WO2020200959A1 - Procédé de lutte contre la maladie ganoderma des palmiers à huile - Google Patents

Procédé de lutte contre la maladie ganoderma des palmiers à huile Download PDF

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Publication number
WO2020200959A1
WO2020200959A1 PCT/EP2020/058358 EP2020058358W WO2020200959A1 WO 2020200959 A1 WO2020200959 A1 WO 2020200959A1 EP 2020058358 W EP2020058358 W EP 2020058358W WO 2020200959 A1 WO2020200959 A1 WO 2020200959A1
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plant
ganoderma
bacillus subtilis
methyl
palm
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PCT/EP2020/058358
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English (en)
Inventor
Christian Arnold
Suprianto SIDIK
Febi ARYANA
Kukuh Ambar WALUYO
Phuah BOON GIAP
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Bayer Aktiengesellschaft
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Publication of WO2020200959A1 publication Critical patent/WO2020200959A1/fr

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    • 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
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
    • A01N63/22—Bacillus

Definitions

  • the present invention relates to biological control of diseases caused by Ganoderma spp. in agricultural plants and, particularly, basal stem rot caused by G. boninense in oil palms.
  • the African oil palm Elaeis guineensis Jacq. is an important oil-food crop.
  • Oil palm plants are monoecious, i.e. single plants produce both male and female flowers. They are characterized by an alternating series of male and female inflorescences.
  • the male inflorescence is made up of numerous spikelets, and can bear well over 100,000 flowers.
  • Oil palm is naturally cross-pollinated by insects and the wind.
  • the female inflorescence is a spadix that contains several thousands of flowers borne on thorny spikelets. A bunch carries 500 to 4,000 fruit.
  • the oil palm fruit is a sessile drupe that is spherical to ovoid or elongated in shape and is composed of an exocarp, a mesocarp containing palm oil, and an endocarp surrounding a kernel.
  • Oil palm is important both because of its high yield and because of the high quality of its oil.
  • yield oil palm is the highest yielding oil-food crop, with a recent average yield of 3.67 tons per hectare per year and with the best progenies known to produce about 10 tons per hectare per year.
  • Oil palm is also the most efficient plant known for harnessing the energy of sunlight for producing oil.
  • the palm kernel oil is more saturated than the mesocarp oil. Both are low in free fatty acids.
  • the current combined output of palm oil and palm kernel oil is about 50 million tonnes per year, and demand is expected to increase substantially in the future with increasing global population and per capita consumption of oils and fats.
  • Basal stem rot is a disease of oil palm plants that is becoming an increasingly common problem among oil palm plantations in Malaysia and Indonesia and that threatens to limit the productivity of these plantations.
  • Basal stem rot is caused by a species of the f mgus Ganoderma, and in particular G. boninense, and is considered to be the most destructive disease in the oil palm industry in Southeast Asia. Infections appear to advance more quickly in subsequent generations of oil palm planting because of accumulation of inoculums, and the possibility of evolution of more virulent strains of the pathogen.
  • the mating system of G. boninense strongly favors outcrossing and this provides a possible mechanism for the selection of pathogen virulence.
  • Infection is believed to be caused by contact of the pathogen with roots.
  • Earliest visual symptoms include wilting of fronds and malnutrition.
  • Diagnostic symptoms for confirmation of basal stem rot include detection of disease lesions (also termed dry rot) at the base of oil palm plants and the appearance of fruiting bodies.
  • Basal stem rot can result in oil palm plants that have small canopies, show signs of loose vigor (the so-called skirting), are impaired with respect to fruit production, and are subject to being toppled by the wind.
  • Basal stem rot threatens to reduce fresh fruit bunch yield in Malaysia and Indonesia, as shown in a recent case study at Johor estate (Roslan & Idris, 2012, Oil Palm Industry Economic Journal, 12(1), 24-30).
  • Affected oil palm plants can die within six to twelve months after the development of symptoms, and up to 80% of plantings may die within the first half of the otherwise expected economic life of the oil palm plants.
  • Recycled fields from coconut plantings and fields left with infected stumps have created a mass of inoculum of Ganoderma boninense in the soil. New plantings of oil palms in these fields will be subject to infection, and thus these fields will have shorter cycles in the next generation of planting.
  • Synthetic chemical treatments may be impractical for large-scale application due to their high cost of implementation.
  • synthetic fungicides often are non-specific and therefore can act on organisms other than the target organisms, including other naturally occurring beneficial organisms.
  • Consumers worldwide are increasingly conscious of the potential environmental and health problems associated with the residuals of chemicals, particularly in food products. This has resulted in growing consumer pressure to reduce the use or at least the quantity of chemical (i.e., synthetic) fungicides.
  • a further problem arising with the use of synthetic fungicides is that the repeated and exclusive application of a fungicide often leads to selection of resistant pathogens. Normally, such strains are also cross-resistant against other active ingredients having the same mode of action. An effective control of the pathogens with said active compounds is then not possible any longer. However, active ingredients having new mechanisms of action are difficult and expensive to develop.
  • Ganoderma without increasing the occurrence of resistant strains of the pathogen are particularly desirable.
  • compositions comprising a biological control agent with activity against Ganoderma.
  • compositions that, when applied to a crop, results in a decreased amount of residues in the crop, thereby reducing the risk of resistance formation and nevertheless providing efficient pest and/or disease control.
  • compositions and methods according to the invention preferably fulfills the above-described needs. It has been discovered surprisingly that the application of the compositions according to the present invention to plants, plant parts, and/or plant’s locus of growth preferably allows efficient control of phytopathogens and, particularly, Ganoderma spp., the causative agents basal stem rot in oil palms.
  • the present invention provides a composition for controlling a fungal infection by Ganoderma spp. in an agricultural plant, the composition comprising Bacillus subtilis QST713 deposited under NRRL Accession No. B-21661 or a fungicidal mutant thereof.
  • the composition comprises a fermentation product of the Bacillus subtilis QST713 deposited under NRRL Accession No. B-21661 or fungicidal mutant thereof.
  • the fermentation product may comprise Bacillus subtilis QST713 cells or cells of a fungicidal mutant of Bacillus subtilis QST713, metabolites and residual fermentation broth.
  • the Bacillus subtilis QST713 cells or cells of a fungicidal mutant of Bacillus subtilis QST713 are spores.
  • the present invention relates to the use of a composition disclosed herein for treating a fungal infection by Ganoderma sp. in an agricultural plant.
  • the present invention relates to a method for controlling a fungal infection by Ganoderma spp. in an agricultural plant, the method comprising applying to an agricultural plant and/or locus for plant growth Bacillus subtilis QST713 deposited under NRRL Accession No. B- 21661 or a fungicidal mutant thereof.
