene glycol, glycerol, mannitol (C6H14O6), sorbitol (C6H14O6),25 xylitol (C5H12O5), erythritol, maltitol (C12H24O11) or a combination thereof. In another aspect, the present invention provides that these lignin-derived aromatic oligomers are 0,5 to 30 wt% by dry weight of aromatic compounds, preferably 1 to 20 wt% by dry weight of aromatic compounds, more preferably 2 to 10 wt% of the 30 composition in dry state. Some of the methods described above may be embodied as promoting induced systemic resistance, for inducing latent host defenses or for priming the intrinsic resistance mechanisms in a plant, comprising applying to a plant or a plant part, the 35 composition described in these methods above. 135
Some of the methods described above may be embodied as promoting induced systemic resistance , for inducing latent host defenses or for priming the intrinsic resistance mechanisms in a plant, comprising by spraying on said plant or contacting the roots of said plant with the lignin-derived aromatic oligomers or a composition 5 therewith. Some of the methods described above may be embodied as these method of present invention for protecting plants against plant pests, comprising applying an effective and substantially non-phytotoxic amount of the lignin-derived aromatic oligomers to 10 said plants, for instance wherein said plant pests are selected from the group comprising: fungi, oomycetes, bacteria, viruses, nematodes and insects. Some of the methods described above may be embodied as the method according to any of statements thereon hereabove described, wherein the composition is 15 applied before harvest or post-harvest to the whole plant, the leaves, the flowers, fruits, seeds, seedlings or seedlings pricking out, propagation material such as tubers or rhizomes, plants pricking out, and/or to the soil or inert substrate wherein the plant is growing or in which it is desired to grow, by spraying, drenching, soaking, dipping, injection or administration through fertilising or irrigation systems. 20 These inventive methods of plant treatment comprising applying to a plant or a plant part, a composition comprising the plant defense elicitor of present invention are suitable for promoting induced systemic resistance in a plant, for inducing latent host defenses of a plant or for priming the intrinsic resistance mechanisms in the plant. 25 By using these inventive methods of plant treatment it is possible activating plant defense against a pathogen stressor and/or abiotic stressor in a plant selected from the group comprising: cotton, flax, vine, fruit, vegetable, major horticultural and forest crops such as: Rosaceae sp., Ribesioidae sp., Juglandaceae sp., Betulaceae 30 sp., Anacardiaceae sp., Fagaceae sp., Moraceae sp., Oleaceae sp., Actinidaceae sp., Lauraceae sp., Musaceae sp., Rubiaceae sp., Theaceae sp., Sterculiceae sp., Rutaceae sp., Solanaceae sp., Vitaceae sp., Liliaceae sp., Asteraceae sp., Umbelliferae sp., Cruciferae sp., Chenopodiaceae sp., Cucurbitaceae sp., Papilionaceae sp., such as Graminae sp., Fabacae sp.. 35 136
These inventive methods of plant treatment by spraying on said plant or contacting the roots of said plant with the plant defense elicitor according to any one of above stated methods are suitable for promoting induced systemic resistance of a plant, for inducing latent host defenses of a plant or for priming the intrinsic resistance 5 mechanisms in a plant. A further embodiment of the invention, concerns use of a lignin-derived aromatic oligomers of present invention and described here above as a plant defense elicitor. With this use the induced systemic resistance can be promoted in a plant or the latent host defenses in a plant can be induced by priming of the intrinsic disease10 resistance mechanisms of a plant. Present invention also concerns the use of the plant defense elicitor of present invention on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of an attack by a phytopathogenic pathogen or pest. Present invention also concerns the use of the plant defense elicitor of present 15 invention on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of an attack by a phytopathogenic pathogen or pest whereby the phytopathogenic pathogen or pest