[go: up one dir, main page]

WO2025210150A1 - Fungicidal composition - Google Patents

Fungicidal composition

Info

Publication number
WO2025210150A1
WO2025210150A1 PCT/EP2025/059132 EP2025059132W WO2025210150A1 WO 2025210150 A1 WO2025210150 A1 WO 2025210150A1 EP 2025059132 W EP2025059132 W EP 2025059132W WO 2025210150 A1 WO2025210150 A1 WO 2025210150A1
Authority
WO
WIPO (PCT)
Prior art keywords
strain
plant
streptomyces
composition
bacillus
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/EP2025/059132
Other languages
French (fr)
Inventor
Stephane BIERI
Dimitrios DRAKOPOULOS
Renu KAPIL
Santiago VALDES
George STAVRIDES
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Syngenta Crop Protection AG Switzerland
Original Assignee
Syngenta Crop Protection AG Switzerland
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Syngenta Crop Protection AG Switzerland filed Critical Syngenta Crop Protection AG Switzerland
Publication of WO2025210150A1 publication Critical patent/WO2025210150A1/en
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P3/00—Fungicides
    • 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
    • A01N63/28—Streptomyces

Definitions

  • the present invention relates to a composition
  • a composition comprising components (A) and (B) as active ingredients, wherein component (A) comprises a Streptomyces chrestomyceticus and component (B) comprises a compound selected from the group consisting of sulphur, phosphonate, copper hydroxide, copper oxychloride, bicarbonate, Bacillus sp, Trichoderma sp, aureobasidin A, jawsamycin, kasugamycin, natamycin, a polysaccharide elicitor comprising laminarin, a composition comprising chitooligosaccharides and oligogalactunorides (COS-OGA), and/or yeast extract, a plant biostimulant (PBS) comprising an algal extract, a plant extract and/or vinasse, orange oil, and / or tea tree oil.
  • component (A) comprises a Streptomyces chrestomyceticus
  • component (B) comprises
  • the present invention relates to a method of controlling or preventing phytopathogenic diseases, preferably phytopathogenic fungi, on a plant or on plant propagation material and / or on harvested food crops, which comprises applying to the plant, on plant propagation material, the locus thereof, and / or on harvested food crops the composition according to the present invention.
  • phytopathogenic diseases preferably phytopathogenic fungi
  • compositions according to the present invention provided effective control against phytopathogenic microorganisms, preferably against phytopathogenic fungi.
  • the benefits provided by certain compositions according to the invention may also include, inter alia, advantageous levels of biological activity for protecting plants against diseases that are caused by pests, for instance fungi, or superior properties for use as agricultural active ingredients (for example, greater biological activity, an advantageous spectrum of activity, an increased safety profile, improved physico-chemical properties, or increased biodegradability).
  • a composition according to the present invention typically is a pesticidal composition.
  • a composition, microorganism, metabolite or compound having “pesticidal activity” or “pesticide” as used herein means a composition, microorganism, metabolite or compound that controls, modifies, or prevents the growth of pests.
  • the term “pesticidally effective amount” or “effective amount of pesticide” where used means the quantity of such a composition, microbial strain, compound, or metabolite that is capable of producing an effect on the growth of pests. Controlling or modifying effects include all deviation from natural development, such as killing, retardation and the like, and prevention includes barrier or other defensive formation in or on a plant to prevent pest infection.
  • the term “pesticidal activity” or “pesticides” includes insecticidal or fungicidal activity or “insecticides” or “fungicides”.
  • the composition according to the present invention may also have activity against oomycetes.
  • a composition according to the present invention typically is a fungicidal composition.
  • fungicide or “fungicidal” as used herein means a composition, metabolite or microorganism or compound that controls, modifies, or prevents the growth of fungi.
  • fungicidally effective amount or “effective amount of fungicide” means the quantity of such a compound, metabolite, microorganism or composition that is capable of producing an effect on the growth of fungi. Controlling or modifying effects include all deviation from natural development, such as killing, retardation and the like, and prevention includes barrier or other defensive formation in or on a plant to prevent fungal infection, eg. the composition according to the present invention may act as a plant growth regulator.
  • the composition according to the present invention comprises a fungicidally effective amount of the component (A) and/or the component (B).
  • composition according to the present invention typically is an agricultural-acceptable composition.
  • composition stands for the various mixtures or combinations of components (A) and (B) (including the above-defined embodiments), for example in a single “ready-mix” form, in a combined spray mixture, or combined slurry mixture, composed from separate formulations of the single active ingredient components, such as a “tank-mix”, and in a combined use of the single active ingredients when applied in a sequential manner, i.e. one after the other with a reasonably short period, such as a few hours or days.
  • the order of applying the components (A) and (B) is not essential for working the present invention.
  • component (A) comprises a Streptomyces chrestomyceticus.
  • the Streptomyces chrestomyceticus preferably comprises a nucleotide sequence which has at least 99.8 %, preferably at least 99.9% consult preferably at least 99.91%, 99.92%, 99.93%, 99.94%, 99.95%, 99.96%, 99.97%, 99.98%, or 99.99% identity to SEQ ID NO: 1.
  • the Streptomyces chrestomyceticus comprises a nucleotide sequence according to SEQ ID NO:1.
  • SEQ ID NO: 1 comprises the 16S RNA gene of Streptomyces sp. Saigon413 deposited with the Westerdijk Institute under accession number CBS149411
  • the component (A) comprises a Streptomyces chrestomyceticus, which comprises a genome sequence which has at least 95% identity, preferably at least 96%, 97%, 98%, 99% identity to the whole genome of Streptomyces sp. Saigon413 deposited with the Westerdijk Institute under accession number CBS149411 .
  • the Streptomyces may comprise a genome sequence which has at least 99.1 %, 99.2%, 99.3%. 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% identity to the whole genome of Streptomyces sp. Saigon413 deposited with the Westerdijk Institute under accession number CBS149411 .
  • component (A) comprises a Streptomyces chrestomyceticus, which is Streptomyces sp. Saigon413 deposited with the Westerdijk Institute under accession number CBS149411.
  • percent identity refers to the relatedness of two or more nucleotide or amino acid sequences, which may be calculated by (i) comparing two optimally aligned sequences over a window of comparison, (ii) determining the number of positions at which the identical nucleic acid base (for nucleotide sequences) or amino acid residue (for proteins) occurs in both sequences to yield the number of matched positions, (iii) dividing the number of matched positions by the total number of positions in the window of comparison, and then (iv) multiplying this quotient by 100 percent to yield the percent identity.
  • the percent identity is being calculated in relation to a reference sequence without a particular comparison window being specified, then the percent identity is determined by dividing the number of matched positions over the region of alignment by the total length of the reference sequence. Accordingly, for purposes of the present invention, when two sequences (query and subject) are optimally aligned (with allowance for gaps in their alignment), the "percent identity" for the query sequence is equal to the number of identical positions between the two sequences divided by the total number of positions in the query sequence over its length (or a comparison window), which is then multiplied by 100 percent.
  • the component (A) comprising Streptomyces as disclosed herein comprises a cell count of the Streptomyces from 1*10° to 1*10 14 cfu / g dry weight, for instance from 1*10 1 to 1*10 13 cfu / g dry weight, from 1*10 2 to 1*10 13 cfu I g dry weight, for instance from 1*10 3 to 1*10 12 cfu / g dry weight, from 2*10 3 to 2*10 11 cfu / g dry weight, from 5*10 3 to 5*10 11 cfu / g dry weight, for instance from 1*10 4 to 1*10 1 ° cfu / g dry weight, from 2*10 4 to 2*10 1 ° cfu / g dry weight, such as from 1 *10 5 to 1 *10 9 cfu / g dry weight, from 2*10 5 to 2*10 9 cfu I g dry weight, from 5*10 5 to
  • the component (A) comprising a Streptomyces chrestomyceticus as disclosed herein comprises a fermentation broth comprising the Streptomyces chrestomyceticus, preferably a spray-dried fermentation broth or a freeze-dried fermentation broth. Spray-drying or freeze-drying of a fermentation broth is known in the art.
  • Component (A) comprising a Streptomyces as disclosed herein also includes a formulation comprising the Streptomyces.
  • a Streptomyces such as Streptomyces chrestomyceticus
  • a fermentation broth is produced during cultivation of or when cultivating a strain of Streptomyces chrestomyceticus.
  • Suitable fermentation conditions for cultivating Streptomyces sp. such as S. chrestomyceticus are known to a person skilled in art.
  • Cultivating a strain of Streptomyces for instance Streptomyces chrestomyceticus strain, as disclosed herein, comprises cultivating the microbial strain under aerobic conditions at a temperature of from 15 degrees Celsius to 45 degrees Celsius, preferably a temperature of from 20 to 35 degree Celsius, preferably a temperature of between 25 to 32 degrees Celsius, in the presence of a carbon source and a nitrogen source.
  • a suitable carbon source may be molasses, such as beet or cane molasses, polysaccharides, flour, starch, sugar, or glucose.
  • a suitable nitrogen source may be casein hydrolysate, tryptone, ammonium sulphate, ammonia, yeast extract, peptone or urea.
  • the process for cultivating a Streptomyces as disclosed herein may be performed in a batch, fed-batch or continuous culture.
  • Component (A) as disclosed herein also includes a formulation comprising the Streptomyces chrestomyceticus.
  • Formulations of microbial strains are known in the art for instance as disclosed in Croda Crop Care, the Nouryon formulator toolbox and in: Formulation of Microbial Biopesticides: Beneficial microorganisms, nematodes and seed treatments (412 p., 6 December 2012) eds. Burges H.D., Springer, ISBN 978-94-011-4926-6.
  • Component (A) comprising a Streptomyces chrestomyceticus as disclosed herein comprises a formulation comprising the Streptomyces chrestomyceticus, wherein the formulation is an oil dispersion (OD), a non-aqueous dispersion (NAD) or a flowable formulation.
  • OD oil dispersion
  • NAD non-aqueous dispersion
  • Component (A) may be a component (A) comprising composition comprising a Streptomyces chrestomyceticus as disclosed herein.
  • component (A) further comprises a metabolite.
  • a metabolite is preferably produced by the Streptomyces chrestomyceticus disclosed herein.
  • a metabolite may be a supernatant comprising a metabolite as disclosed herein.
  • a component (A) comprises a metabolite, preferably wherein the metabolite is malonomicin.
  • Malonomicin (sometimes spelt ‘malonomycin’) is ⁇ [(2S)-2-amino-3-hydroxypropanoyl]amino ⁇ ⁇ 2-[(5S)-5-(aminomethyl)-4-hydroxy-2-oxo-2,5-dihydro-1 H-pyrrol-3-yl]-2-oxoethyl ⁇ malonic acid can be produced as disclosed in Example I of W02006/078939.
  • Malonomicin may also be prepared according to the method disclosed in Example I A and B in EP 1860939, or according to Law et al, 2018 (Nature Catalys is
  • the polyene is further characterized by the spectrum of light absorption as shown in Figure 10.
  • Cyclothiazomycin C is a known compound. The structure of cyclothazomycin C is disclosed on p.
  • Streptimidone is a known compound of Formula III
  • Streptimidone can be synthesised following the method disclosed in Kondo, H., Oritani, T., and Kiyota, H. Synthesis and antifungal activity of the four stereoisomers of streptimidone, a glutarimide antibiotic from Streptomyces rimosus forma paromomycinus. Eur. J. Org. Chem. (20), 3459-3462 (2000).
  • Suitable agricultural adjuvants and/or carriers can be solid or liquid and are substances useful in formulation technology, e.g. natural or regenerated mineral substances, solvents, dispersants, wetting agents, tackifiers, thickeners, binders or fertilizers.
  • a broad range of surface-active agents are advantageously employed in both said liquid and solid compositions, especially those designed to be diluted with carrier before application.
  • These agents when used, normally comprise from 0.1% to 15% by weight of the formulation. They can be anionic, cationic, non-ionic or polymeric in character and can be employed as emulsifying agents, wetting agents, suspending agents or for other purposes.
  • adjuvants commonly utilized in agricultural compositions include crystallisation inhibitors, viscosity modifiers, suspending agents, spray droplet modifiers, pigments, antioxidants, foaming agents, anti-foaming agents, light-blocking agents, compatibilizing agents, sequestering agents, neutralising agents and buffers, corrosion inhibitors, dyes, odorants, spreading agents, penetration aids, micronutrients, emollients, lubricants and sticking agents.
  • composition according to the present invention may be used in unmodified form or, preferably, together with the adjuvants conventionally employed in the art of formulation. To this end they may be conveniently formulated in known manner to emulsifiable concentrates, coatable pastes, directly sprayable or dilutable solutions or suspensions, dilute emulsions, wettable powders, soluble powders, dusts, granulates, and also encapsulations e.g. in polymeric substances. As with the type of the compositions, the methods of application, such as spraying, atomising, dusting, scattering, coating or pouring, are chosen in accordance with the intended objectives and the prevailing circumstances.
  • Suspension concentrates are aqueous formulations in which finely divided solid particles of the active ingredient are suspended. Such formulations include anti-settling agents and dispersing agents and may further include a wetting agent to enhance pesticidal activity as well an anti-foam and a crystal growth inhibitor. In use, these concentrates are diluted in water and normally applied as a spray to the area to be treated. The amount of active ingredient may range from 0.5% to 95% of the concentrate.
  • Wettable powders are in the form of finely divided particles which disperse readily in water or other liquid carriers.
  • the particles contain an active ingredient retained in a solid matrix.
  • Typical solid matrices include fuller’s earth, kaolin clays, silicas and other readily wet organic or inorganic solids. Wettable powders normally contain from 5% to 95% of an active ingredient plus a small amount of wetting, dispersing or emulsifying agent.
  • Emulsifiable concentrates are homogeneous liquid compositions dispersible in water or other liquid and may consist entirely of an active ingredient mixed with a liquid or solid emulsifying agent, or may also contain a liquid carrier, such as xylene, heavy aromatic naphthas, isophorone and other non-volatile organic solvents. In use, these concentrates are dispersed in water or other liquid and normally applied as a spray to the area to be treated. The amount of active ingredient may range from 0.5% to 95% of the concentrate.
  • Granular formulations include both extrudates and relatively coarse particles and are usually applied without dilution to the area in which treatment is required.
  • Typical carriers for granular formulations include sand, fuller’s earth, attapulgite clay, bentonite clays, montmorillonite clay, vermiculite, perlite, calcium carbonate, brick, pumice, pyrophyllite, kaolin, dolomite, plaster, wood flour, ground corn cobs, ground peanut hulls, sugars, sodium chloride, sodium sulphate, sodium silicate, sodium borate, magnesia, mica, iron oxide, zinc oxide, titanium oxide, antimony oxide, cryolite, gypsum, diatomaceous earth, calcium sulphate and other organic or inorganic materials which absorb or which can be coated with the active compound.
  • compositions of the invention may be employed in any conventional form, for example in the form of a twin pack, a powder for dry seed treatment (DS), an emulsion for seed treatment (ES), a flowable concentrate for seed treatment (FS), a solution for seed treatment (LS), a water dispersible powder for seed treatment (WS), a capsule suspension for seed treatment (CF), a gel for seed treatment (GF), an emulsion concentrate (EC), a suspension concentrate (SC), a suspo-emulsion (SE), a capsule suspension (CS), a water dispersible granule (WG), an emulsifiable granule (EG), an emulsion, water in oil (EG), an emulsion, oil in water (EW), a micro-emulsion (ME), an oil dispersion (OD), an oil miscible flowable (OF), an oil miscible liquid (OL), a soluble concentrate (SL), an ultra-low volume suspension (SU), an ultra-low volume liquid (UL), a technical concentrate (TK
  • compositions may be produced in conventional manner, e.g. by mixing the active ingredients with appropriate formulation inerts (diluents, solvents, fillers and optionally other formulating ingredients such as surfactants, biocides, anti-freeze, stickers, thickeners and compounds that provide adjuvancy effects).
  • appropriate formulation inerts diiluents, solvents, fillers and optionally other formulating ingredients such as surfactants, biocides, anti-freeze, stickers, thickeners and compounds that provide adjuvancy effects.
  • conventional slow release formulations may be employed where long lasting efficacy is intended.
  • Particularly formulations to be applied in spraying forms such as water dispersible concentrates (e.g. EC, SC, DC, OD, SE, EW, EO and the like), wettable powders and granules, may contain surfactants such as wetting and dispersing agents and other compounds that provide adjuvancy effects, e.g.
  • a seed dressing formulation is applied in a manner known per se to the seeds employing the combination of the invention and a diluent in suitable seed dressing formulation form, e.g. as an aqueous suspension or in a dry powder form having good adherence to the seeds.
  • suitable seed dressing formulation form e.g. as an aqueous suspension or in a dry powder form having good adherence to the seeds.
  • seed dressing formulations are known in the art.
  • Seed dressing formulations may contain the single active ingredients or the combination of active ingredients in encapsulated form, e.g. as slow release capsules or microcapsules.
  • the present invention relates to a method of controlling or preventing phytopathogenic diseases, preferably phytopathogenic fungi, on a plant or on plant propagation material and / or on harvested food crops, which comprises applying to the plant, on plant propagation material, the locus thereof, and / or on harvested food crops the composition according to the present invention.
  • phytopathogenic diseases preferably phytopathogenic fungi
  • a preferred method of controlling or preventing an infestation of crop plants by phytopathogenic microorganisms, especially fungal organisms comprises the application of a composition according to the present invention is foliar application.
  • the frequency of application and the rate of application will depend on the risk of infestation by the corresponding pathogen.
  • the compounds A and/or B of the composition according to the present invention can also penetrate the plant through the roots via the soil (systemic action) by drenching the locus of the plant with a liquid formulation, or by applying said compounds in solid form to the soil, e.g. in granular form (soil application). In crops of water rice such granulates can be applied to the flooded rice field.
  • Components (A) and (B) of the composition according to the present invention may be applied simultaneously, such as in a ready to mix formulation, or in a sequential manner.
  • the method according to the present invention comprises applying the composition according to the present invention, wherein the components (A) and (B) are applied in a simultaneous or a sequential manner on the plant, the plant propagation material or the locus thereof.
  • composition of the invention may be used to control plant diseases caused by a broad spectrum of plant pathogens in the Basidiomycete, Ascomycete, Oomycete and/or Deuteromycete, Blasocladiomycete, Chrytidiomycete, Glomeromycete, Mucoromycete classes, and/or Oomycete classes.
  • pathogens may include:
  • Oomycetes including Phytophthora diseases such as those caused by Phytophthora capsici, Phytophthora infestans, Phytophthora sojae, Phytophthora fragariae, Phytophthora nicotianae, Phytophthora cinnamomi, Phytophthora citricola, Phytophthora citrophthora and Phytophthora erythroseptica; Pythium diseases such as those caused by Pythium aphanidermatum, Pythium arrhenomanes, Pythium graminicola, Pythium irregulare, Pythium sylvaticum and Pythium ultimum; diseases caused by Peronosporales such as Peronospora destructor, Peronospora parasitica, Plasmopara viticola, Plasmopara halstedii, Pseudoperonospora cubensis, Albugo Candida,
  • Ascomycetes including blotch, spot, blast or blight diseases and/or rots for example those caused by Pleosporales such as Stemphylium solani, Stagonospora tainanensis, Spilocaea oleaginea, Setosphaeria turcica, Pyrenochaeta lycoperisici, Pleospora herbarum, Phoma destructiva, Phaeosphaeria herpotrichoides, Phaeocryptocus gaeumannii, Ophiosphaerella graminicola, Ophiobolus graminis, Leptosphaeria maculans, Hendersonia creberrima, Helminthosporium triticirepentis, Setosphaeria turcica, Drechslera glycines, Didymella bryoniae, Cycloconium oleagineum, Corynespora cassiicola, Cochliobolus sativus, Bi
  • Valsa ceratosperma Valsa ceratosperma, and others such as Actinothyrium graminis, Ascochyta pisi, Aspergillus flavus, Aspergillus fumigatus, Aspergillus nidulans, Asperisporium caricae, Blumeriella jaapii, Candida spp., Capnodium ramosum, Cephaloascus spp., Cephalosporium gramineum, Ceratocystis paradoxa, Chaetomium spp., Hymenoscyphus pseudoalbidus, Coccidioides spp., Cylindrosporium padi, Diplocarpon malae, Drepanopeziza campestris, Elsinoe ampelina, Epicoccum nigrum, Epidermophyton spp., Eutypa lata, Geotrichum candidum, Gibellina cerealis, Gloeocercospora sorghi,
  • Gerlachia nivale Gibberella fujikuroi
  • Gibberella zeae Gibberella zeae
  • Gliocladium spp. Myrothecium verrucaria, Nectria ramulariae, Trichoderma viride, Trichothecium roseum, and Verticillium theobromae;
  • Basidiomycetes including smuts for example those caused by Ustilaginales such as Ustilaginoidea virens, Ustilago nuda, Ustilago tritici, Ustilago zeae, rusts for example those caused by Pucciniales such as Cerotelium fici, Chrysomyxa arctostaphyli, Coleosporium ipomoeae, Hemileia vastatrix, Puccinia arachidis, Puccinia cacabata, Puccinia gram inis, Puccinia recondita, Puccinia sorghi, Puccinia hordei, Puccinia striiformis f.sp.
  • Ustilaginales such as Ustilaginoidea virens, Ustilago nuda, Ustilago tritici, Ustilago zeae
  • rusts for example those caused by Pucciniales such as Cerotelium fici, Chr
  • Puccinia striiformis f.sp. Secalis Pucciniastrum coryli, or Uredinales such as Cronartium ribicola, Gymnosporangium juniperi-viginianae, Melampsora medusae, Phakopsora pachyrhizi, Phragmidium mucronatum, Physopella ampelosidis, Tranzschelia discolor and Uromyces viciae-fabae; and other rots and diseases such as those caused by Cryptococcus spp., Exobasidium vexans, Marasmiellus inoderma, Mycena spp., Sphacelotheca reiliana, Typhula ishikariensis, Urocystis agropyri, Itersonilia perplexans, Corticium invisum, Laetisaria fuciformis, Waitea circinata, Rhizoctonia solani, Tha
  • Blastocladiomycetes such as Physoderma maydis
  • Mucoromycetes such as Choanephora cucurbitarum.; Mucor spp.; Rhizopus arrhizus; as well as diseases caused by other species and genera closely related to those listed above.
  • Colletotrichum lagenarium Colletotrichum orbiculare
  • Botrytis cinerea preferably Zymoseptoria tritici, Puccinia recondita f. sp. tritici (Puccinia triticina), and / or Pyricularia oryzae,
  • composition according to the present invention can be used in the agricultural sector and related fields of use for controlling plant pests or on non-living materials for control of spoilage microorganisms or organisms potentially harmful to man.