  • the applying is preceded by identifying that the agricultural plant and/or the locus for plant growth needs treatment.
  • the agricultural plant comprises propagation material selected from the group consisting of a seedling, a nursery plant, and a seed.
  • the Bacillus subtilis QST713 deposited under NRRL Accession No. B-21661 or fungicidal mutant thereof is applied to the agricultural plant and/or the locus for plant growth prior to planting and/or at planting.
  • the composition can be applied before, during or after the plant or plant part comes into contact with the soil.
  • the methods of the present invention include but are not limited to applying the composition using an application method such as soil surface drench, shanked-in, injected or applied in-furrow.
  • the methods of the present invention include applying the compositions as a seed treatment.
  • the treatment of the plants and plant parts with the compositions according to the invention may be carried out directly or by acting on the environment, habitat or storage space using customary treatment methods, for example by dipping, spraying, atomizing, misting, evaporating, dusting, fogging, scattering, foaming, painting on, spreading, injecting, drenching, trickle irrigation and, in the case of propagation material, in particular in the case of seed, furthermore by the dry seed treatment method, the wet seed treatment method, the slurry treatment method, by encrusting, by coating with one or more coats and the like. It is furthermore possible to apply the active substances by the ultra-low volume method or to inject the active substance preparation or the active substance itself into the soil.
  • the Bacillus subtilis QST713 deposited under NRRL Accession No. B-21661 or fungicidal mutant thereof is applied to soil.
  • the Bacillus subtilis QST713 deposited under NRRL Accession No. B-21661 or fungicidal mutant thereof is applied to soil in contact with roots of the agricultural plant or soil at a base of the agricultural plant.
  • such application results in enhanced yield and/or improved health of the plant. In one embodiment, the application results in a reduction of disease severity.
  • the Bacillus subtilis QST713 deposited under NRRL Accession No. B-21661 or fungicidal mutant thereof is applied at a rate of about 1 c 10 4 to about 1 c 10 8 colony forming units (cfu) per gram of soil.
  • B-21661 or fungicidal mutant thereof is applied at a rate of about 1 x 10 5 to about 1 * 10 8 cfu per gram of soil, at a rate of about 1 * 10 6 to about 1 * 10 8 cfu per gram of soil, at a rate of about 1 * 10 4 to about 1 * 10 7 cfu per gram of soil, or at a rate of about 1 * 10 5 to about 1 x 10 7 cfu per gram of soil.
  • the method further comprises applying to the agricultural plant and/or locus for plant growth an herbicide, insecticide and/or a fungicide.
  • the herbicide, insecticide and/or fungicide may be applied sequentially or simultaneously with the Bacillus subtilis QST713 deposited under NRRL Accession No. B-21661 or a fungicidal mutant thereof.
  • the fungicide is a triazole fungicide.
  • a non-limiting list of triazole fungicides includes azaconazole, bitertanol, bromuconazole, cyproconazole, diclobutrazol, difenoconazole, diniconazole, diniconazole-M, epoxiconazole, etaconazole, fenbuconazole, fluquinconazole, flusilazole, flutriafol, furconazole, furconazole-cis, hexaconazole, imibenconazole, ipconazole, metconazole, myclobutanil, paclobutrazol, penconazole, propiconazole, prothioconazole, quinconazole, simeconazole, tebuconazole, tetraconazole, triadimefon, triadimenol, triticonazole, uniconazole, uniconazole-P, voriconazole, l
  • the fungicidal mutant strain has a genomic sequence with greater than about 90% sequence identity to Bacillus subtilis QST713 deposited under NRRL Accession No. B- 21661 and/or the fungicidal mutant strain has fungicidal activity that is comparable to or better than that oiBacillus subtilis QST713 deposited under NRRL Accession No. B-21661.
  • the Ganoderma spp. are Ganoderma boninense, Ganoderma orbiforme, Ganoderma zonatum, and/or Ganoderma miniatocinctum.
  • the Ganoderma spp. are Ganoderma boninense, Ganoderma orbiforme, Ganoderma zonatum, and/or Ganoderma miniatocinctum.
  • Ganoderma spp. is Ganoderma boninense. It is most preferred that Ganoderma boninense is controlled by Bacillus subtilis QST713.
  • the agricultural plant is a palm tree selected from the group consisting of oil palm, coconut palm, date palm, sago palm, nipa palm, areca palm, and ornamental palm.
  • the agricultural plant is an oil palm.
  • the present invention provides methods of treating a plant to control Ganoderma spp. wherein the methods comprise applying an effective amount of at least one biological control agent selected from Bacillus subtilis QST713 deposited under NRRL Accession No. B-21661, metabolites produced therefrom, a cell -free extract thereof, and combinations thereof to the plant, to a part of the plant and/or to the locus surrounding the plant, such as to a plant’s growth media.
  • FIG. 1 A depicts control of Ganoderma boninense strain PER71 and Ganoderma boninense strain UP by Bacillus subtilis QST713.
  • FIG. IB depicts control of Ganoderma boninense strain PER71 and Ganoderma boninense strain UP with the triazole fungicide, tebuconazole.
  • FIG. 2A depicts control of Ganoderma disease in the roots and internal plant tissue of oil palm trees treated with SERENADE ® ASO ( Bacillus subtilis QST713) or one of four chemical treatments.
  • FIG. 2B depicts control of Ganoderma disease in the stems and foliage of oil palm trees treated with SERENADE ® ASO ( Bacillus subtilis QST713) or one of four chemical treatments.
  • microorganisms and particular strains described herein, unless specifically noted otherwise, are all separated from nature and grown under artificial conditions such as in shake flask cultures or through scaled-up manufacturing processes, such as in bioreactors to maximize bioactive metabolite production, for example. Growth under such conditions leads to strain
  • the SERENADE ® product (EPA Registration No. 69592-12) contains a unique strain of Bacillus subtilis (strain QST713) and many different lipopeptides that work synergistically to destroy disease pathogens and provide superior antimicrobial activity.
  • the SERENADE ® product is used to protect plants such as vegetables, fruit, nut, and vine crops against diseases such as Fire Blight, Botrytis, Sour Rot, Rust, Sclerotinia, Powdery Mildew, Bacterial Spot and White Mold.
  • SERENADE ® products are available as either liquid or dry formulations, which can be applied as a foliar and/or soil treatments. Copies of EPA Master Labels for SERENADE ® products, including SERENADE ® ASO, SERENADE ® MAX, SERENADE ® OPTIMUM (or OPTI), and SERENADE SOIL ® , are publicly available through National Pesticide Information Retrieval System’s (NPIRS ® ) USEPA/OPP Pesticide Product Label System (PPLS).
