Means and methods for controlling pathogens and pests in plants

Abstract

The present invention relates to microbial strains and compositions comprising strains of Streptomyces, processes for producing the microbial strains or compositions, and methods of using the microbial strains and compositions to prevent or control pests in plants.

Description

[Technical Field]

[0001] The present invention relates to novel microbial strains having pesticidal activity. The present invention also relates to compositions comprising Streptomyces, processes for the preparation of the microbial strains or compositions, and methods for using Streptomyces chrestomyceticus or compositions in agriculture or horticulture to prevent or control phytopathogenic infestation of plants, harvested food crops, seeds, or non-living materials. [Background technology]

[0002] Pesticides are widely used in agriculture to protect plants from damage caused by plant pathogenic microorganisms. Pesticides can be of chemical or biological origin. Due to some of the adverse effects of chemical pesticides on the environment, there is an increasing need for pesticides of biological, e.g., microbial, origin. Known microorganisms that produce antibiotics against fungi are actinomycetes, such as Streptomyces sp. A very well-known genus is Streptomyces natalensis, which produces the antifungal compound natamycin, used in food and crop protection. U.S. Patent Application No. 5,356,624 discloses a Streptomyces rimosus strain that was found to be active against several wood-decomposing fungi. WO 2022 / 038180 discloses new Streptomyces species that produce several known antifungal compounds or metabolites, such as streptimidon, natamycin (pimaricin), or albofungin. Extracts of these bacterial strains have been found to be active against well-known plant pests, such as Fusarium graminearum, Zymoseptoria tritici, and Puccinia striiformis. Due to the development of resistance to pesticides by plant pathogenic microorganisms, government regulations and societal pressures, there is a continuing need for improved products, such as microbial strains or compounds of biological origin, that have activity against plant pathogenic microorganisms. Summary of the Invention [Means for solving the problem]

[0003] The present invention relates to an isolated microbial strain comprising a genome sequence having at least 99.8%, 99.9% identity or 100% identity to the complete genome of Streptomyces sp. Saigon413, deposited at the Westerdijk Institute under accession number CBS149411.

[0004] In a second aspect, the present invention relates to an isolated microbial strain, which comprises a nucleotide sequence having at least 99.8% identity to SEQ ID NO:1.

[0005] In a third aspect, the present invention provides a method for producing a medicament for the treatment of cancer, comprising: 53 H 90 N2O 44 and further characterized by the NMR spectra listed in Tables 2 and 3, preferably structural formula I: [ka] a strain of Streptomyces, at least one compound selected from the group consisting of malonomycin and cyclothiazomycin C, and streptimidon, a compound according to compound I, a compound according to formula II: [ka] or a salt thereof, wherein R1 is CH3 or C2H5, and 67 H 115 NO 25wherein the polyene is further characterized by a light absorption spectrum as shown in FIG.

[0006] In a fourth aspect, the present invention relates to a process for producing a microbial strain as disclosed herein or a composition as disclosed herein, comprising culturing a microbial strain or a microbial strain of the genus Streptomyces in a suitable fermentation medium under suitable fermentation conditions, and optionally recovering the microbial strain or composition.

[0007] In a fifth aspect, the present invention relates to a method for controlling or preventing infestation of plants, plant propagation material and / or harvested food crops by plant pathogenic microorganisms by treating the plants, plant propagation material and / or harvested food crops with an effective amount of Streptomyces chrestomyceticus, an isolated microbial strain disclosed herein, or a composition disclosed herein.

[0008] In a sixth aspect, the present invention relates to a plant or plant propagation material treated with a microbial strain or a composition according to the invention.

[0009] In a seventh aspect, the present invention relates to the use of a microbial strain according to the present invention, or S. chrestomyceticus as disclosed herein, or a composition according to the present invention, as a pesticide, preferably as a fungicide.

[0010] In an eighth aspect, the present invention provides a method for producing a medicament for the treatment of a medicament comprising: 53 H 90 N2O 44and further characterized by the NMR spectra listed in Tables 2 and 3, preferably characterized by structural formula I, at least one of compounds selected from the group consisting of malonomycin, cyclothiazomycin C, and streptimidon, an oligosaccharide compound according to compound I, a lipopeptide or a salt thereof according to formula II (wherein R1 is CH3 or C2H5), and C 67 H 115 NO 25 and wherein the polyene is further characterized by an optical absorption spectrum as shown in Figure 10.

[0011] Surprisingly, it has been found that strains of Streptomyces, such as the isolated microbial strains disclosed herein, e.g., Streptomyces sp. Saigon 413, and / or metabolites thereof, are effective in treating phytopathogenic microbial diseases of crops.

[0012] The present invention relates to an isolated microbial strain comprising a genome sequence having at least 99.8%, 99.9% identity or 100% identity to the complete genome of Streptomyces sp. Saigon413, deposited at the Westerdijk Institute under accession number CBS149411.

[0013] The present invention also relates to an isolated microbial strain, the strain comprising a nucleotide sequence having at least 99.9%, 99.91%, 99.92%, 99.93%, 99.94%, 99.95%, 99.96%, 99.97%, 99.98%, 99.99%, or 100% identity to SEQ ID NO:1.

[0014] The isolated microbial strain according to the present invention is preferably a microbial strain comprising a nucleotide sequence having at least 99.9%, 99.91%, 99.92%, 99.93%, 99.94%, 99.95%, 99.96%, 99.97%, 99.98%, 99.99% or 100% identity to SEQ ID NO: 1, and the microbial strain comprises a genome sequence having at least 99.8%, 99.9% identity or 100% identity to the complete genome of Streptomyces sp. Saigon413 deposited at the Westerdijk Institute under accession number CBS149411.

[0015] For example, the microbial strain can comprise a nucleotide sequence having at least 99.9% identity to SEQ ID NO: 1, wherein the microbial strain comprises a genome sequence having at least 99.8%, 99.9% identity, or 100% identity to the entire genome of Streptomyces sp. Saigon413 deposited at the Westerdijk Institute under accession number CBS 149411. As another example, the microbial strain can comprise a nucleotide sequence having at least 99.91% identity to SEQ ID NO: 1, wherein the microbial strain comprises a genome sequence having at least 99.8%, 99.9% identity, or 100% identity to the entire genome of Streptomyces sp. Saigon413 deposited at the Westerdijk Institute under accession number CBS 149411. As another example, the microbial strain can comprise a nucleotide sequence having at least 99.92% identity to SEQ ID NO:1, wherein the microbial strain comprises a genome sequence having at least 99.8%, 99.9% identity, or 100% identity to the entire genome of Streptomyces sp. Saigon413 deposited at the Westerdijk Institute under accession number CBS 149411. As another example, the microbial strain can comprise a nucleotide sequence having at least 99.93% identity to SEQ ID NO:1, wherein the microbial strain comprises a genome sequence having at least 99.8%, 99.9% identity, or 100% identity to the entire genome of Streptomyces sp. Saigon413 deposited at the Westerdijk Institute under accession number CBS 149411. As another example, the microbial strain can comprise a nucleotide sequence having at least 99.94% identity to SEQ ID NO:1, and the microbial strain comprises a genome sequence having at least 99.8%, 99.9% identity, or 100% identity to the complete genome of Streptomyces sp. Saigon413, deposited at the Westerdijk Institute under accession number CBS149411.As another example, the microbial strain can comprise a nucleotide sequence having at least 99.95% identity to SEQ ID NO:1, wherein the microbial strain comprises a genome sequence having at least 99.8%, 99.9% identity, or 100% identity to the entire genome of Streptomyces sp. Saigon413 deposited at the Westerdijk Institute under accession number CBS149411. As another example, the microbial strain can comprise a nucleotide sequence having at least 99.96% identity to SEQ ID NO:1, wherein the microbial strain comprises a genome sequence having at least 99.8%, 99.9% identity, or 100% identity to the entire genome of Streptomyces sp. Saigon413 deposited at the Westerdijk Institute under accession number CBS149411. As another example, the microbial strain can comprise a nucleotide sequence having at least 99.97% identity to SEQ ID NO:1, wherein the microbial strain comprises a genome sequence having at least 99.8%, 99.9% identity, or 100% identity to the entire genome of Streptomyces sp. Saigon413 deposited at the Westerdijk Institute under accession number CBS 149411. As another example, the microbial strain can comprise a nucleotide sequence having at least 99.98% identity to SEQ ID NO:1, wherein the microbial strain comprises a genome sequence having at least 99.8%, 99.9% identity, or 100% identity to the entire genome of Streptomyces sp. Saigon413 deposited at the Westerdijk Institute under accession number CBS 149411. As another example, the microbial strain can comprise a nucleotide sequence having at least 99.99% identity to SEQ ID NO:1, and the microbial strain comprises a genome sequence having at least 99.8%, 99.9% identity, or 100% identity to the complete genome of Streptomyces sp. Saigon413, deposited at the Westerdijk Institute under accession number CBS149411.As another example, the microbial strain can comprise a nucleotide sequence having at least 100% identity to SEQ ID NO:1, and the microbial strain comprises a genome sequence having at least 99.8%, 99.9% identity, or 100% identity to the entire genome of Streptomyces sp. Saigon413, deposited at the Westerdijk Institute under accession number CBS149411.