is selected from the group of a fungi, a bacteria, and insect. Present invention also concerns the use of the plant defense elicitor of present 20 invention on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of an attack by a phytopathogenic of the group consisting of fungi, bacteria, viruses, viroids, mycoplasma-like organisms, protozoa, insects, acari, and nematodes. Present invention also concerns the use of the plant defense elicitor of present 25 invention on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of abiotic stress. Present invention also concerns the use of the plant defense elicitor of present invention on a plant or parts thereof to prime the intrinsic resistance mechanisms of a plant for stronger or faster induced plant defense preventive to or in the event of 30 an attack by a phytopathogenic pathogen or pest or of abiotic stress, as compared to control or other plant defense inducers. Here by the plant defense elicitor can be used in foliar spray agent, in a root drench. 137
Present invention also concerns the use of the plant defense elicitor of present invention in agricultural applications or to protect plants against plant pests. Such plant pests can be selected from the group comprising: phytopestic fungi, oomycetes, bacteria, viruses, nematodes and insects. 5 A further embodiment of the invention, concerns use of a lignin-derived aromatic oligomers of present invention a as a plant defense elicitor to enhance the efficacy of said fungicide in said composition, or to stimulate the plant immune system. Such use thereof can be on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of an attack by a phytopathogenic pathogen 10 or pest or by an abiotic stressor, or such use can be on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of an attack by a phytopathogenic pathogen or pest whereby the phytopathogenic pathogen or pest is selected from the group of a fungi, a bacteria, and insect. It can be on a plant or parts thereof whereby the latent host defenses are activated 15 preventive to or in the event of an attack by a phytopathogenic of the group consisting of fungi, bacteria, viruses, viroids, mycoplasma-like organisms, protozoa, insects, acari, and nematodes, on a plant or parts thereof whereby the latent host defenses are activated preventive to or in the event of abiotic stress or on a plant or parts thereof to prime the intrinsic resistance mechanisms of a plant for stronger or 20 faster induced plant defense preventive to or in the event of an attack by a phytopathogenic pathogen or pest or of abiotic stress, as compared to control or other plant defense inducers. A further embodiment is a method for producing phytopharmaceutical or agrochemical compositionof present invention, the method comprising a. Subjecting 25 lignin or lignocellulose in a liquid phase to solvolytic lignin depolymerisation, b. Fractionating the extract to obtain a purified fraction enriched in lignin oligomers, c. Formulating the purified fraction into a phytopharmaceutically or agrochemically acceptable dosage form. Yet a further embodiment is a method for producing phytopharmaceutical or 30 agrochemical composition of present invention, whereby the liquid phase described here above contains one or more solvents selected from the group comprising water, methanol, ethanol, n-propanol and isopropylalcohol and mixtures of two or more thereof. 138
Yet a further embodiment the a method for producing phytopharmaceutical or agrochemical composition of present invention, whereby the solvolytic lignin depolymerisation and fractioning is of the group consisting of reductive catalytic fractionation (RCF), non-catalytic thermo-solvolytic depolymerisation and oxidative 5 catalytic fractioning (OCF). Particular and preferred aspects of the invention are set out in the accompanying independent and dependent claims. Features from the dependent claims may be combined with features of the independent claims and with features of other dependent claims as appropriate and not merely as explicitly set out in the claims. 