EP4048072A1 - Hydrolysat pour favoriser la croissance des plantes, la biostimulation et la lutte biologique, et son utilisation en agriculture - Google Patents

Hydrolysat pour favoriser la croissance des plantes, la biostimulation et la lutte biologique, et son utilisation en agriculture

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Publication number
EP4048072A1
EP4048072A1 EP20804679.7A EP20804679A EP4048072A1 EP 4048072 A1 EP4048072 A1 EP 4048072A1 EP 20804679 A EP20804679 A EP 20804679A EP 4048072 A1 EP4048072 A1 EP 4048072A1
Authority
EP
European Patent Office
Prior art keywords
hydrolysate
percentage
amino acids
starting
free amino
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
EP20804679.7A
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German (de)
English (en)
Inventor
Roberto Verga
Silvia RAPACIOLI
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Bict Srl
Original Assignee
Bict Srl
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Filing date
Publication date
Application filed by Bict Srl filed Critical Bict Srl
Publication of EP4048072A1 publication Critical patent/EP4048072A1/fr
Pending legal-status Critical Current

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Classifications

    • 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
    • A01N43/14—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 six-membered rings
    • A01N43/16—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 six-membered rings with oxygen as the ring hetero atom
    • 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
    • A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
    • A01N37/44—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a nitrogen atom attached to the same carbon skeleton by a single or double bond, this nitrogen atom not being a member of a derivative or of a thio analogue of a carboxylic group, e.g. amino-carboxylic acids
    • 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
    • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/03—Algae
    • 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
    • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/08—Magnoliopsida [dicotyledons]
    • 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
    • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/08—Magnoliopsida [dicotyledons]
    • A01N65/10—Apiaceae or Umbelliferae [Carrot family], e.g. parsley, caraway, dill, lovage, fennel or snakebed
    • 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
    • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/08—Magnoliopsida [dicotyledons]
    • A01N65/20—Fabaceae or Leguminosae [Pea or Legume family], e.g. pea, lentil, soybean, clover, acacia, honey locust, derris or millettia
    • 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
    • A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
    • A01N65/40—Liliopsida [monocotyledons]
    • A01N65/44—Poaceae or Gramineae [Grass family], e.g. bamboo, lemon grass or citronella grass
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
    • C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F5/00—Fertilisers from distillery wastes, molasses, vinasses, sugar plant or similar wastes or residues, e.g. from waste originating from industrial processing of raw material of agricultural origin or derived products thereof
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
    • Y02A40/20—Fertilizers of biological origin, e.g. guano or fertilizers made from animal corpses
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/141—Feedstock
    • Y02P20/145—Feedstock the feedstock being materials of biological origin
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
    • Y02W30/00—Technologies for solid waste management
    • Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse

Definitions

  • the present invention relates to a hydrolysate from a natural raw material, which has been shown to be significantly effective in stimulating plant growth, or in protecting the plant against phytopathogenic agents.
  • the hydrolysate from a natural raw material therefore finds advantageous application in the agricultural industry.
  • Plant growth is dictated by both internal and external factors.
  • the internal mechanisms originate from the genetic composition of the plant and influence the extent and timing of the growth thereof. These internal mechanisms are regulated by various types of signals transmitted within the plant cells or around the plant itself.
  • the external factors are directly related to the environment surrounding the plant. These external influences on plant growth include factors such as light, temperature, water, and nutrients.
  • the external environment can limit the extent to which internal mechanisms enable the plant to grow and develop, with two of the most important factors being the availability of water and nutrients in the ground.
  • Cell expansion is directly related to water supply and therefore any shortages result in a smaller plant. Mineral nutrients are necessary for the plant's biochemical processes. When nutritional substances are insufficient, growth will be less vigorous and, in extreme cases, will cease altogether.
  • Nutrients needed for plant growth include: primary macronutrients, namely nitrogen (N), phosphorus (P), and potassium (K), secondary macronutrients, namely calcium (Ca), sulphur (S), and magnesium (Mg), and micronutrients or trace minerals, namely boron (B), chlorine (Cl), manganese (Mn), iron (Fe), zinc (Zn), copper (Cu), molybdenum (Mo), and selenium (Se).
  • primary macronutrients namely nitrogen (N), phosphorus (P), and potassium (K
  • secondary macronutrients namely calcium (Ca), sulphur (S), and magnesium (Mg)
  • micronutrients or trace minerals namely boron (B), chlorine (Cl), manganese (Mn), iron (Fe), zinc (Zn), copper (Cu), molybdenum (Mo), and selenium (Se).
  • fertilisers can come in the form of a neat liquid, a suspension, or a solid. They can be administered to plants by fertilising the growth medium or by applying them to the foliage of plants, by - for example - spraying, irrigation, or similar methods. Over the last few years, foliar fertilisers have gradually replaced the soil fertilisers commonly used in farming areas, as they have fewer adverse environmental impacts. Research has shown that the conventional fertilisation process - i.e. soil fertilisation - has contributed to the contamination of surface water and groundwater. This is mainly due to the leaching of soluble fertiliser nutrients, such as nitrogen, into the water table.
  • Foliar fertilisers appear to overcome the drawbacks of the soil fertilisation process, however, incorrect application of said foliar fertilisers to plants, such as -for example - direct application of a high concentration of nutrients to the foliage, can result in leaf damage to the foliage consisting of necrotic areas or leaf burns, resulting in lower crop yields. It has been hypothesised that foliar fertiliser with a lower quantity of nutrients could prevent foliar damage. However, this is not very practical as fertilising plants with low-nutrient foliar fertiliser is a labour-intensive activity.