  • the composition can be used to inhibit or destroy the pests that occur on plants or parts of plants (fruit, blossoms, leaves, stems, tubers, roots) of different crops of useful plants, while at the same time protecting also those parts of the plants that grow later e.g. from phytopathogenic microorganisms.
  • Useful plants and / or target crops in accordance with the invention include conventional as well as genetically enhanced or engineered varieties such as, for example, insect resistant (e.g. Bt. and VIP varieties) as well as disease resistant, herbicide tolerant (e.g. glyphosate- and glufosinate-resistant maize varieties) and nematode tolerant varieties.
  • Useful plants include plants transformed by the use of recombinant DNA techniques, for example, to be capable of synthesising one or more selectively acting toxins, such as are known, for example, from toxin-producing bacteria.
  • toxins which can be expressed include 8-endotoxins, vegetative insecticidal proteins (Vip), insecticidal proteins of bacteria colonising nematodes, and toxins produced by scorpions, arachnids, wasps and fungi.
  • Vip vegetative insecticidal proteins
  • insecticidal proteins of bacteria colonising nematodes include toxins produced by scorpions, arachnids, wasps and fungi.
  • Plants, including target crops and/or useful plants to be protected in a method of the invention typically comprise perennial and annual crops, such as berry plants for example blackberries, blueberries, cranberries, raspberries and strawberries; cereals for example barley, maize (corn), millet, oats, rice, rye, sorghum triticale and wheat; fibre plants for example cotton, flax, hemp, jute and sisal; field crops for example sugar and fodder beet, coffee, hops, mustard, oilseed rape (canola), poppy, sugar cane, sunflower, tea and tobacco; fruit trees for example apple, apricot, avocado, banana, cherry, citrus, nectarine, peach, pear and plum; grasses for example Bermuda grass, bluegrass, bentgrass, centipede grass, fescue, ryegrass, St.
  • perennial and annual crops such as berry plants for example blackberries, blueberries, cranberries, raspberries and strawberries
  • cereals for example barley, maize
  • Augustine grass and Zoysia grass herbs such as basil, borage, chives, coriander, lavender, lovage, mint, oregano, parsley, rosemary, sage and thyme; legumes for example beans, lentils, peas and soya beans; nuts for example almond, cashew, ground nut, hazelnut, peanut, pecan, pistachio and walnut; palms for example oil palm; ornamentals for example flowers, shrubs and trees; other trees, for example cacao, coconut, olive and rubber; vegetables for example asparagus, aubergine, broccoli, cabbage, carrot, cucumber, garlic, lettuce, marrow, melon, okra, onion, pepper, potato, pumpkin, rhubarb, spinach and tomato; and vines for example grapes.
  • herbs such as basil, borage, chives, coriander, lavender, lovage, mint, oregano, parsley, rosemary, sage and thyme
  • legumes for example beans, lentils, peas and soya beans
  • a plant in a method according to the present invention preferably comprises wheat, barley, rice, corn, soya, sugar beet, banana, tomato, cucumber, and / or groundnut.
  • the plant comprises wheat and / or rice.
  • the composition comprising components (A) and (B) as defined herein may also be applied to seeds (coating) by impregnating the seeds or tubers either with a liquid formulation of the composition of the invention or coating them with a solid formulation.
  • a method of controlling or preventing a phytopathogenic disease includes spraying, atomizing, dusting, brushing on, dressing, scattering or pouring - which are to be selected to suit the intended aims of the prevailing circumstances.
  • Typical rates of concentration are between 0.1 and 1000 ppm, preferably between 0.1 and 500 ppm, of active ingredient.
  • the rate of application per hectare is preferably 1 g to 2000 g of active ingredient per hectare, more preferably 10 to 1000 g/ha, most preferably 10 to 600 g/ha.
  • seed drenching agent convenient dosages are from 10mg to 1g of active substance per kg of seeds.
  • rates of 0.001 to 50 g of the composition per kg of seed preferably from 0.01 to 10g per kg of seed are generally sufficient.
  • the method comprises applying an effective amount of component A comprising Streptomyces as disclosed herein above, wherein the effective amount comprises from 2*10 2 to 5*10 17 , from 3*10 2 to 5*10 16 , from 5*10 2 to 5*10 15 , from 2*10 2 to 5*10 14 , from 2*10 2 to 5*10 13 , preferably from 5*10 2 to 5*10 12 , from 1*10 3 to 5*10 11 , from 5*10 3 to 1*10 11 , from 1*10 4 to 5*10 10 , from 5*10 4 to 1*10 10 , from 1*10 5 to 5*10 9 , from 5*10 5 to 1*10 9 , from 1*10 6 to 5*10 8 , from 5*10 6 to 1*10 8 colony forming unit (cfu) of the Streptomyces such as Streptomyces chrestomyceticus, per hectare.
  • the effective amount comprises from 2*10 2 to 5*10 17 , from 3
  • NCAIM (P) B001389) (WO 2013/034938) from Certis USA LLC, a subsidiary of Mitsui & Co.; Bacillus subtilis CX-9060 from Certis USA LLC, a subsidiary of Mitsui & Co.; Paenibacillus polymyxa, in particular strain AC-1 (e.g. TOPSEED® from Green Biotech Company Ltd.); Pseudomonas proradix (e.g. PRORADIX® from Sourcon Padena); Pantoea agglomerans, in particular strain E325 (Accession No. NRRL B-21856) (available as BLOOMTIME BIOLOGICALTM FD BIOPESTICIDE from Northwest Agri Products); and
  • Bacillus subtilis in particular strain QST713/AQ713 (available as SERENADE OPTI or SERENADE ASO from Bayer CropScience LP, US, having NRRL Accession No. B21661 and described in U.S. Patent No. 6,060,051); Bacillus pumilus, in particular strain QST2808 (available as SONATA® from Bayer CropScience LP, US, having Accession No. NRRL B-30087 and described in U.S. Patent No.
  • bacteria for example Bacillus subtilis, in particular strain QST713/AQ713 (available as SERENADE OPTI or SERENADE ASO from Bayer CropScience LP, US, having NRRL Accession No. B21661 and described in U.S. Patent No. 6,060,051)
  • Bacillus pumilus in particular strain QST2808 (available as SONATA® from Bayer CropScience LP, US, having Accession No. NRRL B-30087 and described in U.S. Patent No.
  • Bacillus subtilis strain MBI 600 (available as SUBTILEX from BASF SE), having Accession Number NRRL B-50595, U.S. Patent No. 5,061 ,495; Bacillus subtilis strain GB03 (available as Kodiak® from Bayer AG, DE); Bacillus subtilis var. amyloliquefaciens strain FZB24 having Accession No. DSM 10271 (available from Novozymes as TAEGRO® or TAEGRO® ECO (EPA Registration No. 70127-5)); Bacillus mycoides, isolate J , having Accession No.
  • Trichoderma harzianum strain T-22 e.g. Trianum-P from Andermatt Biocontrol or Koppert
  • strain Cepa SimbT5 from Simbiose Agro
  • Gliocladium roseum also known as Clonostachys rosea f rosea
  • strain 321 U from Adjuvants Plus
  • strain ACM941 as disclosed in Xue (Efficacy of Clonostachys rosea strain ACM941 and fungicide seed treatments for controlling the root tot complex of field pea, Can Jour Plant Sci 83(3): 519-524), or strain IK726 (Jensen DF, et al.
  • Trichoderma atroviride T34 Biocontrol by Biocontrol Technologies S.L., ES) or strain ICC 012 from Isagro; Trichoderma atroviride, strain CNCM 1-1237 (e.g. Esquive® WP from Agrauxine, FR); Trichoderma atroviride, strain no. V08/002387; Trichoderma atroviride, strain NMI no. V08/002388; Trichoderma atroviride, strain NMI no. V08/002389; Trichoderma atroviride, strain NMI no. V08/002390; Trichoderma atroviride, strain LC52 (e.g.
  • Trichoderma atroviride strain ATCC 20476 (IMI 206040); Trichoderma atroviride, strain T11 (IMI352941Z CECT20498); Trichoderma harmatum; Trichoderma harzianum; Trichoderma harzianum rifai T39 (e.g. Trichodex® from Makhteshim, US); Trichoderma asperellum, in particular, strain kd (e.g. T-Gro from Andermatt Biocontrol); Trichoderma harzianum, strain ITEM 908 (e.g. Trianum-P from Koppert); Trichoderma harzianum, strain TH35 (e.g.
  • Trichoderma virens also known as Gliocladium virens
  • strain GL-21 e.g. SoilGard by Certis, US
  • Trichoderma viride e.g. Trianum-P by Koppert
  • Ampelomyces quisqualis in particular strain AQ 10 (e.g.
  • Aureobasidium pullulans in particular blastospores of strain DSM14940; Aureobasidium pullulans, in particular blastospores of strain DSM 14941 ; Aureobasidium pullulans, in particular mixtures of blastospores of strains DSM14940 and DSM 14941 (e.g. Botector® by bio-ferm, CH); Cladosporium cladosporioides, strain H39, having Accession No. CBS122244, US 2010/0291039 (by Stichting Divennewenne Onderzoek); Gliocladium catenulatum (Synonym: Clonostachys rosea f.
  • Trichoderma atroviride strain SKT- 1 (FERM P-16510), JP Patent Publication (Kokai) 11-253151 A; Trichoderma atroviride, strain SKT-2 (FERM P-16511), JP Patent Publication (Kokai) 11-253151 A; Trichoderma atroviride, strain SKT-3 (FERM P-17021), JP Patent Publication (Kokai) 11-253151 A; Trichoderma gamsii (formerly T. viride), strain ICC080 (IMI CC 392151 CABI, e.g. BioDerma by AGROBIOSOL DE MEXICO, S.A.
  • IMI CC 392151 CABI e.g. BioDerma by AGROBIOSOL DE MEXICO, S.A.
  • Trichoderma harzianum strain DB 103 (available as T-GRO® 7456 by Dagutat Biolab); Trichoderma polysporum, strain IMI 206039 (e.g. Binab TF WP by BINAB Bio-Innovation AB, Sweden); Trichoderma stromaticum, having Accession No. Ts3550 (e.g. Tricovab by CEPLAC, Brazil); Ulocladium oudemansii strain U3, having Accession No. NM 99/06216 (e.g., BOTRY-ZEN® by Botry-Zen Ltd, New Zealand and BOTRYSTOP® from BioWorks, Inc.); Verticillium albo-atrum (formerly V.
  • strain WCS850 having Accession No. WCS850, deposited at the Central Bureau for Fungi Cultures (e.g., DUTCH TRIG® by Tree Care Innovations); Verticillium chlamydosporium; mixtures of Trichoderma asperellum strain ICC 012 (also known as Trichoderma harzianum ICC012), having Accession No. CABI CC IMI 392716 and Trichoderma gamsii (formerly T. viride) strain ICC 080, having Accession No. IMI 392151 (e.g., BIO-TAMTM from Isagro USA, Inc. and BIODERMA® by Agrobiosol de Mexico, S.A.
  • IMI 392151 e.g., BIO-TAMTM from Isagro USA, Inc. and BIODERMA® by Agrobiosol de Mexico, S.A.
  • Trichoderma asperelloides JM41 R (Accession No. NRRL B-50759) (TRICHO PLUS® from BASF SE); Aspergillus flavus strain NRRL 21882 (products known as AFLA-GUARD® from Syngenta/ChemChina); Chaetomium cupreum (Accession No. CABI 353812) (e.g. BIOKUPRUMTM by AgriLife); Saccharomyces cerevisiae, in particular strain LASO2 (from Agro-Levures et Derives), strain LAS117 cell walls (CEREVISANE® from Lesaffre; ROMEO® from BASF SE), strains CNCM No. 1-3936, CNCM No. 1-3937, CNCM No.
  • Trichoderma virens strain G-41 formerly known as Gliocladium virens (Accession No. ATCC 20906) (e.g., ROOTSHIELD® PLUS WP and TURFSHIELD® PLUS WP from BioWorks, US); Trichoderma hamatum, having Accession No. ATCC 28012; Ampelomyces quisqualis strain AQ10, having Accession No.
  • CNCM 1-807 e.g., AQ 10® by IntrachemBio Italia); Phlebiopsis gigantea strain VRA 1992 (ROTSTOP® C from Danstar Ferment); Penicillium steckii (DSM 27859; WO 2015/067800) from BASF SE; Chaetomium globosum (available as RIVADIOM® by Rivale); Cryptococcus flavescens, strain 3C (NRRL Y-50378); (B2.2.99) Dactylaria Candida Dilophosphora alopecuri (available as TWIST FUNGUS®); Fusarium oxysporum, strain Fo47 (available as FUSACLEAN® by Natural Plant Protection); Pseudozyma flocculosa, strain PF-A22 UL (available as SPORODEX® L by Plant Products Co., CA); (2.2.103) Trichoderma gamsii (formerly T.
  • strain ICC 080 IMI CC 392151 CABI
  • BIODERMA® AGROBIOSOL DE MEXICO, S.A. DE C.V.
  • Trichoderma fertile e.g. product TrichoPlus from BASF
  • Muscodor roseus in particular strain A3-5 (Accession No. NRRL 30548); Simplicillium lanosoniveum;
  • bacteria selected from the group consisting of Bacillus pumilus, in particular strain QST2808 (having Accession No. NRRL No. B-30087); Bacillus subtilis, in particular strain QST713/AQ713 (having NRRL Accession No. B-21661 and described in U.S. Patent No. 6,060,051 ; available as SERENADE® OPTI or SERENADE® ASO from Bayer CropScience LP, US); Bacillus subtilis, in particular strain AQ30002 (having Accession Nos. NRRL B-50421 and described in U.S. Patent Application No.
  • Bacillus amyloliquefaciens SB3281 ATCC # PTA-7542; WO 2017/205258
  • Bacillus amyloliquefaciens TJ1000 available as QUIKROOTS® from Novozymes
  • Bacillus firmus in particular strain CNMC 1-1582 (e.g. VOTIVO®from BASF SE)
  • Bacillus pumilus in particular strain GB34 (e.g. YIELD SHIELD® from Bayer Crop Science, DE); Bacillus amyloliquefaciens, in particular strain IN937a; Bacillus amyloliquefaciens, in particular strain FZB42 (e.g.
  • Trianum-P from Andermatt Biocontrol or Koppert Myrothecium verrucaria strain AARC-0255 (e.g. DiTeraTM from Valent Biosciences); Penicillium bilaii strain ATCC ATCC20851 ; Pythium oligandrum strain M1 (ATCC 38472; e.g. Polyversum from Bioprepraty, CZ); Trichoderma virens strain GL-21 (e.g. SoilGard® from Certis, USA); Verticillium albo-atrum (formerly V. dahliae) strain WCS850 (CBS 276.92; e.g. Dutch Trig from Tree Care Innovations); Trichoderma atroviride, in particular strain no.
  • AARC-0255 e.g. DiTeraTM from Valent Biosciences
  • Penicillium bilaii strain ATCC ATCC20851 e.g. DiTeraTM from Valent Biosciences
  • Bacillus sphaericus in particular Serotype H5a5b strain 2362 (strain ABTS-1743) (e.g. VECTOLEX® from Valent BioSciences, US); Bacillus thuringiensis subsp. kurstaki strain BMP 123 from Becker Microbial Products, IL (BARITONE from Bayer CropScience); Bacillus thuringiensis subsp. aizawai, in particular serotype H-7 (e.g. FLORBAC® WG from Valent BioSciences, US); Bacillus thuringiensis subsp. kurstaki strain HD-1 (e.g.
  • DIPEL® ES from Valent BioSciences, US); Bacillus thuringiensis subsp. kurstaki strain BMP 123 by Becker Microbial Products, IL; Bacillus thuringiensis israelensis strain BMP 144 (e.g. AQUABAC® by Becker Microbial Products IL); Brevibacillus laterosporus (LATERAL from Ecolibrium Biologicals); Burkholderia spp., in particular Burkholderia rinojensis strain A396 (also known as Burkholderia rinojensis strain MBI 305) (Accession No. NRRL B-50319; WO 2011/106491 and WO 2013/032693; e.g.
  • MBI206 TGAI and ZELTO® from Marrone Bio Innovations Chromobacterium subtsugae, in particular strain PRAA4-1T (MBI-203; e.g. GRANDEVO® from Marrone Bio Innovations); Lecanicillium muscarium Ve6 (MYCOTAL from Koppert); Paenibacillus popilliae (formerly Bacillus popilliae; e.g. MILKY SPORE POWDERTM and MILKY SPORE GRANULARTM from St. Gabriel Laboratories); Pasteuria nishizawae strain Pn1 (CLARIVA from Syngenta/ChemChina); Bacillus thuringiensis subsp.
  • israelensis (serotype H-14) strain AM65-52 (Accession No. ATCC 1276) (e.g. VECTOBAC® by Valent BioSciences, US); Bacillus thuringiensis var. kurstaki strain EVB-113-19 (e.g., BIOPROTEC® from AEF Global); Bacillus thuringiensis subsp. tenebrionis strain NB 176 (SD-5428; e.g. NOVODOR® FC from BioFa DE); Bacillus thuringiensis var. japonensis strain Buibui; Bacillus thuringiensis subsp. kurstaki strain ABTS 351 ; Bacillus thuringiensis subsp.
  • aizawai strain GC-91 Serratia entomophila (e.g. INVADE® by Wrightson Seeds); Serratia marcescens, in particular strain SRM (Accession No. MTCC 8708); Trichoderma asperellum (TRICHODERMAX from Novozymes); and Wolbachia pipientis ZAP strain (e.g., ZAP MALES® from MosquitoMate); and
  • fungi selected from the group consisting of Isaria fumosorosea (previously known as Paecilomyces fumosoroseus) strain apopka 97 PREFERAL from SePRO; ; Beauveria bassiana strain ATCC 74040 (e.g. NATURALIS® from Intrachem Bio Italia); Beauveria bassiana strain GHA (Accession No. ATCC74250; e.g.
  • Viruses selected from the group consisting of Adoxophyes orana (summer fruit tortrix) granulosis virus (GV), Cydia pomonella (codling moth) granulosis virus (GV), Helicoverpa armigera (cotton bollworm) nuclear polyhedrosis virus (NPV), Spodoptera exigua (beet armyworm) mNPV, Spodoptera frugiperda (fall armyworm) mNPV, and Spodoptera littoralis (African cotton leafworm) NPV;
  • Adoxophyes orana sumr fruit tortrix granulosis virus
  • GV Cydia pomonella (codling moth) granulosis virus
  • NPV Helicoverpa armigera (cotton bollworm) nuclear polyhedrosis virus
  • Spodoptera exigua beet armyworm
  • Spodoptera frugiperda fall armyworm
  • mNPV Spodoptera
  • Bacteria and fungi which can be added as ’inoculant’ to plants or plant parts or plant organs and which, by virtue of their particular properties, promote plant growth and plant health selected from the group consisting of Agrobacterium spp., Azorhizobium caulinodans, Azospirillum spp., Azotobacter spp., Bradyrhizobium spp., Burkholderia spp., in particular Burkholderia cepacia (formerly known as Pseudomonas cepacia), Gigaspora spp., or Gigaspora monosporum, Glomus spp., Laccaria spp., LactoBacillus buchneri, Paraglomus spp., Pisolithus tinctorus, Pseudomonas spp., Rhizobium spp., in particular Rhizobium trifolii, Rhizopogon spp., Scleroderma s
  • FIG. 1 Spectrum of light absorption (UV-VIS) 200-400nm of a lipopeptide according to Formula II (a), Formula II (b) or Lipopeptin A
  • Figure 8 The upfield region of the 1 D 1 H NMR spectrum of a lipopeptide according to Formula ll(a) in CD3OD at 600 MHz
  • Example 1.1 Source and fermentation
  • Streptomyces sp. Saigon413, isolated in Vietnam before 1961 was deposited at the Westerdijk institute Uppsalalaan 8, 3584 CT Utrecht, the Netherlands, under accession number CBS149411 .
  • the deposit was made by Syngenta Ltd., Jealott’s Hill Research International Centre, Bracknell, Berkshire, RG42 6EY, UK under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure.
  • the Streptomyces was cultivated in Erlenmeyer flasks with a liquid medium consisting of (g / I) casein hydrolysate 10, glucose 40, K2HPO4 1.25, soytone 2, tryptone, 8 and incubated at 28°C in an incubator shaking 150 rpm with 25 mm throw for 4 days. Large scale fermentations
  • TGAI technical grade active ingredient (unformulated product).
  • Example 1.3 Isolation of 16S rDNA, genomic DNA and species identification
  • cyclothiazomycin C is disclosed on p. 3 of WO2015191789 and can be extracted and analysed according to the method discosed in Wang et al. (2010) Appl. Environmental Microbiology, Vol. 76, No. 7 p.2336. Malonomicin can be extracted and analysed according to the method disclosed in Example I (B) of W02006/078939. Streptimidone can be extracted and isolated according to the method as disclosed in Lee et al. J. of Antibiotics (2020) 73: p. 184-188, including the supplementary information.
  • the compound I was further purified by Hydrophilic Interaction Liquid Chromatography (HILIC) using Mass guided fractionation and ELSD detector. Using for example Waters XBridge Amide, 5 micron, 30x100mm using a gradient of acetonitrile and 10mM Ammonium Acetate.
  • HILIC Hydrophilic Interaction Liquid Chromatography
  • the compound I was determined in the purified fermentation broth according to the methods disclosed below.
  • the molecular composition and total molecular mass was C53H90N2O44, and 1458.487 g, respectively which were determined using MS-MS and NMR spectroscopy as disclosed below.
  • the mass spectrometer was connected to a Vanquish Flex UHPLC from Thermo Scientific using a Vanquish Split Sampler FT, Vanquish Binary Pump F, Vanquish Column Compartment H, Vanquish Diode Array Detector FG and Vanquish Charged Aerosol Detector.
  • NMR Spectroscopy NMR spectra were recorded on a Bruker AVIII 600 NMR spectrometer, equipped with a 5 mm Bruker ( 1 H/ 19 F)/ 13 C/ 15 N TCI cryoprobe fitted with Z gradients, using standard Bruker pulse sequences. Samples were dissolved in D2O, and the spectra were recorded at 300° K and referenced to acetone at 2.225 ppm for 1 H and 31 .07 ppm for 13 C.
  • Figures 1 to 4 show NMR spectra of the compound of the present invention.
  • the one bond 1 H- 13 C correlation spectrum contains peaks corresponding to 1 methyl (CH3) and 40 methine (CH) groups (listed in Table 1) and 9 methylene (CH2) groups (listed in Table 2).
  • the 1 D 13 C spectrum contains signals from 3 quaternary carbons at 104.7, 159.3 and 175.2 ppm ( ⁇ 0.1).
  • the lipopeptides were purified from the ethyl acetate fraction by preparative reverse phase (C18) HPLC. The lipopeptides were relatively aploar and elute in the higher organic fraction in a gradient system with 0.1% formic acid and acetonirile (0.1% formic acid). A gradient of
  • Experiment 1 MS OT (Orbitrap Resolution: 50,000, Scan Range (m/z): 200 to 2000, RF Lens (%): 60, AGO Target: Standard, Maximum Injection Time Mode: Auto, Microscans: 1 , Data Type: Profile, Polarity: Both), Experiment 2: tMS2 OT CID (MSn Level (n): 2, Isolation Window (m/z): 1.0, Activation Type: CID, CID Collision Energy (%): 30, Detector Type: Orbitrap, Orbitrap Resolution: 30,000, RF Lens (%): 60, Polarity: Positive),
  • the mass spectrometer was connected to a Vanquish Flex UHPLC from Thermo Scientific using a Vanquish Split Sampler FT, Vanquish Binary Pump F, Vanquish Column Compartment H, Vanquish Diode Array Detector FG and Vanquish Charged Aerosol Detector.
  • Conidia of the fungus from cryogenic storage were directly mixed into nutrient broth (potato dextrose broth).
  • a DMSO solution of the test compounds was placed into a microtiter plate (96-well format) and the nutrient broth containing the fungal spores was added to it.
  • the test plates were incubated at 24°C and the inhibition of growth was determined after 72 hrs.
  • Streptomyces sp. isolate CBS149411 was fermented, dried to powder and formulated as an oil dispersion, with a loading of 40% of dry matter as disclosed above.
  • CBS149411 formulated as OD40% was sprayed at a use rate of 500g/ha (active ingredient), and a second product at a use rate of 100g/ha (active ingredient), resulting in a ratio of 5:1 (CBS 149411 : 2nd compound).
  • the ratio of volumes for the two products is indicated (volume in liter of product per hectare). Mixtures resulting in 30% or more reduction of disease symptoms were seen as effective to provide a means for disease control or disease reduction.
  • FRAC Code List 2023 provides a comprehensive overview of different mode of action used to control fungal and bacterial diseases on crop plants (FRAC: Fungicide resistance action committee, is a specialist technical group of Crop Life international). Where available, active ingredients are also identified by a CAS number (CAS: Chemical Abstracts Service, a division of the American Chemical Society (2540 Olentangy River Road, Columbus, Ohio 43202 USA).
  • Table 8 List of compounds tested in mixture with Streptomyces sp. CBS149411 . a) also bactericide b) details on manufacturer
  • Syngenta Syngenta Crop Protection AG, Rosentalstr. 67, 4058 Basel, SWITZERLAND
  • Zymoseptoria tritici (EPPO code: SEPTTR): 14 days old wheat plants variety Riband were inoculated by spraying a spore suspension on them one day after application (1.5 Mio spores per ml in water supplemented with 0.01 % Tween20). After an incubation period of 4 days at 22°C/21 °C (day/night) and 95% relative humidity, the inoculated test plants were kept at 22°C/21°C (day/night) and 70% relative humidity in a greenhouse. Efficacy was assessed directly when an appropriate level of disease appeared on untreated check plants (14 - 19 days after application). The isolate used is a recent field isolate expressing tolerance to DMI fungicides, SDHI fungicides and Qol fungicides.
  • Pyricularia oryzae (EPPO code: PYRIOR): 3 week old rice plants cv. Balilla were treated with the formulated test compound in a spray chamber. Two days after application rice plants were inoculated by spraying a spore suspension (1 x 10 5 conidia/ml) on the test plants. After an incubation period of 6 days at 25° C and 95% relative humidity the disease incidence was assessed.
  • Results The results of the foliar experiments of a mixture of Streptomyces chrestomyceticus CBS149411 and a biological product are shown in the following Tables 9 to 25.
  • the biological compound (2 nd product) used in the experiments is indicated in the title of Tables 9 to 25.
  • Treatment of the mixtures is compared with the treatments by a composition of S. chrestomyceticus CBS 149411 alone and the biological compound alone.
  • Table 9 Disease control by S. chrestomyceticus CBS 149411 mixed with orange oil
  • Table 10 Disease control by S. chrestomyceticus CBS 149411 mixed with Tea tree Oil (Melaleuca alternifolia)
  • Table 11 Disease control by S. chrestomyceticus CBS 149411 mixed with sulfur (Seffika)
  • Table 12 Disease control by S. chrestomyceticus CBS 149411 mixed with phosphonate (Pygmalion®)
  • Table 13 Disease control by S. chrestomyceticus CBS 149411 mixed with sulfur (Thiovit Jet)
  • Table 14 Disease control by S. chrestomyceticus CBS 149411 mixed with phosphonate (Redeli)
  • Table 16 Disease control by S. chrestomyceticus CBS 149411 mixed with Laminarin