  • NIRS ® National Pesticide Information Retrieval System
  • SERENADE ® ASO (Aqueous Suspension-Organic) contains 1.34% of dried QST713 as an active ingredient and 98.66% of other ingredients.
  • SERENADE ® ASO is formulated to contain a minimum of 1 x 10 9 cfii/g of QST713 while the maximum amount of QST713 has been determined to be 3.3 x 10 10 cfu/g.
  • Alternate commercial names for SERENADE ® ASO include SERENADE BIOFUNGICIDE ® , SERENADE SOIL ® and SERENADE ® GARDEN DISEASE. For further information, see the U.S. EPA Master Labels for SERENADE ® ASO dated January 4, 2010 and SERENADE SOIL ® , each of which is incorporated by reference herein in its entirety.
  • SERENADE ® MAX contains 14.6% of dried QST713 as an active ingredient and 85.4% of other ingredients. SERENADE ® MAX is formulated to contain a minimum of 7.3 x 10 9 cfu/g of QST713 while the maximum amount of QST713 has been determined to be 7.9 x 10 10 cfu/g. For further information, see the U.S. EPA Master Label for SERENADE ® MAX, which is incorporated by reference herein in its entirety.
  • SERENADE ® OPTIMUM (or OPTI) contains 26.2% of dried QST713 as an active ingredient and 73.8% of other ingredients.
  • SERENADE ® OPTIMUM (or OPTI) is formulated to contain a minimum of 1.31 x 10 10 cfu/g of QST713.
  • U.S. EPA Master Label for SERENADE ® OPTIMUM (or OPTI), which is incorporated by reference herein in its entirety.
  • NRRL is the abbreviation for the Agricultural Research Service Culture Collection, an international depositary authority for the purposes of deposing microorganism strains under the Budapest Treaty on the international recognition of the deposit of microorganisms for the purposes of patent procedure, having the address National Center for Agricultural Utilization Research, Agricultural Research Service, U.S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604, U.S.A.
  • Bacillus subtilis QST713, its mutants, its supernatants, and its lipopeptide metabolites, and methods for their use to control plant pathogens and insects are fully described in U.S. Patent Nos. 6,060,051; 6,103,228; 6,291,426; 6,417,163; and 6,638,910; each of which is specifically and entirely incorporated by reference herein for everything it teaches.
  • the strain is referred to as AQ713, which is synonymous with QST713.
  • Bacillus subtilis QST713 has been deposited with the NRRL on May 7, 1997, under the provisions of the Budapest Treaty on the
  • the QST713 strain was designated as a Bacillus subtilis based on classical, physiological, biochemical and morphological methods. Taxonomy of the Bacillus species has evolved since then, especially in light of advances in genetics and sequencing technologies, such that species designation is based largely on DNA sequence rather than the methods used in 1998. After aligning protein sequences from B. amyloliquefaciens FZB42, B. subtilis 168 and QST713,
  • B. amyloliquefaciens FZB42 approximately 95% of proteins found in B. amyloliquefaciens FZB42 are 85% or greater identical to proteins found in QST713; whereas only 35% of proteins in B. subtilis 168 are 85% or greater identical to proteins in QST713.
  • B. subtilis 168 proteins found in B. subtilis 168 are 85% or greater identical to proteins in QST713.
  • a pesticidal product based on B. subtilis strain FZB24 which is as closely related to QST713 as is FZB42, is classified in documents of the Environmental Protection Agency as B. subtilis var. amyloliquefaciens .
  • B. subtilis strain AQ30002 (aka QST30002) or AQ30004 (aka QST30004), deposited as Accession Nos. NRRL B-50421 and NRRL B-50455 which are described in International Patent Publication No. WO 2012/087980 or mutants of these B. subtilis strains having all of the physiological and morphological characteristics of B. subtilis strain AQ30002 (aka QST30002) or AQ30004 (aka QST30004) can also be used in the method of the invention, either alone or in mixture with B. subtilis QST713.
  • mutant refers to a genetic variant derived from QST713.
  • the mutant has all the identifying characteristics of QST713.
  • the mutant controls plant pathogens (e.g., Ganoderma spp.) at least as well as the parent QST713 strain.
  • mutants are genetic variants having a genomic sequence that has greater than about 85%, greater than about 90%, greater than about 95%, greater than about 98%, or greater than about 99% sequence identity to the QST713 strain. Mutants may be obtained by treating QST713 cells with chemicals or irradiation or by selecting spontaneous mutants from a population of QST713 cells (such as phage resistant mutants) or by other means well known to those practiced in the art.
  • compositions of the present invention can be obtained by culturing Bacillus subtilis QST713 or mutants thereof according to methods well known in the art, including by using the media and other methods described in U.S. Patent No. 6,060,051.
  • Conventional large-scale microbial culture processes include submerged fermentation, solid-state fermentation, or liquid surface culture.
  • Bacillus subtilis cells Towards the end of fermentation, as nutrients are depleted, Bacillus subtilis cells begin the transition from growth phase to sporulation phase, such that the final product of fermentation is largely spores, metabolites and residual fermentation medium. Sporulation is part of the natural life cycle of Bacillus subtilis and is generally initiated by the cell in response to nutrient limitation.
  • Fermentation is configured to obtain high levels of colony forming units of Bacillus subtilis and to promote sporulation.
  • the bacterial cells, spores and metabolites in culture media resulting from fermentation may be used directly or concentrated by conventional industrial methods, such as centrifugation, tangential-flow filtration, depth filtration, and evaporation. Fermentation broth and broth concentrate are both referred to herein as “fermentation products.”
  • Compositions of the present invention include fermentation products.
  • the concentrated fermentation broth is washed, for example, via a diafiltration process, to remove residual fermentation broth and metabolites.
  • the fermentation broth or broth concentrate can be dried with or without the addition of carriers using conventional drying processes or methods such as spray drying, freeze drying, tray drying, fluidized-bed drying, drum drying, or evaporation.
  • the resulting dry products may be further processed, such as by milling or granulation, to achieve a specific particle size or physical format. Carriers, described below, may also be added post-drying.
  • Cell-free preparations of fermentation broth of the novel variants and strains of Bacillus of the present invention can be obtained by any means known in the art, such as extraction, centrifugation and/or filtration of fermentation broth. Those of skill in the art will appreciate that so- called cell-free preparations may not be devoid of cells but rather are largely cell-free or essentially cell- free, depending on the technique used (e.g., speed of centrifugation) to remove the cells.
  • the resulting cell-free preparation may be dried and/or formulated with components that aid in its application to plants or to plant growth media. Concentration methods and drying techniques described above for fermentation broth are also applicable to cell-free preparations.
  • Metabolites of Bacillus subtilis can be obtained according to the methods set forth in U.S. Patent No. 6,060,051.