  • An object of the present invention is therefore to effectively promote plant growth without causing damage to foliage and the plant in general and preserve, in particular, human and animal health, as well as the crops and the environment.
  • Said object has been achieved by a hydrolysate from a natural raw material, as reported in Claim 1, as well as a process for the preparation thereof.
  • the present invention relates to a composition
  • a composition comprising said hydrolysate from a natural raw material and a microbial inoculant.
  • the present invention relates to the use of said hydrolysate from a natural raw material and likewise said composition, to promote plant growth and fruit production in agriculture, or for biocontrol.
  • the present invention relates to an agrochemical product comprising the hydrolysate from a natural raw material or the composition, and agrochemical additives.
  • the present invention relates to a method for promoting plant growth and fruit production, said method comprising the step of applying the hydrolysate from a natural raw material or the composition or the agrochemical product to a plant or a growing medium for plants.
  • plant means any plant or plants that can be grown and harvested for profit or subsistence purposes, i.e. crops, and therefore including cereals, vegetables, fruit, and flowers, as well as those grown and harvested for gardening or personal use.
  • growing medium for plants means the medium in which the plant is grown or in which the plant is sown or in which the plant will be sown, and therefore includes soils, earth, and soil-free media, including hydroculture and hydroponics media.
  • the invention therefore relates to a hydrolysate from a natural raw material comprising 1-25% free amino acids and up to 55% dry residue, wherein said plant raw material is selected from com steep liquor, lupine, algae, molasses, coriander, cocoa, olive residues, and combinations thereof.
  • hydrolysate from a natural raw material according to the present invention not only stimulates plant growth and productivity in terms of fruit production, but also protects plants against phytopathogenic agents and pests.
  • Com steep liquor is a liquid by-product of the wet milling of corn used to make com starch and high fmctose corn symp (HFCS).
  • CSL consists of soluble com concentrates which are extracted by a process during which the shelled and air-purified corn is soaked in water (steeped) and then separated into the main components thereof through a combination of flotation and wet-sieving. During steeping, the soluble materials dissolve, the corn softens, and its structure weakens and breaks, thereby facilitating the milling and further separations of the components thereof.
  • the resulting concentrate is a cmde com steep liquor, which can be further combined with gluten and fibrous materials to be sold as feed or used for other purposes, with or without undergoing further processing.
  • CSL has also been used in the penicillin industry as a culture medium for the production of penicillin.
  • Corn steep liquor (CAS n. 66071-94-1) is commercially available in an aqueous solution (about 50% water), while the rest is made up of the natural nutrients of corn, such as water-soluble proteins, amino acids (e.g. alanine, arginine, aspartic acid, cysteine, glutamic acid, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, threonine, tyrosine, valine), vitamins (e.g. B-complex), carbohydrates, organic acids (e.g. lactic acid), minerals (e.g. Mg, P, K, Ca, Ca, S), enzymes, and other nutrients.
  • amino acids e.g. alanine, arginine, aspartic acid, cysteine, glutamic acid, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, threonine,
  • CSL is a viscous mixture which is a light to dark brown in colour and has a pH of about
  • the lupine bean is a legume originating from the Mediterranean area, East Africa, and the Americas, where it was already grown as far back as 4,000 years ago. Today, however, it is grown in Australia, Europe, and South America for food purposes, both for human and animal diets, and the plant is also used as green manure (i.e. a crop grown to enrich the soil with nutrients which are synthesised by nitrogen-fixing bacteria found on the roots of the lupine plant). In addition to its beans, lupine can also be consumed in the form of a flour obtained by milling the dry beans.
  • lupine features a very high content of soluble and insoluble protein and fibre and a low carbohydrate and fat content.
  • Lupine flour contains at least 40% protein, 31% fibre, 10% sugar, 10% fat, and various micronutrients.
  • the fats contained in lupine beans are, for the most part, "good” fats, since they amount to 67% monounsaturated fatty acids, 19% polyunsaturated fatty acids, and only 14% saturated fatty acids.
  • Lupine beans are a good source of thiamine (vitamin Bl) and folic acid (vitamin B9), as well as certain mineral salts, in particular, manganese, copper, magnesium, phosphorus, zinc, and potassium.
  • bio stimulating natural raw materials include algae and, in particular, algae extracts. They have been used in agriculture for hundreds of years as soil conditioners to enhance soil fertility. Just over 50 years ago, production began on liquid extracts to enhance the bio stimulating properties of algae. Today there are numerous biostimulant products in commerce which are based on algae extracts available on the market.
  • the extracts are obtained from green, red, or brown algae and more specifically Ascophyllum nodosum, Ecklonia maxima, Laminaria digitata, and Fucus spp.
  • the algae is harvested manually or mechanically along the ocean coasts and is washed, cut, and then undergoes extraction. Extraction can be performed in various ways and using solvents of various kinds. Techniques for the production of algae extracts have also been proposed which involve microbial fermentation of the plant-derived starter. The type of algae used, the harvesting period, and the extraction process greatly influence the chemical characteristics of the extract and therefore its biostimulating properties.
  • Algae extracts act as biostimulants by enhancing germination speed, growth, fruit set, production, product quality, and resistance to environmental stress. Furthermore, the algae extracts increase the absorption of macro and micronutrients in various crops. The biostimulating effects are mainly due to the presence of phytohormones, polysaccharides, polyphenols, and other organic molecules. The phytohormones identified in algae extracts that stimulate plant growth are auxins, cytokinins, abscisic acid, gibberellins, etc.
  • molasses is the by-product of the production process of sucrose (commonly known as sugar), which is obtained by extraction from beet ( Beta vulgaris ) in the Mediterranean basin and sugar cane ( Saccharum ojficinarum ) in particularly in Central and South American countries.