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Zoology (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Environmental Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Pest Control & Pesticides (AREA)
  • Plant Pathology (AREA)
  • Health & Medical Sciences (AREA)
  • Agronomy & Crop Science (AREA)
  • Dentistry (AREA)
  • Virology (AREA)
  • Biotechnology (AREA)
  • Mycology (AREA)
  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

The present invention relates to a composition comprising components (A) and (B) as active ingredients, wherein component (A) comprises a Streptomyces and component (B) is a compound selected from the group consisting of sulphur, phosphonate, copper hydroxide, copper oxychloride, bicarbonate, Bacillus sp, Trichoderma sp, aureobasidin A, jawsamycin, kasugamycin, natamycin, a polysaccharide elicitor, a plant biostimulant (PBS) comprising an algal extract, a plant extract and/or vinasse, orange oil, and / or tea tree oil. The invention further relates to a method of controlling or preventing phytopathogenic diseases, preferably phytopathogenic fungi, on a plant or on plant propagation material and / or on harvested food crops, which comprises applying to the plant, on plant propagation material, the locus thereof, and / or on harvested food crops the composition according to the present invention.

Description

COMPOSITION
The present invention relates to novel compositions, to their use in agriculture or horticulture for controlling diseases caused by phytopathogens, especially phytopathogenic fungi and / or oomycetes, and to methods of controlling diseases on useful plants.
BACKGROUND
Whilst many compounds and compositions, belonging to various different chemical classes, or which are from biological origin, for instance bacterial origin, have been/are being developed for use as pesticides in crops of useful plants, crop tolerance and activity against pests, such as phytopathogenic fungi do not always satisfy the needs of agricultural practice in many respects. Therefore, there is a continuing need to find new compounds and compositions having superior biological properties for use in controlling or preventing infestation of plants by phytopathogenic pests such as phytopathogenic microorganisms. For example, compounds or compositions possessing a greater biological activity, an advantageous spectrum of activity, an increased safety profile, improved physico-chemical properties, or increased biodegradability. Or else, compositions possessing a broader spectrum of activity, improved crop tolerance, synergistic interactions or potentiating properties, or compositions which display a more rapid onset of action or which have longer lasting residual activity or which enable a reduction in the number of applications and/or a reduction in the application rate of compounds and compositions required for effective control of a phytopathogen, thereby enabling beneficial resistance-management practices, reduced environmental impact and reduced operator exposure. The use of compositions comprising mixtures of different fungicidal compounds possessing different modes of action can address some of these needs (eg. by combining fungicides with differing spectra of activity).
Fungicidal compositions from bacterial origin such as from Streptomyces are for instance disclosed in W02022/038180 and in Rahila et al, (2023) Current Microbiology, 80: 107. Fungicidal compositions from bacterial origin such as from Streptomyces are for instance disclosed in W02022/038180 and in Rahila et al, (2023) Current Microbiology, 80: 107. In US 5,356,624 a Streptomyces rimosus strain is disclosed that was found active against several wood-degrading fungi. There is a need for mixtures of a composition comprising Streptomyces and a second composition comprising another pesticide, which can solve some of the problems outlined above.
SUMMARY
The present invention relates to a composition comprising components (A) and (B) as active ingredients, wherein component (A) comprises a Streptomyces chrestomyceticus and component (B) comprises a compound selected from the group consisting of sulphur, phosphonate, copper hydroxide, copper oxychloride, bicarbonate, Bacillus sp, Trichoderma sp, aureobasidin A, jawsamycin, kasugamycin, natamycin, a polysaccharide elicitor comprising laminarin, a composition comprising chitooligosaccharides and oligogalactunorides (COS-OGA), and/or yeast extract, a plant biostimulant (PBS) comprising an algal extract, a plant extract and/or vinasse, orange oil, and / or tea tree oil.
In a second aspect the present invention relates to a method of controlling or preventing phytopathogenic diseases, preferably phytopathogenic fungi, on a plant or on plant propagation material and / or on harvested food crops, which comprises applying to the plant, on plant propagation material, the locus thereof, and / or on harvested food crops the composition according to the present invention.
In a third aspect the present invention relates a plant, plant propagation material or the locus thereof, or harvested food crops comprising a composition according to the present invention.
DETAILED DESCRIPTION
The present invention relates to a composition comprising components (A) and (B) as active ingredients, wherein component (A) comprises a Streptomyces chrestomyceticus and component (B) comprises, or is, a compound selected from the group consisting of sulphur, phosphonate, copper hydroxide (Cu(OH)2), copper oxychloride (Cu2(OH)3CI), bicarbonate, Bacillus sp, Trichoderma sp, aureobasidin A, jawsamycin, kasugamycin, natamycin, a polysaccharide elicitor comprising laminarin, a composition comprising chitooligosaccharides and oligogalactunorides (COS-OGA), and/or yeast extract, a plant biostimulant (PBS) comprising an algal extract, a plant extract and/or vinasse, orange oil, and / or tea tree oil.
Surprisingly, it was found that a composition according to the present invention provided effective control against phytopathogenic microorganisms, preferably against phytopathogenic fungi. The benefits provided by certain compositions according to the invention may also include, inter alia, advantageous levels of biological activity for protecting plants against diseases that are caused by pests, for instance fungi, or superior properties for use as agricultural active ingredients (for example, greater biological activity, an advantageous spectrum of activity, an increased safety profile, improved physico-chemical properties, or increased biodegradability).
A composition according to the present invention typically is a pesticidal composition.
A composition, microorganism, metabolite or compound having “pesticidal activity” or “pesticide” as used herein means a composition, microorganism, metabolite or compound that controls, modifies, or prevents the growth of pests. The term “pesticidally effective amount” or “effective amount of pesticide” where used means the quantity of such a composition, microbial strain, compound, or metabolite that is capable of producing an effect on the growth of pests. Controlling or modifying effects include all deviation from natural development, such as killing, retardation and the like, and prevention includes barrier or other defensive formation in or on a plant to prevent pest infection. The term “pesticidal activity” or “pesticides” includes insecticidal or fungicidal activity or “insecticides” or “fungicides”. The composition according to the present invention may also have activity against oomycetes.
A composition according to the present invention typically is a fungicidal composition. The term “fungicide” or “fungicidal” as used herein means a composition, metabolite or microorganism or compound that controls, modifies, or prevents the growth of fungi. The term “fungicidally effective amount” or “effective amount of fungicide” means the quantity of such a compound, metabolite, microorganism or composition that is capable of producing an effect on the growth of fungi. Controlling or modifying effects include all deviation from natural development, such as killing, retardation and the like, and prevention includes barrier or other defensive formation in or on a plant to prevent fungal infection, eg. the composition according to the present invention may act as a plant growth regulator. The composition according to the present invention comprises a fungicidally effective amount of the component (A) and/or the component (B).
The composition according to the present invention typically is an agricultural-acceptable composition.
Throughout this document the expression “composition” stands for the various mixtures or combinations of components (A) and (B) (including the above-defined embodiments), for example in a single “ready-mix” form, in a combined spray mixture, or combined slurry mixture, composed from separate formulations of the single active ingredient components, such as a “tank-mix”, and in a combined use of the single active ingredients when applied in a sequential manner, i.e. one after the other with a reasonably short period, such as a few hours or days. The order of applying the components (A) and (B) is not essential for working the present invention.
In a preferred embodiment, component (A) comprises a Streptomyces chrestomyceticus. The Streptomyces chrestomyceticus preferably comprises a nucleotide sequence which has at least 99.8 %, preferably at least 99.9%„ preferably at least 99.91%, 99.92%, 99.93%, 99.94%, 99.95%, 99.96%, 99.97%, 99.98%, or 99.99% identity to SEQ ID NO: 1. In one embodiment the Streptomyces chrestomyceticus comprises a nucleotide sequence according to SEQ ID NO:1. SEQ ID NO: 1 comprises the 16S RNA gene of Streptomyces sp. Saigon413 deposited with the Westerdijk Institute under accession number CBS149411
In another preferred embodiment the component (A) comprises a Streptomyces chrestomyceticus, which comprises a genome sequence which has at least 95% identity, preferably at least 96%, 97%, 98%, 99% identity to the whole genome of Streptomyces sp. Saigon413 deposited with the Westerdijk Institute under accession number CBS149411 . The Streptomyces may comprise a genome sequence which has at least 99.1 %, 99.2%, 99.3%. 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9% identity to the whole genome of Streptomyces sp. Saigon413 deposited with the Westerdijk Institute under accession number CBS149411 . Preferably component (A) comprises a Streptomyces chrestomyceticus, which is Streptomyces sp. Saigon413 deposited with the Westerdijk Institute under accession number CBS149411.
As used herein, the terms "percent identity," and "percent identical" refer to the relatedness of two or more nucleotide or amino acid sequences, which may be calculated by (i) comparing two optimally aligned sequences over a window of comparison, (ii) determining the number of positions at which the identical nucleic acid base (for nucleotide sequences) or amino acid residue (for proteins) occurs in both sequences to yield the number of matched positions, (iii) dividing the number of matched positions by the total number of positions in the window of comparison, and then (iv) multiplying this quotient by 100 percent to yield the percent identity. If the "percent identity" is being calculated in relation to a reference sequence without a particular comparison window being specified, then the percent identity is determined by dividing the number of matched positions over the region of alignment by the total length of the reference sequence. Accordingly, for purposes of the present invention, when two sequences (query and subject) are optimally aligned (with allowance for gaps in their alignment), the "percent identity" for the query sequence is equal to the number of identical positions between the two sequences divided by the total number of positions in the query sequence over its length (or a comparison window), which is then multiplied by 100 percent.
The component (A) comprising Streptomyces as disclosed herein comprises a cell count of the Streptomyces from 1*10° to 1*1014 cfu / g dry weight, for instance from 1*101 to 1*1013 cfu / g dry weight, from 1*102 to 1*1013 cfu I g dry weight, for instance from 1*103 to 1*1012 cfu / g dry weight, from 2*103 to 2*1011 cfu / g dry weight, from 5*103 to 5*1011 cfu / g dry weight, for instance from 1*104 to 1*101° cfu / g dry weight, from 2*104 to 2*101° cfu / g dry weight, such as from 1 *105 to 1 *109 cfu / g dry weight, from 2*105 to 2*109 cfu I g dry weight, from 5*105 to 5*109 cfu I g dry weight, from 1*106 to 1*108 cfu I g dry weight, such as from 2*106 to 2*108 cfu / g dry weight.
The component (A) comprising a Streptomyces chrestomyceticus as disclosed herein comprises a fermentation broth comprising the Streptomyces chrestomyceticus, preferably a spray-dried fermentation broth or a freeze-dried fermentation broth. Spray-drying or freeze-drying of a fermentation broth is known in the art. Component (A) comprising a Streptomyces as disclosed herein also includes a formulation comprising the Streptomyces.
A Streptomyces, such as Streptomyces chrestomyceticus, may be cultivated in a suitable fermentation medium under suitable fermentation conditions, and optionally comprising a step of recovering the Streptomyces chrestomyceticus strain. Usually, a fermentation broth is produced during cultivation of or when cultivating a strain of Streptomyces chrestomyceticus. Suitable fermentation conditions for cultivating Streptomyces sp. such as S. chrestomyceticus are known to a person skilled in art.
Cultivating a strain of Streptomyces, for instance Streptomyces chrestomyceticus strain, as disclosed herein, comprises cultivating the microbial strain under aerobic conditions at a temperature of from 15 degrees Celsius to 45 degrees Celsius, preferably a temperature of from 20 to 35 degree Celsius, preferably a temperature of between 25 to 32 degrees Celsius, in the presence of a carbon source and a nitrogen source. A suitable carbon source may be molasses, such as beet or cane molasses, polysaccharides, flour, starch, sugar, or glucose. A suitable nitrogen source may be casein hydrolysate, tryptone, ammonium sulphate, ammonia, yeast extract, peptone or urea. The process for cultivating a Streptomyces as disclosed herein may be performed in a batch, fed-batch or continuous culture.
Component (A) as disclosed herein also includes a formulation comprising the Streptomyces chrestomyceticus. Formulations of microbial strains are known in the art for instance as disclosed in Croda Crop Care, the Nouryon formulator toolbox and in: Formulation of Microbial Biopesticides: Beneficial microorganisms, nematodes and seed treatments (412 p., 6 December 2012) eds. Burges H.D., Springer, ISBN 978-94-011-4926-6. Component (A) comprising a Streptomyces chrestomyceticus as disclosed herein comprises a formulation comprising the Streptomyces chrestomyceticus, wherein the formulation is an oil dispersion (OD), a non-aqueous dispersion (NAD) or a flowable formulation.
Component (A) may be a component (A) comprising composition comprising a Streptomyces chrestomyceticus as disclosed herein.
In one embodiment component (A) further comprises a metabolite. A metabolite is preferably produced by the Streptomyces chrestomyceticus disclosed herein. As disclosed herein a metabolite may be a supernatant comprising a metabolite as disclosed herein.
In one embodiment, a component (A) comprises a metabolite, preferably wherein the metabolite is malonomicin. Malonomicin (sometimes spelt ‘malonomycin’) is {[(2S)-2-amino-3-hydroxypropanoyl]amino} {2-[(5S)-5-(aminomethyl)-4-hydroxy-2-oxo-2,5-dihydro-1 H-pyrrol-3-yl]-2-oxoethyl}malonic acid can be produced as disclosed in Example I of W02006/078939. Malonomicin may also be prepared according to the method disclosed in Example I A and B in EP 1860939, or according to Law et al, 2018 (Nature Catalys is | VOL 1 | DECEMBER 2018 | 977-984).
Component (A) may comprise a further metabolite, such as at least one, at least two, at least three, at least four or at least five of the metabolite(s), selected from the group consisting of cyclothiazomycin C, streptimidone, an oligosaccharide compound which comprises a molecular formula according to C53H90N2O44, further characterised by the NMR spectra listed in Table 1 and Table 2, further characterized by a structural Formula I, Formula (I), a lipopeptide according to Formula II, or a salt thereof wherein R1 = CHs or C2H5
Formula (II) and a polyene compound characterized by a molecular formula according to C67H115NO25, wherein the polyene is further characterized by a spectrum of light absorption with absorbance maxima at a wavelength of 235.5 nm, 301.1 nm, 315.8 nm, 330.9 nm and 348.3 nm when measured in an aqueous acetonitrile solution. Preferably, the polyene is further characterized by the spectrum of light absorption as shown in Figure 10.
Suitably, component (A) comprise a further metabolite wherein the metabolite is selected from the group consisting of cyclothiazomycin C, streptimidone, an oligosaccharide compound which comprises a molecular formula according to C53H90N2O44, further characterised by the NMR spectra listed in Table 1 and Table 2, further characterized by a structural Formula I, Formula (I), and a lipopeptide according to Formula II, or a salt thereof wherein R1 = CHs or C2H5
Formula (II)
Cyclothiazomycin C is a known compound. The structure of cyclothazomycin C is disclosed on p.
3 of WO2015191789 and can be produced as disclosed in Example 4 of WO2015/191789.
Streptimidone is a known compound of Formula III
Streptimidone can be synthesised following the method disclosed in Kondo, H., Oritani, T., and Kiyota, H. Synthesis and antifungal activity of the four stereoisomers of streptimidone, a glutarimide antibiotic from Streptomyces rimosus forma paromomycinus. Eur. J. Org. Chem. (20), 3459-3462 (2000).
The metabolites disclosed herein above are preferably produced by Streptomyces chrestomyceticus as disclosed herein.
The composition according to the present invention comprises a component (B) wherein component (B) comprises a compound selected from the group consisting of sulphur (S), phosphonate, copper hydroxide (Cu(OH)2), copper oxychloride (Cu2(OH)3CI), bicarbonate, Bacillus sp, Trichoderma sp, aureobasidin A, jawsamycin, kasugamycin, natamycin, a polysaccharide elicitor comprising laminarin, a composition comprising chitooligosaccharides and oligogalactunorides (COS-OGA), and/or yeast extract, a plant biostimulant (PBS) comprising an algal extract, a plant extract and/or vinasse, orange oil, and / or tea tree oil. Phosphonate as used herein comprises potassium phosphosnate (K2HPO3) and/or sodium phosphonate (Na2HPOs).
Bicarbonate as used herein may comprise potassium bicarbonate (KHCO3) or sodium bicarbonate (NaHCO3)
In one embodiment component (B) comprises a Bacillus sp., wherein the Bacillus sp. comprises Bacillus amyloliquefaciens, and/or Bacillus velezensis. Bacillus amyloliquefaciens may also be annotated as Bacillus velezenis. Bacillus velezensis comprises several Bacillus strains that were previously registered as B. subtilis or B. amyloliquefaciens. In addition, Bacillus amyloliquefaciens has been previously also known as a subspecies of Bacillus subtilis: B. subtilis subsp. amyloliquefaciens. According to a recent reclassification based on phylogenetic analyses at the whole genome level, many commercialized Bacillus strains belong to the B. velezensis species (Dunlap et al., 2016, doi: 10.1099/ijsem.0.000858 Anckaert et al., 2021 , doi: 10.19103/AS.2021 .0093.10). These include, but are not limited to, B. subtilis strain GBO3, B. amyloliquefaciens strain MBI600, B. subtilis strain QST713, B. amyloliquefaciens strain FZB24, B. velezensis strain FZB24, B. subtilis strain D747, and B. amyloliquefaciens strain D747 (Fan et al., 2017, doi: 10.3389/fmicb.2017.00022; Dunlap, 2019, doi: 10.1016/j. biocontrol.2019.04.011 ; Anckaert et al., 2021). Other bacterial strains may also fall under the Bacillus velezensis classification, including those that are yet to be (re-)classified.
Preferably, component B as disclosed herein comprises a bacterium B.velezensis and / or B. amyloliquefaciens, Preferably, the component B comprises a B. amyloliquefaciens strain FZB24 or B. velezensis strain FZB24, Bacillus velezensis CNPSo 3602, and / or Bacillus amyloliquefaciens MBI600
In one embodiment component (B) comprises a Trichoderma sp., for instance Trichoderma sp. disclosed in Woo, S.L., et al, “The open Mycology Journal, 2014, Vol. 8 (Suppl.-1 , M4), p. 71-126,
Suitable Trichoderma species are for instance T. asperellum, T. atroviride, T. gamsii, T. hamatum. T. harzianum, T. polysporum, T. viren, T. viride. Preferably, the Trichoderma sp. comprises Trichoderma afroharzianum, syn. Trichoderma harzianum,
Component (B) may comprise a polysaccharide elicitor, for instance chitin, pectin or dextran. Preferably a polysaccharide elicitor comprises laminarin, a composition comprising chitooligosaccharides and oligogalactunorides (COS-OGA), and/or yeast extract. A polysaccharide elicitor as used herein is a polysaccharide that triggers or stimulates a defense mechanism in a plant, and/or kill or limit the development of pathogens.
The yeast extract as disclosed herein comprises an extract from any suitable yeast, preferably the yeast extract comprises a Saccharomyces cerevisiae yeast extract.
Component (B) may comprise a plant biostimulant comprising an algal and / or a plant extract. Any suitable algal extract may be used for instance from macroalgae and/or microalgae. Preferably, the algal extract comprises an extract from macroalgae such as a seaweed, for instance red, brown or green seaweed. Brown seaweed may be selected from: Ascophyllum nodosum, Ecklonia maxima, Laminaria saccharina, Laminaria digitata, Fucus spiralis, Fucus serratus, F. vesiculosus, Macrocystis spp., Pelvetia canaliculata, Himantalia elongata, Undaria pinnatifida, Sargassum spp. Red seaweed may be selected from Kappaphycus spp., Chondrus spp., Palmaria spp., Gracilaria spp., Porphyra spp., Porphyridium spp., Mastocarpus spp., Polysiphonia spp. Green seaweed as disclosed herein may be selected from: Ulva spp., Caulerpa spp., Codium spp., Halimeda spp, Acetabularia spp., Cladophora spp. Preferably the algal extract is from Ascophyllum nodosum.
Microalgae as disclosed herein may be selected from: Spirulina, Scenedesmus, Nannochloropsis, Haematococcus, Chlorella, Phaeodactylum, Arthrospyra, Tetraselmis, Isochrysis, Synechocystis, Clamydomonas, Parietochloris, Desmodesmus, Neochloris, Dunaliella, Thalassiosira, Pavlova, Navicula, Chaetocerous.
A suitable plant extract may be an extract from a plant which is selected from: beet, sugar cane, alfalfa, maize, brassica, halophytes, soya, wheat, yucca, quillaja, hop, coffee, citrus, olive, lupine, bean, pea, lentils, mushrooms, carrot, apple, tomato and combinations thereof. The plant extract may be an extract from the whole plant or any part thereof, for instance from leaves, roots, stems, fruits, flowers, seeds, seedlings, bark, berries, skins, and combinations thereof.