  • the term“metabolites” as used herein may refer to semi-pure and pure or essentially pure metabolites or to metabolites that have not been separated from Bacillus subtilis.
  • the metabolites may be purified by size exclusion filtration such as the SEPHADEX ® resins including LH- 20, G10, and G15 and G25 that group metabolites into different fractions based on molecular weight cut-off, such as molecular weight of less than about 2000 Daltons, less than about 1500 Daltons, less than about 1000 Daltons and so on, as the lipopeptides are between 800 Daltons and 1600 Daltons.
  • size exclusion filtration such as the SEPHADEX ® resins including LH- 20, G10, and G15 and G25 that group metabolites into different fractions based on molecular weight cut-off, such as molecular weight of less than about 2000 Daltons, less than about 1500 Daltons, less than about 1000 Daltons and so on, as the lipopeptides are between 800 Daltons and 1600 Daltons.
  • compositions of the present invention may include formulation inerts added to compositions comprising cells, cell-free preparations or metabolites to improve efficacy, stability, and usability and/or to facilitate processing, packaging and end-use application.
  • formulation inerts and ingredients may include carriers, stabilization agents, nutrients, or physical property modifying agents, which may be added individually or in combination.
  • the carriers may include liquid materials such as water, oil, and other organic or inorganic solvents and solid materials such as minerals, polymers, or polymer complexes derived biologically or by chemical synthesis.
  • the carrier is a binder or adhesive that facilitates adherence of the composition to a plant part, such as a seed or root.
  • the stabilization agents may include anti-caking agents, anti-oxidation agents, desiccants, protectants or preservatives.
  • the nutrients may include carbon, nitrogen, and phosphors sources such as sugars, polysaccharides, oil, proteins, amino acids, fatty acids and phosphates.
  • the physical property modifiers may include bulking agents, wetting agents, thickeners, pH modifiers, rheology modifiers, dispersants, adjuvants, surfactants, antifreeze agents or colorants.
  • the composition comprising cells, cell -free preparation or metabolites produced by fermentation can be used directly with or without water as the diluent without any other formulation preparation.
  • the formulation inerts are added after concentrating fermentation broth and during and/or after drying.
  • compositions of the present invention may include carriers, which are inert formulation ingredients added to compositions comprising a lipopeptide -containing fermentation product, cell -free preparations of lipopeptide s or purified, semi -purified or crude extracts of lipopeptides to improve recovery, efficacy, or physical properties and/or to aid in packaging and administration.
  • carriers which are inert formulation ingredients added to compositions comprising a lipopeptide -containing fermentation product, cell -free preparations of lipopeptide s or purified, semi -purified or crude extracts of lipopeptides to improve recovery, efficacy, or physical properties and/or to aid in packaging and administration.
  • Such carriers may be added individually or in combination.
  • inventive compositions can be used as such or, depending on their particular physical and/or chemical properties, in the form of their formulations or the use forms prepared therefrom, such as aerosols, capsule suspensions, cold-fogging concentrates, warm-fogging
  • the inventive compositions are liquid formulations.
  • liquid formulations include suspension concentrations and oil dispersions.
  • inventive compositions are solid formulations.
  • solid formulations include freeze-dried powders and spray-dried powders.
  • compositions of the present invention may include formulation ingredients added to compositions of the present invention to improve recovery, efficacy, or physical properties and/or to aid in processing, packaging and administration.
  • formulation ingredients may be added individually or in combination.
  • the formulation ingredients may be added to compositions comprising cells, cell- free preparations, isolated compounds, and/or metabolites to improve efficacy, stability, and physical properties, usability and/or to facilitate processing, packaging and end-use application.
  • Such formulation ingredients may include agriculturally acceptable carriers, inerts, stabilization agents, preservatives, nutrients, or physical property modifying agents, which may be added individually or in combination.
  • the carriers may include liquid materials such as water, oil, and other organic or inorganic solvents and solid materials such as minerals, polymers, or polymer complexes derived biologically or by chemical synthesis.
  • the formulation ingredient is a binder, adjuvant, or adhesive that facilitates adherence of the composition to a plant part, such as leaves, seeds, or roots.
  • the stabilization agents may include anti-caking agents, anti-oxidation agents, anti-settling agents, antifoaming agents, desiccants, protectants or preservatives.
  • the nutrients may include carbon, nitrogen, and phosphorus sources such as sugars, polysaccharides, oil, proteins, amino acids, fatty acids and phosphates.
  • the physical property modifiers may include bulking agents, wetting agents, thickeners, pH modifiers, rheology modifiers, dispersants, adjuvants, surfactants, film-formers, hydrotropes, builders, antifreeze agents or colorants.
  • the composition comprising cells, cell-free preparation and/or metabolites produced by fermentation can be used directly with or without water as the diluent without any other formulation preparation.
  • a wetting agent, or a dispersant is added to a fermentation solid, such as a freeze-dried or spray-dried powder.
  • the formulation inerts are added after concentrating fermentation broth and/or during and/or after drying.
  • a wetting agent increases the spreading and penetrating properties, or a dispersant increases the dispersability and solubility of the active ingredient (once diluted) when it is applied to surfaces.
  • Exemplary wetting agents are known to those of skill in the art and include sulfosuccinates and derivatives, such as MULTIWETTM MO-70R (Croda Inc., Edison, NJ); siloxanes such as BREAK-THRU ® (Evonik, Germany); nonionic compounds, such as ATLOXTM 4894 (Croda Inc., Edison, NJ); alkyl
  • polyglucosides such as TERWET ® 3001 (Huntsman International LLC, The Woodlands, Texas); C12- C14 alcohol ethoxylate, such as TERGITOL ® 15-S-15 (The Dow Chemical Company, Midland, Michigan); phosphate esters, such as RHODAFAC ® BG-510 (Rhodia, Inc.); and alkyl ether carboxylates, such as EMULSOGENTM LS (Clariant Corporation, North Carolina).
  • TERWET ® 3001 Heuntsman International LLC, The Woodlands, Texas
  • C12- C14 alcohol ethoxylate such as TERGITOL ® 15-S-15 (The Dow Chemical Company, Midland, Michigan)
  • phosphate esters such as RHODAFAC ® BG-510 (Rhodia, Inc.
  • alkyl ether carboxylates such as EMULSOGENTM LS (Clariant Corporation, North Carolina).
  • compositions of the present invention for use in controlling Ganoderma spp. may be mixed with and/or used in rotation with other chemical and non-chemical additives, adjuvants and/or treatments, wherein such treatments include but are not limited to chemical and non-chemical fungicides, insecticides, miticides, nematicides, fertilizers, nutrients, minerals, auxins, growth stimulants and the like.