10 Thus, the claims following the detailed description are hereby expressly incorporated into this detailed description, with each claim standing on its own as a separate embodiment of this invention. Tables Description Number Primer name Sequence 1 PFIN760 5’ – GTG TCG CAC ACT GTA CCC ATT TAT – 3’ 2 PFIN761 5’ – ATC TTC ATC ATG TAG TCG GTC AAG T – 3’ 3 PFIN762 5’ – AAT CAC AGC ACT TGC ACC A – 3’ 4 PFIN763 5’- GAG GGA AGC AAG AAT GGA AC – 3’ Table 1 15 Drawing Description BRIEF DESCRIPTION OF THE DRAWINGS A figure can contain different panels for instance figure 1 contains panel A, B, C.. which can be marked Fig. 1A, Fig. 1B and Fig 1C. ‘Pine’ refers to biomass from the Pine family (Pinaceae). 20 The present invention will become more fully understood from the detailed description given herein below and the accompanying drawings which are given by way of illustration only, and thus are not limitative of the present invention, and wherein: FIG. 1 is a schematic diagram showing depolymerized lignin extraction from biomass 25 through reductive catalytic fractionation (RCF) (Fig 1A), with use of heterogenous 139
catalysts such as ruthenium or palladium on carbon with use of an organic solvent (or mixture) in a reductive environment, and non-catalytic thermo-solvolytic fractionation (Fig. 1B) without catalyst and under inert atmosphere. Subsequent fractionation of the lignin oil by liquid-liquid extraction is depicted in Fig. 1C. The RCF 5 and non-catalytic lignin oil (fractions) are named after i) the biomass used (e.g., poplar, Fig. 2A and 2B; or pine, Fig. 2C, 2D, 2E) ii) the presence of a catalyst used during the lignin extraction process or not as indicated by the acronym of the catalyst (e.g. Ru (Fig. 2A and 2C) and Pd (Fig. 2B and 2D) for heterogenous catalyst containing ruthenium or palladium, respectively), iii) the final solvent composition 10 used during subsequent liquid-liquid extraction, being a mixture of heptane and ethyl acetate in a certain ratio (% v/v, e.g., 80 % heptane and 20 % ethyl acetate, H80E20) and iv) whether the obtained fraction is either the dissolved liquid (‘L’) or the residue (‘R’) when using the respective solvent composition. For example, the lignin oil obtained using pine biomass with ruthenium catalyst in a RCF reaction and 15 the obtained residue thereof by subsequent fractionation with 100 % heptane is called PineRuH100R. FIG. 2 are graphics showing the gel permeation chromatography (GPC) of depolymerized lignin oil and fractions thereof obtained through liquid-liquid extraction from pine and poplar biomass obtained through RCF in panel A, B, C and 20 D or in a reductive environment or a non-catalytic thermo-solvolytic fractionation of pine biomass without catalyst and under inert atmosphere in panel E. FIG. 3 are graphics that show in panel A the relative pathogen proliferation of Hyaloperonospora arabidopsidis in A. thaliana plants treated with reductive catalytic fractionation (RCF) lignin oil or fractions thereof (H100R or H100L) from two different 25 biomass types (poplar and pine) using two different heterogenous catalysts (Ruthenium, Ru, and Paladium, Pd), in panel B a dose study on the plant disease resistance inducing capabilities ) of the lignin-derived di/triphenolics fraction heptane insoluble fraction (PineRuH100R) from RCF of pine biomass. Panel C is a graphic display of a dose-response study on the plant disease resistance inducing 30 (capabilities of PineRuH100R = compared to the PimeRu80E20L fraction obtained through liquid-liquid extraction of PineRuH100R (Fig. 1C). On A. thaliana thaliana after the treatment with the lignin derivative H. arabidopsidis pathogen spores were used as shown in panel A) to identify the most active lignin fraction; B) to optimize the treatment concentration; and in C) identify the active IR 140
eliciting dimers within the PineRuH100R fraction. Therefore A. thaliana seedlings were treated with RCF extracts 8 days post seeding by spraying plant leaves till run- off. . Three days post treatment, plants were harvested, subsequently genomic DNA was extracted and qPCR was done to determine the disease index. Treatment 5 concentration in panel A was 1 mg/ml. The concentration of the PineRuH80E20L subfraction of the PineRuH100R (Fig. 1C) is adjusted based on the relative weight of this fractions obtained after liquid-liquid extraction from the PineRuH100R, in this case being 0.238 mg/ml. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. 141