Processes for preparing plant and / or algal extracts are known in the art. Usually a process for preparing a plant and / or an algal extracts comprises providing a sample of a plant and / or an algae and contacting the plant and / or algal sample with an aqueous solution comprising an extraction agent. An extraction agent may be solvents, acids, bases, and / or enzymes. A chemical means may also be used. An extraction process is usually carried out at a temperature of from -20 °C to 120°C, for instance from 0°C to 100°C. An extraction process may be followed by a step of removing or separating a solid fraction if desired. Alternatively, a suspension of a plant and / or an algal extract may be used.
A plant biostimulant as disclosed herein may comprise a plant extract as disclosed herein and vinasse. As disclosed herein vinasse is usually a byproduct from the sugar and/or ethanol industry.
A plant biostimulant may be a composition comprising an algal extract, a plant extract as disclosed herein and vitamins. A suitable vitamin may be selected from: vitamin B1 , B2, B3, B5, B6, B9, B12, E, A, D, C, PP, H, K1 , K2, K3 and combinations thereof. The plant biostimulant may further comprise betaines.
A plant biostimulant as disclosed herein may further comprise macro-, or micronutrients, such as a nitrogen source, a potassium source, a phosphorus source, a manganese source, a zinc source, an iron source, a copper source, a boron source, a molybdenum source, a calcium source, or a magnesium source.
Component (B) as disclosed herein may comprise orange oil. Orange oil as used herein usually is a blend of cold-pressed orange oil.
Component (B) as disclosed herein may comprise tea tree oil. Tea tree oil is an extract of the tea tree plant Melaluca alternifolia.
Certain compositions comprising a component (A) and a component (B) as disclosed herein may show a synergistic effect. This occurs whenever the action of an active ingredient combination is greater than the sum of the actions of the individual components. The action to be expected E for a given active ingredient combination obeys the so-called COLBY formula and can be calculated as follows (COLBY, S.R. "Calculating synergistic and antagonistic responses of herbicide combination". Weeds, Vol. 15, pages 20- 22; 1967): ppm = milligrams of active ingredient (= a.i.) per liter of spray mixture
X = % action by active ingredient A) using p ppm of active ingredient
Y = % action by active ingredient B) using q ppm of active ingredient.
According to COLBY, the expected (additive) action of active ingredients A)+B) using p+q ppm of active ingredient is:
If the action actually observed (O) is greater than the expected action (E), then the action of the combination is super-additive, i.e. there is a synergistic effect. In mathematical terms, synergism corresponds to a positive value for the difference of (O-E). In the case of purely complementary addition of activities (expected activity), said difference (O-E) is zero. A negative value of said difference (O-E) signals a loss of activity compared to the expected activity.
In one embodiment the composition according to the present invention comprises a component (B) as disclosed herein, wherein the Bacillus sp. comprises Bacillus amyloliquefaciens, and/or Bacillus velezensis, the Trichoderma sp. comprises Trichoderma afroharzianum , the polysaccharide elicitor comprises laminarin, a composition comprising chitooligosaccharides and oligogalactunorides (COS-OGA), and/or yeast extract, wherein preferably the yeast extract comprises Saccharomyces cerevisiae yeast extract, and / or the algal extract comprises an extract from Ascophyllum nodosum.
In one embodiment the composition according to the present invention comprises a component (B) wherein component (B) comprises a compound selected from the group consisting of sulfur, phosphonate, Bacillus amyloliquefaciens, preferably Bacillus amyloliquefaciens FZB24, a composition comprising chitooligosaccharides and oligogalactunorides (COS-OGA), jawsamycin, laminarin, and orange oil.
It was found that a composition as disclosed herein comprising a component (B) which comprises a compound selected from the group consisting of sulfur, phosphonate, Bacillus amyloliquefaciens, a composition comprising chitooligosaccharides and oligogalactunorides (COS-OGA), jawsamycin, laminarin, and orange oil resulted in a surprising synergistic activity against phytopathogenic fungi.
Components (A) and (B) as active ingredient may be used at any suitable ratio. Preferably, a composition of the present invention comprises a component (A) and component (B) at a weight ratio of component (A) to component (B) of from 100:1 to 1 :100, for instance a weight ratio of from 90:1 to 1 :90, for instance a weight ratio of from 80:1 to 1 :80, for instance a weight ration of from 70:1 to 1 :70, for instance a weight ration of from 60:1 to 1 :60, for instance a weight ratio of compound (A) to compound (B) of from 50:1 to 1 :50, for instance a weight ratio of from 40:1 to 1 :40, for instance weight ratio of from 30:1 to 1 :30, for instance a weight ration of from 20:1 to 1 :20, for instance a weight ration of from 10:1 to 1 : 10, for instance a weight ration of from 5:1 to 1 :5.. In one embodiment a composition comprising components (A) and (B) as active ingredients, is a composition wherein component (A) comprises a Streptomyces chrestomyceticus as disclosed herein above, and wherein component (B) comprises sulfur at a weight ratio of 1 :1 to 1 :100, preferably at a weight ratio of 1 :2 to 1 :80, preferably at a weight ratio of 1 :4 to 1 : 75. A composition according to this embodiment was surprisingly effective against Zymoseptoria tritici and Puccinia recondita f. sp. tritici.
In one embodiment a composition comprising components (A) and (B) as active ingredients, is a composition wherein component (A) comprises a Streptomyces chrestomyceticus as disclosed herein above, and wherein component (B) comprises phosphonate, for instance potassium or sodium phosphonate, at a weight ratio of 1 :1 to 1 :100, preferably at a weight ratio of 1 :2 to 1 :90, preferably at a weight ratio of 1 :4 to 1 :80. A composition according to this embodiment was surprisingly effective against Zymoseptoria tritici and Puccinia recondita f. sp. tritici
In one embodiment a composition comprising components (A) and (B) as active ingredients, is a composition wherein component (A) comprises a Streptomyces chrestomyceticus as disclosed herein above, and wherein component (B) comprises Bacillus amyloliquefaciens, preferably Bacillus amyloliquefaciens FZB24, for instance at a weight ratio of 50:1 to 1 :20, preferably at a weight ratio of 40:1 to 1 :10, preferably at a weight ratio of 30:1 to 1 :5. A composition according to this embodiment was surprisingly effective against Zymoseptoria tritici and Puccinia recondita f. sp. tritici.
In one embodiment a composition comprising components (A) and (B) as active ingredients, is a composition wherein component (A) comprises a Streptomyces chrestomyceticus as disclosed herein above, and wherein component (B) comprises orange oil at a weight ratio of 50:1 to 1 :20, preferably at a weight ratio of 40:1 to 1 :10, preferably at a weight ratio of 30:1 to 1 :5. A composition according to this embodiment was surprisingly effective against Zymoseptoria tritici, Puccinia recondita f. sp. tritici and Pyricularia oryzae.
In one embodiment a composition comprising components (A) and (B) as active ingredients, is a composition wherein component (A) comprises a Streptomyces chrestomyceticus as disclosed herein above, and wherein component (B) comprises laminarin at a weight ratio of 20:1 to 1 :10, preferably at a weight ratio of 10:1 to 1 :5, preferably at a weight ratio of 5:1 to 1 :2. A composition according to this embodiment was surprisingly effective against Zymoseptoria tritici
In one embodiment a composition comprising components (A) and (B) as active ingredients, is a composition wherein component (A) comprises a Streptomyces chrestomyceticus as disclosed herein above, and wherein component (B) comprises jawsamycin at a weight ratio of 30:1 to 1 :20, preferably at a weight ratio of 25:1 to 1 :10, preferably at a weight ratio of 20:1 to 1 :5. A composition according to this embodiment was surprisingly effective against Zymoseptoria tritici and Puccinia recondita f. sp. tritici
In one embodiment a composition comprising components (A) and (B) as active ingredients, is a composition wherein component (A) comprises a Streptomyces chrestomyceticus as disclosed herein above, and wherein component (B) comprises a composition comprising chitooligosaccharides and oligogalactunorides (COS-OGA) at a weight ratio of 75:1 to 1 :20, preferably at a weight ratio of 60:1 to 1 :10, preferably at a weight ratio of 50:1 to 1 :5. A composition according to this embodiment was surprisingly effective against Zymoseptoria tritici and Puccinia recondita f. sp. tritici
A composition according to the present invention is an agriculturally acceptable composition.
In one embodiment, the composition according to the present invention further comprises an agriculturally acceptable carrier and, optionally, a surfactant and/or formulation adjuvants.
Suitable agricultural adjuvants and/or carriers can be solid or liquid and are substances useful in formulation technology, e.g. natural or regenerated mineral substances, solvents, dispersants, wetting agents, tackifiers, thickeners, binders or fertilizers.
Suitable solvents and liquid carriers include, for example water, organic solvents, oils of vegetable or animal origin, cyclic and aromatic hydrocarbons, alcohols, esters, fatty acids, a glycol or any other suitable liquid carrier known in the art. The solvent or liquid carrier may be water or DMSO.
Suitable solid carriers include, for example ammonium salts, talc, titanium dioxide, pyrophyllite clay, silica, attapulgite clay, kieselguhr, chalk, diatomaxeous earth, lime, calcium carbonate, bentonite clay, fuller’s earth, cotton seed hulls, wheat flour, soybean flour, pumice, wood flour, walnut shell flour and lignin. Water is generally the carrier of choice for the dilution of concentrates. The amount of carrier may typically range from 0.9% to 99.99% by weight of the composition.
A broad range of surface-active agents are advantageously employed in both said liquid and solid compositions, especially those designed to be diluted with carrier before application. These agents, when used, normally comprise from 0.1% to 15% by weight of the formulation. They can be anionic, cationic, non-ionic or polymeric in character and can be employed as emulsifying agents, wetting agents, suspending agents or for other purposes.
Other adjuvants commonly utilized in agricultural compositions include crystallisation inhibitors, viscosity modifiers, suspending agents, spray droplet modifiers, pigments, antioxidants, foaming agents, anti-foaming agents, light-blocking agents, compatibilizing agents, sequestering agents, neutralising agents and buffers, corrosion inhibitors, dyes, odorants, spreading agents, penetration aids, micronutrients, emollients, lubricants and sticking agents.
The composition according to the present invention may be used in unmodified form or, preferably, together with the adjuvants conventionally employed in the art of formulation. To this end they may be conveniently formulated in known manner to emulsifiable concentrates, coatable pastes, directly sprayable or dilutable solutions or suspensions, dilute emulsions, wettable powders, soluble powders, dusts, granulates, and also encapsulations e.g. in polymeric substances. As with the type of the compositions, the methods of application, such as spraying, atomising, dusting, scattering, coating or pouring, are chosen in accordance with the intended objectives and the prevailing circumstances. The compositions may also contain further adjuvants such as stabilizers, antifoams, viscosity regulators, binders or tackifiers as well as fertilizers, micronutrient donors or other formulations for obtaining special effects. Component (A) as disclosed herein may be a formulation, wherein Streptomyces chrestomyceticus as disclosed herein is formulated as an oil dispersion (OD), a non-aqueous dispersion (NAD) or a flowable formulation.
An oil dispersion (OD) is a solid active ingredient dispersed in oil, which is a water immiscible carrier. An oil dispersion typically comprises (in wt%) active ingredient (5-60%) non-aqueous dispersants (1-10% of solids) aqueous dispersants (1-10% of solids) emulsifier (2-10% of iol) rheology modifier (0.2- 5%), and an oil carrier to make up 100%. Suitable oils in an oil dispersion can be mineral oils, paraffinic oils, vegetable oils or methylated oils.
Suitable adjuvants, dispersants, emulsifiers and rheology modifiers in an oil dispersion depend on the type of oil and are known in the art. Suitable emulsifiers can be alcohol ethoxylates/alcoxylates, such as C16/18 ethoxylates, or C16/C18 alcoxylate, or block co-polymers. An adjuvant can be an alkyl polyglucoside. Suitable rheology modifiers can be clay, hydrogenated castor oil and derivatives, fumed silicam polyamides, polyesters or agrilan CDS.
A non-aqueous dispersion (NAD) is a liquid formulation wherein a solid ingredient is dispersed in a water-miscible carrier. The solid ingredient is uniformly suspended in the carrier but not dissolved.
Suspension concentrates are aqueous formulations in which finely divided solid particles of the active ingredient are suspended. Such formulations include anti-settling agents and dispersing agents and may further include a wetting agent to enhance pesticidal activity as well an anti-foam and a crystal growth inhibitor. In use, these concentrates are diluted in water and normally applied as a spray to the area to be treated. The amount of active ingredient may range from 0.5% to 95% of the concentrate.
Wettable powders are in the form of finely divided particles which disperse readily in water or other liquid carriers. The particles contain an active ingredient retained in a solid matrix. Typical solid matrices include fuller’s earth, kaolin clays, silicas and other readily wet organic or inorganic solids. Wettable powders normally contain from 5% to 95% of an active ingredient plus a small amount of wetting, dispersing or emulsifying agent.
Emulsifiable concentrates are homogeneous liquid compositions dispersible in water or other liquid and may consist entirely of an active ingredient mixed with a liquid or solid emulsifying agent, or may also contain a liquid carrier, such as xylene, heavy aromatic naphthas, isophorone and other non-volatile organic solvents. In use, these concentrates are dispersed in water or other liquid and normally applied as a spray to the area to be treated. The amount of active ingredient may range from 0.5% to 95% of the concentrate.
Granular formulations include both extrudates and relatively coarse particles and are usually applied without dilution to the area in which treatment is required. Typical carriers for granular formulations include sand, fuller’s earth, attapulgite clay, bentonite clays, montmorillonite clay, vermiculite, perlite, calcium carbonate, brick, pumice, pyrophyllite, kaolin, dolomite, plaster, wood flour, ground corn cobs, ground peanut hulls, sugars, sodium chloride, sodium sulphate, sodium silicate, sodium borate, magnesia, mica, iron oxide, zinc oxide, titanium oxide, antimony oxide, cryolite, gypsum, diatomaceous earth, calcium sulphate and other organic or inorganic materials which absorb or which can be coated with the active compound.
The compositions of the invention may be employed in any conventional form, for example in the form of a twin pack, a powder for dry seed treatment (DS), an emulsion for seed treatment (ES), a flowable concentrate for seed treatment (FS), a solution for seed treatment (LS), a water dispersible powder for seed treatment (WS), a capsule suspension for seed treatment (CF), a gel for seed treatment (GF), an emulsion concentrate (EC), a suspension concentrate (SC), a suspo-emulsion (SE), a capsule suspension (CS), a water dispersible granule (WG), an emulsifiable granule (EG), an emulsion, water in oil (EG), an emulsion, oil in water (EW), a micro-emulsion (ME), an oil dispersion (OD), an oil miscible flowable (OF), an oil miscible liquid (OL), a soluble concentrate (SL), an ultra-low volume suspension (SU), an ultra-low volume liquid (UL), a technical concentrate (TK), a dispersible concentrate (DC), a wettable powder (WP) or any technically feasible formulation in combination with agriculturally acceptable adjuvants.
Such compositions may be produced in conventional manner, e.g. by mixing the active ingredients with appropriate formulation inerts (diluents, solvents, fillers and optionally other formulating ingredients such as surfactants, biocides, anti-freeze, stickers, thickeners and compounds that provide adjuvancy effects). Also conventional slow release formulations may be employed where long lasting efficacy is intended. Particularly formulations to be applied in spraying forms, such as water dispersible concentrates (e.g. EC, SC, DC, OD, SE, EW, EO and the like), wettable powders and granules, may contain surfactants such as wetting and dispersing agents and other compounds that provide adjuvancy effects, e.g. the condensation product of formaldehyde with naphthalene sulphonate, an alkylarylsulphonate, a lignin sulphonate, a fatty alkyl sulphate, and ethoxylated alkylphenol and an ethoxylated fatty alcohol.
A seed dressing formulation is applied in a manner known per se to the seeds employing the combination of the invention and a diluent in suitable seed dressing formulation form, e.g. as an aqueous suspension or in a dry powder form having good adherence to the seeds. Such seed dressing formulations are known in the art. Seed dressing formulations may contain the single active ingredients or the combination of active ingredients in encapsulated form, e.g. as slow release capsules or microcapsules.
In general, the formulations include from 0.01 to 95% by weight of active ingredients, from 0 to 20% agriculturally acceptable surfactant and 10 to 99.99% solid or liquid formulation inerts and adjuvant(s), the active ingredients consisting of at least the components (A) and (B) as defined herein, and optionally other active agents, particularly microbiocides or conservatives or the like. Concentrated forms of compositions generally contain in between about 2 and 80%, preferably between about 5 and 70% by weight of active ingredients. Application forms of formulation may for example contain from 0.01 to 20% by weight, preferably from 0.01 to 5% by weight of active ingredients. Whereas commercial products will preferably be formulated as concentrates, the end user will normally employ diluted formulations.
In a one aspect, the present invention relates to a method of controlling or preventing phytopathogenic diseases, preferably phytopathogenic fungi, on a plant or on plant propagation material and / or on harvested food crops, which comprises applying to the plant, on plant propagation material, the locus thereof, and / or on harvested food crops the composition according to the present invention.
Controlling or preventing means reducing infestation by phytopathogenic or spoilage microorganisms, especially fungal organisms, to such a level that an improvement is demonstrated.
A preferred method of controlling or preventing an infestation of crop plants by phytopathogenic microorganisms, especially fungal organisms, comprises the application of a composition according to the present invention is foliar application. The frequency of application and the rate of application will depend on the risk of infestation by the corresponding pathogen. However, the compounds A and/or B of the composition according to the present invention can also penetrate the plant through the roots via the soil (systemic action) by drenching the locus of the plant with a liquid formulation, or by applying said compounds in solid form to the soil, e.g. in granular form (soil application). In crops of water rice such granulates can be applied to the flooded rice field.
Components (A) and (B) of the composition according to the present invention may be applied simultaneously, such as in a ready to mix formulation, or in a sequential manner. The method according to the present invention comprises applying the composition according to the present invention, wherein the components (A) and (B) are applied in a simultaneous or a sequential manner on the plant, the plant propagation material or the locus thereof.
Any phytopathogenic disease can be controlled or prevented in the method according to the present invention. The composition of the invention may be used to control plant diseases caused by a broad spectrum of plant pathogens in the Basidiomycete, Ascomycete, Oomycete and/or Deuteromycete, Blasocladiomycete, Chrytidiomycete, Glomeromycete, Mucoromycete classes, and/or Oomycete classes. These pathogens may include:
Oomycetes, including Phytophthora diseases such as those caused by Phytophthora capsici, Phytophthora infestans, Phytophthora sojae, Phytophthora fragariae, Phytophthora nicotianae, Phytophthora cinnamomi, Phytophthora citricola, Phytophthora citrophthora and Phytophthora erythroseptica; Pythium diseases such as those caused by Pythium aphanidermatum, Pythium arrhenomanes, Pythium graminicola, Pythium irregulare, Pythium sylvaticum and Pythium ultimum; diseases caused by Peronosporales such as Peronospora destructor, Peronospora parasitica, Plasmopara viticola, Plasmopara halstedii, Pseudoperonospora cubensis, Albugo Candida, Sclerophthora macrospora and Bremia lactucae' and others such as Aphanomyces spp., Labyrinthula zosterae, Peronosclerospora sorghi and Sclerospora graminicola'