[0016] Suitably, the microbial strain may comprise a nucleotide sequence having at least 99.99% identity to SEQ ID NO: 1, and the microbial strain comprises a genome sequence having at least 99.9% identity to the complete genome of Streptomyces sp. Saigon413 deposited at the Westerdijk Institute under accession number CBS149411. The term "isolated" in relation to a microbial strain means that the microbial strain has been isolated from its natural environment.

[0017] Surprisingly, it has been found that the microbial strains according to the present invention exhibit pesticidal activity against plant pathogenic microorganisms found in various crops. Surprisingly, the microbial strains disclosed herein produce one or more novel metabolites or combinations of metabolites that have pesticidal activity, preferably fungicidal activity. Surprisingly, the microbial strains disclosed herein produce one or more novel metabolites or combinations of metabolites that have pesticidal activity, preferably fungicidal activity. 53 H 90 N2O 44 and further characterized by the NMR spectra listed in Tables 1 and 2, preferably characterized by structural formula I, malonomycin, cyclothiazomycin C, and streptimidon, an oligosaccharide compound according to compound I, a lipopeptide according to formula II, or a salt thereof (wherein R is CH or C H ), and C 67 H 115 NO 25wherein the polyene is further characterized by a spectrum of optical absorption as shown in FIG. 10.

[0018] Preferably, the microbial strains disclosed herein are C. 53 H 90 N2O 44 and further characterized by the NMR spectra listed in Tables 1 and 2, and preferably characterized by structural formula I disclosed herein, as well as cyclothiazomycin C, streptimidon, an oligosaccharide compound according to compound I, a lipopeptide or a salt thereof according to formula II disclosed herein (wherein R is CH or C H ), and C 67 H 115 NO 25 wherein the polyene is further characterized by a spectrum of optical absorption as shown in FIG. 10.

[0019] Preferably, the microbial strains disclosed herein are C. 53 H 90 N2O 44 and at least one, at least two, or at least three compounds selected from malonomycin, cyclothiazomycin C, and streptimidon, which comprise a molecular formula according to the formula I and are further characterized by the NMR spectra listed in Tables 1 and 2, and are preferably characterized by structural formula I disclosed herein; at least one, at least two, or at least three compounds selected from oligosaccharide compounds according to compound I; a lipopeptide according to formula II disclosed herein, or a salt thereof, wherein R is CH or C H; 67 H 115 NO25 wherein the polyene produces or is capable of producing a polyene compound characterized by a molecular formula according to the formula:

[0020] As used herein, the term "microbial strain, compound, metabolite, or pesticidal composition" or "pesticide" refers to a microbial strain, compound, metabolite, or composition that controls, modifies, or prevents the growth of pests. The term "pesticidally effective amount," when used, refers to the amount of such a microbial strain, compound, metabolite, or composition, or combination of such compounds, that is capable of producing an effect on the growth of pests. A controlling or modifying effect includes any deviation from natural occurrence, such as killing, delaying, etc., and prevention includes the formation of a barrier or other defense in the plant to prevent pest infection.

[0021] As used herein, the terms "fungicidally active microbial strain, compound, metabolite, or composition" or "fungicide" refer to a microbial strain, compound, metabolite, or composition that controls, modifies, or prevents fungal growth. The term "fungicidally effective amount," when used, refers to the amount of such a microbial strain, compound, metabolite, or composition, or combination of such compounds, that is capable of producing an effect against fungal growth. A control or modification effect includes any deviation from natural occurrence, such as killing, delaying, etc., and prevention includes the formation of a barrier or other defense in plants to prevent fungal infection.

[0022] Preferably, the microbial strains disclosed herein are C. 53 H 90 N2O 44and at least one compound selected from malonomycin, cyclothiazomycin C, streptimidon, and an oligosaccharide compound according to compound I, which comprises a molecular formula according to the formula: 67 H 115 NO 25 wherein the polyene produces or is capable of producing a polyene compound characterized by a molecular formula according to the formula:

[0023] Suitably, the microbial strains disclosed herein are C. 53 H 90 N2O 44 and further characterized by the NMR spectra listed in Tables 1 and 2, preferably characterized by structural formula I, malonomycin, cyclothiazomycin C, streptimidon, an oligosaccharide compound according to compound I, a lipopeptide or a salt thereof according to formula II (wherein R is CH or C H ), and C 67 H 115 NO 25 wherein the polyene produces or is capable of producing a polyene compound characterized by a molecular formula according to the formula:

[0024] As used herein, the terms "percent identity" and "percent identity" 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 in both sequences where the same nucleic acid base (for nucleotide sequences) or amino acid residue (for proteins) occurs to obtain the number of matched positions, (iii) dividing the number of matched positions by the total number of positions in the comparison window, and then (iv) multiplying this quotient by 100 percent to obtain the percent identity. When "percent identity" is calculated relative to a reference sequence where a specific comparison window is not designated, the percent identity is determined by dividing the number of matched positions over the alignment region by the total length of the reference sequence. Thus, for the purposes of the present invention, when two sequences (query and subject) are optimally aligned (gaps in their alignment are allowed), the "percent identity" of the query sequence equals the number of identical positions between the two sequences divided by the total number of positions in the query sequence over its length (or comparison window), then multiplied by 100 percent.

[0025] In one embodiment, the microbial strain according to the invention is Streptomyces chrestomyceticus, Streptomyces sp. Saigon413, deposited at the Westerdijk Institute under accession number CBS149411.

[0026] The present invention also provides a method for producing a 53 H 90 N2O 44 and further characterized by the NMR spectra listed in Tables 2 and 3, preferably structural formula I: [ka] and at least one compound selected from the group consisting of malonomycin, cyclothiazomycin C, and streptimidon, and an oligosaccharide compound according to compound I, formula II: [ka] or a salt thereof, wherein R1 is CH3 or C2H5, and 67 H 115 NO 25 wherein the polyene is further characterized by a light absorption spectrum as shown in FIG. 67 H 115 NO 25 A polyene having a molecular formula of has a molecular weight of 1333.7758g.

[0027] Preferably, the present disclosure provides 53 H 90 N2O 44 and further characterized by the NMR spectra listed in Tables 2 and 3, preferably characterized by structural formula I, and at least one compound selected from the group consisting of malonomycin, cyclothiazomycin C, and streptimidon, and at least one compound selected from the group consisting of oligosaccharide compounds according to compound I, a lipopeptide according to formula II, or a salt thereof, wherein R is CH or C H , and 67 H 115 NO 25wherein the polyene is further characterized by a light absorption spectrum as shown in FIG.

[0028] The compositions according to the invention are effective against phytopathogenic microorganisms, in particular those of the genera Zymoseptoria, Puccinia, Mycorsphearella, Pyricularia, Rhizoctonia, Blumeria, Alternaria, Colletotrichum, Ramularia, Parastagonospora, Rhynchosporium, Oculimacula, Fusarium, Gaeumannomyces, and the like. It has been found that the compounds have a surprising level of biological activity against plant pathogenic fungi such as Botrytis sp., Botrytis or Sclerotinia, plant pathogenic bacteria Xanthomonas, and plant pathogenic oomycetes, e.g. Aphanomyces sp.

[0029] Cyclothiazomicin C is a known compound. The structure of cyclothiazomycin C is disclosed on page 3 of WO2015191789 and can be produced as disclosed in Example 4 of WO2015 / 191789.

[0030] Malonomycin (sometimes spelled "malonomycin") is {[(2S)-2-amino-3-hydroxypropanoyl]amino}{2-[(5S)-5-(aminomethyl)-4-hydroxy-2-oxo-2,5-dihydro-1H-pyrrol-3-yl]-2-oxoethyl}malonic acid and can be produced as disclosed in Example I of WO 2006 / 078939. Malonomycin can also be prepared according to the methods disclosed in Examples IA and B of EP 1 860 939 or according to Law et al., 2018 (Nature Catalyses | VOL 1 | DECEMBER 2018 | 977-984).

[0031] Streptimidon is a compound of formula III [ka] is a known compound.

[0032] Strepimidone can be synthesized according to 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).

[0033] The composition according to the invention preferably comprises 53 H 90 N2O 44and at least one, at least two, or at least three of the compounds selected from the group consisting of malonomycin, cyclothiazomycin C, and streptimidon, and an oligosaccharide compound according to compound I, which comprises a molecular formula according to formula I and is further characterized by the NMR spectra listed in Tables 1 and 2, and is preferably characterized by structural formula I; a lipopeptide according to formula II, or a salt thereof (wherein R is CH or C H ); and C 67 H 115 NO 25 wherein the polyene is further characterized by an optical absorption spectrum as shown in FIG.

[0034] The compositions disclosed herein are non-naturally occurring compositions.

[0035] Preferably, the composition according to the invention comprises 53 H 90 N2O 44 and further characterized by the NMR spectra listed in Tables 1 and 2, preferably characterized by structural formula I, malonomycin, cyclothiazomycin C, and streptimidon, an oligosaccharide compound according to compound I, a lipopeptide according to formula II, or a salt thereof (wherein R is CH or C H ), and C 67 H 115 NO 25 wherein the polyene is further characterized by a light absorption spectrum as shown in FIG. 10.

[0036] The lipopeptides according to Formula II disclosed herein have the formula II(a): [ka] Contains lipopeptides by.

[0037] The lipopeptide according to formula II(a) has the molecular formula C 55 H 85 N 11 O 19 and an exact mass of 1203.602 g. The lipopeptide according to formula I(a) has a solubility in DMSO of greater than 10,000 ppm.

[0038] The lipopeptides according to Formula II disclosed herein have the formula II(b): [ka] Contains lipopeptides by.