  • sucrose commonly known as sugar
  • sugar cane Saccharum ojficinarum
  • molasses is frequently, but improperly, used to mean other by-products or residues of extraction processes for obtaining sugars other than sucrose.
  • the term 'black-strap molasses' means molasses in general, regardless of its origin. More specifically though, molasses alone refers to beet molasses, while black-strap molasses refers to cane molasses.
  • Molasses is an example of a raw material, which is widely used on an industrial scale as a source of carbon in numerous microbiological processes and in animal feed.
  • This raw material is a brown viscous liquid, which is obtained by concentrating the mother liquors left over after sucrose extraction, within a process involving beet being cut into strips or cane being subjected to pressure.
  • the molasses undergoes physical-chemical pre-treatments before being used.
  • the composition of molasses can vary considerably.
  • Molasses obtained from beet and that obtained from cane have a similar total sugar content, but they differ specifically in the sucrose content and the content of inverted sugar, i.e. sucrose hydrolysed into glucose and fructose.
  • cane molasses has a higher content of vitamins, in particular, biotin content (1-3 pg/g), than beet molasses (0.04-0.15 pg/g). For this reason, beet molasses is frequently supplemented with cane molasses, regardless of the geographical area of production and therefore of transport costs, in order to increase the biotin levels in the microbial cultures.
  • molasses include other residues such as those from the extraction of sucrose from sorghum (50% sucrose content) and corn molasses or hydrol (sugar content: 50-60%), and mother liquors from the production of glucose from com starch.
  • High-test molasses (15 to 35% sucrose and 40 to 60% inverted sugar), on the other hand, is not a residue or by-product of sucrose production but rather the main product of the process (without the sucrose crystallisation step) as it is directly obtained through evaporation of the cane juice and partial inversion with invertase. This product can also originate from the processing of citrus fruits.
  • the product can have the following characteristics:
  • Coriander Coriandrum sativum, L. 1753 or Chinese parsley or with the Spanish name cilantro, is an annual herbaceous plant from the Apiaceae (or Umbelliferae ) family. It belongs to the same family as cumin, dill, fennel, and parsley.
  • Coriandrum is a Latin word used by Pliny (in his Naturalis Historia ) whose roots lie in the Greek word corys or korios (meaning 'cimicid', or 'stink bug') followed by the suffix -ander (meaning 'similar'), referring to the supposed similarity of the smell released by its unripe fruits or when its leaves are rubbed with that released by said insect.
  • coriander can be used as seed flour but the agricultural waste originating from the preparation thereof also offers a cost-effective active starting substrate, albeit with fewer properties. Seventeen components have been identified, which make up 91.84% of the residual essential oil of Egyptian coriander.
  • Trans-anethole has been identified as the main compound of spent coriander oil, measuring 29.29%, followed by linalool (20.06%), butanoic acid, esters of 2-methyl- ,2- methoxy-4-(2-propyl)phenyl (14.17%), estragon (10.25%), longifolene (6.82%), and carvacrol (5.1%)
  • 'Cocoa' means the plant and beans of Theobroma cacao Linn, an arboreal plant from the Sterculiacee family grown in tropical and subtropical regions.
  • various products are obtained from the processing of cocoa beans, which differ in terms of quantity and quality and include various by-products such as the shell, the pulp, the integuments, and the germ.
  • these by-products include vast amounts of material that could be used for new production chains and could be of help to developing countries, where the largest cocoa producers are located.
  • the international cocoa market is split in two, i.e.
  • the producing countries consisting of the Southern Hemisphere
  • the processing and consumer countries consisting of the Northern Hemisphere, in particular the Netherlands and North America.
  • the undisputed leader among the producing countries is the Ivory Coast, which accounts for about 40% of the world's production, followed by Ghana and Indonesia. Next come Ecuador, Cameroon, Nigeria, and Brazil, which account for about 37% in total. The remaining 23% is divided between the producing countries in the tropical belt.
  • cocoa needs deep, clayey soils for growth and a temperature of between 25 °C and 35 °C. It also needs abundant rainfall.
  • the cocoa tree reaches about 8 m in height, in the most extreme cases it can even reach 18 m, with green leaves and reddish flowers arranged in panicles.
  • An individual fruit weighs about 400-500 g and is yellow or red in colour when ripe and brown when dry.
  • a white mucilaginous acidulous pulp containing the seeds, which are enveloped by an integument consisting of a film coating around the bean.
  • the species farmed belongs to the Teobroma Cacao genus and the most commercially widespread cultivars are the Criollo, Forastero, and Trinitario varieties.
  • they are picked from the tree using a hook knife.
  • the fruits are broken and the cocoa beans are spread on wooden pallets or on the ground on top of a layer of banana leaves, in order to facilitate the fermentation triggered by the yeasts.
  • green cocoa is obtained, which is then left to dry at ambient temperature until the humidity decreases.
  • the beans are then roasted at a temperature of between 130 °C and 140 °C.
  • special machines detach the integument from the germ.
  • By-products of the processing consist of the shells, the pulp, the integuments, and the germ.
  • the shells are the outermost part of the fruit and are the most important by product. Research in Japan has shown that they have an anticariogenic action, attributable to the polyphenolic compounds and unsaturated fatty acids. Furthermore, the high mineral content makes the shells suitable for the production of soap (especially black soap) and detergents, while the potassium makes them good fertilisers. Mixed with other plant by-products, they can also be used for pig and sheep feed. During fermentation the pulp is removed. The following can be obtained from the pulp: pectin for jams, alcohol for industrial and hospital use, vinegar, and soft drinks.
  • the pulp can be frozen and then used for the production of ice cream, to give yogurt flavour, but it is only marketed locally due to the high cost.