FIG. 4shows the IR activity of the heptane insoluble fraction from a depolymerized lignin obtained from the catalytic RCF process (PineRuH100R) in comparison to the heptane insoluble fraction and a non-catalytic thermo-solvolytic fractioning process (PineH100R) both emulsified in 1 % v/v DMSO at a concentration of 1 mg/ml prior 5 to immediate testing (Panel A) or after an accelerated stability test at higher temperatures for a shorter period of time (Panel B, 30 days 54 °C) and stability test at lower temperature for a prolonged period of time (Panel C, 58 days, 4 °C). It concerns stability tests with lignin oil in solution/emulsified form in 1 % v/v DMSO at 54 °C (accelerated) and at 4 °C with subsequent testing of the ISR activity . A. 10 thaliana is treated with the Hyaloperonospora arabidopsidis pathogen spores. Panel A shows induced plant disease resistance activity of the depolymerized lignin fraction of the non-catalytic thermo-solvolytic fractioning process (PineH100R) and of the RCF process (PineRuH100R); Panel B shows the difference in stability of the PineRuH100R and PineH100R with the lignin-derived di/triphenolics after storage for 30 days at 54 15 °C and panel C 58 days at 4 °C. It can be concluded that the disease resistance inducing activity was initially observed, however in solution/emulsification of 1 % v/v DMSO at 4 °C for 58 days or at 54 °C for 30 days, the plant disease resistance potential is clearly absent in the lignin oil fraction obtained from the non-catalytic thermo-solvolytic fractioning process (PineH100R) compared to the one from the 20 catalytic RCF process (PineRuH100R) which maintained its activity. A repolymerization stabilizer needs to be incorporated in the final formulation to maintain the plant disease resistance potential of the depolymerized lignin oil from the non-catalytic thermo-solvolytic fractioning process. PineRuH100R shows higher stability than the non-catalytic thermo-solvolytic fractioning equivalent PineH100R,25 when kept at similar conditions at 54°C for 30 days and 4°C for 58 days. FIG. 5 is a graphic display that shows infection of A. thaliana by Hyaloperonospora arabidopsidis pathogen to determine optimal time between treatment by the plant disease resistance inducer comprises lignin-derived di/triphenolics (PineRuH100R 30 from RCF of pine biomass) and infection. Based on this data we concluded that the biggest relative reduction in pathogen proliferation is observed when plants are treated 24h before exposure to H. arabidopsidis FIG.6 is a graphic display that in panel A shows the plant disease resistance activity 35 of various tomato (Solanum lycopersicum) cultivars after treatment with PineRuH100R (from RCF of pine biomass), which comprises lignin-derived 142
diphenolics, triphenolics and tetraphenolics, and infection with Botrytis cinerea . Solid,darker bars are the infected tomato cultivars that were not treated with the PineRuH100R which comprises lignin-derived diphenolics, triphenolics and tetraphenolics. Panel B shows that IR induction is at least maintained for 12 days 5 post PineRuH100R treatment (dpt) in S. lycopersicum when infected with B. cinerea. Briefly, 24 days post seeding, S. lycopersicum plants were treated with PineRuH100R by spraying the leaves with compound solution until run-off. Treatment with the solvent 1% v/v DMSO was included as mock treatment (indicated with . Three, twelve and seventeen days after treatment, five leaflets per plant were inoculated 10 with 5 μl droplets of a B. cinerea R16 strain spore suspension of 5 x 105 spores/ml in potato dextrose broth . The hydroponics tanks were placed inside an infection box, containing a moist mat to obtain high humidity, in the growth chamber. The disease symptoms were quantified by measuring the diameter parallel to the midrib of the developing necrotic lesions at 2 dpi. The lesion area was calculated using the formula15 below. Lesion area = (Lesion diameter / 2)^2 * π. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. 