[0039] The lipopeptide according to formula II(b) has the molecular formula C 56 H 87 N 11 O 19 and an exact mass of 1217.618 g. The lipopeptide according to formula I(b) has a solubility in DMSO of greater than 10,000 ppm.

[0040] Preferably, the composition comprises a strain of Streptomyces, the strain being Streptomyces chrestomyceticus. Preferably, the composition comprises a strain of Streptomyces, preferably Streptomyces chrestomyceticus, which comprises a genomic sequence having at least 91%, 92%, 93%, 94%, preferably at least 95%, 96%, 97%, 98%, or at least 99% identity to the complete genome of Streptomyces chrestomyceticus NRRL-3672 or the complete genome of Streptomyces sp. Saigon413 deposited at the Westerdijk Institute under accession number CBS149411. In one embodiment, a composition comprises Streptomyces chrestomyceticus comprising a genome sequence having 100% identity to the entire genome of Streptomyces sp. Saigon413, deposited at the Westerdijk Institute under accession number CBS 149411. In one embodiment, a composition comprises Streptomyces chrestomyceticus that is Streptomyces sp. Saigon413, deposited at the Westerdijk Institute under accession number CBS 149411.

[0041] The microbial strain according to the invention, or the strain of Streptomyces disclosed herein, preferably comprises at least one nucleotide sequence encoding a protein having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity or 100% identity to the amino acid sequence set forth in SEQ ID NOs: 71 to 115, preferably at least one nucleotide sequence encoding a protein having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity or 100% identity to the amino acid sequence set forth in SEQ ID NO: 91 and / or SEQ ID NO: 92.

[0042] In one embodiment, a microbial strain according to the invention or a Streptomyces strain disclosed herein comprises at least one nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity, or 100% identity to at least one nucleotide sequence of SEQ ID NOs: 2 to 46. The nucleotide sequences set forth in SEQ ID NOs: 2 to 46 comprise a gene cluster as shown in Figure 11 for the production of a lipopeptide according to Formula II.

[0043] Preferably, the microbial strains according to the present invention and / or strains of Streptomyces, such as Streptomyces chrestomyceticus, disclosed herein are selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, At least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, or at least one of SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16; SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, or SEQ ID NO:46 at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, at least 34, at least 35, at least 36, at least 37, at least 38, at least 39, at least 40, at least 41, at least 42, at least 43, at least 44, at least 45 nucleotide sequences to which the nucleotide sequences share / have at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity.Preferably, the microbial strain according to the invention comprises a nucleotide sequence which has at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity or 100% identity to at least one of the nucleotide sequences set forth in SEQ ID NO: 22 and / or 23. Suitably, the microbial strain according to the invention comprises a nucleotide sequence which has at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity or 100% identity to the nucleotide sequence set forth in SEQ ID NO: 22. Suitably, the microbial strain according to the invention comprises a nucleotide sequence which has at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity or 100% identity to the nucleotide sequence set forth in SEQ ID NO: 23.

[0044] In another embodiment, a microbial strain according to the invention and / or a strain of Streptomyces, such as Streptomyces chrestomyceticus, disclosed herein comprises at least one nucleotide sequence encoding a protein having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity, or 100% identity to at least one of the amino acid sequences set forth in SEQ ID NOs: 116-139.

[0045] Preferably, the microbial strain according to the present invention and / or Streptomyces crestomyceticus Strains of Streptomyces such as Streptomyces sp. (e.g., Streptomyces chrestomyceticus) may have an amino acid sequence having at least 80% identity, preferably at least 85%, preferably at least 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, preferably at least 99%, preferably 100% identity to the amino acid sequence set forth in SEQ ID NO:116, SEQ ID NO:117, SEQ ID NO:118, SEQ ID NO:119, SEQ ID NO:120, SEQ ID NO:121, SEQ ID NO:122, SEQ ID NO:123, SEQ ID NO:124, SEQ ID NO:125, SEQ ID NO:126, SEQ ID NO:127, SEQ ID NO:128, SEQ ID NO:129, SEQ ID NO:130, SEQ ID NO:131, SEQ ID NO:132, SEQ ID NO:133, SEQ ID NO:134, SEQ ID NO:135, SEQ ID NO:136, SEQ ID NO:137, SEQ ID NO:138, and / or SEQ ID NO:139. It comprises or contains at least one, preferably at least two, preferably at least three, preferably at least four, preferably at least five, preferably at least six, preferably at least seven, preferably at least eight, preferably at least nine, preferably at least ten, preferably at least eleven, preferably at least 12, preferably at least 13, preferably at least 14, preferably at least 15, preferably at least 16, preferably at least 17, preferably at least 18, preferably at least 19, preferably at least 20, preferably at least 21, preferably at least 22, preferably at least 23, preferably all 24 nucleotide sequences encoding proteins that have / have identity thereto.

[0046] Preferably, the microbial strain according to the invention and / or the strain of Streptomyces such as Streptomyces chrestomyceticus disclosed herein comprises at least one nucleotide sequence encoding a protein having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity, or 100% identity to at least one of the amino acid sequences set forth in SEQ ID NO: 136 and / or SEQ ID NO: 137. Suitably, the microbial strain according to the invention and / or the strain of Streptomyces, such as Streptomyces chrestomyceticus disclosed herein comprises a nucleotide sequence encoding a protein having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 136. Suitably, the microbial strain according to the invention and / or the strain of Streptomyces, such as Streptomyces chrestomyceticus disclosed herein comprises a nucleotide sequence encoding a protein having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 137.

[0047] In another embodiment, a microbial strain according to the invention and / or a strain of Streptomyces, such as Streptomyces chrestomyceticus, disclosed herein comprises at least one nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to at least one of the nucleotide sequences set forth in SEQ ID NOs: 47-70. 67 H 115 NO 2512 for the production of a polyene having a molecular formula according to the present invention, wherein the polyene is further characterized by an optical absorption spectrum as shown in Figure 10. Preferably, a microbial strain according to the present invention and / or a strain of Streptomyces, such as Streptomyces chrestomyceticus disclosed herein, comprises a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to at least one of the nucleotide sequences set forth in SEQ ID NO: 67 and / or 68. Suitably, a microbial strain according to the present invention comprises a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the nucleotide sequence set forth in SEQ ID NO: 67. Preferably, the microbial strain according to the invention comprises a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity or 100% identity to the nucleotide sequence set forth in SEQ ID NO: 68.

[0048] Preferably, the microbial strains according to the present invention and / or strains of Streptomyces, such as Streptomyces chrestomyceticus, are selected from the group consisting of SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52. or comprises at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24 nucleotide sequence of SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, or SEQ ID NO:70, or which nucleotide sequence has at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity thereto.

[0049] The microbial strains or strains of Streptomyces or Streptomyces chrestomyceticus disclosed herein can be naturally occurring or recombinant microorganisms. Recombinant microorganisms can be produced by methods known to those skilled in the art. Recombinant microorganisms can be produced by transforming a microorganism with one of the nucleotide sequences encoding the proteins with amino acids set forth in SEQ ID NOS: 71-115 and / or 116-139, preferably the nucleotide sequences set forth in SEQ ID NOS: 2-46, preferably SEQ ID NOS: 22 and / or 23, and / or the nucleotide sequences set forth in SEQ ID NOS: 47-70, preferably SEQ ID NOS: 67 and 68.

[0050] Preferably, the compositions according to the present invention comprise an adjuvant, preferably an agriculturally acceptable adjuvant. The compositions disclosed herein are preferably agriculturally acceptable compositions.

[0051] Suitable adjuvants are known in the art and include, for example, solvents, liquid carriers, solid carriers or fillers, surfactants, dispersants, emulsifiers, wetting agents, adjuvants, solubilizers, penetration enhancers, protective colloids, adhesives, thickeners, humectants, repellents, attractants, nutrient stimulants, compatibilizers, bactericides, antifreeze agents, antifoaming agents, colorants, tackifiers, and binders.

[0052] Suitable solvents and liquid carriers include, for example, water, organic solvents, oils of plant or animal origin, cyclic and aromatic hydrocarbons, alcohols, esters, fatty acids, glycols, or any other suitable liquid carrier known in the art. The solvent or liquid carrier can be water or DMSO.

[0053] Suitable solid carriers include, for example, ammonium salts, talc, titanium dioxide, pyrophyllite clay, silica, attapulgite clay, kieselguhr, chalk, diatomaceous earth, lime, calcium carbonate, bentonite clay, fuller's earth, cottonseed hulls, wheat flour, soy flour, pumice, wood flour, walnut hulls, and lignin.

[0054] The amount of carrier may typically range anywhere from 0.9% to 99.99% by weight of the composition.

[0055] Adjuvants can be surfactants, crystallization inhibitors, viscosity modifiers, suspending agents, spray droplet modifiers, pigments, antioxidants, foaming agents, antifoaming agents, light-blocking agents, compatibilizers, sequestering agents, neutralizing and buffering agents, corrosion inhibitors, dyes, odorants, spreading agents, penetration aids, micronutrients, emollients, lubricants, and adhesives. Adjuvants can include, for example, alkyl polyglucosides and / or polyoxyethylene (6) C9-C11 alcohols, or methylcellulose.

[0056] Dispersants include, but are not limited to, surfactants and wetting agents. Generally, dispersants have low toxicity to the microorganisms in the inoculant composition and to the plant parts to which the inoculant composition is applied.