  • a pigment used as a food colouring can be extracted from the integuments, i.e. polyflavone glucoside.
  • antioxidants such as flavonoids can be extracted using solvents.
  • the germ is removed to enhance the finished product. Its oil is extracted for use in the food and cosmetic industry. Summing up, by-products are obtained from cocoa processing that can be used in other fields and can also be useful for economic development. The amount of by-products that originate is considerable and these can be placed on the market and be used to develop new production chains.
  • the waste product used was cocoa husks, exemplificative analysis of which identified the following characteristic components:
  • Olive residues consist of the solid residues (stones, skin films, parts of the flesh) left after the pressing of the olive paste. It represents 30-50% of the olives processed. Its composition is:
  • the acidity of the oil extracted from the olive residues ranges from 15 to 80%; high acidity should be avoided as it complicates the adjustment process, making it more expensive. Acidity increases during the time spent at the oil mill due to hydrolysis caused by lipase, which is active in the presence of thO, and to auto-oxidation caused by contact with air and the considerable surface area. Anti-fermentation agents such as NaHS0 3 (2-3%) are not recommended because they cause drawbacks upon extraction. The best preservation is achieved by drying in special rotary ovens, which are generally fed with olive residues; however, these ovens must never be used to obtain anhydrous olive residues because the cell membranes would be denatured and, as they would form an impermeable layer, the oil could not be extracted.
  • the optimal humidity value is 7% and in industrial practice it is checked by listening to the noise produced by squeezing a handful of olive residues in one's hand. If the humidity is higher, the lipase may cause an exothermic reaction which can also lead to fires due to self-combustion of the stationary product awaiting extraction.
  • Particularly preferred embodiments are those wherein said hydrolysate from a natural raw material is hydrolysate of com steep liquor.
  • said hydrolysate from a natural raw material is:
  • the present invention relates to a process for preparing said hydrolysate from a natural raw material, comprising the following steps: i) providing a natural raw material, and optionally adding water, ii) adding an enzyme complex comprising at least one peptide bond hydrolase and at least one fibre and/or carbohydrate hydrolase, and iii) obtaining a liquid mixture, i.e. the hydrolysate from a natural raw material.
  • a liquid mixture i.e. the hydrolysate from a natural raw material.
  • water is added to the natural raw material.
  • the temperature is kept at 4-70 °C for a period of 2-20 hours, more preferably the temperature is kept at 10-50 °C for a period of 4-16 hours.
  • enzymes which act on the protein component for example, enzymes in class EC 3.4, i.e. enzymes that act on the peptide bond, such as, for example, mono-, di- , or tripeptidases, or mixtures thereof, proteases, and proteinases.
  • enzymes which act on fibres and/or carbohydrates for example, enzymes in class EC 3.2 - Glycosidases, i.e. enzymes that act on O- and S-glycosyl compounds or N-glycosyl compounds, such as, for example, amylases, cellulases, hemicellulases, laccases, and xylanases.
  • enzymes which act on lipids for example, enzymes in class EC 3.1, i.e. enzymes that act on the ester bond, such as, for example, lipases.
  • the hydrolase is selected from mono-, di-, or tripeptidases, proteases, proteinases, cellulases, hemicellulases, pectinases, xylanases, amylases, laccases, lipases, and mixtures thereof.
  • the enzymatic complex comprises a mixture of at least two hydrolases, i.e. at least one hydrolase which acts on fibres and/or carbohydrates and at least one hydrolase which acts on the protein component. Indeed, it has been observed that this mixture acts synergistically, allowing to achieve surprising results as compared to the use of a single enzyme.
  • a pH adjuster is added, preferably an organic or inorganic base, such as NaOH or KOH, to adjust the pH to about 6.
  • an inorganic or organic acid such as phosphoric acid, can also be used in order to reach the optimal reaction pH of the specific enzymatic mix.
  • the natural raw material was treated with an anti-fermentation agent and a mixture of EC 3.2 enzymes for 6 hours at about 52 °C and at about pH 4. Subsequently, KOH was added to bring the pH up to 6, followed by a mixture of EC 3.2 and 3.4 enzymes for a further 6 hours.
  • this step also included the addition of EC 3.1 enzymes, i.e. enzymes that act on the ester bond, such as lipases.
  • the natural raw material was treated with an anti-fermentation agent and a mixture of EC 3.2 and E 3.4 enzymes for 6 hours at about 52 °C and at about pH 4.
  • this step also included the addition of EC 3.1 enzymes, i.e. enzymes that act on the ester bond, such as lipases.
  • KOH was added to bring the pH up to 5.5-6.
  • the present invention relates to a hydrolysate from a natural raw material obtainable by the process described above, wherein said plant raw material is selected from com steep liquor, lupine, algae, molasses, coriander, cocoa, olive residues, and combinations thereof, in which the percentage of free amino acids in the hydrolysate is at least 0.5% higher than the percentage in the starting plant raw material, and in which the dry residue in the hydrolysate is at least 2% less than the percentage in the starting plant raw material.
  • said hydrolysate from a natural raw material obtainable by the process described above is: - hydrolysate of com steep liquor, in which the percentage of free amino acids in the hydrolysate is 0.5-11% higher than the percentage in the starting corn steep liquor, and in which the dry residue in the hydrolysate is 10-15% lower than the percentage in the starting com steep liquor,
  • the present invention also relates to the use of said hydrolysate from a natural raw material to promote plant growth and fmit production in agriculture, or for biocontrol.
  • this hydrolysate should preferably be used at a rate of 0.5-50 litres per hectare of soil if liquid, or in equivalent proportions for other liquid or solid formulations, such as, for example, solid granules which may be either dispersible in water or non-dispersible in water.