Ascomycetes, including blotch, spot, blast or blight diseases and/or rots for example those caused by Pleosporales such as Stemphylium solani, Stagonospora tainanensis, Spilocaea oleaginea, Setosphaeria turcica, Pyrenochaeta lycoperisici, Pleospora herbarum, Phoma destructiva, Phaeosphaeria herpotrichoides, Phaeocryptocus gaeumannii, Ophiosphaerella graminicola, Ophiobolus graminis, Leptosphaeria maculans, Hendersonia creberrima, Helminthosporium triticirepentis, Setosphaeria turcica, Drechslera glycines, Didymella bryoniae, Cycloconium oleagineum, Corynespora cassiicola, Cochliobolus sativus, Bipolaris cactivora, Venturia inaequalis, Pyrenophora teres, Pyrenophora tritici-repentis, Alternaria alternata, Alternaria brassicicola, Alternaria solani and Alternaria tomatophila, Capnodiales such as Septoria tritici, Septoria nodorum, Septoria glycines, Cercospora arachidicola, Cercospora sojina, Cercospora zeae-maydis, Cercosporella capsellae and Cercosporella herpotrichoides , Cladosporium carpophilum, Cladosporium effusum, Passalora fulva, Cladosporium oxysporum, Dothistroma septosporum, Isariopsis clavispora, Mycosphaerella fijiensis, Mycosphaerella graminicola, Mycovellosiella koepkeii, Phaeoisariopsis bataticola, Pseudocercospora vitis, Pseudocercosporella herpotrichoides, Ramularia beticola, Ramularia collo-cygni, Magnaporthales such as Gaeumannomyces graminis, Magnaporthe grisea, Pyricularia oryzae, Diaporthales such as Anisogramma anomala, Apiognomonia errabunda, Cytospora platani, Diaporthe phaseolorum, Discula destructiva, Gnomonia fructicola, Greeneria uvicola, Melanconium juglandinum, Phomopsis viticola, Sirococcus clavigignenti-juglandacearum, Tubakia dryina, Dicarpella spp. , Valsa ceratosperma, and others such as Actinothyrium graminis, Ascochyta pisi, Aspergillus flavus, Aspergillus fumigatus, Aspergillus nidulans, Asperisporium caricae, Blumeriella jaapii, Candida spp., Capnodium ramosum, Cephaloascus spp., Cephalosporium gramineum, Ceratocystis paradoxa, Chaetomium spp., Hymenoscyphus pseudoalbidus, Coccidioides spp., Cylindrosporium padi, Diplocarpon malae, Drepanopeziza campestris, Elsinoe ampelina, Epicoccum nigrum, Epidermophyton spp., Eutypa lata, Geotrichum candidum, Gibellina cerealis, Gloeocercospora sorghi, Gloeodes pomigena, Gloeosporium perennans; Gloeotinia temulenta, Griphospaeria corticola, Kabatiella lini, Leptographium microsporum, Leptosphaerulinia crassiasca, Lophodermium seditiosum, Marssonina graminicola, Microdochium nivale, Monilinia fructicola, Monographella albescens, Monosporascus cannonballus, Naemacyclus spp., Ophiostoma novo-ulmi, Paracoccidioides brasiliensis, Penicillium expansum, Pestalotia rhododendri, Petriellidium spp., Pezicula spp., Phialophora gregata, Phyllachora pomigena, Phymatotrichum omnivora, Physalospora abdita, Plectosporium tabacinum, Polyscytalum pustulans, Pseudopeziza medicaginis, Pyrenopeziza brassicae, Ramulispora sorghi, Rhabdocline pseudotsugae, Rhynchosporium secalis, Sacrocladium oryzae, Scedosporium spp., Schizothyrium pomi, Sclerotinia sclerotiorum , Sclerotinia minor, Sclerotium spp., Typhula ishikariensis, Seimatosporium mariae, Lepteutypa cupressi, Septocyta ruborum, Sphaceloma perseae, Sporonema phacidioides, Stigmina palmivora, Tapesia yallundae, Taphrina bullata, Thielviopsis basicola, Trichoseptoria fructigena, Zygophiala jamaicensis; powdery mildew diseases for example those caused by Erysiphales such as Blumeria graminis, Erysiphe polygon!, Uncinula necator, Sphaerotheca fuligena, Podosphaera leucotricha, Podospaera macularis Golovinomyces cichoracearum , Leveillula taurica, Microsphaera diffusa, Oidiopsis gossypii, Phyllactinia guttata and Oidium arachidis; molds for example those caused by Botryosphaeriales such as Dothiorella aromatica, Diplodia seriata, Guignardia bidwellii, Botrytis cinerea, Botryotinia allii, Botryotinia fabae, Fusicoccum amygdali, Lasiodiplodia theobromae, Macrophoma theicola, Macrophomina phaseolina, Phyllosticta cucurbitacearum; anthracnoses for example those caused by Glommerelales such as Colletotrichum gloeosporioides, Colletotrichum lagenarium, Colletotrichum gossypii, Glomerella cingulata, and Colletotrichum graminicola; and wilts or blights for example those caused by Hypocreales such as Acremonium strictum, Claviceps purpurea, Fusarium culmorum, Fusarium graminearum, Fusarium virguliforme, Fusarium oxysporum, Fusarium subglutinans, Fusarium oxysporum f.sp. cubense, Gerlachia nivale, Gibberella fujikuroi, Gibberella zeae, Gliocladium spp., Myrothecium verrucaria, Nectria ramulariae, Trichoderma viride, Trichothecium roseum, and Verticillium theobromae;
Basidiomycetes, including smuts for example those caused by Ustilaginales such as Ustilaginoidea virens, Ustilago nuda, Ustilago tritici, Ustilago zeae, rusts for example those caused by Pucciniales such as Cerotelium fici, Chrysomyxa arctostaphyli, Coleosporium ipomoeae, Hemileia vastatrix, Puccinia arachidis, Puccinia cacabata, Puccinia gram inis, Puccinia recondita, Puccinia sorghi, Puccinia hordei, Puccinia striiformis f.sp. Hordei, Puccinia striiformis f.sp. Secalis, Pucciniastrum coryli, or Uredinales such as Cronartium ribicola, Gymnosporangium juniperi-viginianae, Melampsora medusae, Phakopsora pachyrhizi, Phragmidium mucronatum, Physopella ampelosidis, Tranzschelia discolor and Uromyces viciae-fabae; and other rots and diseases such as those caused by Cryptococcus spp., Exobasidium vexans, Marasmiellus inoderma, Mycena spp., Sphacelotheca reiliana, Typhula ishikariensis, Urocystis agropyri, Itersonilia perplexans, Corticium invisum, Laetisaria fuciformis, Waitea circinata, Rhizoctonia solani, Thanetephorus cucurmeris, Entyloma dahliae, Entylomella microspora, Neovossia moliniae and Tilletia caries;
Blastocladiomycetes, such as Physoderma maydis;
Mucoromycetes, such as Choanephora cucurbitarum.; Mucor spp.; Rhizopus arrhizus; as well as diseases caused by other species and genera closely related to those listed above.
Phytopatogenic diseases that are advantageously controlled or prevented are phytopathogenic fungi belonging to a genus belonging to Zymoseptoria (Septoria), Puccinia, Pyricularia, Fusarium, Pythium, Monographella, Microdochium, Glomerella, Colletotrichum, and / or Botrytis, preferably belonging to a species belonging to Zymoseptoria tritici, Puccinia recondita f. sp. tritici (Puccinia triticina), Pyricularia oryzae, Pythium ultimum, Fusarium culmorum, Monographella nivalis syn. Microdochium nivale, Fusarium nivale, Septoria tritici, Glomerella lagenarium syn. Colletotrichum lagenarium (Colletotrichum orbiculare), and/ or Botrytis cinerea, preferably belonging to a species belonging to Zymoseptoria (Septoria) tritici, Puccinia recondita f. sp. tritici (Puccinia triticina), Pyricularia oryzae, Fusarium culmorum, Monographella nivalis syn. Microdochium nivale, Fusarium nivale, Glomerella lagenarium syn. Colletotrichum lagenarium (Colletotrichum orbiculare), and/ or Botrytis cinerea, preferably Zymoseptoria tritici, Puccinia recondita f. sp. tritici (Puccinia triticina), and / or Pyricularia oryzae,
The composition according to the present invention can be used in the agricultural sector and related fields of use for controlling plant pests or on non-living materials for control of spoilage microorganisms or organisms potentially harmful to man. The composition can be used to inhibit or destroy the pests that occur on plants or parts of plants (fruit, blossoms, leaves, stems, tubers, roots) of different crops of useful plants, while at the same time protecting also those parts of the plants that grow later e.g. from phytopathogenic microorganisms.
In the scope of the present invention, a plant includes a useful plant and/or crop. The term “locus” as used herein means fields in or on which plants are growing, or where seeds of cultivated plants are sown, or where seed will be placed into the soil. It includes soil, seeds, and seedlings, as well as established vegetation.
The term “plant propagation material” is understood to denote generative parts of the plant, such as seeds, which can be used for the multiplication of the latter, and vegetative material, such as cuttings or tubers, for example potatoes. There may be mentioned for example seeds (in the strict sense), roots, fruits, tubers, bulbs, rhizomes and parts of plants. Germinated plants and young plants which are to be transplanted after germination or after emergence from the soil, may also be mentioned. These young plants may be protected before transplantation by a total or partial treatment by immersion. Preferably “plant propagation material” is understood to denote seeds.
Useful plants and / or target crops in accordance with the invention include conventional as well as genetically enhanced or engineered varieties such as, for example, insect resistant (e.g. Bt. and VIP varieties) as well as disease resistant, herbicide tolerant (e.g. glyphosate- and glufosinate-resistant maize varieties) and nematode tolerant varieties. Useful plants include plants transformed by the use of recombinant DNA techniques, for example, to be capable of synthesising one or more selectively acting toxins, such as are known, for example, from toxin-producing bacteria. Examples of toxins which can be expressed include 8-endotoxins, vegetative insecticidal proteins (Vip), insecticidal proteins of bacteria colonising nematodes, and toxins produced by scorpions, arachnids, wasps and fungi.
Plants, including target crops and/or useful plants to be protected in a method of the invention typically comprise perennial and annual crops, such as berry plants for example blackberries, blueberries, cranberries, raspberries and strawberries; cereals for example barley, maize (corn), millet, oats, rice, rye, sorghum triticale and wheat; fibre plants for example cotton, flax, hemp, jute and sisal; field crops for example sugar and fodder beet, coffee, hops, mustard, oilseed rape (canola), poppy, sugar cane, sunflower, tea and tobacco; fruit trees for example apple, apricot, avocado, banana, cherry, citrus, nectarine, peach, pear and plum; grasses for example Bermuda grass, bluegrass, bentgrass, centipede grass, fescue, ryegrass, St. Augustine grass and Zoysia grass; herbs such as basil, borage, chives, coriander, lavender, lovage, mint, oregano, parsley, rosemary, sage and thyme; legumes for example beans, lentils, peas and soya beans; nuts for example almond, cashew, ground nut, hazelnut, peanut, pecan, pistachio and walnut; palms for example oil palm; ornamentals for example flowers, shrubs and trees; other trees, for example cacao, coconut, olive and rubber; vegetables for example asparagus, aubergine, broccoli, cabbage, carrot, cucumber, garlic, lettuce, marrow, melon, okra, onion, pepper, potato, pumpkin, rhubarb, spinach and tomato; and vines for example grapes.
A plant in a method according to the present invention preferably comprises wheat, barley, rice, corn, soya, sugar beet, banana, tomato, cucumber, and / or groundnut. Preferably the plant comprises wheat and / or rice. The composition comprising components (A) and (B) as defined herein may also be applied to seeds (coating) by impregnating the seeds or tubers either with a liquid formulation of the composition of the invention or coating them with a solid formulation.
A method of controlling or preventing a phytopathogenic disease includes spraying, atomizing, dusting, brushing on, dressing, scattering or pouring - which are to be selected to suit the intended aims of the prevailing circumstances. Typical rates of concentration are between 0.1 and 1000 ppm, preferably between 0.1 and 500 ppm, of active ingredient. The rate of application per hectare is preferably 1 g to 2000 g of active ingredient per hectare, more preferably 10 to 1000 g/ha, most preferably 10 to 600 g/ha. When used as seed drenching agent, convenient dosages are from 10mg to 1g of active substance per kg of seeds. When the composition of the present invention is used for treating seed, rates of 0.001 to 50 g of the composition per kg of seed, preferably from 0.01 to 10g per kg of seed are generally sufficient.
In one embodiment, the method comprises applying an effective amount of component A comprising Streptomyces as disclosed herein above, wherein the effective amount comprises from 2*102 to 5*1017 , from 3*102 to 5*1016, from 5*102 to 5*1015 , from 2*102 to 5*1014, from 2*102 to 5*1013, preferably from 5*102 to 5*1012, from 1*103to 5*1011 , from 5*103 to 1*1011 , from 1*104to 5*1010, from 5*104 to 1*1010 , from 1*105 to 5*109, from 5*105 to 1*109 , from 1*106 to 5*108, from 5*106 to 1*108 colony forming unit (cfu) of the Streptomyces such as Streptomyces chrestomyceticus, per hectare.
An effective amount of component A comprising Streptomyces as disclosed herein above, comprises from 1 g to 10 kg / per hectare (ha), such as from 5 g to 5 kg, such as from 10 g to 1 kg / ha, such as from 50 g to 800 g / ha, such as from 100 g to 700 g / ha, such as from 200 to 600 g / ha. The weight in g and kg is dry weight of the Streptomyces.
In one embodiment the method according to the present invention comprises treating plant propagation material wherein the plant propagation material is seed and the effective amount of component (A) comprises from 5x102 to 5x1015, from 2x103 to 5x1014, from 5x103 to 5x1013, from 2x105 to 5x1012, preferably from 5*102 to 5*1012, from 1*103to 5*1011 , from 5*103 to 1*1011, from 1*104to 5*101°, from 5*104 to 1 *101° , from 1 *105 to 5*109, from 5*105 to 1 *109 , from 1 *106 to 5*108, from 5*106 to 1 *108 colony forming unit (cfu) of the Streptomyces chrestomyceticus as disclosed herein per kg of seed.
When the plant propagation material is seed, an effective amount of component (A) comprising Streptomyces chrestomyticus thereof as disclosed herein above, may also comprise from 0.001 g to 100 g / per kg of seeds, such as from 0.005 g to 80 g / kg seeds, such as from 0.01 g to 50 g / kg seeds, such as from 0.5 g to 10 g / kg seeds, The weight in g dry weight of component (A) comprising Streptomyces chrestomyceticus per kg dry weight of seeds.
Suitably, the composition according to the present invention can be applied either preventative, meaning prior to disease development or curative, meaning after disease development.
In one further aspect the present invention relates to a plant, plant propagation material, the locus thereof and/or a harvested food crop comprising a composition according to the present invention. The description related to the features disclosed herein above are also applicable to this aspect of the invention. Disclosed herein is also a composition comprising component (A) and a component (B), wherein component (A) comprises a strain of Streptomyces chrestomyceticus , preferably Streptomyces sp. Saigon413 deposited with the Westerdijk Institute under accession number CBS149411 , and component (B) comprises any of the agents, fungicides, plant extracts and products according to the list herein below:
(1) antibacterial agents selected from the group of
(1.1) bacteria, such as Bacillus subtilis, in particular strain QST713/AQ713 (available as SERENADE OPTI or SERENADE ASO from Bayer CropScience LP, US, having NRRL Accession No. B21661 , U.S. Patent No. 6,060,051); Bacillus sp., in particular strain D747 (available as DOUBLE NICKEL® from Kumiai Chemical Industry Co., Ltd.), having Accession No. FERM BP-8234, U.S. Patent No. 7,094,592; Bacillus pumilus, in particular strain BU F-33, having NRRL Accession No. 50185 (available as part of the CARTISSA® product from BASF, EPA Reg. No. 71840-19); Bacillus subtilis var. amyloliquefaciens strain FZB24 having Accession No. DSM 10271 (available from Novozymes as TAEGRO® or TAEGRO® ECO (EPA Registration No. 70127-5)); a PaeniBacillus sp. strain having Accession No. NRRL B-50972 or Accession No. NRRL B-67129, WO 2016/154297; Bacillus subtilis strain BU1814, (available as VELONDIS® PLUS, VELONDIS® FLEX and VELONDIS® EXTRA from BASF SE); Bacillus mojavensis strain R3B (Accession No. NCAIM (P) B001389) (WO 2013/034938) from Certis USA LLC, a subsidiary of Mitsui & Co.; Bacillus subtilis CX-9060 from Certis USA LLC, a subsidiary of Mitsui & Co.; Paenibacillus polymyxa, in particular strain AC-1 (e.g. TOPSEED® from Green Biotech Company Ltd.); Pseudomonas proradix (e.g. PRORADIX® from Sourcon Padena); Pantoea agglomerans, in particular strain E325 (Accession No. NRRL B-21856) (available as BLOOMTIME BIOLOGICAL™ FD BIOPESTICIDE from Northwest Agri Products); and
(1.2) fungi, such as Aureobasidium pullulans, in particular blastospores of strain DSM14940, blastospores of strain DSM 14941 or mixtures of blastospores of strains DSM14940 and DSM14941 (e.g., BOTECTOR® and BLOSSOM PROTECT® from bio-ferm, CH); Pseudozyma aphidis (as disclosed in WO2011/151819 by Yissum Research Development Company of the Hebrew University of Jerusalem); Saccharomyces cerevisiae, in particular strains CNCM No. 1-3936, CNCM No. 1-3937, CNCM No. 1-3938 or CNCM No. 1-3939 (WO 2010/086790) from Lesaffre et Compagnie, FR;
(2) biological fungicides selected from the group of: (2.1) bacteria, for example Bacillus subtilis, in particular strain QST713/AQ713 (available as SERENADE OPTI or SERENADE ASO from Bayer CropScience LP, US, having NRRL Accession No. B21661 and described in U.S. Patent No. 6,060,051); Bacillus pumilus, in particular strain QST2808 (available as SONATA® from Bayer CropScience LP, US, having Accession No. NRRL B-30087 and described in U.S. Patent No. 6,245,551); Bacillus pumilus, in particular strain GB34 (available as Yield Shield® from Bayer AG, DE); Bacillus pumilus, in particular strain BU F-33, having NRRL Accession No. 50185 (available as part of the CARTISSA product from BASF, EPA Reg. No. 71840-19); Bacillus amyloliquefaciens, in particular strain D747 (available as Double Nickel™ from Kumiai Chemical Industry Co., Ltd., having accession number FERM BP-8234, US Patent No. 7,094,592); Bacillus subtilis Y1336 (available as BIOBAC® WP from Bion-Tech, Taiwan, registered as a biological fungicide in Taiwan under Registration Nos. 4764, 5454, 5096 and 5277); Bacillus subtilis strain MBI 600 (available as SUBTILEX from BASF SE), having Accession Number NRRL B-50595, U.S. Patent No. 5,061 ,495; Bacillus subtilis strain GB03 (available as Kodiak® from Bayer AG, DE); Bacillus subtilis var. amyloliquefaciens strain FZB24 having Accession No. DSM 10271 (available from Novozymes as TAEGRO® or TAEGRO® ECO (EPA Registration No. 70127-5)); Bacillus mycoides, isolate J , having Accession No. B-30890 (available as BMJ TGAI® or WG and LifeGard™ from Certis USA LLC, a subsidiary of Mitsui & Co.); Bacillus licheniformis, in particular strain SB3086 , having Accession No. ATCC 55406, WO 2003/000051 (available as ECOGUARD® Biofungicide and GREEN RELEAF™ from Novozymes); a Paenibacillus sp. strain having Accession No. NRRL B-50972 or Accession No. NRRL B-67129, WO 2016/154297; Bacillus subtilis strain BU1814, (available as VELONDIS® PLUS, VELONDIS® FLEX and VELONDIS® EXTRA from BASF SE); Bacillus subtilis CX-9060 from Certis USA LLC, a subsidiary of Mitsui & Co.; Bacillus amyloliquefaciens strain F727 (also known as strain MB1110) (NRRL Accession No. B- 50768; WO 2014/028521) (STARGUS® from Marrone Bio Innovations); Bacillus amyloliquefaciens strain FZB42, Accession No. DSM 23117 (available as RHIZOVITAL® from ABiTEP, DE); Bacillus licheniformis FMCH001 and Bacillus subtilis FMCH002 (QUARTZO® (WG) and PRESENCE® (WP) from FMC Corporation); Bacillus mojavensis strain R3B (Accession No. NCAIM (P) B001389) (WO 2013/034938) from Certis USA LLC, a subsidiary of Mitsui & Co.; Paenibacillus polymyxa ssp. plantarum (WO 2016/020371) from BASF SE; Paenibacillus epiphyticus (WO 2016/020371) from BASF SE; Pseudomonas chlororaphis strain AFS009, havingf Accession No. NRRL B-50897, WO 2017/019448 (e.g., HOWLER™ and ZIO® from AgBiome Innovations, US); Pseudomonas chlororaphis, in particular strain MA342 (e.g. CEDOMON®, CERALL®, and CEDRESS® by Bioagri and Koppert); Streptomyces lydicus strain WYEC108 (also known as Streptomyces lydicus strain WYCD108US) (ACTINO-IRON® and ACTINOVATE® from Novozymes); Agrobacterium radiobacter strain K84 (e.g. GALLTROL-A® from AgBioChem, CA); Agrobacterium radiobacter strain K1026 (e.g. NOGALL™ from BASF SE); Bacillus subtilis KTSB strain (FOLIACTIVE® from Donaghys); Bacillus subtilis IAB/BS03 (AVIV™ from STK Bio-Ag Technologies); Bacillus subtilis strain Y1336 (available as BIOBAC® WP from Bion-Tech, Taiwan, registered as a biological fungicide in Taiwan under Registration Nos. 4764, 5454, 5096 and 5277); Bacillus amyloliquefaciens isolate B246 (e.g. AVOGREEN™ from University of Pretoria); Bacillus methylotrophicus strain BAC-9912 (from Chinese Academy of Sciences’ Institute of Applied Ecology); Pseudomonas proradix (e.g. PRORADIX® from Sourcon Padena); Streptomyces griseoviridis strain K61 (also known as Streptomyces galbus strain K61) (Accession No. DSM 7206) (MYCOSTOP® from Verdera; PREFENCE® from BioWorks; cf. Crop Protection 2006, 25, 468-475); Pseudomonasfluorescens strain A506 (e.g. BLIGHTBAN® A506 by NuFarm); and
(2.2) fungi, for example: Coniothyrium minitans, in particular strain CON/M/91-8 (Accession No. DSM9660; e.g. Contans ® from Bayer CropScience Biologies GmbH); Metschnikowia fructicola, in particular strain NRRL Y-30752; (B2.2.3) Microsphaeropsis ochracea; Trichoderma atroviride, in particular strain SC1 (having Accession No. CBS 122089, WO 2009/116106 and U.S. Patent No. 8,431 ,120 (from Bi- PA)), strain 77B (T77 from Andermatt Biocontrol) or strain LU132 (e.g. Sentinel from Agrimm Technologies Limited); Trichoderma harzianum strain T-22 (e.g. Trianum-P from Andermatt Biocontrol or Koppert) or strain Cepa SimbT5 (from Simbiose Agro); Gliocladium roseum (also known as Clonostachys rosea f rosea), in particular strain 321 U from Adjuvants Plus, strain ACM941 as disclosed in Xue (Efficacy of Clonostachys rosea strain ACM941 and fungicide seed treatments for controlling the root tot complex of field pea, Can Jour Plant Sci 83(3): 519-524), or strain IK726 (Jensen DF, et al. Development of a biocontrol agent for plant disease control with special emphasis on the near commercial fungal antagonist Clonostachys rosea strain ’IK726’; Australas Plant Pathol. 2007;36:95-101); Talaromyces flavus, strain V117b; Trichoderma viride, in particular strain B35 (Pietr et al., 1993, Zesz. Nauk. A R w Szczecinie 161 : 125-137); Trichoderma asperellum, in particular strain SKT-1 , having Accession No. FERM P-16510 (e.g. ECO-HOPE® from Kumiai Chemical Industry), strain T34 (e.g. T34 Biocontrol by Biocontrol Technologies S.L., ES) or strain ICC 012 from Isagro; Trichoderma atroviride, strain CNCM 1-1237 (e.g. Esquive® WP from Agrauxine, FR); Trichoderma atroviride, strain no. V08/002387; Trichoderma atroviride, strain NMI no. V08/002388; Trichoderma atroviride, strain NMI no. V08/002389; Trichoderma atroviride, strain NMI no. V08/002390; Trichoderma atroviride, strain LC52 (e.g. Tenet by Agrimm Technologies Limited); Trichoderma atroviride, strain ATCC 20476 (IMI 206040); Trichoderma atroviride, strain T11 (IMI352941Z CECT20498); Trichoderma harmatum; Trichoderma harzianum; Trichoderma harzianum rifai T39 (e.g. Trichodex® from Makhteshim, US); Trichoderma asperellum, in particular, strain kd (e.g. T-Gro from Andermatt Biocontrol); Trichoderma harzianum, strain ITEM 908 (e.g. Trianum-P from Koppert); Trichoderma harzianum, strain TH35 (e.g. Root-Pro by Mycontrol); Trichoderma virens (also known as Gliocladium virens), in particular strain GL-21 (e.g. SoilGard by Certis, US); Trichoderma viride, strain TV1 (e.g. Trianum-P by Koppert); Ampelomyces quisqualis, in particular strain AQ 10 (e.g. AQ 10® by IntrachemBio Italia); Aureobasidium pullulans, in particular blastospores of strain DSM14940; Aureobasidium pullulans, in particular blastospores of strain DSM 14941 ; Aureobasidium pullulans, in particular mixtures of blastospores of strains DSM14940 and DSM 14941 (e.g. Botector® by bio-ferm, CH); Cladosporium cladosporioides, strain H39, having Accession No. CBS122244, US 2010/0291039 (by Stichting Dienst Landbouwkundig Onderzoek); Gliocladium catenulatum (Synonym: Clonostachys rosea f. catenulate) strain J1446 (e.g. Prestop ® by Lallemand); Lecanicillium lecanii (formerly known as Verticillium lecanii) conidia of strain KV01 (e.g. Vertalec® by Koppert/Arysta); Penicillium vermiculatum;P/ch/a anomala, strain WRL-076 (NRRL Y-30842), U.S. Patent No. 7,579,183; Trichoderma atroviride, strain SKT- 1 (FERM P-16510), JP Patent Publication (Kokai) 11-253151 A; Trichoderma atroviride, strain SKT-2 (FERM P-16511), JP Patent Publication (Kokai) 11-253151 A; Trichoderma atroviride, strain SKT-3 (FERM P-17021), JP Patent Publication (Kokai) 11-253151 A; Trichoderma gamsii (formerly T. viride), strain ICC080 (IMI CC 392151 CABI, e.g. BioDerma by AGROBIOSOL DE MEXICO, S.A. DE C.V.); Trichoderma harzianum, strain DB 103 (available as T-GRO® 7456 by Dagutat Biolab); Trichoderma polysporum, strain IMI 206039 (e.g. Binab TF WP by BINAB Bio-Innovation AB, Sweden); Trichoderma stromaticum, having Accession No. Ts3550 (e.g. Tricovab by CEPLAC, Brazil); Ulocladium oudemansii strain U3, having Accession No. NM 99/06216 (e.g., BOTRY-ZEN® by Botry-Zen Ltd, New Zealand and BOTRYSTOP® from BioWorks, Inc.); Verticillium albo-atrum (formerly V. dahliae), strain WCS850 having Accession No. WCS850, deposited at the Central Bureau for Fungi Cultures (e.g., DUTCH TRIG® by Tree Care Innovations); Verticillium chlamydosporium; mixtures of Trichoderma asperellum strain ICC 012 (also known as Trichoderma harzianum ICC012), having Accession No. CABI CC IMI 392716 and Trichoderma gamsii (formerly T. viride) strain ICC 080, having Accession No. IMI 392151 (e.g., BIO-TAM™ from Isagro USA, Inc. and BIODERMA® by Agrobiosol de Mexico, S.A. de C.V.); Trichoderma asperelloides JM41 R (Accession No. NRRL B-50759) (TRICHO PLUS® from BASF SE); Aspergillus flavus strain NRRL 21882 (products known as AFLA-GUARD® from Syngenta/ChemChina); Chaetomium cupreum (Accession No. CABI 353812) (e.g. BIOKUPRUM™ by AgriLife); Saccharomyces cerevisiae, in particular strain LASO2 (from Agro-Levures et Derives), strain LAS117 cell walls (CEREVISANE® from Lesaffre; ROMEO® from BASF SE), strains CNCM No. 1-3936, CNCM No. 1-3937, CNCM No. 1-3938, CNCM No. 1-3939 (WO 2010/086790) from Lesaffre et Compagnie, FR; Trichoderma virens strain G-41 , formerly known as Gliocladium virens (Accession No. ATCC 20906) (e.g., ROOTSHIELD® PLUS WP and TURFSHIELD® PLUS WP from BioWorks, US); Trichoderma hamatum, having Accession No. ATCC 28012; Ampelomyces quisqualis strain AQ10, having Accession No. CNCM 1-807 (e.g., AQ 10® by IntrachemBio Italia); Phlebiopsis gigantea strain VRA 1992 (ROTSTOP® C from Danstar Ferment); Penicillium steckii (DSM 27859; WO 2015/067800) from BASF SE; Chaetomium globosum (available as RIVADIOM® by Rivale); Cryptococcus flavescens, strain 3C (NRRL Y-50378); (B2.2.99) Dactylaria Candida Dilophosphora alopecuri (available as TWIST FUNGUS®); Fusarium oxysporum, strain Fo47 (available as FUSACLEAN® by Natural Plant Protection); Pseudozyma flocculosa, strain PF-A22 UL (available as SPORODEX® L by Plant Products Co., CA); (2.2.103) Trichoderma gamsii (formerly T. viride), strain ICC 080 (IMI CC 392151 CABI) (available as BIODERMA® by AGROBIOSOL DE MEXICO, S.A. DE C.V.); Trichoderma fertile (e.g. product TrichoPlus from BASF); Muscodor roseus, in particular strain A3-5 (Accession No. NRRL 30548); Simplicillium lanosoniveum;