[0057] The surfactants may be ionic (cationic or anionic) or non-ionic surfactants such as ionic or non-ionic emulsifiers, foam formers known in the art, e.g., acids such as polyacrylic acids, esters, ethers, and the like.

[0058] Suitable surfactants include polysorbates, for example polysorbate 20, such as Tween 20®. The compositions disclosed herein comprise polysorbate 20 in an amount of surfactant of 0.0005 wt / wt% to 0.5 wt / wt%, preferably 0.001 to 0.1 wt / wt%, preferably 0.002 to 0.08 wt / wt%, preferably 0.004 to 0.06 wt / wt%, preferably 0.005 to 0.04 wt / wt%, preferably 0.006 to 0.025% wt / wt%.

[0059] In one embodiment, the composition disclosed herein is a fermentation broth, preferably a spray-dried or freeze-dried fermentation broth, or a blend. Spray-drying or freeze-drying of fermentation broths is known in the art.

[0060] The compositions disclosed herein can be used to increase the cell count of Streptomyces chrestomyceticus to 1 x 10 cells per gram of dry weight. 2 ~1×10 13 cfu, e.g., 1 x 10 per gram of dry weight 3 ~1×10 12 cfu, 2 × 10 per g dry weight 3 ~2×10 11 cfu, 5 × 10 per g dry weight 3 ~5×10 11 cfu, e.g., 1 x 10 per gram of dry weight 4 ~1×10 10 cfu, 2 × 10 per g dry weight4 ~2×10 10 cfu, e.g., 1 x 10 per gram of dry weight 5 ~1×10 9 cfu, 2 × 10 per g dry weight 5 ~2×10 9 cfu, 5 × 10 per g dry weight 5 ~5×10 9 cfu, 1 × 10 per g dry weight 6 ~1×10 8 cfu, e.g., 2 x 10 per gram of dry weight 6 ~2×10 8 Contains cfu.

[0061] The compositions disclosed herein include formulations. Thus, the formulations include the compositions disclosed herein. The formulations may be any suitable composition for incorporating the microbial strains disclosed herein, such as Streptomyces sp., for example, Streptomyces chrestomyceticus, for example, Streptomyces sp. Saigon 413, deposited at the Westerdijk Institute under accession number CBS149411.

[0062] Formulations of microbial strains are known in the art, as disclosed, for example, in Croda Crop Care, the Nouryon formulator toolbox, and Formulation of Microbial Biopesticides: Beneficial microorganisms, nematodes, and seed treatments (412 p., 6 December 2012) eds. Burgess HD, Springer, ISBN 978-94-011-4926-6. Preferably, the compositions disclosed herein are formulations in which the microbial strains disclosed herein, such as Streptomyces chrestomyceticus, are formulated as an oil dispersion (OD), a non-aqueous dispersion (NAD), or a flowable formulation.

[0063] Formulations known in the art are, for example, emulsifiable concentrates, coatable pastes, sprayable or dilutable solutions or suspensions, powders, dusts, granules and encapsulations.

[0064] Suspension concentrates are aqueous formulations in which finely divided solid particles of the active ingredient are suspended. Such formulations contain anti-settling and dispersing agents and may further contain wetting agents, as well as anti-foaming and crystal growth inhibitors to enhance pesticidal activity. When used, these concentrates are diluted in water and typically applied as a spray to the area to be treated. The amount of active ingredient can range from 0.5% to 95% of the concentrate.

[0065] Wettable powders are in the form of finely divided particles that disperse readily in water or other liquid carriers. The particles contain the active ingredient held in a solid matrix. Typical solid matrices include fuller's earth, kaolin clay, silica, and other readily wet organic or inorganic solids. Wettable powders usually contain 5% to 95% of the active ingredient and small amounts of wetting agents, dispersing agents, or emulsifying agents.

[0066] Emulsifiable concentrates are homogeneous liquid compositions that are dispersible in water or other liquids and may consist solely of the active ingredient and a liquid or solid emulsifier, or may also contain a liquid carrier such as xylene, high-boiling aromatic naphtha, isophorone, and other non-volatile organic solvents. In use, these concentrates are dispersed in water or other liquid and typically applied as a spray to the area to be treated. The amount of active ingredient can range from 0.5% to 95% of the concentrate.

[0067] Granular formulations include both extrudates and relatively coarse particles, and are usually applied undiluted to the area where treatment is required. Typical carriers for granular formulations include sand, fuller's earth, attapulgite clay, bentonite clay, montmorillonite clay, vermiculite, perlite, calcium carbonate, brick, pumice, pyrophyllite, kaolin, dolomite, gypsum, wood flour, ground corn cobs, ground peanut shells, sugar, sodium chloride, sodium sulfate, sodium silicate, sodium borate, magnesia, mica, iron oxide, zinc oxide, titanium oxide, antimony oxide, cryolite, gypsum, diatomaceous earth, calcium sulfate, and other organic or inorganic materials that can absorb or be coated with the active compound. Granular formulations usually contain 5% to 25% active ingredient, which may include surfactants or vegetable oils such as high-boiling aromatic naphtha, kerosene, and other petroleum fractions; and / or adhesives such as dextrin, glue, or synthetic resins.

[0068] An oil dispersion is a solid active ingredient dispersed in a water-immiscible carrier, oil. Oil dispersions typically contain (by weight): active ingredient (5-60%), non-aqueous dispersant (1-10% solids), aqueous dispersant (1-10% solids), emulsifier (2-10% isol), rheology modifier (0.2-5%), and oil carrier, making up 100%. Suitable oils in oil dispersions can be mineral oil, paraffinic oil, vegetable oil, or methylated oil.

[0069] Suitable adjuvants, dispersants, emulsifiers, and rheology modifiers for oil dispersions depend on the type of oil and are known in the art. Suitable emulsifiers can be alcohol ethoxylates / alkoxylates, such as C16 / 18 ethoxylates or C16 / C18 alkoxylates, or block copolymers. Adjuvants can be alkyl polyglucosides. Suitable rheology modifiers can be clays, hydrogenated castor oil and derivatives, fumed silica polyamides, polyesters, or Agrylan ODS.

[0070] The oil dispersion may comprise an adjuvant, the adjuvant comprising an alkyl polyglucoside and / or a polyoxyethylene (6) C9-C11 alcohol. In the therapeutic treatment of Zymoseptoria tritici and Puccinia recondita, compositions according to the invention formulated as an oil dispersion and an adjuvant, for example an alkyl polyglucoside and / or a polyoxyethylene (6) C9-C11 alcohol, have been found to provide improved reduction of fungal diseases.

[0071] A non-aqueous dispersion is a liquid formulation in which solid ingredients are dispersed in a water-miscible carrier. The solid ingredients are uniformly suspended in the carrier but are not dissolved.

[0072] Flowable formulations contain solid particles in a liquid, usually water. Flowable concentrates also include suspension concentrates. Flowable formulations generally contain (wt%) the active ingredient (5-60%), dispersing / wetting agents (1-10%), rheology modifiers (0.1-0.1%), biocides (0.1%), antifreeze agents (5-10%), antifoaming agents (0.2%), adjuvants (up to 25%), and 100% water.

[0073] As disclosed herein, the isolated microbial strain or composition of the present invention can advantageously be in the form of a soluble concentrate (SL), or a flowable concentrate (FS) for seed treatment, or a suspension concentrate (SC), more preferably a seed treatment slurry to be applied to seeds. Slurries for seed treatment applications are well known in the art. For example, the slurry can contain an active ingredient, such as the isolated microbial strain or composition of the present invention, in the form of a commercially available product or not, mixed with water and optionally at least one polymer to optimize adhesion around the plant propagation material.

[0074] Each active ingredient can be applied to the plant propagation material from a different composition, or the active ingredients can be combined in the same composition (a pre-mixed composition). The composition can contain from about 0.001% to about 99% by weight of the active ingredient, based on the total weight of the composition. Preferably, the composition contains from about 0.001% to about 60% by weight of the active ingredient, based on the total weight of the composition.

[0075] The solid carrier can be a natural or synthetic solid material that is insoluble in water.This carrier is generally inert and acceptable in agriculture, especially in the treatment of seeds or other propagation materials.It can be selected from, for example, clay, diatomaceous earth, natural or synthetic silicate, titanium dioxide, magnesium silicate, aluminum silicate, talc, pyrophyllite clay, silica, attapulgite clay, kieselguhr, chalk, limestone, calcium carbonate, calcium montmorillonite, bentonite clay, fuller's earth, cottonseed husk, wheat flour, soybean flour, pumice, wood flour, crushed walnut shell, lignin, etc.

[0076] Thus, the isolated microbial strains and / or compositions according to the invention can preferably be attached to propagation material such as seeds.

[0077] Compositions according to the present invention include dressing compositions which refer to liquid compositions useful for at least partially or totally coating and / or wetting plant propagation material, more preferably seeds.

[0078] The compositions according to the invention are particularly suitable for dressing application to plant propagation material, especially seeds.

[0079] Typically, tank-mix formulations for seed treatment applications comprise 0.25% to 80% by weight, particularly 1 to 75% by weight, of an active ingredient, such as an isolated microbial strain or composition disclosed herein, and 99.75% to 20% by weight, particularly 99% to 25% by weight, of a solid or liquid adjuvant (including, for example, a solvent such as water), which adjuvant may be a surfactant in an amount of 0 to 40% by weight, particularly 0.5 to 30% by weight, based on the total weight of the tank-mix formulation.