20 FIG. 7 concerns induction by RCF lignin of increased resistance in tomato (Solanum lycopersicum) against insects and it a graphic display that quantifies that number of necrotic rings caused by Nesidiocoris tenuis (also known as the tomato bug, tobacco leaf bug, tomato mirid, or green tobacco capsid) feeding post mock (1% v/v DMSO) and PineRuH100R (1 mg/ml, emulsified in 1% v/v DMSO) treatment in accordance 25 with Example 9. It is clearly demonstrated that the treatment with the depolymerized lignin is protective against insect feeding on tomato plants. FIG. 8 is a graphic display that demonstrates the difference in tomato transpiration rate between mock and PineRuH100R sprayed plants under heat stress conditions30 (38 °C), in a treatment in accordance with Example 10. FIG. 9 is a display that shows the seedling survival post mock (1% v/v DMSO) and PineRuH100R (1 mg/ml in 1% v/v DMSO) six days after heat stress exposure. The PineRuH100R renders that the seedling are more protected to heat stress had a35 higher survival rate. 143
FIG. 10 is a schematic diagram showing depolymerized lignin extraction from biomass through oxidative catalytic fractionation (OCF), with use of heterogenous catalysts with use of an alkaline aqueous solution in an oxidative environment. In example 11 the heterogenous catalysts, CuO, is used as heterogenous catalysts for 5 OCF on birch wood at 160°C. FIG. 11 shows the A. thaliana (ecotype: Columbia-0) plants seedlings in little pots (20 well-developed, freestanding seedlings) treated after eight days after sowing treated with mock (1% v/v DMSO) (Fig 11A.), and depolymerized lignin monomers 10 from OCF (OCF 4.1, 1 mg/ml) (Fig 11 B.) fraction by spraying the leaves with compound solution until run-off. The Figures clearly show the toxic impact of the monomers from OCF (OCF 4.1) on the plants. FIG. 12 shows the relative pathogen proliferation of Hyaloperonospora arabidopsidis 15 in A. thaliana plants treated with the heptane insoluble fraction from a depolymerized lignin obtained from the catalytic RCF process of pine wood (PineRuH100R) or OCF oligomers (OCF 4.2) obtained from birch wood using a catalytic OCF process demonstrating equal elicitor activity of depolymerized lignin oligomers (DP ≥ 2) from RCF and OCF. 20 Abbreviations in these figures: H80E20L = the dissolved liquid (‘L’) subfraction (of the H100R) obtained in the fractioning of lignin oil through liquid-liquid extraction from mixture of heptane and 25 ethyl acetate in a certain ratio (% v/v, e.g., 80 % heptane and 20 % ethyl acetate), see figure 1C. H80E20R = the residue (‘R’) subfraction (of the H100R) obtained in the fractioning of lignin oil through liquid-liquid extraction from mixture of heptane and ethyl acetate in a certain ratio (% v/v, e.g., 80 % heptane and 20 % ethyl acetate), see30 figure 1C. PineH100R = the depolymerized lignin (lignin oil) residue (‘R’) fraction from the 100% heptane extraction of the non-catalytic thermo-solvolytic fractioning process using pine biomass, see figure 1C. PineH100L = the depolymerized lignin (lignin oil) dissolved liquid (‘L’) fraction from 35 the 100% heptane extraction of the non-catalytic thermo-solvolytic fractioning process using pine biomass. 144
PineRuH100R = the depolymerized lignin (lignin oil) residue (‘R’) fraction from the 100% heptane extraction of the reductive catalytic fractionation (RCF) process using pine biomass with ruthenium catalyst, see figure 1C. PineCdH100R = the depolymerized lignin (lignin oil) residue (‘R’) fraction from the 5 100% heptane extraction of the reductive catalytic fractionation (RCF) process using pine biomass with cadmium catalyst, see figure 1C. PineRuH80E20L = the dissolved liquid (‘L’) subfraction (of the PineRuH100R) obtained in the fractioning of lignin oil through liquid-liquid extraction from mixture of heptane and ethyl acetate in a certain ratio (% v/v, e.g., 80 % heptane and 20 % 10 ethyl acetate), see figure 1C. PineCdH80E20L = the dissolved liquid (‘L’) subfraction (of the PineCdH100R) obtained in the fractioning of lignin oil through liquid-liquid extraction from mixture of heptane and ethyl acetate in a certain ratio (% v/v, e.g., 80 % heptane and 20 % ethyl acetate), see figure 1C. 15 PopH100R = the depolymerized lignin (lignin oil) residue (‘R’) fraction from the 100% heptane extraction of the non-catalytic thermo-solvolytic