(3) biological control agents having an effect for improving plant growth and/or plant health which may be combined in the compound combinations as disclosed herein including
(3.1) bacteria selected from the group consisting of Bacillus pumilus, in particular strain QST2808 (having Accession No. NRRL No. B-30087); Bacillus subtilis, in particular strain QST713/AQ713 (having NRRL Accession No. B-21661 and described in U.S. Patent No. 6,060,051 ; available as SERENADE® OPTI or SERENADE® ASO from Bayer CropScience LP, US); Bacillus subtilis, in particular strain AQ30002 (having Accession Nos. NRRL B-50421 and described in U.S. Patent Application No. 13/330,576); Bacillus subtilis, in particular strain AQ30004 (and NRRL B-50455 and described in U.S. Patent Application No. 13/330,576); Sinorhizobium meliloti strain NRG-185-1 (NITRAGIN® GOLD from Bayer CropScience); Bacillus subtilis strain BU1814, (available as TEQUALIS® from BASF SE); Bacillus subtilis rm303 (RHIZOMAX® from Biofilm Crop Protection); Bacillus amyloliquefaciens pm414 (LOLI-PEPTA® from Biofilm Crop Protection); Bacillus mycoides BT155 (NRRL No. B-50921), Bacillus mycoides EE118 (NRRL No. B-50918), Bacillus mycoides EE141 (NRRL No. B-50916), Bacillus mycoides BT46-3 (NRRL No. B- 50922), Bacillus cereus family member EE128 (NRRL No. B-50917), Bacillus thuringiensis BT013A (NRRL No. B-50924) also known as Bacillus thuringiensis 4Q7, Bacillus cereus family member EE349 (NRRL No. B-50928), Bacillus amyloliquefaciens SB3281 (ATCC # PTA-7542; WO 2017/205258), Bacillus amyloliquefaciens TJ1000 (available as QUIKROOTS® from Novozymes); Bacillus firmus, in particular strain CNMC 1-1582 (e.g. VOTIVO®from BASF SE); Bacillus pumilus, in particular strain GB34 (e.g. YIELD SHIELD® from Bayer Crop Science, DE); Bacillus amyloliquefaciens, in particular strain IN937a; Bacillus amyloliquefaciens, in particular strain FZB42 (e.g. RHIZOVITAL® from ABiTEP, DE); Bacillus amyloliquefaciens BS27 (Accession No. NRRL B-5015); a mixture of Bacillus licheniformis FMCH001 and Bacillus subtilis FMCH002 (available as QUARTZO® (WG), PRESENCE® (WP) from FMC Corporation); Bacillus cereus, in particular strain BP01 (ATCC 55675; e.g. MEPICHLOR® from Arysta Lifescience, US); Bacillus subtilis, in particular strain MBI 600 (e.g. SUBTILEX® from BASF SE); Bradyrhizobium japonicum (e.g. OPTIMIZE® from Novozymes); Mesorhizobium cicer (e.g., NODULATOR from BASF SE); Rhizobium leguminosarium biovar viciae (e.g., NODULATOR from BASF SE); Delftia acidovorans, in particular strain RAY209 (e.g. BIOBOOST® from Brett Young Seeds); Lactobacillus sp. (e.g. LACTOPLANT® from LactoPAFI); Paenibacillus polymyxa, in particular strain AC-1 (e.g. TOPSEED® from Green Biotech Company Ltd.); Pseudomonas proradix (e.g. PRORADIX® from Sourcon Padena); Azospirillum brasilense (e.g., VIGOR® from KALO, Inc.); Azospirillum lipoferum (e.g., VERTEX-IF™ from TerraMax, Inc.); a mixture of Azotobacter vinelandii and Clostridium pasteurianum (available as INVIGORATE® from Agrinos); Pseudomonas aeruginosa, in particular strain PN1 ; Rhizobium leguminosarum, in particular bv. viceae strain Z25 (Accession No. CECT 4585); Azorhizobium caulinodans, in particular strain ZB-SK-5; Azotobacter chroococcum , in particular strain H23; Azotobacter vinelandii, in particular strain ATCC 12837; Bacillus siamensis, in particular strain KCTC 13613T; Bacillus tequilensis, in particular strain NII-0943; Serratia marcescens, in particular strain SRM (Accession No. MTCC 8708); Thiobacillus sp. (e.g. CROPAID® from Cropaid Ltd UK); and
(3.2) fungi selected from the group consisting of Purpureocillium lilacinum (previously known as Paecilomyces lilacinus) strain 251 (AGAL 89/030550; e.g. BioActfrom Bayer CropScience Biologies GmbH) Penicillium bilaii, strain ATCC 22348 (e.g. JumpStart® from Acceleron BioAg), Talaromyces flavus, strain V117b; Trichoderma atro viride strain CNCM 1-1237 (e.g. Esquive® WP from Agrauxine, FR), Trichoderma viride, e.g. strain B35 (Pietr et al., 1993, Zesz. Nauk. A R w Szczecinie 161 : 125-137); Trichoderma atroviride strain LC52 (also known as Trichoderma atroviride strain LU132; e.g. Sentinel from Agrimm Technologies Limited); Trichoderma atroviride strain SC1 described in International Application No. PCT/IT2008/000196); Trichoderma asperellum strain kd (e.g. T-Gro from Andermatt Biocontrol); Trichoderma asperellum strain Eco-T (Plant Health Products, ZA); Trichoderma harzianum strain T-22 (e.g. Trianum-P from Andermatt Biocontrol or Koppert); Myrothecium verrucaria strain AARC-0255 (e.g. DiTera™ from Valent Biosciences); Penicillium bilaii strain ATCC ATCC20851 ; Pythium oligandrum strain M1 (ATCC 38472; e.g. Polyversum from Bioprepraty, CZ); Trichoderma virens strain GL-21 (e.g. SoilGard® from Certis, USA); Verticillium albo-atrum (formerly V. dahliae) strain WCS850 (CBS 276.92; e.g. Dutch Trig from Tree Care Innovations); Trichoderma atroviride, in particular strain no. V08/002387, strain no. NMI No. V08/002388, strain no. NMI No. V08/002389, strain no. NMI No. V08/002390; Trichoderma harzianum strain ITEM 908; Trichoderma harzianum, strain TSTh20; Trichoderma harzianum strain 1295- 22; Pythium oligandrum strain DV74; Rhizopogon amylopogon (e.g. comprised in Myco-Sol from Helena Chemical Company); Rhizopogon fulvigleba (e.g. comprised in Myco-Sol from Helena Chemical Company); or; Trichoderma virens strain GI-3;
(4) insecticidally active biological control agents selected from
(4.1) bacteria selected from the group consisting of Agrobacterium radiobacter strain K84 (Galltrol from AgBiochem Inc.), Bacillus amyloliquefaciens, in particular strain PTS-4838 (e.g. AVEO from Valent Biosciences, US); Bacillus firmus, in particular strain CNMC 1-1582 (e.g. VOTIVO® from BASF SE); Bacillus thuringiensis subsp. aizawai, in particular strain ABTS-1857 (SD-1372; e.g. XENTARI® from Valent BioSciences); Bacillus mycoides, isolate J. (e.g. BmJ from Certis USA LLC, a subsidiary of Mitsui & Co.); Bacillus sphaericus, in particular Serotype H5a5b strain 2362 (strain ABTS-1743) (e.g. VECTOLEX® from Valent BioSciences, US); Bacillus thuringiensis subsp. kurstaki strain BMP 123 from Becker Microbial Products, IL (BARITONE from Bayer CropScience); Bacillus thuringiensis subsp. aizawai, in particular serotype H-7 (e.g. FLORBAC® WG from Valent BioSciences, US); Bacillus thuringiensis subsp. kurstaki strain HD-1 (e.g. DIPEL® ES from Valent BioSciences, US); Bacillus thuringiensis subsp. kurstaki strain BMP 123 by Becker Microbial Products, IL; Bacillus thuringiensis israelensis strain BMP 144 (e.g. AQUABAC® by Becker Microbial Products IL); Brevibacillus laterosporus (LATERAL from Ecolibrium Biologicals); Burkholderia spp., in particular Burkholderia rinojensis strain A396 (also known as Burkholderia rinojensis strain MBI 305) (Accession No. NRRL B-50319; WO 2011/106491 and WO 2013/032693; e.g. MBI206 TGAI and ZELTO® from Marrone Bio Innovations); Chromobacterium subtsugae, in particular strain PRAA4-1T (MBI-203; e.g. GRANDEVO® from Marrone Bio Innovations); Lecanicillium muscarium Ve6 (MYCOTAL from Koppert); Paenibacillus popilliae (formerly Bacillus popilliae; e.g. MILKY SPORE POWDER™ and MILKY SPORE GRANULAR™ from St. Gabriel Laboratories); Pasteuria nishizawae strain Pn1 (CLARIVA from Syngenta/ChemChina); Bacillus thuringiensis subsp. israelensis (serotype H-14) strain AM65-52 (Accession No. ATCC 1276) (e.g. VECTOBAC® by Valent BioSciences, US); Bacillus thuringiensis var. kurstaki strain EVB-113-19 (e.g., BIOPROTEC® from AEF Global); Bacillus thuringiensis subsp. tenebrionis strain NB 176 (SD-5428; e.g. NOVODOR® FC from BioFa DE); Bacillus thuringiensis var. japonensis strain Buibui; Bacillus thuringiensis subsp. kurstaki strain ABTS 351 ; Bacillus thuringiensis subsp. kurstaki strain PB 54; Bacillus thuringiensis subsp. kurstaki strain SA 11 (JAVELIN from Certis, US); Bacillus thuringiensis subsp. kurstaki strain SA 12 (THURICIDE, Certis, USA)); Bacillus thuringiensis subsp. kurstaki strain EG 2348 (LEPINOX, Certis USA); Bacillus thuringiensis subsp. kurstaki strain EG 7841 (CRYMAX from Certis, US); Bacillus thuringiensis var. Colmeri (e.g. TIANBAOBTC by Changzhou Jianghai Chemical Factory); Bacillus thuringiensis subsp. aizawai strain GC-91 ; Serratia entomophila (e.g. INVADE® by Wrightson Seeds); Serratia marcescens, in particular strain SRM (Accession No. MTCC 8708); Trichoderma asperellum (TRICHODERMAX from Novozymes); and Wolbachia pipientis ZAP strain (e.g., ZAP MALES® from MosquitoMate); and
(4.2) fungi selected from the group consisting of Isaria fumosorosea (previously known as Paecilomyces fumosoroseus) strain apopka 97 PREFERAL from SePRO; ; Beauveria bassiana strain ATCC 74040 (e.g. NATURALIS® from Intrachem Bio Italia); Beauveria bassiana strain GHA (Accession No. ATCC74250; e.g. BOTANIGUARD® ES and MYCONTROL-O® from Laverlam International Corporation); Zoophtora radicans; Metarhizium robertsii 15013-1 (deposited under NRRL accession number 67073), Metarhizium robertsii 23013-3 (deposited under NRRL accession number 67075), and Metarhizium anisopliae 3213-1 (deposited under NRRL accession number 67074) (WO 2017/066094; Pioneer Hi-Bred International); Paecilomyces lilacinus strain 251 (MELOCON from Certis, US); Beauveria bassiana strain ATP02 (Accession No. DSM 24665).
(5) Viruses selected from the group consisting of Adoxophyes orana (summer fruit tortrix) granulosis virus (GV), Cydia pomonella (codling moth) granulosis virus (GV), Helicoverpa armigera (cotton bollworm) nuclear polyhedrosis virus (NPV), Spodoptera exigua (beet armyworm) mNPV, Spodoptera frugiperda (fall armyworm) mNPV, and Spodoptera littoralis (African cotton leafworm) NPV;
(6) Bacteria and fungi which can be added as ’inoculant’ to plants or plant parts or plant organs and which, by virtue of their particular properties, promote plant growth and plant health selected from the group consisting of Agrobacterium spp., Azorhizobium caulinodans, Azospirillum spp., Azotobacter spp., Bradyrhizobium spp., Burkholderia spp., in particular Burkholderia cepacia (formerly known as Pseudomonas cepacia), Gigaspora spp., or Gigaspora monosporum, Glomus spp., Laccaria spp., LactoBacillus buchneri, Paraglomus spp., Pisolithus tinctorus, Pseudomonas spp., Rhizobium spp., in particular Rhizobium trifolii, Rhizopogon spp., Scleroderma spp., Suillus spp., and Streptomyces spp.;
(7) Plant extracts and products formed by microorganisms including proteins and secondary metabolites which can be used as biological control agents, such as Allium sativum (NEMGUARD from Eco-Spray; BRALIC from ADAMA), Artemisia absinthium, azadirachtin (e.g. AZATIN XL from Certis, US), Biokeeper WP, Cassia nigricans, Celastrus angulatus, Chenopodium anthelminticum, chitin, Armour-Zen, Dryopteris filix-mas, Equisetum arvense, Fortune Aza, Fungastop, Heads Up (Chenopodium quinoa saponin extract), PROBLAD (naturally occurring Blad polypeptide from Lupin seeds); Certis EU, FRACTURE (naturally occurring Blad polypeptide from Lupin seeds); FMC, Pyrethrum/Pyrethrins, Quassia amara, Quercus, Quillaja extract (QL AGRI 35 from BASF), Reynoutria sachalinensis extract (REGALLIA / REGALIA MAXX from Marrone Bio), "Requiem ™ Insecticide", rotenone, ryan/a/ryanodine, Symphytum officinale, Tanacetum vulgare, thymol Thymol mixed with Geraniol (CEDROZ from Eden Research); Thymol mixed with Geraniol and Eugenol (MEVALONE from Eden Research);, Triact 70, TriCon, Tropaeulum majus, Melaleuca alternifolia extract (TIMOREX GOLD from STK), Urtica dioica, Veratrin, Viscum album, Brassicaceae extract, in particular oilseed rape powder or mustard powder. FIGURES
Figure 1. 1 D 1H NMR spectrum of the compound I in D2O at 600 MHz
Figure 2. 1 D 13C NMR spectrum of the compound I in D2O at 600 MHz
Figure 3. 2D Dept Edited 1 H-13C HSQC NMR Spectrum of the compound I in D2<D.at 600 MHz showing the positive (CH) signals
Figure 4. 2D Dept Edited 1 H-13C HSQC NMR Spectrum of the compound I in D2O.at 600 MHz showing the negative (CH2) signals
Figure 5. Spectrum of light absorption (UV-VIS) 200-400nm of a lipopeptide according to Formula II (a), Formula II (b) or Lipopeptin A
Figure 6. LC-ESI-MS/MS spectrum of precursor 1204.6 m/z (M+H)+ for a lipopeptide of Formula ll(a) depicting fragment peaks consistent with amino acids: aspartic acid, hydroxy-glutamine, serine, methylasparagine, methyl-phenylalanine
Figure 7. LC-ESI-MS/MS/MS spectrum of precursor 294.2 m/z for a lipoeptide of Formula 11 (a) depicting peaks consistent with the molecule C14H25-OH2-C4H5ON
Figure 8. The upfield region of the 1 D 1H NMR spectrum of a lipopeptide according to Formula ll(a) in CD3OD at 600 MHz
Figure 9. The downfield region of the 1 D 1H NMR spectrum of a lipopeptide according to Formula 11 (a) in CD3OD at 600 MHz
Figure 10. Spectrum of light absorption (UV-VIS) 200-500nm of polyene compound
EXAMPLES
Example 1.1. Source and fermentation
Fermentation of Streptomyces sp.
Streptomyces sp. Saigon413, isolated in Vietnam before 1961 , was deposited at the Westerdijk institute Uppsalalaan 8, 3584 CT Utrecht, the Netherlands, under accession number CBS149411 . The deposit was made by Syngenta Ltd., Jealott’s Hill Research International Centre, Bracknell, Berkshire, RG42 6EY, UK under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure.
The Streptomyces was cultivated in Erlenmeyer flasks with a liquid medium consisting of (g / I) casein hydrolysate 10, glucose 40, K2HPO4 1.25, soytone 2, tryptone, 8 and incubated at 28°C in an incubator shaking 150 rpm with 25 mm throw for 4 days. Large scale fermentations
For large scale production, standard procedures were applied for cultivating Streptomyces chrestomyceticus CBS149411 to high cell density using fed-batch fermentation. After harvesting, the broth was spray dried or freeze dried according to methods known to a person skilled in the art.
The final product (TGAI) after spray- or freeze drying had a cell count of 1*105 to 1*1013 CFU/g dry mass. TGAI: technical grade active ingredient (unformulated product).
For the in vitro tests (Example 2) with a second compound, spray-dried or freeze-dried
S. chrestomyceticus CBS149411 (Streptomyces Saigon 413) TGAI at 107 to 1010 cfu / g TGAI was used.
Example 1 .2. OD formulation
For foliar treatments of diseases on different crops (Example 3), an oil dispersion (OD) of spray-dried or freeze dried fermentation broth Streptomyces Saigon 413 TGAI was prepared.
An OD40% w/w was prepared of 40% dried fermentation broth in a vegetable oil containing an emulsifier, such as a surfactant, using general formulation technology as disclosed on Croda Crop Care, the Nouryon formulator toolbox and in: Formulation of Microbial Biopesticides: Beneficial microorganisms, nematodes and seed treatments (412 p., 6 December 2012) eds. Burges H.D., Springer, ISBN 978-94-011-4926-6.
Example 1.3. Isolation of 16S rDNA, genomic DNA and species identification
Genomic DNA was isolated from Streptomyces sp. Saigon413 using the method described in Kutchma et al. (1998) Biotechniques 24(3):452-457. The 16S rRNA gene was amplified using universal 16S primers and sequenced using Sanger sequencing. The 16S rRNA of Streptomyces sp. Saigon413 is shown in SEQ ID NO: 1.
Whole genome sequencing, using the genomic DNA from Streptomyces sp. Saigon413, was completed using both Pacific Biosciences and Illumina sequencing technologies. The genome was assembled using HFAP4 and polished with Pilon using the Illumina reads
Example 1.4. Analysis of Streptimidone, cyclothiazomycin C and malonomicin
The structure of cyclothiazomycin C is disclosed on p. 3 of WO2015191789 and can be extracted and analysed according to the method discosed in Wang et al. (2010) Appl. Environmental Microbiology, Vol. 76, No. 7 p.2336. Malonomicin can be extracted and analysed according to the method disclosed in Example I (B) of W02006/078939. Streptimidone can be extracted and isolated according to the method as disclosed in Lee et al. J. of Antibiotics (2020) 73: p. 184-188, including the supplementary information.
Example 1.5. Oligosaccharide compound I
Purification and characterisation of the oligosaccharide compound I
Whole broth Streptomyces chrestomyceticus CBS149411 (Streptomyces sp. Saigon413), was centrifuged to produce an aqueous extract and a pellet. The aqueous extract was freeze dried. The material was resuspended in a minimal volume of water and partitioned with ethyl acetate to remove lipophilic components. The aqueous suspension was retained and freeze dried and resuspended in a minimal volume of water before being applied to an activated charcoal column.
The column was washed with water and eluted with water:acetone (50:50).
The compound I was further purified by Hydrophilic Interaction Liquid Chromatography (HILIC) using Mass guided fractionation and ELSD detector. Using for example Waters XBridge Amide, 5 micron, 30x100mm using a gradient of acetonitrile and 10mM Ammonium Acetate.
Characterisation of oligosaccharide compound I
The compound I was determined in the purified fermentation broth according to the methods disclosed below.
Molecular composition and total molecular mass
The molecular composition and total molecular mass was C53H90N2O44, and 1458.487 g, respectively which were determined using MS-MS and NMR spectroscopy as disclosed below.
Solubility
The solubility of the oligosaccharide compound I in water, pH 7.01 , was >10’000 ppm, and in DMSO it was > 9772 ppm.
MS-MS Spectrometry and liquid chromatrography
Spectra were recorded on an Orbitrap ID-X Tribrid Mass Spectrometer from Thermo Scientific equipped with an OptaMax NG Heated Electrospray Source (Spray Voltage: Static, Polarity Ion (V): 3400 (Positive ion mode) & 2400 (Negative ion mode), Sheath Gas (Arb): 40, Aux Gas (Arb): 5, Sweep Gas (Arb): 1 , Ion Transfer Tube Temperature: 350 °C, Vaporizer Temperature: 350 °C). The Scan Parameters were as follows;
Experiment 1 : MS OT (Orbitrap Resolution: 60,000, Scan Range (m/z): 200 to 2000, RF Lens (%): 60, AGO Target: Standard, Maximum Injection Time Mode: Auto, Microscans: 1 , Data Type: Profile, Polarity: Both), Experiment 2: tMS2 OT CID (MSn Level (n): 2, Isolation Window (m/z): 1.6, Activation Type: CID, CID Collision Energy (%): 30, Detector Type: Orbitrap, Orbitrap Resolution: 30,000, RF Lens (%): 60, Polarity: Negative). The mass spectrometer was connected to a Vanquish Flex UHPLC from Thermo Scientific using a Vanquish Split Sampler FT, Vanquish Binary Pump F, Vanquish Column Compartment H, Vanquish Diode Array Detector FG and Vanquish Charged Aerosol Detector. Liquid Chromatography Conditions included: Thermo Scientific Hypercarb™ Porous Graphitic Carbon column 5pm 4.6x50mm, P.N. 35005-054630. Temp: 40°C, DAD wavelength range: 250 to 260nm, Solvent gradient: Solvent A: H2O with 0.1% formic acid, Solvent B: CH3CN with 0.1% formic acid, gradient: 0 min 1% B, 99% A; 4. OOmin 50% B, 50% A; 4.25min 100% B; 4.50min 100% B; 4.95min 1% B, 99% A; 6. OOmin 1% B, 99% A, Flow rate: 1.0ml/min, Injection volume: 2 uL, Total run time: 6.0min.
NMR Spectroscopy NMR spectra were recorded on a Bruker AVIII 600 NMR spectrometer, equipped with a 5 mm Bruker (1H/19F)/13C/15N TCI cryoprobe fitted with Z gradients, using standard Bruker pulse sequences. Samples were dissolved in D2O, and the spectra were recorded at 300° K and referenced to acetone at 2.225 ppm for 1H and 31 .07 ppm for 13C. Figures 1 to 4 show NMR spectra of the compound of the present invention. The one bond 1H-13C correlation spectrum contains peaks corresponding to 1 methyl (CH3) and 40 methine (CH) groups (listed in Table 1) and 9 methylene (CH2) groups (listed in Table 2).
In addition, the 1 D 13C spectrum contains signals from 3 quaternary carbons at 104.7, 159.3 and 175.2 ppm (± 0.1).
Table 1. Methyl and methine signals in the one bond 1H-13C correlation spectrum of the oligosaccharide compound I together with multiplicity information for protons resolved in the 1 D 1 H spectrum.