  • the compositions of the present invention when mixed with or used in rotation with other chemical and non-chemical additives, adjuvants and/or treatments produce a synergistic or superadditive effect in controlling Ganoderma spp.
  • the Bacillus subtilis QST713, fungicidal mutant thereof, metabolites produced therefrom, and/or cell-free extract thereof are mixed with and/or used in rotation with a fungicide.
  • preferred fungicides are selected from the group consisting of:
  • Inhibitors of the ergosterol biosynthesis for example (1.001) cyproconazole,
  • Mefentrifluconazole (1.056) 2- ⁇ [3-(2-chlorophenyl)-2-(2,4-difluorophenyl)oxiran-2-yl]methyl ⁇ -2,4- dihydro-3H-l,2,4-triazole-3-thione, (1.057) 2- ⁇ [rel(2R,3R)-3-(2-chlorophenyl)-2-(2,4- difluorophenyl)oxiran-2-yl]methyl ⁇ -2,4-dihydro-3H-l,2,4-triazole-3-thione, (1.058) 2- ⁇ [rel(2R,3S)-3- (2-chlorophenyl)-2-(2,4-difluorophenyl)oxiran-2-yl]methyl ⁇ -2,4-dihydro-3H-l,2,4-triazole-3-thione, (1.059) 5-(4-chlorobenzyl)-2-(chloromethyl)-2 -methyl- 1-(1H
  • Inhibitors of the respiratory chain at complex I or II for example (2.001) benzovindiflupyr, (2.002) bixafen, (2.003) boscalid, (2.004) carboxin, (2.005) fluopyram, (2.006) flutolanil, (2.007) fluxapyroxad, (2.008) furametpyr, (2.009) Isofetamid, (2.010) isopyrazam (anti- epimeric enantiomer 1R,4S,9S), (2.011) isopyrazam (anti-epimeric enantiomer 1S,4R,9R), (2.012) isopyrazam (anti-epimeric racemate 1RS,4SR,9SR), (2.013) isopyrazam (mixture of syn-epimeric racemate 1RS,4SR,9RS and anti-epimeric racemate 1RS,4SR,9SR), (2.014) isopyrazam (syn-epimeric enantiomer 1R,
  • Inhibitors of the respiratory chain at complex III for example (3.001) ametoctradin, (3.002) amisulbrom, (3.003) azoxystrobin, (3.004) coumethoxystrobin, (3.005) coumoxystrobin, (3.006) cyazofamid, (3.007) dimoxystrobin, (3.008) enoxastrobin, (3.009)
  • famoxadone (3.010) fenamidone, (3.011) flufenoxystrobin, (3.012) fluoxastrobin, (3.013) kresoxim- methyl, (3.014) metominostrobin, (3.015) orysastrobin, (3.016) picoxystrobin, (3.017) pyraclostrobin, (3.018) pyrametostrobin, (3.019) pyraoxystrobin, (3.020) trifloxystrobin, (3.021) (2E)-2- ⁇ 2-[( ⁇ [(lE)-l- (3- ⁇ [(E)-l-fluoro-2-phenylvinyl]oxy ⁇ phenyl)ethylidene]amino ⁇ oxy)methyl]phenyl ⁇ -2-(methoxyimino)- N-methylacetamide, (3.022) (2E,3Z)-5- ⁇ [l-(4-chlorophenyl)-lH-pyrazol-3-
  • Inhibitors of the amino acid and/or protein biosynthesis for example (7.001) cyprodinil, (7.002) kasugamycin, (7.003) kasugamycin hydrochloride hydrate, (7.004) oxytetracy cline, (7.005) pyrimethanil, (7.006) 3 -(5 -fluoro-3 ,3 ,4,4-tetramethyl-3 ,4-dihydroisoquinobn- 1 -yl)quinoline .
  • Inhibitors of the ATP production for example (8.001) silthiofam.
  • Inhibitors of the cell wall synthesis for example (9.001) benthiavalicarb, (9.002) dimethomorph, (9.003) flumorph, (9.004) iprovabcarb, (9.005) mandipropamid, (9.006) pyrimorph, (9.007) valifenalate, (9.008) (2E)-3-(4-tert-butylphenyl)-3-(2-chloropyridin-4-yl)-l-(morpholin-4- yl)prop-2-en- 1 -one, (9.009) (2Z)-3 -(4-tert-butylphenyl)-3 -(2-chloropyridin-4-yl)- 1 -(morpholin-4- yl)prop-2-en- 1 -one .
  • Inhibitors of the lipid and membrane synthesis for example (10.001) propamocarb, (10.002) propamocarb hydrochloride, (10.003) tolclofos-methyl.
  • Inhibitors of the signal transduction for example (13.001) fludioxonil, (13.002) iprodione, (13.003) procymidone, (13.004) proquinazid, (13.005) quinoxyfen, (13.006) vinclozolin.
  • the fungicide is a synthetic fungicide.
  • synthetic defines a compound that has not been obtained from a biological control agent. Especially a synthetic insecticide or fungicide is no metabolite of the biological control agents according to the present invention.
  • the Bacillus subtilis QST713 or fungicidal mutant thereof and the fungicide are applied in a synergistically effective amount.
  • the synergistically effective amount may be calculated as a synergistic weight ratio and/or range of synergistic weight ratios as described further below.
  • the biological control agent i.e., Bacillus subtilis QST713 or fungicidal mutant thereof
  • the fungicide may be used or employed in a synergistic weight ratio.
  • the skilled person understands that these ratios refer to the ratio within a combined-formulation as well as to the calculative ratio of the biological control agent described herein and the fungicide when both components are applied as mono-formulations to a plant to be treated.
  • the skilled person can calculate this ratio by simple mathematics since the volume and the amount of the biological control agent and fungicide, respectively, in a mono-formulation is known to the skilled person.
  • the ratio can be calculated based on the amount of the fungicide, at the time point of applying said component of a combination according to the invention to a plant, plant part, or locus for plant growth and the amount of a biological control agent shortly prior (e.g., 48 h, 24 h, 12 h, 6 h, 2 h, 1 h) or at the time point of applying said component of a combination according to the invention to a plant, plant part, or locus for plant growth.
  • the application of the biological control agent and the fungicide to a plant, plant part, or locus for plant growth can take place simultaneously or at different times as long as both components are present on or in the plant after the application(s).
  • the skilled person can determine the concentration of fungicide on/in a plant by chemical analysis known in the art, at the time point or shortly before the time point of applying the biological control agent.
  • the concentration of a biological control agent can be determined using tests, which are also known in the art, at the time point or shortly before the time point of applying the fungicide.