  • the present invention relates to a composition
  • a composition comprising a microbial inoculant and hydrolysate from a natural raw material as described above, in which said microbial inoculant is selected from a bacterial inoculant, a fungal inoculant, or a combination thereof.
  • Microbial inoculants also known as soil inoculants or bio-inoculants, are agricultural remedies that use beneficial rhizospheric or endophytic microbes to promote plant health. Many of the microbes involved form symbiotic relationships with the target crops which are beneficial for both parties (mutualism). While microbial inoculants are applied to improve plant nutrition, they can also be used to promote plant growth by stimulating the production of plant hormones. When added to seeds, growth media, or leaves, microbial inoculants have been shown to be useful in all crops, both in open fields and in greenhouses. Microbial inoculants can also be used to initiate systemic acquired resistance (SAR) in plant species to several common crop diseases.
  • SAR systemic acquired resistance
  • the microbial inoculant is chosen from a bacterial inoculant, a fungal inoculant, or a combination thereof.
  • Preferred genera of bacterial micro-organisms are Azospirillum, Rhizobium, Bacillus, Pseudomonas, Streptomyces, Zooglia, Agrobacterium, and combinations thereof.
  • the bacterial inoculant belongs to one of the following species: Bacillus subtilis, Bacillus megaterium, Bacillus velezensis, Bacillus amyloliquefaciens, Azotobacter vinelandi, Azospirillum brasilense, Ensifer meliloti, Pseudomonas vancouverensis, Paenibacillus polymyxa, or combinations thereof.
  • Rhizobium is a genus of soil bacteria that fix nitrogen and form symbiotic associations within the nodules on the roots of pulses. This increases nitrogen nutrition and is important for the cultivation of soybeans, chickpeas, and many other pulses.
  • Bacillus, Pseudomonas and Streptomyces provide some, if not all, of the following benefits: increased plant growth, decomposition of organic matter and pesticide residues, increased nutrient cycle and nitrogen fixation, increased resistance to biotic and abiotic stresses, greater solubility of meso- and microelements, greater production of natural hormones which are useful for plant growth, better soil structure, and better germination and vitality of seeds.
  • PBS phosphate solubilising bacteria
  • the composition according to the invention comprises forms of micro-organisms in a concentration of said bacterial inoculant of lxlO 5 CFU/ml to lxlO 10 CFU/ml.
  • Preferred genera of fungal micro-organisms are ascomycetes and basidiomycetes.
  • the fungal inoculant belongs to the species Trichoderma asperellum, Trichoderma longibrachiatum, Metarhizium anisopliae, Pochonia chlamidospora, and combinations thereof. These fungi provide many of the same plant health benefits as those offered by the above bacteria, including increasing the plant's resistance to environmental stresses and the production of natural hormones.
  • the composition according to the invention comprises forms of micro-organisms in a concentration of said fungal inoculant of lxlO 5 CFU/ml to lxlO 10 CFU/ml.
  • said microbial inoculant comprises up to five different micro-organisms .
  • said microbial inoculant comprises Bacillus subtilis, Bacillus megaterium, Bacillus velezensis, Bacillus amyloliquefaciens, Azotobacter vinelandi, Azospirillum brasilense, Ensifer meliloti, Pseudomonas vancouverensis, or a combination thereof.
  • the composition consists essentially of a hydrolysed microbial inoculant of natural raw material, as described above.
  • the expression " consists essentially of” means that said microbial inoculant and said corn steep liquor hydrolysate are the only active ingredients that act as plant growth and fruit production promoters present in the compositions, since the other possible components show different activities which do not interfere with those of the microbial and hydrolysed inoculant, or are simply co-formulants.
  • the composition consists of a hydrolysed microbial inoculant of natural raw material, as described above.
  • composition according to the invention further comprises corn steep liquor, lupine, algae, molasses, coriander, cocoa, olive residues, and combinations thereof.
  • composition according to the invention further comprises additional ingredients such as glycerol, humates (from humic acid), fulvates (from fulvic acid), acetic acid, propionic acid, citric acid, lactic acid, or combinations thereof.
  • additional ingredients such as glycerol, humates (from humic acid), fulvates (from fulvic acid), acetic acid, propionic acid, citric acid, lactic acid, or combinations thereof.
  • composition according to the invention further comprises additional ingredients such as botanical extracts, fermented plant extracts, or combinations thereof.
  • additional ingredients such as amino acids.
  • Amino acids can act as an energy source to increase plant metabolism and improve plant nutrient absorption.
  • Said additional amino acids may be tryptophan, asparagine, glutamine, glycine, selenocysteine, serine, ornithine, taurine, or combinations thereof.
  • the composition according to the invention further includes additional ingredients such as vitamins and minerals.
  • Vitamins act as catalysts for beneficial enzymes and improve plant metabolism.
  • Folic acid and biotin two components of the vitamin B complex
  • vitamins and minerals may be boron, copper, iron, manganese, zinc, molybdenum, chlorine, phosphorus, potassium, calcium, magnesium, sulphur, or combinations thereof.
  • the composition according to the invention further comprises additional ingredients such as carbohydrates.
  • Said carbohydrates may be glucose, galactose, galactose, fructose, arabinose, xylose, sucrose, lactose, maltose, amylose, amylopectin, glycogen, glyceraldehyde, ribose, or combinations thereof.
  • composition according to the invention further comprises additional ingredients such as enzymes.
  • enzymes may be phospholipases, lipases, proteases, amylases, cellulases, catalases, laccases, or combinations thereof.
  • the composition may be applied at a rate of 0.5-50 litres/hectare, more preferably with periodic treatments, depending on the crop and the season.