[0080] Typically, premix formulations for seed treatment applications comprise 0.5 to 99.9% by weight, particularly 1 to 95% by weight, of an active ingredient, such as an isolated microbial strain or composition disclosed herein, and 99.5 to 0.1% by weight, particularly 99% to 5% by weight, of a solid or liquid adjuvant (e.g., including a solvent such as water), which adjuvant may be a surfactant in an amount of 0 to 50% by weight, particularly 0.5 to 40% by weight, based on the total weight of the premix formulation.

[0081] Commercially available products will preferably be formulated as concentrates (e.g., premix compositions), while end users will typically employ dilute formulations (e.g., tank-mix compositions).

[0082] Preferably, compositions such as formulations of Streptomyces chrestomyceticus disclosed herein, for example, Streptomyces sp. Saigon 413 deposited at the Westerdijk Institute under accession number CBS 149411, comprise 10-60 wt / wt% per gram of dry weight of spray-dried or freeze-dried fermentation broth of Streptomyces chrestomyceticus, such as Streptomyces sp. Saigon 413 deposited at the Westerdijk Institute under accession number CBS 149411, and preferably 20-50 wt / wt% per gram of dry weight of the spray-dried or freeze-dried fermentation broth.

[0083] In one embodiment, the isolated microbial strain and / or composition of the present invention further comprises at least one additional active ingredient used in the methods of the present invention adjacent to a Streptomyces strain, such as Streptomyces chrestomyceticus, or the isolated microbial strain and composition disclosed herein, applied simultaneously or sequentially with the microbial strain and / or composition of the present invention. Active ingredients, as defined herein, have fungicidal and / or insecticidal and / or herbicidal activity, or have activity as a plant growth regulator. The isolated microbial strain or composition of the present invention may be mixed with one or more additional ingredients having pesticidal activity, such as a fungicide, insecticide, herbicide, bactericide, miticide, or nematicide, and / or the additional ingredient optionally includes a plant growth regulator. Pesticides are referred to herein by their common names and are known, for example, from "The Pesticide Manual," 19th Ed., British Crop Protection Council, 2021.

[0084] Additional components with pesticide activity, such as fungicide activity, can produce unexpected synergistic activity.Additional components with pesticide activity and / or plant growth regulator activity can be used in the method of the present invention in combination with the microbial strain or composition of the present invention, and can be applied simultaneously or sequentially with the composition of the present invention.When applied simultaneously, these additional components can be formulated together with the composition of the present invention, or can be mixed, for example, in a spray tank.As an alternative to directly mixing these additional components with pesticide activity, the components can be used in separate fungicide, insecticide or herbicide applications as part of a fungus, insect or grass control program that extends over part or all of the growing season.

[0085] The at least one additional ingredient having pesticidal activity and / or plant growth regulator can be any suitable known fungicide, insecticide, herbicide and / or plant growth regulator. The at least one additional ingredient having pesticidal activity and / or plant growth regulator can be of chemical or biological origin, for example, plant or microbial origin.

[0086] Additionally, the compositions of the present invention can be applied in conjunction with one or more systemic acquired resistance inducers ("SAR" inducers). SAR inducers are known and are described, for example, in U.S. Patent No. 6,919,298, and include, for example, salicylates and the commercially available SAR inducer acibenzolar-S-methyl.

[0087] The isolated microbial strain or composition according to the present invention may induce plant resistance priming, a mechanism that leads to a physiological state that allows plants to respond more quickly and / or more robustly after exposure to a biological or biotic stress, as described, for example, in the review article: P. Aranega-Bou et al., "Priming of plant resistance by natural compounds. Hexanoic acid as a model." Front. Plant. Sci. 1, October 2014.

[0088] The composition of the invention and the at least one additional active ingredient are preferably mixed in a ratio of 100:1 to 1:6000, in particular 50:1 to 1:50, more particularly 20:1 to 1:20, more particularly 10:1 to 1:10, in particular 5:1 and 1:5, with a ratio of 2:1 to 1:2 being particularly preferred, and a ratio of 4:1 to 2:1 being likewise preferred, in particular 1:1, or 5:1, or 5:2, or 5:3, or 5:4, or 4:1, or 4:2, or 4:3, or 3:1, or 3:2, or or 2:1, or 1:5, or 2:5, or 3:5, or 4:5, or 1:4, or 2:4, or 3:4, or 1:3, or 2:3, or 1:2, or 1:600, or 1:300, or 1:150, or 1:35, or 2:35, or 4:35, or 1:75, or 2:75, or 4:75, or 1:6000, or 1:3000, or 1:1500, or 1:350, or 2:350, or 4:350, or 1:750, or 2:750, or 4:750. These mixing ratios are by weight. These mixtures may be used in methods for controlling pests, which methods comprise applying to the pest or its environment a composition comprising the mixture as described above, excluding methods for treating the human or animal body by surgery or therapy or diagnostic methods performed on the human or animal body.

[0089] Compositions comprising mixtures of isolated microbial strains or compositions of the present invention and one or more active ingredients as described above can be applied, for example, in the form of a single "ready-mix"; in combined spray mixtures made up of separate formulations of single active ingredient components, such as "tank mixes"; and in combined use of single active ingredients when applied in a sequential manner, i.e., one after the other, separated by a reasonably short period of time, such as a few hours or days.

[0090] In another aspect, the present invention also relates to a process for producing a microbial strain according to the invention as disclosed hereinabove or a composition according to the invention as disclosed hereinabove, comprising culturing a microbial strain or strain of Streptomyces, such as Streptomyces chrestomyceticus, under suitable fermentation conditions in a suitable fermentation medium, and optionally recovering the microbial strain or composition. Typically, a fermentation broth is produced during or at the time of culturing the microbial strain or strain of Streptomyces, such as a Streptomyces chrestomyceticus strain. Suitable fermentation conditions for culturing Streptomyces sp. are known to those skilled in the art.

[0091] Culturing the microbial strains disclosed herein, or the Streptomyces strains disclosed herein, such as Streptomyces chrestomyceticus strains, includes culturing the microbial strains under aerobic conditions in the presence of a carbon source and a nitrogen source at a temperature between 15°C and 45°C, preferably between 20°C and 35°C, and preferably between 25°C and 32°C. Suitable carbon sources may be molasses, such as beet molasses or sugarcane molasses, polysaccharides, wheat flour, starch, sugar, or glucose. Suitable nitrogen sources may be casein hydrolysate, tryptone, ammonium sulfate, ammonia, yeast extract, peptone, or urea. The process for producing the microbial strains or S. chrestomyceticus disclosed herein may be carried out in batch, fed-batch, or continuous culture.

[0092] Preferably, the process according to the invention comprises culturing a microbial strain or strain of Streptomyces as disclosed herein, preferably Streptomyces chrestomyceticus, wherein the microbial strain or strain of Streptomyces, preferably Streptomyces chrestomyceticus, is selected from the group consisting of C. 53 H 90 N2O 44 and further characterized by the NMR spectra listed in Tables 1 and 2, preferably wherein Compound I is further characterized by structural formula I, a compound selected from malonomycin, cyclothiazomycin C, streptimidon, an oligosaccharide compound according to Compound I, a lipopeptide according to Formula II, or a salt thereof (wherein R is CH or C H ), and C 67 H 115 NO 25 wherein the polyene is further characterized by a light absorption spectrum as shown in FIG. 10.

[0093] Preferably, the process according to the invention comprises culturing a microorganism or strain of Streptomyces, such as Streptomyces chrestomyceticus, the microorganism or strain being selected from the group consisting of C. 53 H 90 N2O 44and further characterized by the NMR spectra listed in Tables 1 and 2, preferably compound I being further characterized by structural formula I, at least one, at least two, or at least three of compounds selected from the group consisting of malonomycin, cyclothiazomycin C, streptimidon, and at least one, at least two, or at least three of compounds selected from oligosaccharide compounds according to compound I, and a lipopeptide according to formula II, or a salt thereof, wherein R is CH or C H , and C 67 H 115 NO 25 wherein the polyene is further characterized by a spectrum of optical absorption as shown in Figure 10. Preferably, the microbial strain or strain of Streptomyces, such as Streptomyces chrestomyceticus, produces a polyene compound characterized by a molecular formula according to the formula: 53 H 90 N2O 44 and further characterized by the NMR spectra listed in Tables 1 and 2, preferably Compound I being further characterized by structural formula I, a compound selected from the group consisting of malonomycin, cyclothiazomycin C, streptimidon and a compound according to Compound I, and a lipopeptide or a salt thereof according to Formula II (wherein R is CH or C H ), and C 67 H 115 NO 25 The polyene compound is further characterized by its optical absorption spectrum as shown in FIG.

[0094] The process according to the invention may further comprise the step of recovering the microbial strain or composition according to the invention. Recovering the microbial strain or composition according to the invention may comprise centrifuging or filtering the fermentation broth. Preferably, the recovery comprises drying the fermentation broth, for example by spray drying or freeze drying. Spray drying or freeze drying are methods known to those skilled in the art. The composition may be prepared by the method of C. 53 H 90 N2O 44 and further characterized by the NMR spectra listed in Tables 1 and 2, preferably compound I being further characterized by structural formula I, compounds malonomycin, cyclothiazomycin C, streptimidon, oligosaccharide compounds according to compound I, lipopeptides or salts thereof according to formula II (wherein R1 is CH3 or C2H5), and C 67 H 115 NO 25 wherein the polyene is further characterized by an optical absorption spectrum as shown in FIG. 10, which may be recovered by suitable methods known in the art, for example, by crystallization or chromatography, e.g., HPLC.