  • the synergistic weight ratio of the biological control agent/spore preparation and the fungicide lies in the range of 1 : 500 to 1000 : 1, preferably in the range of 1 : 500 to 500 : 1, more preferably in the range of 1 : 500 to 300 : 1. It has to be noted that these ratio ranges refer to the biological control agent/spores preparation (to be combined with at least one fungicide (I) or a preparation of at least one fungicide (I)) of around 10 10 cells/spores per gram preparation of said cells/spores.
  • a ratio of 100: 1 means 100 weight parts of a biological control agent/spore preparation having a cell/ spore concentration of about 10 9 or 10 10 cells/spores per gram preparation and 1 weight part of fungicide are combined (either as a solo formulation, a combined formulation or by separate applications to plants so that the combination is formed on the plant).
  • the synergistic weight ratio of the biological control agent/spore preparation to fungicide is in the range of 1 : 100 to 20.000 : 1, preferably in the range of 1 :50 to 10.000: 1 or even in the range of 1 :50 to 1000: 1.
  • the mentioned ratios ranges refer to biological control agent/spore preparations of biological control agents of around 10 9 or 10 10 cells or spores per gram preparation of said biological control agent.
  • the synergistic weight ratio of the biological control agent/spore preparation to the fungicide is in the range of 1 :0.0001 to 1 : 1, preferably in the range of 1 :0.0005 to 1 :0.1 or even in the range of 1 : 0.001 to 1:0.05.
  • the mentioned ratio ranges refer to the amount in ppm of the biological control agent/spore preparation and the fungicide, wherein the amount of the biological control agent refers to the dried content of the biological control agent/spore preparation solution.
  • the cell/spore concentration of preparations can be determined by applying methods known in the art. To compare weight ratios of the biological control agent/ spore preparation to fungicide, the skilled person can easily determine the factor between a preparation having a biological control agent/spore concentration different from 10 9 or 10 10 cells/spores per gram cell/spore preparation and a preparation having a biological control agent/ spore concentration of 10 9 or 10 10 cells/spores per gram preparation to calculate whether a ratio of a biological control agent/spore preparation to fungicide is within the scope of the above listed ratio ranges. [0083] In another embodiment, the Bacillus subtilis QST713, fungicidal mutant thereof, metabolites produced therefrom, and/or cell-free extract thereof are mixed with and/or used in rotation with an herbicide.
  • Examples of active compounds which may be mentioned as herbicides which are known from the literature and which can be combined with the Bacillus subtilis QST713, fungicidal mutant thereof, metabolites produced therefrom, and/or cell-free extract thereof are the following (compounds are either described by“common name” in accordance with the International Organization for Standardization (ISO) or by chemical name or by a customary code number), and always comprise all applicable forms such as acids, salts, ester, or modifications such as isomers, like stereoisomers and optical isomers. As an example at least one applicable form and/or modifications can be mentioned.
  • herbicides are:
  • prosulfocarb prosulfuron, pyraclonil, pyraflufen, pyraflufen- ethyl, pyrasulfotole, pyrazolynate (pyrazolate), pyrazosulfuron, pyrazosulfuron-ethyl, pyrazoxyfen, pyribambenz, pyribambenz-isopropyl, pyribambenz-propyl, pyribenzoxim, pyributicarb, pyridafol, pyridate, pyriftalid, pyriminobac, pyriminobac-methyl
  • the Bacillus subtilis QST713, fungicidal mutant thereof, metabolites produced therefrom, and/or cell-free extract thereof are mixed with and/or used in rotation with indaziflam, glyphosate, or a sulfonylurea.
  • the herbicide is glyphosate.
  • the herbicide is indaziflam.
  • the Bacillus subtilis QST713, fungicidal mutant thereof, metabolites produced therefrom, and/or cell-free extract thereof are mixed with and/or used in rotation with a fertilizer.
  • compositions and strains of the present invention onto plant propagation material, especially seeds, are known in the art, and include dressing, coating, pelleting and soaking application methods of the propagation material.
  • the application methods according to the invention part for the protection of a plant propagation material, which, in accordance with the invention, is any plant material capable of developing complete plants after planting or sowing to the site of planting or sowing, for example seedlings, rhizomes, nursery plants, cuttings or, in particular, seed (seeds), such as fruits, tubers, kernels or bulbs, against attack by pests are characterized in that, for example, suitable compositions are applied in such a manner that they are applied in close spatial proximity to, or spatially together with, planting or sowing the propagation material to the site of planting or sowing.
  • suitable compositions are applied in such a manner that they are applied in close spatial proximity to, or spatially together with, planting or sowing the propagation material to the site of planting or sowing.
  • compositions in close spatial proximity to planting or sowing the propagation material to the site of planting or sowing takes place in accordance with the invention, preferably prior to planting or sowing the propagation material, by applying the compositions by soil application directly to the site where the propagation material has been planted or sown, for example preferably prior to sowing into the seed furrow or to a closely delimited area around the site of planting or sowing the propagation material.
  • the Bacillus subtilis QST713, and/or metabolites produced therefrom are added to the seed prior to sowing in a seed-dressing apparatus and the composition is distributed uniformly over the seed, for example by stirring the contents of the seed-dressing apparatus and/or by rotating and/or shaking the entire seed dressing apparatus.
  • seed-dressing treatment comprise, for example, immersing the seed in a liquid composition, coating the seed with a solid composition (seed coating) or by achieving penetration of the active ingredient into the seed by adding the composition to the water used for pre-soaking the seed (seed soaking).
  • compositions and strains of the present invention can be applied to the seeds using conventional treating techniques and machines, such as fluidized bed techniques, the roller mill method, rotostatic seed treaters, and drum coaters. Other methods, such as spouted beds may also be usefiil.
  • the seeds may be pre-sized before coating. After coating, the seeds are typically dried and then transferred to a sizing machine for sizing. Such sizing and treating procedures are known in the art.
  • compositions and strains of the present invention can be applied or treated on to the plant propagation material by a method such that the germination is not induced; generally seed soaking induces germination because the moisture content of the resulting seed is too high.
  • suitable methods for applying (or treating) plant propagation material, such as a seed are seed dressing, seed coating or seed pelleting and the like.
  • the plant propagation material is seed.
  • the present method can be applied to a seed in any physiological state, it is preferred that the seed be in a sufficiently durable state that it incurs no damage during the treatment process.
  • the seed would be a seed that had been harvested from the field; removed from the plant; and separated from any cob, stalk, outer husk, and surrounding pulp or other non-seed plant material.
  • the seed would preferably also be biologically stable to the extent that the treatment would cause no biological damage to the seed. It is believed that the treatment can be applied to the seed at any time between harvest of the seed and sowing of the seed or during the sowing process (seed directed applications).
  • the seed may also be primed according to techniques understood by those skilled in the art either before or after the treatment.