fractioning process using poplar biomass, see figure 1C. PopH100L = the depolymerized lignin (lignin oil) dissolved liquid (‘L’) fraction from the 100% heptane extraction of the non-catalytic thermo-solvolytic fractioning20 process using poplar biomass, see figure 1C. PopRuH100R = the depolymerized lignin (lignin oil) residue (‘R’) fraction from the 100% heptane extraction of the reductive catalytic fractionation (RCF) process using poplar biomass with ruthenium catalyst, see figure 1C. PopCdH100R = the depolymerized lignin (lignin oil) residue (‘R’) fraction from the 25 100% heptane extraction of the reductive catalytic fractionation (RCF) process using poplar biomass with cadmium catalyst, see figure 1C.. PopRuH80E20L = the dissolved liquid (‘L’) subfraction (of the PopRuH100R) obtained in the fractioning of lignin oil through liquid-liquid extraction from mixture of heptane and ethyl acetate in a certain ratio (% v/v, e.g., 80 % heptane and 20 % 30 ethyl acetate), see figure 1C. PopRuOil = the lignin oil obtained by lignin depolymerized lignin by reductive catalytic fractionation (RCF) process using poplar biomass with ruthenium catalyst. PopRuOil = the lignin oil obtained by lignin depolymerized lignin by reductive catalytic fractionation (RCF) process using poplar biomass with ruthenium catalyst. 35 PineRuOil = the lignin oil obtained by lignin depolymerized lignin by reductive catalytic fractionation (RCF) process using pine biomass with ruthenium catalyst. 145
PineRuOil = the lignin oil obtained by lignin depolymerized lignin by reductive catalytic fractionation (RCF) process using pine biomass with ruthenium catalyst. PineOil = The lignin oil obtained by lignin depolymerized lignin by non-catalytic thermo-solvolytic depolymerisation.dpt = days post treatment. 5 References to this application - Angelova, Z., Georgiev, S., & Roos, W. (2006). Elicitation of plants. Biotechnology & Biotechnological Equipment, 20(2), 72-83. 10 - Bektas, Y. (2022). Fytosol, a promising plant defense elicitor, controls early blight (alternaria solani) disease in the tomato by inducing host resistance- associated gene expression. Horticulturae, 8(6), 484. https://doi.org/10.3390/horticulturae8060484 - Chalal, M., Winkler, J., Gourrat, K., Trouvelot, S., Adrian, M., Schnitzler, J., … 15 & Daire, X. (2015). Sesquiterpene volatile organic compounds (vocs) are markers of elicitation by sulfated laminarine in grapevine. Frontiers in Plant Science, 6. - Hatami, M., Badi, H., & Ghorbanpour, M. (2019). Nano-elicitation of secondary pharmaceutical metabolites in plant cells: a review. Journal of Medicinal20 Plants, 3(71), 6-36. https://doi.org/10.29252/jmp.3.71.6 - Maffei, M., Arimura, G., & Mithöfer, A. (2012). Natural elicitors, effectors and modulators of plant responses. Natural Product Reports, 29(11), 1288. - Shinya, T., Ménard, R., Kozone, I., Matsuoka, H., Shibuya, N., Kauffmann, S., … & Saito, M. (2006). Novel β-1,3-, 1,6-oligoglucan elicitor from alternaria25 alternata 102 for defense responses in tobacco. Febs Journal, 273(11), 2421- 2431. - Sudhamoy, M. (2010). Induction of phenolics, lignin and key defense enzymes in eggplant (solanum melongena l.) roots in response to elicitors. African Journal of Biotechnology, 9(47), 8038-8047. 30 https://doi.org/10.5897/ajb10.984 Wang, J., Cai, Y., Gou, J., Mao, Y., Xu, Y., Jiang, W., … & Chen, X. (2004). Vdnep, an elicitor from verticillium dahliae, induces cotton plant wilting. Applied and Environmental Microbiology, 70(8), 4989-4995. 146

Claims

PLANT DEFENSE INDUCER 5 Claims What is claimed is: . 1) A method of for improving abiotic and/or biotic stress tolerance in plants, comprising contacting a plant or a plant’s organ with a composition comprising 10 an effective amount of a lignin oligomer with a degree of polymerization (DP) of 2 to 8, preferably of 2 to 4. 2) The method according to claim 1, whereby the lignin oligomers comprising, or essentially consisting of or consisting of I) at least one aromatic compound 15 , - wherein each R1, R3, and R4 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 20 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, 147