[0095] The microbial strains and strains of Streptomyces of the invention, such as Streptomyces chrestomyceticus, embodiments thereof as disclosed hereinabove are also applicable to the microbial strains, strains of Streptomyces, and Streptomyces chrestomyceticus in the processes and methods of the invention.

[0096] The process according to the present invention may further comprise the step of formulating the microbial strain or the composition according to the present invention into a suitable formulation or composition as disclosed herein above.

[0097] In another aspect, the present invention relates to a method for controlling or preventing infestation of plants, plant propagation material, and / or harvested food, non-food, and feed crops by phytopathogenic microorganisms by treating the plants, plant propagation material, and / or harvested food crops with an effective amount of Streptomyces chrestomyceticus, a microbial strain, or a composition described herein. Surprisingly, Streptomyces chrestomyceticus, such as a microbial strain according to the present invention, or a composition according to the present invention, has been found to be highly effective in treating phytopathogenic microorganisms on plants. Streptomyces chrestomyceticus, such as a microbial strain according to the present invention, or a composition according to the present invention, was surprisingly active as a pesticide and against fungi, bacteria, and oomycetes.

[0098] Methods according to the invention exclude methods of treatment of the human or animal body by surgery or therapy and diagnostic methods practised on the human or animal body.

[0099] The term "plant" refers to any physical part of a plant, including seeds, seedlings, saplings, roots, tubers, stems, stalks, leaves, and fruits. It also includes germinated plants and young plants that are to be transplanted after germination or emergence from the soil. These young plants can be protected by a complete or partial immersion treatment before transplanting.

[0100] The term "plant propagation material" is understood to refer to reproductive parts of plants, such as seeds, and vegetative parts, such as cuttings or tubers (e.g., potatoes), roots, fruits, bulbs, rhizomes, or parts of plants, that can be used for their propagation. The plant propagation material can be treated with the isolated microbial strain or composition of the present invention before the material is sown or planted. Alternatively, the plant propagation material may be treated with the isolated microbial strain or composition of the present invention during sowing or planting. Furthermore, the isolated microbial strain or composition of the present invention may be applied to previously treated propagation material before or during its planting. The isolated microbial strain or composition of the present invention may be applied during sowing of seeds. The isolated microbial strain or composition may also be used on plant propagation material derived from plants grown in greenhouses and / or during transplanting. More preferably, the plant propagation material is a seed.

[0101] The term plants includes "useful plants" or "crops". The expressions "useful plants" and "crops" are used interchangeably herein. "Useful plants" and "crops" include, for example, berry plants such as blackberries, blueberries, cranberries, raspberries and strawberries; cereals such as barley, maize (corn), millet, oats, rice, rye, sorghum, triticale and wheat; fiber plants such as cotton, flax, hemp, jute and sisal; field crops such as sugar and fodder beets, coffee, hops, mustard, oilseed rape (canola), poppy, sugarcane, sunflower, tea and tobacco; fruit trees such as apple, apricot, avocado, banana, cherry, citrus fruit, nectarine, peach, pear and plum; pasture grasses such as bermudagrass, strawberry vine, bentgrass, centipedegrass, fescue, rye, lawn grass and wild grass; basil, borage These include herbs such as bay leaves, chives, coriander, lavender, lovage, mint, oregano, parsley, rosemary, sage, and thyme; legumes such as beans, lentils, peas, and soybeans; nuts such as almonds, cashews, peanuts, hazelnuts, peanuts, pecans, pistachios, and walnuts; palms such as oil palm; ornamental plants such as flowers, shrubs, and trees; other trees such as cocoa, coconuts, olives, and rubber trees; vegetables such as asparagus, eggplant, broccoli, cabbage, carrots, cucumber, garlic, lettuce, squash, melon, okra, onion, pepper, potato, pumpkin, rhubarb, spinach, and tomato; and perennial and annual crops such as vines, grapes, etc. The term "plant" also includes timber crops, such as pine trees, or woody plants.

[0102] The term "useful plants" should also be understood to include useful plants that have been rendered tolerant to herbicides such as bromoxynil or to certain classes of herbicides (e.g., HPPD inhibitors, ALS inhibitors such as primisulfuron, prosulfuron and trifloxysulfuron, EPSPS (5-enol-pyrobyl-shikimate-3-phosphate-synthase) inhibitors, GS (glutamine synthetase) inhibitors or PPO (protoporphyrinogen-oxidase) inhibitors, etc.) by conventional breeding or genetic engineering methods.

[0103] It should be understood that the term "useful plants" also includes useful plants that have been transformed using recombinant DNA techniques so as to be able to synthesize one or more selectively acting toxins, such as those known from toxin-producing bacteria, particularly bacteria of the genus Bacillus.

[0104] As used herein, the term "habitat" means the field in which the plant is growing or in which the seeds of the plant to be cultivated have been sown or will be sown in the soil. It includes the soil, seeds and seedlings, and established vegetation.

[0105] Any suitable plant, plant propagation material or food or feed crop may be treated with the method according to the invention as defined herein. Preferably, the plant, plant propagation material or food crop comprises or is wheat, barley, rice, maize, soybean, sugar beet, banana, tomato, cucumber, and / or groundnut.

[0106] The phytopathogenic microorganisms affected by the isolated microbial strains or compositions according to the invention are fungi and fungal vectors of diseases as well as phytopathogenic bacteria and viruses. The phytopathogenic microorganisms in the method according to the invention include the following fungi and fungal vectors of diseases and phytopathogenic bacteria: Alternaria spp. including Absidia corymbifera, Albugo candida, and A. solani; Aphanomyces spp.; Ascochyta spp.; Aspergillus spp. including A. flavus, A. fumigatus, A. nidulans, A. niga, and A. terrus; Aureobasidium spp. including A. pullulans; Bacillus subtilis, Blastomyces dermatidis, Blumeria graminis, Blumeriella jaapii, Botryosphaeria spp. including B. dothidea and B. obtusa, Botrytis spp. including B. cinerea and Bremia lactucae, Cadophora gregata Candida spp., including C. gregata, C. albicans, C. glabrata, C. krusei, C. lusitaniae, C. parapsilosis, and C. tropicalis; Cephaloascus fragrans; Ceratocystis spp.; Cercospora spp., including C. arachidicola, C. beticola, C. kikuchii, and C. sojina.), Cercosporidium personatum, Cladosporium spp., Clarireedia homoeocarpa, Clavibacter spp., Claviceps purpurea, Coccidioides immitis, Cochliobolus spp., Colletotrichum spp. including C. dematium, C. lindemuthianum, C. musae, C. orbiculare, and C. truncatum spp., Corynespora cassiicola, Cryptococcus neoformans, Diaporthe spp., Dickeya zeae, Didymella spp., Drechslera spp., Elsinoe spp., Epidermophyton spp., Erwinia spp. including Eremothecium gossypiim, E. amylovora, and E. carotovora. Erysiphe spp., including E. cichoracearum, E. necator, Eutypa lata, F. culmorum, F. graminearum, F. langsethiae, F. moniliforme, F. oxysporum, F. poae, F. proliferatum, F.Several Fusarium species, including F. proliferatum, F. pseudograminearum, F. sacchari, F. sambucinum, F. subglutinans, F. solani, F. sporotrichioides, F. tricinctum, and F. virguliforme, and Gaeumannomyces graminis, Gibberella spp., including G. graminis, G. avenacea, G. fujikuroi, G. intricans, G. moniliformis, and G. zeae, Gloeodes pomigena, Gloeosporium musarum, Glomerella cingulate, Golovinomyces cichoracearum, Gymnosporangium juniperi-virginianae, and Guignardia bidwellii. bidwellii, Gymnosporangium juniperi-virginianae, Helminthosporium spp., Hemileia spp., Helminthosporium spp., Hemileia spp., Histoplasma spp. including H. capsulatum.), Hyaloperonospora parasitica, Kabatiella zeae, Laetisaria fuciformis, Leptographium lundbergii, Leveilla taurica, Lophodermium seditiosum, Microdochium majus, Microdochium nivale, multiple species of Microsporum, multiple species of Monilinia including M. fructicola, multiple species of Monilinia including M. nivalis, and multiple species of Monographella including M. nivalis. spp., Mucor spp., Mycosphaerella spp. including M. arachidis, M. fijiensis, M. graminicola, and M. pomi, Nakataea oryzae, Neopseudocercosporella spp., Oculimacula spp., Oncobasidium theobromaeon, Ophiostoma spp., Pantoea stewartia, and Paracoccidioides spp. spp., Parastagonospora nodorum, Pectobacterium spp., P. digitatum, Penicillium spp. including P. italicum, Petriellidium spp., P. maydis, P.Peronosclerospora spp. including P. philippinensis and P. sorghi, Peronospora spp. including P. destructor, Phaeosphaeria nodorum, Phakopsora pachyrhizi, Phellinus igniarus, and Phialophora spp. Phlyctema vagabunda, Phoma spp., Phomopsis viticola, Phyllachora pomigena, Phyllosticta spp., Physoderma maydis, Phytophthora spp. including P. capsica and P. infestans, Plasmopara spp. including Plasmodiophora brassicae, P. halstedii, and P. viticola, and Plenodomus spp. Pleospora spp., Podosphaera spp. including P. leucotricha, Polymyxa graminis, Polymyxa betae, Pseudocercospora fijiensis, Pseudocercosporella herpotrichoides, Pseudomonas spp. including P. syringae, Pseudoperonospora spp. including P. cubensis and P. humuli Several species of Puccinia, including Pseudopeziza tracheiphila, Pseudopyrenochaeta lycopersici, P. hordei, P. recondita, P. striiformis, and P. triticina, are also found.), Pyrenopeziza spp., Pyrenophora spp., Pyricularia spp. including P. oryzae, Pythium spp. including P. ultimum, Ralstonia solanacearum, Ramularia spp., Rathayibacter spp., Remotididymella destructiva, Rhizoctonia spp., Rhizomucor pusillus, Rhizopus arizos arrhizus, Rhynchosporium spp., Robbsia andropogonis, Sarocladium oryzae, Scedosporium spp. including S. apiospermum and S. prolificans, Schizothyrium pomi, Sclerophthora macrospora, Sclerotinia spp. including S. sclerotiorum, Sclerotium spp., S. nodorum, S.Septoria species including Setosphaeria turcica, Setosphaeria turcica, Sphaerotheca macularis, Sphaerotheca fusca, Spiroplasma kunkelii, Sporothorix species, Stagonospora nodorum, Stagonosporopsis cucurbitacearum, Stemphylium species, Stenocarpella macrospora, Stereum hirsutum hirsutum, Streptomyces spp., Thanatephorus cucumeris, Thielaviopsis basicola, Tilletia spp., Tranzschelia discolor, Trichoderma spp. including T. harzianum, T. pseudokoningii, and T. viride, Trichophyton spp., Typhula spp., Uncinula necator, and Urocystis spp. spp.), multiple species of Uromyces, multiple species of Ustilago, multiple species of Venturia including Vibrio inaequalis.), Verticillium spp., Wilsononomyces carpophilus, or Xanthomonas spp. including Xanthomonas oryzae and Xanthomonas campestris, Xylella spp., Zymoseptoria tritici.