  • Treatment could vary from a thin film (dressing) of the formulation containing the compositions and strains of the present invention on a plant propagation material, such as a seed, where the original size and/or shape are recognizable to an intermediary state (such as a coating) and then to a thicker film (such as pelleting) with many layers of different materials (such as carriers, for example, clays; different formulations, such as of other active ingredients;
  • the seed treatment occurs to an unsown seed.
  • the term “unsown seed” is meant to include seed at any period between the harvest of the seed and the sowing of the seed in the ground for the purpose of germination and growth of the plant.
  • Treatment to an unsown seed is not meant to include those practices in which the active ingredient is applied to the soil, but would include any application practice that would target the seed during the planting process.
  • treatment occurs before sowing of the seed so that the sown seed has been pre-treated with the compositions and strains of the present invention.
  • seed coating or seed pelleting are preferred in the treatment with the compositions and strains described herein.
  • the compositions and strains of the present invention are adhered on to the surface of the seed and therefore available for pest and/or disease control.
  • the treated seeds can be stored, handled, sowed and tilled in the same manner as any other active ingredient treated seed.
  • strains and compositions of the present invention are applied at a rate of about 1 x 10 2 to about 1 x 10 7 cfu/seed, depending on the size of the seed. In some embodiments, the application rate is about 1 x 10 3 to about 1 x 10 6 cfu per seed.
  • compositions of the present invention can be applied as a soil surface drench, shanked-in, injected and/or applied in-furrow or by mixture with irrigation water.
  • the rate of application for drench soil treatments which may be applied at planting, during or after seeding, or after transplanting and at any stage of plant growth, is about 4 x 10 7 to about 8 x 10 14 cfu per acre (1.62 x 10 7 to 3.24 x 10 14 cfu per hectare) or about 4 x 10 9 to about 8 x 10 13 cfu per acre (1.62 x 10 9 to 3.24 x 10 13 cfu per hectare) or about 4 x 10 11 to about 8 x 10 12 cfu per acre (1.62 x 10 11 to 3.24 x 10 12 cfu per hectare) or about 2 x 10 12 to about 6 x 10 13 cfu per acre (8.09 x 10 11 to 2.43 x
  • the rate of application is about 1 x 10 12 to about 6 x 10 12 cfu per acre (4.05 x 10 11 to 2.43 x 10 12 cfu per hectare) or about 1 x 10 13 to about 6 x 10 13 cfu per acre (4.05 x 10 12 to 2.43 xlO 13 cfu per hectare).
  • the rate of application for in-furrow treatments, applied at planting is about 2.5 x 10 10 to about 5 x 10 11 cfu per 1000 row feet (7.6 x 10 9 to 1.52 x 10 11 cfu per 100 row meter).
  • the rate of application is about 6 x 10 10 to about 3 x 10 12 cfu per 1000 row feet (1.83 x 10 10 to 9.1 x 10 11 cfu per 100 row meter) or about 6 x 10 10 to about 4 x 10 11 cfu per 1000 row feet (1.83 x 10 10 to 1.22 x 10 11 cfu per 100 row meter) or about 6 x 10 11 to about 3 x 10 12 cfu per 1000 row feet (1.83 x 10 12 to 9.1 x 10 11 cfu per 100 row meter) or about 6 x 10 11 to about 4 x 10 12 cfu per 1000 row feet (1.83 x 10 11 to 1.22 x 10 12 cfu per 100 row meter).
  • Those of skill in the art will understand how to adjust rates for broadcast treatments and other less common soil treatments.
  • compositions of the present invention can be introduced to the soil before planting or before germination of the seed.
  • the compositions of the present invention can also be introduced to the soil in contact with plant roots, to soil at the base of the plant, or to the soil around the base of the plant (e.g., within a distance of about 5 cm, about 10 cm, about 15 cm, about 20 cm, about 25 cm, about 30 cm, about 35 cm, about 40 cm, about 45 cm, about 50 cm, about 55 cm, about 60 cm, about 65 cm, about 70 cm, about 75 cm, about 80 cm, about 85 cm, about 90 cm, about 95 cm, about 100 cm, or more around or below the base of the plant).
  • the compositions may be applied by utilizing a variety of techniques including, but not limited to, drip irrigation, sprinklers, soil injection or soil drenching.
  • compositions may also be applied to soil and/or plants in plug trays or to seedlings prior to transplanting to a different plant locus.
  • the composition When applied to the soil in contact with the plant roots, to the base of the plant, or to the soil within a specific distance around the base of the plant, including as a soil drench treatment, the composition may be applied as a single application or as multiple applications.
  • compositions may be applied at the rates set forth above for drench treatments or a rate of about 1 x 10 5 to about 1 x 10 8 cfu per gram of soil, 1 x 10 5 to about 1 x 10 7 cfu per gram of soil, 1 x 10 5 to about lxlO 6 cfu per gram of soil, 7 x 10 5 to about 1 x 10 7 cfu per gram of soil, 1 x 10 6 to about 5 x 10 6 cfu per gram of soil, or 1 x 10 5 to about 3 x 10 6 cfu per gram of soil.
  • compositions of the present invention are applied as a single application at a rate of about 7 x 10 5 to about 1 x 10 7 cfu per gram of soil. In another embodiment, the compositions of the present invention are applied as a single application at a rate of about 1 x 10 6 to about 5 x 10 6 cfu per gram of soil. In other embodiments, the compositions of the present invention are applied as multiple applications at a rate of about 1 x 10 5 to about 3 x 10 6 cfu per gram of soil.
  • a composition comprising Bacillus subtilis QST713 or a fungicidal mutant thereof can be applied to any plant or any part of any plant grown in any type of media used to grow plants (e.g., soil, vermiculite, shredded cardboard, and water) or applied to plants or the parts of plants grown aerially, such as orchids or staghorn ferns.
  • the composition may, for instance, be applied by spraying, atomizing, vaporizing, scattering, dusting, watering, squirting, sprinkling, pouring or fumigating.
  • application may be carried out at any desired location where the plant of interest is positioned, such as agricultural, horticultural, forest, plantation, orchard, nursery, organically grown crops, turfgrass and urban environments.
  • the applying is preceded by identifying that the agricultural plant and/or the locus for plant growth needs treatment.
  • Identification of the presence of Ganoderma sp. in infected plants can occur various ways described in the art. Detection of Ganoderma sp. can occur with a colorimetric method, using, for example, ethylenediamine-tetraacetic acid (EDTA), a semi- selective media for Ganoderma cultures from oil palms, or Ganoderma- selective media (GSM) which can detect the pathogen from any infected tissues.