- wherein R2 is –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer, or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R5 is selected of –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, an end-unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, - CH=CHCH3, -(CH2)2CH2OH, -(CH2)2CHO, -CH=CHCH2OH, - (CH2)2CH2OCH3, -CH=CHCH2OCH3, -(CH2)2CH2OCH2CH3, - CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, -CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, -CH=CHCH2OCH(CH3)2, - (CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, a carbon linkage to an aromatic monomer or aromatic oligomer; and/or II) wherein the aromatic compounds comprise at least one aromatic 148
- wherein each R12, R13, R15 and R16 is independently chosen from –H, - OH, O-CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon 5 linkage to an aromatic monomer or aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein R11 and R14 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an 10 aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, and/or III) wherein at the aromatic compounds comprise at least one aromatic compound selected from the formula (viii) , (xi) 15 149
(xviii) each with at least one linkage to an aromatic monomer or aromatic oligomer and - wherein R21 is independently chosen from –H, a β-O-4 linkage to an 5 aromatic monomer or aromatic oligomer, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, an α-O-4 linkage to an aromatic monomer or aromatic oligomer or a carbon-oxygen linkage to an aromatic monomer or aromatic oligomer, - wherein each R22 and R23 is dependently chosen from -H, -OH, O-CH3 10 or from a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or an aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, 15 - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein the alkyl group is derived from the alcohol solvent of the process, - wherein each R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer or aromatic oligomer, a 5-5 linkage to an aromatic monomer or aromatic oligomer, a β-5 20 linkage to an aromatic monomer or aromatic oligomer, a carbon linkage to an aromatic monomer or an aromatic oligomer, or a carbon- oxygen linkage to an aromatic monomer or aromatic oligomer, wherein R27 is independently chosen from –H, a β-O-4 linkage to an aromatic monomer or aromatic oligomer, a β-5 linkage to an aromatic monomer or 150
arom- atic oligomer, a β-β linkage to an aromatic monomer or aromatic oligomer, a β-1 linkage to an aromatic monomer or aromatic oligomer, end- unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, - (CH2)2CH2OH, -(CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, - 5 CH=CHCH2OCH3, -(CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, - (CH2)2CH2O(CH2)2CH3, -CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, - CH=CHCH2OCH(CH3)2, -(CH2)2CH2O(CH2)3CH3, -CH=CHCH2O(CH2)3CH3, a carbon linkage to an aromatic monomer or an aromatic oligomer. 10 3) The method according to any one of the claims claim 1 or 2, whereby lignin oligomers comprising, or essentially consisting of or consisting of I) at least one aromatic compound selected from the formulae 151
and - wherein each R1, R3, and R4 is independently chosen from –H, -OH, O- CH3, 5 - wherein R2 is –H, - wherein R5 is selected of –H, an end-unit selected of CH3, -CH2CH3, - (CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, -(CH2)2CH2OH, -(CH2)2CHO, - CH=CHCH2OH, -(CH2)2CH2OCH3, -CH=CHCH2OCH3, - (CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, -10 CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, - CH=CHCH2OCH(CH3)2, -(CH2)2CH2O(CH2)3CH3, - CH=CHCH2O(CH2)3CH3, - and/or II) wherein the aromatic compounds comprise at least one aromatic compound selected from the formulae (v) 152
and - wherein each R12, R13, R15 and R16 is independently chosen from –H, - OH, O-CH3, 5 - wherein R11 and R14 is independently chosen from –H, and/or III) wherein at the aromatic compounds comprise at least one aromatic compound selected from the formula (viii) 153