Table 2. Methylene signals in the one bond 1H-13C correlation spectrum of the compound I together with multiplicity information for protons resolved in the 1 D 1H spectrum. Example 1.6. Lipopeptide compound of Formula II
Purification of a lipopeptide according to Formula II (Formula ll(a) and Formula ll(b))
The mycelia from fermentation broth from Streptomyces sp. Saigon 413, was separated via centrifugation and the supernatant was treated with butanol. The butanol was removed and the extract partitioned between water and ethyl acetate. The lipopeptides were purified from the ethyl acetate fraction by preparative reverse phase (C18) HPLC. The lipopeptides were relatively aploar and elute in the higher organic fraction in a gradient system with 0.1% formic acid and acetonirile (0.1% formic acid). A gradient of
60% Aqueous to 40% Aqueous with the above solvents allowed separation of a lipopeptide compound according to Formula 11 (a) and Formula I l(b).
Compounds were detected by UV-VIS (Figure 5), mass spectrometry (Figures 6 and 7) and NMR spectrometry (Figures 8 ad 9). Characterisation of a lipopeptide of Formula II
Liquid Chromatography and High-Resolution Mass Spectrometry
Spectra were recorded on an Orbitrap ID-X Tribrid Mass Spectrometer from Thermo Scientific equipped with an OptaMax NG Heated Electrospray Source (Spray Voltage: Static, Polarity Ion (V): 3400 (Positive ion mode) & 2400 (Negative ion mode), Sheath Gas (Arb): 40, Aux Gas (Arb): 5, Sweep Gas (Arb): 1 , Ion Transfer Tube Temperature: 350 °C, Vaporizer Temperature: 350 °C). The Scan Parameters were as follows;
Experiment 1 : MS OT (Orbitrap Resolution: 50,000, Scan Range (m/z): 200 to 2000, RF Lens (%): 60, AGO Target: Standard, Maximum Injection Time Mode: Auto, Microscans: 1 , Data Type: Profile, Polarity: Both), Experiment 2: tMS2 OT CID (MSn Level (n): 2, Isolation Window (m/z): 1.0, Activation Type: CID, CID Collision Energy (%): 30, Detector Type: Orbitrap, Orbitrap Resolution: 30,000, RF Lens (%): 60, Polarity: Positive),
Experiment 3: tMS2 OT HCD (MSn Level (n): 2, Isolation Window (m/z): 1 .0, Activation Type: HCD, HCD Collision Energy (%): 30, Detector Type: Orbitrap, Orbitrap Resolution: 30,000, RF Lens (%): 60, Polarity: Positive), Experiment 4: tMS3 OT HCD (MSn Level (n): 3, Isolation Window (m/z): 1.6, Activation Type: HCD, HCD Collision Energy (%): 30, MS2 Isolation Window (m/z): 2, MS2 Activation Type: HCD, MS2 HCD Collision Energy (%): 30, Detector Type: Orbitrap, Orbitrap Resolution: 30,000, RF Lens (%): 60, Polarity: Positive). The mass spectrometer was connected to a Vanquish Flex UHPLC from Thermo Scientific using a Vanquish Split Sampler FT, Vanquish Binary Pump F, Vanquish Column Compartment H, Vanquish Diode Array Detector FG and Vanquish Charged Aerosol Detector.
Liquid Chromatography Conditions included: Waters ACQUITY UPLC C18 column 1 .7pm 3.0x50mm, P.N. 186004660. Temp: 40°C, DAD wavelength range: 250 to 260nm, Solvent gradient: Solvent A: H2O with 0.1 % formic acid, Solvent B: CH3CN with 0.1 % formic acid, gradient: 0 min 10% B, 90% A; 4. OOmin 90% B, 10% A; 4.25min 90% B, 10% A; 4.50min 10% B, 90% A; 5.00min 10% B, 90% A, Flow rate: 1.0ml/min, Injection volume: 2 uL, Total run time: 5.0min. A purified fermentation broth as described above was injected.
Figures 6 and 7 show LC-ESI-MS/MS/MS spectra of a compound of Formula II (a).
NMR Spectroscopy
NMR spectra were recorded on a Bruker AVIII 600 NMR spectrometer, equipped with a 5 mm Bruker (1H/19F)/13C/15N TCI cryoprobe fitted with Z gradients, using standard Bruker pulse sequences. Samples were dissolved in CDsOD, and the spectra were recorded at 300° K and referenced to the residual solvent signal at 3.31 ppm for 1H. Figures 8 and 9 each cover half of the 1 H NMR spectrum of a compound according to Formula I l(a). Molecular composition and mass
The molecular composition and mass of a lipopeptide according to Formula I l(a) and Formula I l(b) was determined using the results of liquid chromatography and high-resolution mass spectrometry as disclosed above. The lipopeptide compounds of Formula 11 (a) and 11 (b) have the following composition.
Formula II (a) Lipopeptide 1204: Molecular composition C55H85N11 O19 and exact mass of 1203.602.
Formula II (b) Lipopeptide 1218: Molecular composition C56H87N11 O19 and exact mass of 1217.618.
Solubility
The solubility of a compound of Formula 11(a), Formula II (b) and Lipopeptin A was determined in water and DMSO:
Compound _ solvent (pH) solubility (ppm)
Formula II (a) Lipopeptide 1204 water (2.08) 21 .4
Formula II (a) Lipopeptide 1204 water (5.55) >10’000
Formula II (a) Lipopeptide 1204 DMSO >10’000
Formula II (b) Lipopeptide 1218 DMSO >10’000
Example 1.7. Polyene compound
Purification of polyene compound
A spray dried sample from a culture of S. chrestomycetiucs CBS149411 (Streptomyces sp. Saigon 413) was washed with water. The solid residue was extracted twice with isopropanol and the isopropanol was removed. The resulting solid was purified by preparative reverse phase (C18) HPLC using an acetonitrile:water gradient. Further purification was conducted by preparative reverse phase HPLC using a Zorbax C8 column and eluting with an acetonitrile:water gradient.
The compound was detected by UV-VIS (Figure 10).
Characterisation of polyene compound
Liquid Chromatography and High-Resolution Mass Spectrometry
Spectra were recorded on an Orbitrap ID-X Tribrid Mass Spectrometer from Thermo Scientific equipped with an OptaMax NG Heated Electrospray Source (Spray Voltage: Static, Polarity Ion (V): 3400 (Positive ion mode) & 2400 (Negative ion mode), Sheath Gas (Arb): 40, Aux Gas (Arb): 5, Sweep Gas (Arb): 1 , Ion Transfer Tube Temperature: 350 °C, Vaporizer Temperature: 350 °C). The Scan Parameters were as follows;
Experiment 1 : MS OT (Orbitrap Resolution: 50,000, Scan Range (m/z): 200 to 2000, RF Lens (%): 60, AGO Target: Standard, Maximum Injection Time Mode: Auto, Microscans: 1 , Data Type: Profile, Polarity: Both), Experiment 2: tMS2 OT CID (MSn Level (n): 2, Isolation Window (m/z): 1.0, Activation Type: CID, CID Collision Energy (%): 30, Detector Type: Orbitrap, Orbitrap Resolution: 30,000, RF Lens (%): 60, Polarity: Positive), Experiment 3: tMS2 OT HCD (MSn Level (n): 2, Isolation Window (m/z): 1 .0, Activation Type: HCD, HCD Collision Energy (%): 30, Detector Type: Orbitrap, Orbitrap Resolution: 30,000, RF Lens (%): 60, Polarity: Positive), Experiment 4: tMS3 OT HCD (MSn Level (n): 3, Isolation Window (m/z): 1.6, Activation Type: HCD, HCD Collision Energy (%): 30, MS2 Isolation Window (m/z): 2, MS2 Activation Type: HCD, MS2 HCD Collision Energy (%): 30, Detector Type: Orbitrap, Orbitrap Resolution: 30,000, RF Lens (%): 60, Polarity: Positive).
The mass spectrometer was connected to a Vanquish Flex UHPLC from Thermo Scientific using a Vanquish Split Sampler FT, Vanquish Binary Pump F, Vanquish Column Compartment H, Vanquish Diode Array Detector FG and Vanquish Charged Aerosol Detector.
Liquid Chromatography Conditions included: Waters ACQUITY UPLC C18 column 1 .7pm 3.0x50mm, P.N. 186004660. Temp: 40°C, DAD wavelength range: 250 to 260nm, Solvent gradient: Solvent A: H2O with 0.1 % formic acid, Solvent B: CH3CN with 0.1 % formic acid, gradient: 0 min 10% B, 90% A; 4. OOmin 90% B, 10% A; 4.25min 90% B, 10% A; 4.50min 10% B, 90% A; 5.00min 10% B, 90% A, Flow rate: 1.0ml/min, Injection volume: 2 uL, Total run time: 5.0min.
A purified fermentation broth as described under section 1.8.1 was injected.
The key peaks observed were:
Negative ion: C67H114NO25 [M-H]’ Expected: 1332.7685, Observed: 1332.7679
Positive ion: C67H114NO24 [M-H2O+H]+ Expected: 1316.7725, Observed: 1316.7709
Positive ion: C67H115NO24 [M-H2O+2H]2+ Expected: 658.8899, Observed: 658.8895 Positive ion: C67H113NO23 [M-2(H2O)+2H]2+ Expected: 649.8846, Observed: 649.8843 Positive ion: C67H111 NO22 [M-3(H2O)+2H]2+ Expected: 640.8793, Observed: 640.8790
In a second experiment, Liquid Chromatography Conditions included: Kinetex Polar C18 column 100A 4.6x100mm, P.N. H17-055453. Temp: 40°C, DAD wavelength range: 250 to 260nm, Solvent gradient: Solvent A: H2O with 0.1 % formic acid, Solvent B: CH3CN with 0.1 % formic acid, gradient: Omin 10% B, 90% A; 1min 10% B, 90% A; 6.50min 95% B, 5% A; 8.00min 95% B, 5% A; 9.00min 10% B, 90% A; 10. OOmin 10% B, 90% A, Flow rate: 1.0ml/min, Injection volume: 5 uL, Total run time: 10. Omin.
Under these conditions, the polyene compound had a retention time of 5.55-5.57 minutes.
The observed mass of the polyene compound was the same as in the previous Liquid chromatography run (results not shown).
Molecular composition and mass
The molecular composition and mass of the polyene compound was determined using the results of liquid chromatography and high-resolution mass spectrometry as disclosed above. The polyene compound has a molecular composition C67H115NO25 and exact mass of 1333.7758. Solubility
The solubility of a compounds was determined in DMSO:
Compound solvent (pH) solubility (ppm)
Polyene compound DMSO >10’000
Example 2a). In vitro activity of a mixture of Streptomyces chrestomyceticus CBS149411 and a second compound against Fusarium culmorum, Monographella nivalis syn. Microdochium nivale, Fusarium nivale, Septoria tritici, Glomerella lagenarium syn. Colletotrichum lagenarium, and Botrytis cinerea
The following examples show the efficacy of a mixture of spray-dried or freeze-dried material of Streptomyces chrestomyceticus CBS149411 and a second compound against, Fusarium culmorum, Monographella nivalis syn. Microdochium nivale, Fusarium nivale, Zymoseptoria (Septoria) tritici, Glomerella lagenarium syn. Colletotrichum lagenarium, and Botrytis cinerea.
The second compound was a product from the following list:
• Product RIZODERMA, A23624A SC32.4 (3.2% ai), ai: Trichoderma afroharzianum Th2RI99
• Product CeraMax®, FS150 (15% ai), ai: Natamycin
• Product ArvaticoO/BIONEMA, A24401A SC150 (15% ai), ai: Bacillus velezensis CNPSo 3602
• Product Integral® Pro, A22761A FS68 (6.8% ai), ai: Bacillus amyloliquefaciens MBI600
• Product DERANCE, A21849C SL755 (75.5% ai), ai: potassium phosphonate
• Product Epivio Energy, A21814A, calculated as 100% ai. Epivio energy is a biostimulant comprising a seaweed extract and vinasse.
• Jawsamycin was produced by cultivating Streptoverticillium fervens and isolating jawsamycin according to the methods disclosed in Masaru et al, The Journal of Antibiotics, 1990, Volume 43, Issue 7, Pages 748-754.
An efficacy of 0 means that the growth level of the pathogen corresponds to that of the untreated control; an efficacy of 100 means that the growth level of the pathogen was fully inhibited.
Efficacy against Fusarium culmorum
Conidia of the fungus from cryogenic storage were directly mixed into nutrient broth (potato dextrose broth). A DMSO solution of the test compounds was placed into a microtiter plate (96-well format) and the nutrient broth containing the fungal spores was added to it. The test plates were incubated at 24°C and the inhibition of growth was determined after 48 hrs. The following mixture compositions (A:B) (Table 3a) at the reported concentration (in ppm) gave at least 80% efficacy in this test:
Table 3a.
Example 2b): Activity against Fusarium culmorum (root rot):
Conidia of the fungus from cryogenic storage were directly mixed into nutrient broth (potato dextrose broth). A DMSO solution of the test compounds was placed into a microtiter plate (96-well format) and the nutrient broth containing the fungal spores was added to it. The test plates were incubated at 24°C and the growth inhibition of the pathogen was determined after 7 days.
Table 3b: Disease control by S. chrestomyceticus CBS 149411 mixed with Jawsamycin, (Emulsifiable concentrate) in microtiter plates
Efficacy against Monographella nivalis syn. Microdochium nivale, Fusarium nivale
Conidia of the fungus from cryogenic storage were directly mixed into nutrient broth (potato dextrose broth). A DMSO solution of the test compounds was placed into a microtiter plate (96-well format) and the nutrient broth containing the fungal spores was added to it. The test plates were incubated at 24°C and the inhibition of growth was determined after 72 hrs.
The following mixture compositions (A:B) (Table 4) at the reported concentration (in ppm) gave at least 80% efficacy in this test.
Table 4.
Efficacy against Zymoseptoria (Septoria) tritici
Conidia of the fungus from cryogenic storage were directly mixed into nutrient broth (potato dextrose broth). A DMSO solution of the test compounds was placed into a microtiter plate (96-well format) and the nutrient broth containing the fungal spores was added to it. The test plates were incubated at 24°C and the inhibition of growth was determined after 72 hrs.
The following mixture compositions (A:B) (Table 5) at the reported concentration (in ppm) gave at least 80% efficacy in this test:
Table 5. Efficacy against Glomerella laqenarium syn. Colletotrichum laqenarium
Conidia of the fungus from cryogenic storage were directly mixed into nutrient broth (potato dextrose broth). A DMSO solution of the test compounds was placed into a microtiter plate (96-well format) and the nutrient broth containing the fungal spores was added to it. The test plates were incubated at 24°C and the inhibition of growth was determined after 72 hrs.
The following mixture compositions (A:B) (Table 6) at the reported concentration (in ppm) gave at least 80% efficacy in this test:
Table 6 Efficacy against Botrytis cinerea
Conidia of the fungus from cryogenic storage were directly mixed into nutrient broth (Vogel’s minimal media). A DMSO solution of the test compounds was placed into a microtiter plate (96-well format) and the nutrient broth containing the fungal spores was added to it. The test plates were incubated at 24°C and the inhibition of growth was determined after 72 hrs.
The following mixture compositions (A:B) (Table 7) at the reported concentration (in ppm) gave at least 80% efficacy in this test:
Table 7 Example 3. Disease control of a mixture of Streptomyces chrestomyceticus CBS149411 and a second compound against Zymoseptoria tritici, Puccinia recondita sp. tritici and Pyricularia oryzae on planta.
In the following examples, Streptomyces sp. isolate CBS149411 was fermented, dried to powder and formulated as an oil dispersion, with a loading of 40% of dry matter as disclosed above.
Wheat or rice seedlings 14days old were sprayed with only CBS149411 formulated as OD40%, or CBS149411 formulated as OD40% mixed with a second product before spraying the plants and subsequent infection with a fungal pathogen. The efficacy of the mixture was also compared to the efficacy of the solo 2nd compound product. The mixture of the two components was done in water before the mixture was loaded into the tank of the spray equipment. The spray volume was 200L/ha, the application was executed with a boom sprayer-type equipment in a glasshouse setting. Both components were mixed at different use rates, resulting in a multitude of different mixture ratios. For examples, CBS149411 formulated as OD40% was sprayed at a use rate of 500g/ha (active ingredient), and a second product at a use rate of 100g/ha (active ingredient), resulting in a ratio of 5:1 (CBS 149411 : 2nd compound). In absence of information on the content of active ingredient for tested mixture product, the ratio of volumes for the two products is indicated (volume in liter of product per hectare). Mixtures resulting in 30% or more reduction of disease symptoms were seen as effective to provide a means for disease control or disease reduction.
Mixtures were also analyzed for the presence of a synergistic interaction between the two components to control the target pathogen. Synergy of two products was assumed when the observed disease control was at least 30%, and higher than the expected disease control from the same mixture based on the prediction using efficacy of solo components and the equation from COLBY.
Compounds tested in mixtures with Streptomyces sp. CBS149411 were chosen to cover a large diversity of different mode of action. The FRAC Code List 2023 provides a comprehensive overview of different mode of action used to control fungal and bacterial diseases on crop plants (FRAC: Fungicide resistance action committee, is a specialist technical group of Crop Life international). Where available, active ingredients are also identified by a CAS number (CAS: Chemical Abstracts Service, a division of the American Chemical Society (2540 Olentangy River Road, Columbus, Ohio 43202 USA).
Table 8: List of compounds tested in mixture with Streptomyces sp. CBS149411 . a) also bactericide b) details on manufacturer
Agrauxine, 137 rue Gabriel Peri, 59700 Marcq en Baroeul, FRANCE
Biota GmbH, Rudolf-Diesel-Str. 2, D-72525 Munsingen, GERMANY
De Sangosse, Bonnel CS 10005, 47480 Pont-du-Casse, France
Fytofend, Parc Scientifique Crealys, Rue Georges Legrand 6, 5032 Isnes, Belgium
Laboratoires Goemar SAS, CS 41908, Parc Technopolitain Atalante, 35435 Saint Malo cedex, France Stabler Suisse SA, Henzmannstrasse 17 A, 4800 Zofingen, Switzerland
Syngenta: Syngenta Crop Protection AG, Rosentalstr. 67, 4058 Basel, SWITZERLAND
UPL Limited, UPL House, 610 B/2, Bandra Village, Off Western Express Highway, Bandra (East), Mumbai 400 051 INDIA c) Also an insecticide d) used as plant biostimulant (anti-stress and growth activator) e) not listed in FRAC
- Aureobasidin A and its synthesis are known from Takesako et al., The Journal of Antibiotics, 1991 , 44, 919-924. Aureobasidin A is separated from the fermentation broth by extraction with ethyl acetate, followed by extraction of the ethyl acetate concentrate with a mixture of MeOH:H2O (80% by volume) and cyclohexane (20% by volume), and purified by silica gel column chromatography (silica-gel, elution with hexane:ethyl acetate) followed by reverse phase column chromatography (RP18, elution with acetonitrile:H2O).
- Jawsamycin was produced by cultivating Streptoverticillium fervens and isolating jawsamycin according to the methods disclosed in Masaru et al, The Journal of Antibiotics, 1990, Volume 43, Issue 7, Pages 748- 754.
Methods:
Puccinia recondita (EPPO code: PUCCRE) 14 days old wheat plants variety Arina were inoculated by spraying them with a spore suspension one day after application (spore suspension at 80,000 spores per ml in water supplemented with Tween20 at 0.1 %). After an incubation period of 1 day at 20° C and 95% relative humidity, the inoculated test plants were kept at 20° C and 60% relative humidity in a greenhouse. The percentage leaf area covered by disease was assessed visually when an appropriate level of disease appeared on untreated check plants (9 - 12 days after infection).
Zymoseptoria tritici (EPPO code: SEPTTR): 14 days old wheat plants variety Riband were inoculated by spraying a spore suspension on them one day after application (1.5 Mio spores per ml in water supplemented with 0.01 % Tween20). After an incubation period of 4 days at 22°C/21 °C (day/night) and 95% relative humidity, the inoculated test plants were kept at 22°C/21°C (day/night) and 70% relative humidity in a greenhouse. Efficacy was assessed directly when an appropriate level of disease appeared on untreated check plants (14 - 19 days after application). The isolate used is a recent field isolate expressing tolerance to DMI fungicides, SDHI fungicides and Qol fungicides.
Pyricularia oryzae (EPPO code: PYRIOR): 3 week old rice plants cv. Balilla were treated with the formulated test compound in a spray chamber. Two days after application rice plants were inoculated by spraying a spore suspension (1 x 105 conidia/ml) on the test plants. After an incubation period of 6 days at 25° C and 95% relative humidity the disease incidence was assessed.
Results: The results of the foliar experiments of a mixture of Streptomyces chrestomyceticus CBS149411 and a biological product are shown in the following Tables 9 to 25. The biological compound (2nd product) used in the experiments is indicated in the title of Tables 9 to 25. Treatment of the mixtures is compared with the treatments by a composition of S. chrestomyceticus CBS 149411 alone and the biological compound alone. Table 9: Disease control by S. chrestomyceticus CBS 149411 mixed with orange oil Table 10: Disease control by S. chrestomyceticus CBS 149411 mixed with Tea tree Oil (Melaleuca alternifolia) Table 11 : Disease control by S. chrestomyceticus CBS 149411 mixed with sulfur (Seffika)
Table 12: Disease control by S. chrestomyceticus CBS 149411 mixed with phosphonate (Pygmalion®)
Table 13: Disease control by S. chrestomyceticus CBS 149411 mixed with sulfur (Thiovit Jet) Table 14: Disease control by S. chrestomyceticus CBS 149411 mixed with phosphonate (Redeli)
Table 15: Disease control by S. chrestomyceticus CBS 149411 mixed with Bacillus amyloliquefaciens FZB24 (Taegro®)
Table 16: Disease control by S. chrestomyceticus CBS 149411 mixed with Laminarin
Table 17: Disease control by S. chrestomyceticus CBS 149411 mixed with Jawsamycin, (Emulsifiable concentrate)
Table 18: Disease control by S. chrestomyceticus CBS 149411 mixed with bicarbonate (Armicarb®)
Table 19: Disease control by S. chrestomyceticus CBS 149411 mixed with copper hydroxide (Coprantol Hi Bio 2.0) Table 20: Disease control by S. chrestomyceticus CBS 149411 mixed with Aureobasidin A, (Emulsifiable concentrate) Table 21 : Disease control by S. chrestomyceticus CBS 149411 mixed with copper oxychloride
(Coprantol 30WG)
Table 22: Disease control by S. chrestomyceticus CBS 149411 mixed with Kasugamycin (Kasumin®)
Table 23: Disease control by S. chrestomyceticus CBS 149411 mixed with plant and algal extract
(Megafol®, including seaweed extract), The ratio is based on the volume of both products (L/ha). Table 24: Disease control by S. chrestomyceticus CBS 149411 mixed with chitooligosaccharides and oligogalactunorides (COS-OGA, Fytosave®) Table 25: Disease control by S. chrestomyceticus CBS 149411 mixed with yeast extract (Saccharomyces cerevisiae)
Conclusions
The results presented in Tables 3-7 and Tables 9-25 indicate that mixtures of a formulation based on Streptomyces chrestomyceticus CBS149411 biological products result in efficient control of several phytopathogenic fungi. All mixtures tested for the control of Puccinia recondita, Pyricularia oryzae or Zymoseptoria tritici provided at least 30% disease control, on at least one target and in at least one mixture ratio. A broad range of mixture ratios was found to be effective, such as from 100:1 to 1 :100 (ratio CBS 149411 : 2nd compound, in g/ha). The disease control level for a majority of the mixtures indicated a synergistic interaction of Streptomyces chrestomyceticus CBS149411 with the mixture partner in at least one mixture ratio. Synergistic interaction is suggested for the mixtures with Bacillus amyloliquefaciens, COS-OGA, jawsamycin, laminarin, orange oil (Citrus sp.), phosphonate, and sulphur.
(Original in Electronic Form) (This sheet is not part of and does not count as a sheet of the international application)
FOR RECEIVING OFFICE USE ONLY
FOR INTERNATIONAL BUREAU USE ONLY -5 This form was re international Bur -5-1 Authorized officer