  • EDTA ethylenediamine-tetraacetic acid
  • GSM Ganoderma- selective media
  • Other non-limiting examples of methods to detect Ganoderma sp. in infected plants include advanced molecular techniques such as the use of polyclonal or monoclonal antibodies recognizing Ganoderma sp. antigens and using an enzyme-linked
  • ELISA immunosorbent assay
  • PCR polymerase chain reaction
  • device systems such as remote sense systems or e-nose systems can be used to detect the Ganoderma sp. pathogen. See Naher et al, AJCS 7(11): 1723-1727 (2013).
  • agricultural plants refers to crop plants which are employed as plants for obtaining foodstuffs, feedstuffs, fuels or for industrial purposes.
  • planting material and“plant propagule” are to be understood to denote all the generative parts of the plant such as seeds and vegetative plant material such as cuttings and tubers (e.g., potatoes), which can be used for the multiplication of the plant. This includes seeds, roots, fruits, tubers, bulbs, rhizomes, shoots, sprouts and other parts of plants. Seedlings and young plants, which are to be transplanted after germination or after emergence from soil, may also be mentioned. These young plants may also be treated totally or partially by immersion or pouring before transplantation.
  • vegetative plant material such as cuttings and tubers (e.g., potatoes), which can be used for the multiplication of the plant. This includes seeds, roots, fruits, tubers, bulbs, rhizomes, shoots, sprouts and other parts of plants. Seedlings and young plants, which are to be transplanted after germination or after emergence from soil, may also be mentioned. These young plants may also be treated totally or partially by immersion or pouring before transplantation.
  • locus is to be understood as any type of environment, soil, area or material where the plant is growing or intended to grow as well as the environmental conditions (such as temperature, water availability, radiation) that have an influence on the growth and development of the plant and/or its propagules.
  • locus is to be understood as a plant, seed, soil, area, material or environment in which a pest is growing or may grow.
  • the agricultural plants which can be treated and/or improved with the compositions and methods of the present invention include for example the following types of plants: turf, vines, cereals, for example wheat, barley, rye, oats, rice, maize and millet/sorghum; beet, for example sugar beet and fodder beet; fruits, for example pome fruit, stone fruit and soft fruit, for example apples, pears, plums, peaches, almonds, cherries and berries, for example strawberries, raspberries, blackberries; legumes, for example beans, lentils, peas and soybeans; oil crops, for example oilseed rape, mustard, poppies, olives, sunflowers, coconuts, castor oil plants, cacao and peanuts; cucurbits, for example pumpkin/squash, cucumbers and melons; fibre plants, for example cotton, flax, hemp and jute; citrus fruit, for example oranges, lemons, grapefruit and tangerines; vegetables, for example spinach, lettuce, asparagus, cabbage species,
  • Examples of agricultural plants, which are trees, that can be can be treated and/or improved with the compositions and methods of the present invention include: Abies sp., Areca sp., Elaeis sp. Eucalyptus sp., Picea sp., Pinus sp., Aesculus sp., Platanus sp., Tilia sp., Acer sp., Tsuga sp., Fraxinus sp., Sorbus sp., Betula sp., Crataegus sp., Ulmus sp., Quercus sp., Fagus sp., Salix sp., and Populus sp.
  • Preferred trees which can be treated and/or improved in accordance with the compositions and methods according to the invention are: African oil palm ⁇ Elaeis guineensis Jacq.) and American oil palm ⁇ Elaeis oleifera (Kunth) Cortes).
  • Agar plate-based assay was used to evaluate the antifungal activity of Bacillus subtilis QST713, tebuconazole, and fosetyl-aluminum against two strains of Ganoderma boninense : G. boninense strain PER71 and G. boninense strain UP.
  • Agar plates were prepared containing each antifungal agent at the following concentrations: 10 ppm, 1 ppm, 0.1 ppm, 0.01 ppm, 0.001 ppm and 0 ppm (i.e.,“Control”).
  • FIGs. 1A and IB are representative of five replicates tested for each application rate of the antifungal agents.
  • Bacillus subtilis QST713 demonstrated robust control of both G. boninense strain PER71 and G. boninense strain UP at all of the concentrations tested (see FIG. 1A). Tebuconazole controlled G. boninense strain PER71 at concentrations of 10 ppm and 1 ppm and G. boninense strain UP at a concentration of 10 ppm (see FIG. IB). The agar plates containing fosetyl -aluminum did not show any control of either strain of G. boninense (data not shown).
  • Oil palm seedlings were inoculated with Ganoderma sp. and then checked several weeks later to confirm that the Ganoderma sp. mycelia were actively infecting the developing seedlings. Twenty-one days after inoculation, each oil palm seedling was treated with SERENADE ® ASO containing Bacillus subtilis QST713 at an application rate equivalent to 2000 mL/hectare or with one of four different chemical treatments (i.e., Chemical Treatment A, Chemical Treatment B, Chemical Treatment C, and Chemical Treatment D) at application rates similar to those generally used in the field. Each chemical treatment contained a mixture of chemical fungicides. Chemical Treatment A and Chemical Treatment B included tebuconazole in the chemical mixtures.
  • Leaf color was also determined in each group of oil palm seedlings at 1, 2, 3, and 4 months after treatment using the leaf color chart (LCC) developed by the International Rice Research Institute (IRRI).
  • LCC leaf color chart
  • IRRI International Rice Research Institute
  • Oil palm seedlings treated with SERENADE ® ASO ⁇ Bacillus subtilis QST713 generally had better leaf color ratings than the non-inoculated, untreated control plants whereas the oil palm seedlings treated with chemical fungicides generally had leaf color ratings worse than those of the non-inoculated, untreated control plants.

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Abstract

La présente invention concerne une composition pour lutter contre Ganoderma spp. dans une plante agricole, la composition comprenant du Bacillus subtilis QST713 déposé sous le numéro d'accès NRRL B-21661 ou un fongicide mutant de celui-ci. La présente invention concerne également un procédé de lutte contre une infection fongique par Ganoderma spp. dans une plante agricole, le procédé consistant à appliquer à une plante agricole et/ou à un lieu pour la croissance de la plante le Bacillus subtilis QST713 déposé sous le numéro d'accès NRRL B-21661 ou un fongicide mutant de celui-ci.
PCT/EP2020/058358 2019-03-29 2020-03-25 Procédé de lutte contre la maladie ganoderma des palmiers à huile WO2020200959A1 (fr)

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WO2022128812A1 (fr) 2020-12-17 2022-06-23 Basf Se Compositions de spores, leur production et leurs utilisations
WO2023036938A1 (fr) 2021-09-10 2023-03-16 Basf Se Micro-organismes de production d'exopolysaccharides et leurs utilisations

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WO2023036938A1 (fr) 2021-09-10 2023-03-16 Basf Se Micro-organismes de production d'exopolysaccharides et leurs utilisations

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