  • the present invention relates to an agrochemical product comprising the composition and agrochemical additives.
  • Suitable additives include pH regulators, acidity regulators, water hardness regulators, mineral oils, plant oils, fertilisers, natural leaf fertilisers, and combinations thereof.
  • the natural raw material was analysed to establish the content of organic nitrogen and starches and fibres, in order to assess what kind of enzymatic digestion of the components performs best. On the basis of the results, different hydrolases which act on the components were tested, deciding on the amounts required according to the composition.
  • enzymes which act on the protein component for example, enzymes in class EC 3.4, i.e. enzymes that act on the peptide bond, such as, for example, mono-, di- , or tripeptidases, or mixtures thereof, proteases, and proteinases.
  • enzymes which act on fibres and carbohydrates for example, enzymes in class EC 3.2 - Glycosidases, i.e. enzymes that act on O- and S-glycosyl compounds or N-glycosyl compounds, such as, for example, amylase, cellulase and xylanase.
  • Each hydrolase was used within the optimum pH and temperature ranges, by developing different processes which could all lead to the hydrolysis of the different components present, regardless of the pathway.
  • the CSL was treated with an anti fermentation agent and a mixture of EC 3.2 enzymes for 6 hours at about 52 °C and at about pH 4. Subsequently, KOH was added to bring the pH up to 6, followed by a mixture of EC 3.2 and 3.4 enzymes for a further 6 hours.
  • the CSL was treated with an anti-fermentation agent and a mixture of enzymes EC 3.2 and 3.4 for 6 hours at about 52 °C and at pH about 4. Subsequently, KOH was added to bring the pH up to 5.5-6.
  • Amino acid analysis showed a general increase in free amino acids of between 0.5 and 11% and, more specifically, that these are the amino acids which become more bioavailable.
  • the lupine was treated with an anti fermentation agent and a mixture of EC 3.2 enzymes for 6 hours at about 52 °C and at about pH 4. Subsequently, KOH was added to bring the pH up to 6, followed by a mixture of EC 3.2 and 3.4 enzymes for a further 6 hours.
  • the lupine was treated with an anti-fermentation agent and a mixture of EC 3.2 and E 3.4 enzymes for 6 hours at about 52 °C and at about pH 4. Subsequently, KOH was added to bring the pH up to 5.5-6.
  • Amino acid analysis showed a general increase in free amino acids of 2 to 15%.
  • the formulation is as follows:
  • the algae is treated with an anti-fermentation agent and a mixture of EC 3.2 enzymes for 6 hours at about 52 °C and at about pH 4. Subsequently, KOH was added to bring the pH up to 6, followed by a mixture of EC 3.2 and 3.4 enzymes for a further 6 hours.
  • the algae was treated with an anti-fermentation agent and a mixture of EC 3.2 and E 3.4 enzymes for 6 hours at about 52 °C and at about pH 4. Subsequently, KOH was added to bring the pH up to 5.5-6. Following treatment with enzymes from class 3.2 (which act on fibres and complex carbohydrates), the dry residue - which indicates the presence of insoluble material - decreased and the amount of polysaccharides or simple sugars present in the mixture increased. Viscosity decreased and, probably due to this, the protein part became more available for the action of proteolytic enzymes.
  • Amino acid analysis showed a general increase in free amino acids of 4 to 6%.
  • the formulation is as follows: According to a first preparation procedure, the molasses was treated with an anti fermentation agent and a mixture of EC 3.2 enzymes for 6 hours at about 52 °C and at about pH 4. Subsequently, KOH was added to bring the pH up to 6, followed by a mixture of EC 3.2 and 3.4 enzymes for a further 6 hours. According, however, to a second preparation procedure, the CSL was treated with an anti-fermentation agent and a mixture of EC 3.2 and E 3.4 enzymes for 6 hours at about 52 °C and at about pH 4. Subsequently, KOH was added to bring the pH up to 5.5-6.
  • the coriander was treated with an anti fermentation agent and a mixture of EC 3.2 enzymes for 6 hours at about 52 °C and at about pH 4. Subsequently, KOH was added to bring the pH up to 6, followed by a mixture of EC 3.2 and 3.4 enzymes for a further 6 hours.
  • the coriander was treated with an anti-fermentation agent and a mixture of EC 3.2 and E 3.4 enzymes for 6 hours at about 52 °C and at about pH 4. Subsequently, KOH was added to bring the pH up to 5.5-6.
  • the cocoa was treated with an anti- fermentation agent and a mixture of EC 3.2 enzymes for 6 hours at about 52 °C and at about pH 4. Subsequently, KOH was added to bring the pH up to 6, followed by a mixture of EC 3.2 and 3.4 enzymes for a further 6 hours.
  • the cocoa was treated with an anti-fermentation agent and a mixture of EC 3.2 and E 3.4 enzymes for 6 hours at about 52 °C and at about pH 4. Subsequently, KOH was added to bring the pH up to 5.5-6.
  • Amino acid analysis showed a general increase in free amino acids of 0.5 to 1.5%.
  • the olive residues were treated with an anti fermentation agent and a mixture of EC 3.2 enzymes for 6 hours at about 52 °C and at about pH 4. Subsequently, KOH was added to bring the pH up to 6, followed by a mixture of EC enzymes 3.2 and 3.4 and 3.1 for a further 6 hours.
  • the olive residues were treated with an anti-fermentation agent and a mixture of EC 3.2 and E 3.4 enzymes for 6 hours at about 52 °C and at about pH 4. Subsequently, KOH was added to bring the pH up to 5.5-6.
  • a greenhouse test was carried out on cyclamens; 18 plants were tested for each test mixture.