Claims

Claims
1. A composition comprising components (A) and (B) as active ingredients, wherein component (A) comprises Streptomyces chrestomyceticus and component (B) comprises a compound selected from the group consisting of sulphur, phosphonate, copper hydroxide, copper oxychloride, bicarbonate, Bacillus sp, Trichoderma sp, aureobasidin A, jawsamycin, kasugamycin, natamycin, a polysaccharide elicitor, a plant biostimulant (PBS) comprising an algal extract, a plant extract and/or vinasse, orange oil, and / or tea tree oil.
2. The composition according to claim 1 , wherein the Streptomyces chrestomyceticus comprises a nucleotide sequence which has at least 99.8 % identity to SEQ ID NO: 1 .
3. The composition according to claim 1 or 2, wherein the Streptomyces chrestomyceticus comprises a genome sequence which has at least 95% identity to the whole genome of Streptomyces sp. Saigon413 deposited with the Westerdijk Institute under accession number CBS149411 , preferably the Streptomyces chrestomyceticus comprises Streptomyces sp. Saigon413 deposited with the Westerdijk Institute under accession number CBS149411
4. The composition according to any one of the claims 1 to 3, wherein the component (A) comprises a cell count of the Streptomyces chrestomyceticus of 10° to 1014 cfu / g dry weight.
5. The composition according to any one of the claims 1 to 4, wherein component (A) further comprises a metabolite, preferably wherein the metabolite comprises malonomicin.
6. The composition according to claim 5, wherein the fermentation product comprises at least one further metabolite selected from the group consisting of cyclothiazomycin C, streptimidone, an oligosaccharide compound which comprises a molecular formula according to C53H90N2O44, further characterised by the NMR spectra listed in Table 1 and Table 2, further characterized by a structural Formula I,
Formula (I), a lipopeptide according to Formula II, or a salt thereof wherein R1 = CHs or C2H5
Formula (II) and a polyene compound characterized by a molecular formula according to C67H115NO25, wherein the polyene is further characterized by the spectrum of light absorption as shown in Figure 10.
7. The composition according to any one of the claims 1 to 6, wherein the the Bacillus sp. comprises Bacillus amyloliquefaciens, and/or Bacillus velezensis, the Trichoderma sp. comprises Trichoderma afroharzianum , the polysaccharide elicitor comprises laminarin, a composition comprising chitooligosaccharides and oligogalactunorides (COS-OGA), and/or yeast extract, wherein preferably the yeast extract comprises Saccharomyces cerevisiae yeast extract, and / or the algal extract comprises a seaweed extract, preferably an extract from Ascophyllum nodosum.
8. The composition according to any one of the claims 1 to 6, wherein component (B) comprises a compound selected from the group consisting of sulfur, phosphonate, Bacillus amyloliquefaciens FZB24, a composition comprising chitooligosaccharides and oligogalactunorides (COS-OGA), jawsamycin, laminarin, and orange oil.
9. The composition according to any one of the claims 1 to 8, wherein the weight ratio of compound (A) to compound (B) is from 100:1 to 1 :100.
10. The composition according to any one of the claims 1 to 9 wherein the weight ratio of compound (A) to compound (B) is from 50:1 to 1 :50.
11. The composition according to any one of the preceding claims, wherein the composition further comprises an agriculturally acceptable carrier and, optionally, a surfactant and/or formulation adjuvants.
12. A method of controlling or preventing phytopathogenic diseases, preferably phytopathogenic fungi, on a plant or on plant propagation material and / or on harvested food crops, which comprises applying to the plant, on plant propagation material, the locus thereof, and / or on harvested food crops the composition as defined in any one of claims 1 to 11 .
13. The method according to claim 12, wherein the phytopathogenic fungi belong to a genus belonging to Zymoseptoria (Septoria), Puccinia, Pyricularia, Fusarium, Monographella, Microdochium, Glomerella, Colletotrichum, and / or Botrytis, preferably the phytopathogenic fungi belong to a species belonging to Zymoseptoria tritici, Puccinia recondita f. sp. tritici, Pyricularia oryzae, Fusarium culmorum, Monographella nivalis syn. Microdochium nivale, Fusarium nivale, Septoria tritici, Glomerella lagenarium syn. Colletotrichum lagenarium, and/ or Botrytis cinerea.
14. The method according to claim 12 or 13, wherein the plant comprises wheat, barley, rice, corn, soya, sugar beet, banana, tomato, cucumber, and / or groundnut.
15. A plant, plant propagation material or the locus thereof, or harvested food crops comprising a composition according to any one of the claims 1 to 11 .
PCT/EP2025/059132 2024-04-04 2025-04-03 Fungicidal composition Pending WO2025210150A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP24168350 2024-04-04
EP24168350.7 2024-04-04

Publications (1)

Publication Number Publication Date
WO2025210150A1 true WO2025210150A1 (en) 2025-10-09

Family

ID=90717676

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2025/059132 Pending WO2025210150A1 (en) 2024-04-04 2025-04-03 Fungicidal composition

Country Status (1)

Country Link
WO (1) WO2025210150A1 (en)

Citations (35)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5061495A (en) 1988-03-07 1991-10-29 Agricultural Genetics Company Limited Antibiotic derived from b. subtilis
US5356624A (en) 1993-04-23 1994-10-18 The United States Of America As Represented By The Secretary Of Agriculture Biological treatment for controlling wood deteriorating fungi
JPH11253151A (en) 1997-11-13 1999-09-21 Kumiai Chem Ind Co Ltd Rice seedling raising disease control agent
US6060051A (en) 1997-05-09 2000-05-09 Agraquest, Inc. Strain of bacillus for controlling plant diseases and corn rootworm
US6245551B1 (en) 1999-03-30 2001-06-12 Agraquest, Inc. Strain of Bacillus pumilus for controlling plant diseases caused by fungi
WO2003000051A2 (en) 2001-06-22 2003-01-03 Drahos David J Novel biofungicide
WO2006078939A1 (en) 2005-01-21 2006-07-27 Dow Agrosciences Llc Use of malonomicin and analogs in fungicidal applications
US7094592B2 (en) 2001-11-26 2006-08-22 Kumiai Chemical Industry Co., Ltd. Bacillus sp. D747 strain, plant disease controlling agents and insect pest controlling agents using the same and control method using the agents
US7579183B1 (en) 2006-12-01 2009-08-25 The United States Of America As Represented By The Secretary Of Agriculture Saprophytic yeast, Pichia anomala
WO2009116106A1 (en) 2008-03-21 2009-09-24 Trentino Sviluppo S.P.A. Trichoderma atroviride sc1 for biocontrol of fungal diseases in plants
WO2010086790A1 (en) 2009-01-27 2010-08-05 Lesaffre Et Compagnie Saccharomyces cerevisiae strains with phytosanitary capabilities
US20100291039A1 (en) 2007-12-14 2010-11-18 Kohl Jurgen Anton Novel micro-organisms controlling plant pathogens
CN101497870B (en) * 2009-03-02 2011-04-13 北京市京圃园生物工程有限公司 Microbial complex bacterial agent and use thereof
WO2011106491A2 (en) 2010-02-25 2011-09-01 Marrone Bio Innovations, Inc. Isolated bacterial strain of the genus burkholderia and pesticidal metabolites therefrom
WO2011151819A2 (en) 2010-06-01 2011-12-08 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Pseudozyma aphidis as a biocontrol agent against various plant pathogens
WO2013032693A2 (en) 2011-08-27 2013-03-07 Marrone Bio Innovations, Inc. Isolated bacterial strain of the genus burkholderia and pesticidal metabolites therefrom-formulations and uses
WO2013034938A2 (en) 2011-09-08 2013-03-14 Szegedi Tudományegyetem A copper resistant, fengycin-producing bacillus mojavensis strain for controlling vegetable pathogens, its use and compositions containing it
WO2014028521A1 (en) 2012-08-14 2014-02-20 Marrone Bio Innovations, Inc. Bacillus sp. strain with antifungal, antibacterial and growth promotion activity
WO2014160827A1 (en) * 2013-03-28 2014-10-02 Novozymes Bioag A/S Compositions and methods for enhancing microbial stability
WO2015067800A1 (en) 2013-11-11 2015-05-14 Basf Se Antifungal penicillium strains, fungicidal extrolites thereof, and their use
WO2015191789A2 (en) 2014-06-10 2015-12-17 The Board Of Trustees Of The University Of Illinois Reactivity-based screening for natural product discovery
KR101588562B1 (en) * 2015-02-25 2016-01-26 주식회사 코코스토리 surfur removing poison for feedstuff and its product method
WO2016014386A1 (en) * 2014-07-23 2016-01-28 Novozymes Bioag A/S Compositions containing lipo-chitooligosaccharide and microorganism for enhancing plant growth
WO2016020371A1 (en) 2014-08-04 2016-02-11 Basf Se Antifungal paenibacillus strains, fusaricidin-type compounds, and their use
CN105794860A (en) * 2016-04-07 2016-07-27 山东省农业科学院农业质量标准与检测技术研究所 Microbial agent for controlling quarantine fungus diseases of apples and promoting yield increase
WO2016154297A1 (en) 2015-03-26 2016-09-29 Bayer Cropscience Lp A novel paenibacillus strain, antifungal compounds, and methods for their use
WO2017019448A1 (en) 2015-07-24 2017-02-02 AgBiome, Inc. Modified biological control agents and their uses
WO2017066094A1 (en) 2015-10-12 2017-04-20 Pioneer Hi-Bred International, Inc. Biologicals and their use in plants
WO2017205258A1 (en) 2016-05-26 2017-11-30 Novozymes Bioag A/S Bacillus and lipochitooligosaccharide for improving plant growth
CN107586183A (en) * 2016-07-08 2018-01-16 潍坊盛泉生物科技有限公司 It is a kind of to replant special bacteria agent of disease and preparation method thereof for apple tree
CN107815478B (en) * 2016-09-12 2021-07-09 牡丹江佰佳信生物科技有限公司 Method for producing kasugamycin by fermenting streptomyces aureofaciens BJX007
WO2022038180A1 (en) 2020-08-18 2022-02-24 Aphea.Bio Nv Means and methods for controlling pathogens and pests in plants
CN114868770A (en) * 2022-06-25 2022-08-09 石河子大学 Agricultural composition containing bactericide and gamma-polyglutamic acid
CN115505615A (en) * 2022-11-09 2022-12-23 淮北师范大学 A method for synthesizing natamycin based on Streptomyces mixed bacteria fermentation
CN117481150A (en) * 2023-10-26 2024-02-02 江门市新会区林业科学研究所（江门市新会区林业科技推广站） Fermentation liquor for preventing and treating citrus canker as well as preparation method and application thereof

Patent Citations (37)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5061495A (en) 1988-03-07 1991-10-29 Agricultural Genetics Company Limited Antibiotic derived from b. subtilis
US5356624A (en) 1993-04-23 1994-10-18 The United States Of America As Represented By The Secretary Of Agriculture Biological treatment for controlling wood deteriorating fungi
US6060051A (en) 1997-05-09 2000-05-09 Agraquest, Inc. Strain of bacillus for controlling plant diseases and corn rootworm
JPH11253151A (en) 1997-11-13 1999-09-21 Kumiai Chem Ind Co Ltd Rice seedling raising disease control agent
US6245551B1 (en) 1999-03-30 2001-06-12 Agraquest, Inc. Strain of Bacillus pumilus for controlling plant diseases caused by fungi
WO2003000051A2 (en) 2001-06-22 2003-01-03 Drahos David J Novel biofungicide
US7094592B2 (en) 2001-11-26 2006-08-22 Kumiai Chemical Industry Co., Ltd. Bacillus sp. D747 strain, plant disease controlling agents and insect pest controlling agents using the same and control method using the agents
WO2006078939A1 (en) 2005-01-21 2006-07-27 Dow Agrosciences Llc Use of malonomicin and analogs in fungicidal applications
EP1860939A1 (en) 2005-01-21 2007-12-05 Dow Agrosciences LLC Use of malonomicin and analogs in fungicidal applications
US7579183B1 (en) 2006-12-01 2009-08-25 The United States Of America As Represented By The Secretary Of Agriculture Saprophytic yeast, Pichia anomala
US20100291039A1 (en) 2007-12-14 2010-11-18 Kohl Jurgen Anton Novel micro-organisms controlling plant pathogens
US8431120B2 (en) 2008-03-21 2013-04-30 Trentino Sviluppo S.P.A. Trichoderma atroviride SC1 for biocontrol of fungal diseases in plants
WO2009116106A1 (en) 2008-03-21 2009-09-24 Trentino Sviluppo S.P.A. Trichoderma atroviride sc1 for biocontrol of fungal diseases in plants
WO2010086790A1 (en) 2009-01-27 2010-08-05 Lesaffre Et Compagnie Saccharomyces cerevisiae strains with phytosanitary capabilities
CN101497870B (en) * 2009-03-02 2011-04-13 北京市京圃园生物工程有限公司 Microbial complex bacterial agent and use thereof
WO2011106491A2 (en) 2010-02-25 2011-09-01 Marrone Bio Innovations, Inc. Isolated bacterial strain of the genus burkholderia and pesticidal metabolites therefrom
WO2011151819A2 (en) 2010-06-01 2011-12-08 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Pseudozyma aphidis as a biocontrol agent against various plant pathogens
WO2013032693A2 (en) 2011-08-27 2013-03-07 Marrone Bio Innovations, Inc. Isolated bacterial strain of the genus burkholderia and pesticidal metabolites therefrom-formulations and uses
WO2013034938A2 (en) 2011-09-08 2013-03-14 Szegedi Tudományegyetem A copper resistant, fengycin-producing bacillus mojavensis strain for controlling vegetable pathogens, its use and compositions containing it
WO2014028521A1 (en) 2012-08-14 2014-02-20 Marrone Bio Innovations, Inc. Bacillus sp. strain with antifungal, antibacterial and growth promotion activity
WO2014160827A1 (en) * 2013-03-28 2014-10-02 Novozymes Bioag A/S Compositions and methods for enhancing microbial stability
WO2015067800A1 (en) 2013-11-11 2015-05-14 Basf Se Antifungal penicillium strains, fungicidal extrolites thereof, and their use
WO2015191789A2 (en) 2014-06-10 2015-12-17 The Board Of Trustees Of The University Of Illinois Reactivity-based screening for natural product discovery
WO2016014386A1 (en) * 2014-07-23 2016-01-28 Novozymes Bioag A/S Compositions containing lipo-chitooligosaccharide and microorganism for enhancing plant growth
WO2016020371A1 (en) 2014-08-04 2016-02-11 Basf Se Antifungal paenibacillus strains, fusaricidin-type compounds, and their use
KR101588562B1 (en) * 2015-02-25 2016-01-26 주식회사 코코스토리 surfur removing poison for feedstuff and its product method
WO2016154297A1 (en) 2015-03-26 2016-09-29 Bayer Cropscience Lp A novel paenibacillus strain, antifungal compounds, and methods for their use
WO2017019448A1 (en) 2015-07-24 2017-02-02 AgBiome, Inc. Modified biological control agents and their uses
WO2017066094A1 (en) 2015-10-12 2017-04-20 Pioneer Hi-Bred International, Inc. Biologicals and their use in plants
CN105794860A (en) * 2016-04-07 2016-07-27 山东省农业科学院农业质量标准与检测技术研究所 Microbial agent for controlling quarantine fungus diseases of apples and promoting yield increase
WO2017205258A1 (en) 2016-05-26 2017-11-30 Novozymes Bioag A/S Bacillus and lipochitooligosaccharide for improving plant growth
CN107586183A (en) * 2016-07-08 2018-01-16 潍坊盛泉生物科技有限公司 It is a kind of to replant special bacteria agent of disease and preparation method thereof for apple tree
CN107815478B (en) * 2016-09-12 2021-07-09 牡丹江佰佳信生物科技有限公司 Method for producing kasugamycin by fermenting streptomyces aureofaciens BJX007
WO2022038180A1 (en) 2020-08-18 2022-02-24 Aphea.Bio Nv Means and methods for controlling pathogens and pests in plants
CN114868770A (en) * 2022-06-25 2022-08-09 石河子大学 Agricultural composition containing bactericide and gamma-polyglutamic acid
CN115505615A (en) * 2022-11-09 2022-12-23 淮北师范大学 A method for synthesizing natamycin based on Streptomyces mixed bacteria fermentation
CN117481150A (en) * 2023-10-26 2024-02-02 江门市新会区林业科学研究所（江门市新会区林业科技推广站） Fermentation liquor for preventing and treating citrus canker as well as preparation method and application thereof

Non-Patent Citations (17)

* Cited by examiner, † Cited by third party
Title
"Croda Crop Care", THE NOURYON FORMULATOR TOOLBOX
"Formulation of Microbial Biopesticides: Beneficial microorganisms, nematodes and seed treatments", 6 December 2012, SPRINGER, pages: 412
CAN JOUR PLANT SCI, vol. 83, no. 3, pages 519 - 524
COLBY, S.R.: "Calculating synergistic and antagonistic responses of herbicide combination", WEEDS, vol. 15, 1967, pages 20 - 22, XP001112961
CROP PROTECTION, vol. 25, 2006, pages 468 - 475
JENSEN DF ET AL.: "Development of a biocontrol agent for plant disease control with special emphasis on the near commercial fungal antagonist Clonostachys rosea strain 'IK726", AUSTRALAS PLANT PATHOL, vol. 36, 2007, pages 95 - 101
KONDO, H.ORITANI, T.KIYOTA, H.: "Synthesis and antifungal activity of the four stereoisomers of streptimidone, a glutarimide antibiotic from Streptomyces rimosus forma paromomycinus", EUR. J. ORG. CHEM., no. 20, 2000, pages 3459 - 3462, XP072101339, DOI: 10.1002/1099-0690(200010)2000:20<3459::AID-EJOC3459>3.0.CO;2-F
KUTCHMA ET AL., BIOTECHNIQUES, vol. 24, no. 3, 1998, pages 452 - 457
LAW ET AL., NATURE CATALYS IS, vol. 1, December 2018 (2018-12-01), pages 977 - 984
LEE ET AL., J. OF ANTIBIOTICS, vol. 73, 2020, pages 184 - 188
MASARU ET AL., THE JOURNAL OF ANTIBIOTICS, vol. 43, 1990, pages 748 - 754
MICHEL-ACEVES A. C. ET AL: "Control Alternatives for Damping-Off in Tomato Seedling Production", vol. 88, no. 3, 1 January 2019 (2019-01-01), pages 325 - 333, XP093202467, ISSN: 1851-5657, Retrieved from the Internet <URL:https://dx.doi.org/10.32604/phyton.2019.06777> DOI: 10.32604/phyton.2019.06777 *
PIETR ET AL., ZESZ. NAUK. A R W SZCZECINIE, vol. 161, 1993, pages 125 - 137
RAHILA ET AL., CURRENT MICROBIOLOGY, vol. 80, 2023, pages 107
TAKESAKO ET AL., THE JOURNAL OF ANTIBIOTICS, vol. 44, 1991, pages 919 - 924
WANG ET AL., APPL. ENVIRONMENTAL MICROBIOLOGY, vol. 76, no. 7, 2010, pages 2336
WOO, S.L. ET AL., THE OPEN MYCOLOGY JOURNAL, vol. 8, 2014, pages 71 - 126

Similar Documents

Publication Publication Date Title
CN114717164B (en) A kind of Bacillus Velez ZLP-101 and its application
Chaurasia et al. Actinomycetes: an unexplored microorganisms for plant growth promotion and biocontrol in vegetable crops
Cawoy et al. Bacillus-based biological control of plant diseases
Shan et al. Biocontrol of rice blast by the phenaminomethylacetic acid producer of Bacillus methylotrophicus strain BC79
Mishra et al. Bioformulations for plant growth promotion and combating phytopathogens: a sustainable approach
Meng et al. Dry flowable formulations of antagonistic Bacillus subtilis strain T429 by spray drying to control rice blast disease
Shanmugam et al. Biocontrol of vascular wilt and corm rot of gladiolus caused by Fusarium oxysporum f. sp. gladioli using plant growth promoting rhizobacterial mixture
CN107960103A (en) Microbial compositions and methods of use for promoting plant growth and treating plant diseases
Kazerooni et al. Talaromyces pinophilus inhibits Pythium and Rhizoctonia-induced damping-off of cucumber
CN107820392A (en) Bacillus amyloliquefaciens RTI472 compositions and for beneficial to plant growth and treatment plant disease method
TW201428097A (en) Bacillus sp. strain with antifungal, antibacterial and growth promotion activity
Malandrakis et al. A Fusarium solani endophyte vs fungicides: Compatibility in a Fusarium oxysporum f. sp. radicis-lycopersici–tomato pathosystem
JP2023553717A (en) Spore composition, its production and use
CN116323911A (en) Novel agrochemical formulation for fusaricidin-producing bacteria
CN115418333A (en) A kind of Bacillus subtilis ZLP-121 and its application
EP4580409A1 (en) Bacillus strain and variants thereof for inhibition of plant diseases
CN115322928A (en) Bacillus amyloliquefaciens ZLP-01 and application thereof
EP4580410A1 (en) Bacillus strain, variants thereof, fermentation products and compositions thereof for inhibition of plant diseases
KR20100017997A (en) A novel streptomyces lavendulae cmc0992[kctc18169p] active against plant fungal pathogens and preparation of microbial pesticide
Musheer et al. Role of microbiotic factors against the soil-borne phytopathogens
CN115369058A (en) Bacillus licheniformis ZLP-81 and its application
Dart et al. Biopesticide commercialization in Australia: potential and challenges
Theresa et al. Microbial biocontrol formulations for commercial applications
WO2025210150A1 (en) Fungicidal composition
Afandhi et al. The diversity of entomopathogenic fungi collected from leaves and rhizospheres of rice implementing integrated pest management

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 25716429

Country of ref document: EP

Kind code of ref document: A1