[0107] Surprisingly, Streptomyces crestomyceticus (Streptomyces Phytopathogenic microorganisms that have been found to be affected by the chrestomyceticus, microbial strains or compositions according to the invention are fungi, for example fungi belonging to the genera Zymoseptoria, Puccinia, Mycorsphearella, Pyricularia, Rhizoctoonia, Blumeria, Alternaria, Colletotrichum, Ramularia, Parastagonospora, Rhynchosporium, Oculimacula, Fusarium, Gaeumannomyces, Botrytis or Sclerotinia, preferably Zymoseptoria tritici. tritici, Puccinia recondita, Puccinia striiformis, Mycorsphearella fijiensis, Mycorsphearella arachidis, Pyricularia oryzae, Rhizoctonia solani, Blumeria graminis f.sp.tritici, Alternaria solani, Colletotrichum lagenarium, Ramularia collo-Cygni, Parastagonospora nodorum, Rhynchosporium secalis, Oculimacula yallandae, Fusarium avenaceum Fungi belonging to the species F. avenaceum, F. graminearum, F. culmorum, F. virguliforme, F. subglutinans, F. pseudograminearum, F. verticillioides, F. fujikuroi, F. subglutinans, and F. oxysporum, such as F. oxysporum f.sp. cubense and F. oxysporum melonis. f.sp.melonis, F.oxysporum f.sp.vasinfectum, F.oxysporum f.sp.lycopesici, Gaeumannomyces graminis, Botrytis cinerea, or Sclerotinia sclerotiorum, or a bacterium, for example a bacterium belonging to the genus Xanthomonas, preferably Xanthomonas oryzae pv.oryzae, or oomycetes, such as fungi belonging to the genus Aphanomyces, preferably Aphanomyces cochlioides.

[0108] Control or prevention means the reduction of infestation by phytopathogenic microorganisms, especially fungi, to the extent that an improvement can be demonstrated.

[0109] Preferably, the Streptomyces chrestomyceticus or isolated microbial strain or composition of the present invention is preventative (meaning before disease onset) or curative (meaning after disease onset). Surprisingly, it has been found that both preventative and curative application of the composition of the present invention results in a reduction of infestation by plant pathogenic microorganisms.

[0110] A preferred method for controlling or preventing plant infestation by phytopathogenic microorganisms, particularly fungi or insects, involves foliar application of the Streptomyces chrestomyceticus or isolated microbial strains or compositions disclosed herein. The frequency and rate of application depend on the risk of infestation by the corresponding pathogen or insect. However, the Streptomyces chrestomyceticus or isolated microbial strains or compositions disclosed herein can also be applied to the plant roots via the soil (systemic action) by irrigating the plant habitat with a liquid formulation or by applying the Streptomyces chrestomyceticus or isolated microbial strains or compositions disclosed herein in solid form, e.g., granular form, to the soil (soil application). In rice crops, such granules can be applied to flooded rice fields.

[0111] Surprisingly, phytopathogenic microorganisms which have been found to be affected by Streptomyces chrestomyceticus, microbial strains or compositions according to the invention, for example in foliar application, are Zymoseptoria, Puccinia, Mycorsphearella, Pyricularia, Rhizoctoonia, Blumeria, Alternaria, Colletotrichum, Ramularia, Parastagonospora, Rhynchosporium, Oculimacula, Fusarium, Gaeumannomyces sp. sp.), Botrytis, or Sclerotinia, preferably Zymoseptoria tritici, Puccinia recondita, Puccinia striiformis, Mycorsphearella fijiensis, Mycorsphearella arachidis, Pyricularia oryzae, Rhizoctonia solani, Xanthomonas oryzae pv. oryzae, Blumeria graminis f.sp.tritici, Alternaria solani, Colletotrichum lagenarium, Ramularia collo-Cygni, Parastagonospora nodorum, Rhynchosporium secalis, Oculimacula yallandae, Fusarium avenaceum, F. graminearum, and bacteria belonging to the genus Xanthomonas, preferably Xanthomonas oryzae.

[0112] In one embodiment, there is provided a method for controlling or preventing infestation of plants, plant propagation material and / or harvested food crops by plant pathogenic microorganisms by treating the plants, plant propagation material and / or harvested food crops with an effective amount of Streptomyces crestomyceticus. chrestomyceticus, the microbial strain of the invention, or the composition disclosed herein is applied to a plant, part thereof, or its habitat, plant propagation material and / or harvested food crops, and the plant pathogenic microorganism is selected from the group consisting of Zymoseptoria, Puccinia, Mycorsphearella, Pyricularia, Rhizoctoonia, Xanthomonas, Blumeria, Alternaria, Colletotrichum, Ramularia, Parastagonospora, Rhynchosporium, Oculimacula, Fusarium, Gaeumannomyces spp. sp.), Botrytis, or Sclerotinia, preferably Zymoseptoria tritici, Puccinia recondita, Puccinia striiformis, Mycorsphearella fijiensis, Mycorsphearella arachidis, Pyricularia oryzae, Rhizoctonia solani, Xanthomonas oryzae pv. oryzae, Blumeria graminis f.sp.The method is disclosed in which the fungus is a fungus belonging to the genus F. tritici, Alternaria solani, Colletotrichum lagenarium, Ramularia collo-Cygni, Parastagonospora nodorum, Rhynchosporium secalis, Oculimacula yallandae, Fusarium avenaceum, or F. graminearum, and the plant includes wheat, barley, rice, corn, soybean, sugar beet, banana, tomato, cucumber, and / or groundnut.

[0113] In one embodiment, the method for controlling or preventing infestation of plants by phytopathogenic microorganisms comprises application of Streptomyces chrestomyceticus as disclosed herein or an isolated microbial strain or composition according to the invention, wherein the plant is wheat and the phytopathogenic microorganism is Zymoseptoria, Puccinia, Blumeria, Parastagonospora Oculimacula, preferably Zymoseptoria tritici, Puccinia recondita, Puccinia striiformis, Blumeria graminis f.sp. tritici, Parastagonospora nodorum, It is a fungus belonging to the genus Fusarium nodorum, Oculimacula yallandae, Fusarium avenaceum, or F. graminearum.

[0114] In one embodiment, a method for controlling or preventing infestation of plants by phytopathogenic microorganisms comprises application of Streptomyces chrestomyceticus or an isolated microbial strain or composition according to the invention as disclosed herein, wherein the plant is barley and the phytopathogenic microorganism is a fungus belonging to the genus Ramularia sp., Rhynchosporium sp., preferably Ramularia collo-Cygni or Rhynchosporium secalis.

[0115] In one embodiment, a method for controlling or preventing infestation of plants by phytopathogenic microorganisms comprises application of Streptomyces chrestomyceticus disclosed herein or an isolated microbial strain or composition according to the invention, wherein the plant is a banana and the phytopathogenic microorganism is a fungus belonging to the genus Mycorsphearella, preferably Mycosphearella arachidis or Mycosphearella fijiensis.

[0116] In one embodiment, a method for controlling or preventing infestation of plants by plant pathogenic microorganisms comprises application of Streptomyces chrestomyceticus disclosed herein or an isolated microbial strain or composition according to the invention, wherein the plant is rice and the plant pathogenic microorganism is a fungus belonging to the genus Pyricularia, Rhizoctonia, preferably Pyricularia oryzae, Rhizoctonia solani, or a bacterium belonging to the genus Xanthomonas, preferably Xanthomonas oryzae pv. oryzae.