  • Each text mixture was treated in fertigation, with about 25 ml per plant, corresponding to the doses/hectare specified below.
  • the treatment was carried out three times over three weeks, one treatment per week.
  • the following fertigation protocol was applied 45PPM N - NPK (1:0.7: 1.5) - 400ML/PLANT x DAY (3 IRR.) - PH 7
  • a greenhouse test was carried out on cyclamens; 18 plants were tested for each test mixture.
  • Each text mixture was treated in fertigation, with about 25 ml per plant, corresponding to the doses/hectare specified below.
  • the treatment was carried out three times over three weeks, one treatment per week.
  • the following fertigation protocol was applied 45PPM N - NPK (1:0.7: 1.5) - 400ML/PLANT x DAY (3 IRR.) - PH 7 1.
  • algae extract - 2.51/ha 2.
  • Example 6 Synergistic mixture of hydrolysate of lupine, molasses, and algae A greenhouse test was carried out on cyclamens; 18 plants were tested for each test mixture. Each text mixture was treated in fertigation, with about 25 ml per plant, corresponding to the doses/hectare specified below. The treatment was carried out three times over three weeks, one treatment per week. The following fertigation protocol was applied: 45PPM N - NPK (1 :0.7: 1.5) - 400ML/PLANT xD A Y (3 IRR.) - PH 7
  • TREATMENTS, DOSE The compounds and the hydrolysates were diluted within a range of 1:100 to 1:2000. 16 PLANTS/TEST MIXTURES were used and the saline stress was provided by means of a WATER + NACL 300 mm solution IN A SAUCER, to which the biostimulant mixtures were added during the treatment. TREATMENTS, FREQUENCY: IRRIGATIONS WITH 2.5 1/SAUCER
  • PLANTS purchased from a NURSERY, were REPOTTED IN PEAT + PERLITE POTS IRRIGATION was performed WITH 2.5L NACL 300 mm (TAP WATER) +
  • NPK N 50ppm, 1:0.5:1.7
  • the growth temperature was the following: 15-25 °C measured in the GREENHOUSE
  • the test was carried out on stressed tomatoes in a climatic growth chamber, on 5-week- old seedlings transplanted into a soil composed of 40% sand.
  • TREATMENTS FREQUENCY: once a week TREATMENTS, NUMBER: 4 weeks
  • the seedlings were phenotyped according to the height of the aerial part and root development.
  • results are summarised in the following table: The results show how the hydrolysates according to formulations such as Ascophyllum algae, hydrolysed coriander, hydrolysed lupine, hydrolysed molasses and hydrolysed CSL, give significant results in phenotyping, confirming the quality of the process developed and the effectiveness of the products produced in accordance with the claims.
  • the test was carried out on stressed tomatoes in a climatic growth chamber, on 2- month-old seedlings transplanted into a growth medium made up of 40% sand.
  • TREATMENTS, FREQUENCY once upon repotting, once one week later TREATMENTS, NUMBER: 2 weeks
  • the seedlings were phenotyped according to the height of the aerial part and root development.
  • the aerial part showed no significant variations, while the root part, showed the following significant results in sandy growth medium.
  • the positive results of root phenotyping are summarised in the table below: The results show how the hydrolysates according to formulations such as Ascophyllum algae, cocoa, and hydrolysed cocoa, hydrolysed molasses and hydrolysed olive residues, give significant results in phenotyping, confirming the quality of the process developed and the effectiveness of the products produced according to the claims.
  • the corn field was divided as follows:
  • Zone al dose: 4 g/kg seed Zone a2: dose: 16 g/kg seed Zone a2: dose: 20 g/kg seed
  • Zone bl dose: 4 g/kg seed
  • Zone b2 dose: 16 g/kg seed
  • Zone b3 dose: 20 g/kg seed
  • Zone cl dose: 4 g/kg CSL + 16 g/kg Trichoderma asperellum
  • Zone c2 dose: 16 g/kg Trichoderma asperellum + 4 g/kg CSL -Area D, Azospirillum brasilense
  • Zone dl dose: 4 g/kg seed
  • Zone d2 dose: 16 g/kg seed
  • Zone d3 dose: 20 g/kg seed
  • Zone el dose: 4 g/kg CSL + 16 g/kg Azo spirillum brasilense
  • Zone e2 dose: 16 g/kg Azo spirillum brasilense + 4 g/kg CSL
  • Zone/ dose: 4 g/kg seed Zone/2: dose: 16 g/kg seed Zona f3: dose 20 g/kg seme
  • Zone gl dose: 4 g/kg CSL + 16 g/kg Bacillus megaterium
  • Zone g2 dose: 16 g/kg Bacillus megaterium, + 4 g/kg CSL
  • Zone hi dose: 4 g/kg seed
  • Zone h2 dose: 16 g/kg seed
  • Zone /z5 dose: 20 g/kg seed
  • Zone il dose: 4 g/kg CSL + 16 g/kg Glomus intraradices
  • Zone z2 dose: 16 g/kg Glomus intraradices + 4 g/kg CSL
  • Zone 11 dose: 4 g/kg Trichoderma asperellum seed, + 4 g/kg Azospirillum brasilense seed, + 4 g/kg Bacillus megaterium seed + 4 g/kg Glomus intraradices seed
  • Zone 12 dose: 4 g/kg Trichoderma asperellum seed, + 4 g/kg Azospirillum brasilense seed, + 4 g/kg Bacillus megaterium seed + 4 g/kg Glomus intraradices seed + 4 g/kg CSL
  • the microbial cultures used were brought to a concentration of 1c10 L 8 CLU/ml, apart from the Glomus intraradices, which were employed in a concentration of 500 spores/g. 150 kg/ha of N, in the form of urea, was distributed in coverage over the study areas. No irrigation was necessary during the growth cycle.