and (xiv) each with at least one linkage to an aromatic monomer or aromatic oligomer and - wherein R21 is independently chosen from –H, a β-O-4 linkage to an 5 aromatic monomer, a 4-O-5 linkage to an aromatic monomer, an α-O- 4 linkage to an aromatic monomer or a carbon-oxygen linkage to an aromatic monomer, - wherein each R22 and R23 is dependently chosen from -H, -OH, O-CH3 or where at least R22 or R23 is independently chosen from a 4-O-5 10 linkage to an aromatic monomer, a 5-5 linkage to an aromatic monomer, a β-5 linkage to an aromatic monomer, a carbon linkage to an aromatic monomer, or a carbon-oxygen linkage to an aromatic monomer, - wherein R24 is independently chosen from –H, OH, or -O-Alkyl wherein 15 the alkyl group is derived from the alcohol solvent of the process, - wherein each R25 and R26 is independently chosen from –H, -OH, O- CH3, a 4-O-5 linkage to an aromatic monomer, a 5-5 linkage to an aromatic monomer, a β-5 linkage to an aromatic monomer, a carbon linkage to an aromatic monomer, or a carbon-oxygen linkage to an 20 aromatic monomer, - wherein R27 is independently chosen from –H, end-unit selected of CH3, -CH2CH3, -(CH2)2CH3, -CH2CH=CH2, -CH=CHCH3, -(CH2)2CH2OH, - (CH2)2CHO, -CH=CHCH2OH, -(CH2)2CH2OCH3, -CH=CHCH2OCH3, - (CH2)2CH2OCH2CH3, -CH=CHCH2OCH2CH3, -(CH2)2CH2O(CH2)2CH3, - 155
CH=CHCH2O(CH2)2CH3, -(CH2)2CH2OCH(CH3)2, - CH=CHCH2OCH(CH3)2, -(CH2)2CH2O(CH2)3CH3, - CH=CHCH2O(CH2)3CH3. 5 4) The method according to any one of the claims claim 1 or 3, whereby the lignin oligomers comprising, or essentially consisting of or consisting of at least one aromatic compound selected from the formulae (i) 10 , (v) 156
or a combination thereof. 5) The method according to any one of the claims claim 1 or 4, whereby the 5 lignin oligomers are phenolics comprising two benzene rings directly bridged or bridged with a common bridging group of the group consisting of aliphatic 158
chains, alkene groups, carbonyl groups and ether linkages or wherein the phenolic oligomers have two aromatic groups. 6) The method according to any one of the claims claim 1 or 5, whereby the lignin 5 oligomers are phenolics comprising two benzene rings directly bridged or bridged with a common bridging group of the group consisting of -CH₂- groups, -CH=CH- , -C(=O)-and -O-. 7) The method according to any one of the claims 1 to 6, whereby the composition10 contains less than 0,1%, of each acetic acid, methanol and ethanol. 8) The method according to any one of the claims 1 to 7, whereby in the lignin oligomers are comprised in a composition with a pH in the range of 4 to 10, preferably in the range of 5 to 8 or in origin have a pH in the range of 4.0 to 6.0. 15 9) The method according to any one of the claims 1 to 8, whereby the lignin oligomers are comprised in a composition, further comprising one or more one or more anionic, non-ionic, amphoteric, or cationic surfactant, or a combination thereof and one or more polar aprotic solvent. 20 10)The method according to any one of the claims 1 to 9, whereby the dry weight of the composition contains between 0,5 to 30 wt% by dry weight of lignin- oligomers, preferably 1 to 20 wt% by dry weight of lignin-oligomers, more preferably 2 to 10 wt% by dry weight of lignin-oligomers. 25 11)The method according to any one of the claims 1 to 10, whereby the composition further comprising a salicylic acid pathway activator and/or jasmonic acid pathway activator, wherein the lignin oligomer elicitor and the activator of salicylic acid pathway activator component or the lignin oligomer elicitor and jasmonic 30 acid pathway activator exhibit a synergistic effect in enhancing plant defense mechanisms. 12)The method according to any one of the claims 1 to 11, for inducing systemic resistance in plants against a pathogen stressor or against an abiotic stressor. 35 13)The method according to any one of the claims 1 to 12, wherein said plant stressor is a pathogen selected from the group consisting of fungi, bacteria, 159
viruses, viroids, mycoplasma-like organisms, protozoa, insects, acari, and nematodes. 14) The method according to any one of the claims 1 to 13, wherein said plant 5 stressor is an abiotic stress selected from the group consisting of hydric stress, drought and osmotic stress. 15)The method according to any one of the claims 1 to 14, comprising spraying, drenching, soaking, dipping, injection, and any combination thereof of said plant 10 or a plant’s organ with said composition. 160
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