  • the data was collected at three seasonal measurement times, namely at stem elongation, at flowering, and at the ripening of the cobs (hard dough stage).
  • Leaf physiology Plant physiology was measured directly in the field by choosing 10 treated plants per area at random. The measurements were taken using a leaf which had been exposed to the sun and was located on the stalk above the highest cob.
  • ADC pro-SD model 2- Portable gas exchange analyser (ADC pro-SD model), i.e. an instrument that measures changes in the concentration of carbon dioxide caused by photosynthetic activity through the "Assimilation rate” parameter (A).
  • the photosynthetic efficiency (given by the average of several leaves) was calculated using the ratio between ETR (Electron Transport Rate) and A (Assimilation rate). This ratio expresses the number of pmoles of electrons required by PII (Photosystem II) to organise one pmole of CO2.
  • the leaf ecophysiology measured in the field shows appreciable differences in values in plants from different areas.
  • the tests were performed on PCA (0.5% peptone and 0.25% yeast extract) for bacteria, bacteria with hydrolysed citrus molasses, and hydrolysed citrus molasses alone, and on PDA for fungi, fungi with hydrolysed citrus molasses, and hydrolysed citrus molasses alone.
  • PCA peptone and 0.25% yeast extract
  • PDA fungi, fungi with hydrolysed citrus molasses, and hydrolysed citrus molasses alone.
  • the dual tests were performed placing the F. oxysporum strain on one side of the petri dish on the surface of the growth medium while the strain or product for which the biocontrol activity was to be assessed was placed on the other side. The surface of the dish occupied by F. oxysporum was measured at 10 days of growth at 27 °C.
  • the percentage of inhibition was calculated using the ratio between the surface area occupied by F. oxysporum in the presence of the test strains and the surface area occupied by F. oxysporum grown in the absence of a biocontrol agent. Tests were performed with the bacterial cultures alone, with the bacterial cultures supplemented with hydrolysed citrus molasses, and with hydrolysed citrus molasses alone by placing the phytopathogen in the centre of the petri dish on the surface of the growth medium, while the strain or product for which the biocontrol activity was to be assessed was streaked onto the two opposite sides of the dish. The surface area of the dish occupied by F. oxysporum was measured at 10 days of growth at 27 °C and the percentage of inhibition calculated as described earlier.
  • Botrytis cinerea biocontrol Test method Tests were performed on PCA (0.5% peptone and 0.25% yeast extract) for bacteria, bacteria with hydrolysed algae extract, and hydrolysed algae extract alone and on PDA for fungi, fungi with hydrolysed algae extract, and hydrolysed algae extract alone.
  • PCA peptone and 0.25% yeast extract
  • the dual tests were performed placing the Botrytis cinerea strain on one side of the petri dish on the surface of the growth medium while the strain or product for which the biocontrol activity was to be assessed was placed on the other side.
  • the surface of the dish occupied by Botrytis cinerea was measured at 10 days of growth at 27 °C.
  • the percentage of inhibition was calculated using the ratio between the surface area occupied by Botrytis cinerea in the presence of the test strains and the surface area occupied by Botrytis cinerea grown in the absence of a biocontrol agent.
  • Tests were performed with the bacterial cultures alone, with the bacterial cultures supplemented with hydrolysed algae extracts, and with hydrolysed algae extracts alone by placing the phytopathogen in the centre of the petri dish on the surface of the growth medium, while the strain or product for which the biocontrol activity was to be assessed was streaked onto the two opposite sides of the dish. Furthermore, commercially available products Serenade® and Amylo-x® products were also tested for comparison purposes.
  • the surface area of the dish occupied by Botrytis cinerea was measured at 10 days of growth at 27 °C and the percentage of inhibition calculated as described earlier.
  • Concentration of use of bacterial cultures 1c10 L 9 CFU/ml
  • Concentration of use of fungal cultures 5c10 L 7 CFU/ml
  • Hydrolysed algae extract/microbial culture mixture ratio 50%:50%
  • Botrytis cinerea
  • the tests were performed on PC A (0.5% peptone and 0.25% yeast extract) for bacteria, bacteria with polyphenols, and polyphenols alone, and on PDA for fungi, fungi with polyphenols, and polyphenols alone.
  • the dual tests were performed placing the F. fujikuroi strain on one side of the petri dish on the surface of the growth medium while the strain or product for which the biocontrol activity was to be assessed was placed on the other side.
  • the surface of the dish occupied by F. fujikuroi was measured at 10 days of growth at 27 °C.
  • the percentage of inhibition was calculated using the ratio between the surface area occupied by F.
  • the table below shows the results for the strains that have a biocontrol activity against Fusarium oxysporum cubense (FOC).
  • Trichoderma asperellum + 94 polyphenols

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Abstract

L'invention décrit un hydrolysat à partir d'une matière première naturelle qui s'est révélé significativement efficace dans la stimulation de la croissance des plantes, ou dans la protection de plantes contre des agents phytopathogènes. L'hydrolysat à partir d'une matière première naturelle trouve donc une application avantageuse dans l'agriculture.
EP20804679.7A 2019-10-21 2020-10-21 Hydrolysat pour favoriser la croissance des plantes, la biostimulation et la lutte biologique, et son utilisation en agriculture Pending EP4048072A1 (fr)

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PCT/IB2020/059877 WO2021079276A1 (fr) 2019-10-21 2020-10-21 Hydrolysat pour favoriser la croissance des plantes, la biostimulation et la lutte biologique, et son utilisation en agriculture

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