[0117] In one embodiment, a method for controlling or preventing infestation of a plant by a plant pathogenic microorganism comprises application of Streptomyces chrestomyceticus disclosed herein or an isolated microbial strain or composition according to the invention, wherein the plant is a tomato, and the plant pathogenic microorganism is a fungus belonging to the genus Alternaria (Alternaria sp.), preferably Alternaria solani.

[0118] In one embodiment, a method for controlling or preventing infestation of a plant by a plant pathogenic microorganism comprises application of Streptomyces chrestomyceticus disclosed herein or an isolated microbial strain or composition according to the invention, wherein the plant is a cucumber and the plant pathogenic microorganism is a fungus belonging to the genus Colletotrichum, preferably Colletotrichum lagenarium.

[0119] In one embodiment, a method for controlling or preventing infestation of plants by phytopathogenic microorganisms comprises application of Streptomyces chrestomyceticus as disclosed herein or an isolated microbial strain or composition according to the invention, wherein the plant is groundnut and the phytopathogenic microorganism is a fungus belonging to the genus Mycorsphearella, preferably Mycorsphearella arachidis.

[0120] Surprisingly, it has also been found that the Streptomyces chrestomyceticus or isolated microbial strain or composition according to the present invention disclosed herein is mobile throughout the stem or leaf and exhibits fungicidal activity against other parts of the plant than where it is applied.

[0121] For protection against fungal infections and phytopathogenic pests occurring in the soil, the isolated microbial strains and / or compositions according to the present invention may also be used as dressings for the treatment of plant propagation material, such as seeds, such as fruits, tubers, or grains, or plant cuttings. The propagation material may be treated with the isolated microbial strains and / or compositions according to the present invention before planting: for example, seeds may be dressed before sowing. The isolated microbial strains and / or compositions according to the present invention may also be applied to seeds (coatings) by impregnating the seeds in a liquid formulation or by coating the seeds with a solid formulation. The isolated microbial strains and / or compositions according to the present invention may also be applied to the planting site, for example, in the sowing furrow during sowing, when the propagation material is to be planted. Such methods for treating plant propagation material, and plant propagation material treated in this way, are disclosed herein.

[0122] The Streptomyces chrestomyceticus or isolated microbial strains disclosed herein, or compositions according to the present invention, can also be applied to seeds by impregnating the seeds or tubers with a liquid formulation or coating them with a solid formulation. Surprisingly, it has been found that the Streptomyces chrestomyceticus or isolated microbial strains disclosed herein, or compositions according to the present invention, are effective in reducing fungal and oomycete infections in seeds.

[0123] In one embodiment, there is provided a method for controlling or preventing infestation of plants by phytopathogenic microorganisms, comprising treating the seeds with an effective amount of Streptomyces chrestomyceticus, a microbial strain of the present invention, or a composition disclosed herein, wherein the phytopathogenic microorganism is selected from the group consisting of Fusarium avenaceum, Fusarium spp., and Fusarium spp. avenaceum, F. graminearum, F. culmorum, F. virguliforme, F. subglutinans, F. pseudograminearum, F. verticillioides, F. fujikuroi, F. subglutinans, F. oxysporum, e.g., F. oxysporum f.sp. cubense, F. oxysporum f.sp. melonis, F. oxysporum basinfectum f.sp.vasinfectum, F.oxysporum f.sp.lycopesici, Gaeumannomyces graminis, Botrytis cinerea, or Sclerotinia sclerotiorum, and oomycetes, for example, oomycetes belonging to the genus Aphanomyces, preferably Aphanomyces cochlioides.

[0124] Preferably, the seeds are from barley, wheat, corn, sugarcane or soybean. The expressions "soya", "soy" or "soybean" are used interchangeably herein.

[0125] Seed treatment may be applied to seeds before sowing, and the term "seeds before sowing" is meant to include seeds at any time from seed harvest to sowing the seeds in the ground for germination and plant growth. Pre-sowing seed treatment is not meant to include the application of a microbial strain or presenil composition to the soil, but includes any application that targets seeds during the sowing / planting process.

[0126] In one aspect, the present invention relates to a plant or plant propagation material treated with an isolated microbial strain or composition according to the present invention.

[0127] The treated plant propagation material of the present invention can be treated in the same manner as conventional plant propagation material. The treated propagation material can be stored, handled, sown, and cultivated in the same manner as other pesticide-treated material.

[0128] It is believed that the present method can be applied to seeds in any physiological state, but it is preferable that the seeds are sufficiently hardy so that no damage occurs during the treatment process. Typically, the seeds will be harvested in the field; removed from the plant; and separated from the core, stem, outer bracts, and any surrounding pulp or other non-seed plant material. The seeds will also preferably be biologically stable to the extent that the treatment does not cause biological damage to the seeds. It is believed that the treatment can be applied to the seeds at any time between harvesting the seeds and sowing the seeds or during the sowing process (direct seed application).

[0129] The techniques of applying seed treatments are well known to those skilled in the art and can be readily used in the context of the present invention.

[0130] One method of applying the compositions according to the invention consists of spraying or wetting the plant propagation material with an aqueous liquid formulation, or mixing the plant material with such a liquid formulation. Prior to application, the compositions of the invention can also be diluted with water by simple mixing at ambient temperature to prepare on-field seed treatment formulations.

[0131] In one embodiment, the method comprises applying an effective amount of Streptomyces chrestomyceticus, or isolated microbial strain disclosed herein above, or a composition according to the present invention, wherein the effective amount is 2×10 per hectare or more. 2 ~5×10 17 , 3×10 2 ~5×10 16 , 5×10 2 ~5×10 15 , 2 × 10 2 ~5×10 14 , 2 × 10 2 ~5×10 13 , preferably 5 x 10 2 ~5×10 12 , 1×10 3 ~5×10 11 , 5×10 3 ~1×10 11 , 1×10 4 ~5×10 10 , 5×10 4 ~1×10 10 , 1×10 5 ~5×10 9 , 5×10 5 ~1×10 9 , 1×10 6 ~5×10 8 , 5×10 6 ~1×10 8 The present invention includes colony forming units (cfu) of Streptomyces chrestomyceticus, or an isolated microbial strain or composition.

[0132] An effective amount of Streptomyces chrestomyceticus, or isolated microbial strain, or composition according to the present invention disclosed hereinabove comprises from 0.1 g to 10 kg per hectare (ha), such as from 0.5 g to 5 kg, for example from 1 g to 1 kg per ha, for example from 5 g to 500 g per ha, for example from 10 g to 200 g per ha, for example from 50 to 100 g per ha. Weights in g and kg are the dry weights of Streptomyces chrestomyceticus, microbial strains, or compositions.

[0133] In one embodiment, the method according to the invention comprises treating plant propagation material, wherein the plant propagation material is seed, and the effective amount is 5 x 10 per kg of seed. 2 ~5×10 15 , 2 × 10 3 ~5×10 14 , 5×10 3 ~5×10 13 , 2 × 10 5 ~5×10 12 , preferably 5 x 10 2 ~5×10 12 , 1×10 3 ~5×10 11 , 5×10 3 ~1×10 11 , 1×10 4 ~5×10 10 , 5×10 4 ~1×10 10 , 1×10 5 ~5×10 9 , 5×10 5 ~1×10 9 , 1×10 6 ~5×10 8 , 5×10 6 ~1×10 8 The present invention also provides a method for the preparation of a composition of matter comprising the steps of: providing a composition of matter containing colony forming units (cfu) of Streptomyces chrestomyceticus;

[0134] When the plant propagation material is a seed, the effective amount of Streptomyces chrestomyceticus, or isolated microbial strain disclosed herein above, or composition according to the invention may also comprise 0.0001 g to 100 g per kg of seeds, such as 0.0005 g to 80 g per kg of seeds / kg, for example 0.001 g to 50 g per kg of seeds, for example 0.005 g to 10 g per kg of seeds / kg of Streptomyces chrestomyceticus or microbial strain, or composition, per kg dry weight of seeds (g dry weight).

[0135] In another aspect, the present invention relates to the use of Streptomyces chrestomyceticus or an isolated microbial strain or a composition according to the invention as a pesticide, preferably a fungicide.

[0136] In another aspect, the present invention provides a method for producing a medicament for the treatment of cancer, comprising: 53 H 90 N2O 44 and at least one, at least two, at least three, at least four, or at least five of the compounds selected from the group consisting of malonomycin and cyclothiazomycin C, streptimidon, which have a molecular formula according to the formula I and are further characterized by the NMR spectra listed in Tables 2 and 3, and are preferably characterized by structural formula I; an oligosaccharide compound according to formula II; a lipopeptide or a salt thereof according to formula II (wherein R is CH or C H ); and C 67 H 115 NO 25and wherein the polyene is further characterized by an optical absorption spectrum as shown in FIG. 10 .

[0137] The characteristics relating to the isolated microbial strain, the composition according to the invention, the strains of Streptomyces, and Streptomyces chrestomyceticus are as disclosed herein above and are applicable to the uses thereof. [Brief explanation of the drawings]

[0138] [Figure 1] 1D 1H NMR spectrum of compound I in DO at 600 MHz [Figure 2] 1D 13C NMR spectrum of compound I in DO at 600 MHz [Figure 3] 2D Dept-edited 1H-13C HSQC NMR spectrum of compound I in DO at 600 MHz showing a positive (CH) signal. [Figure 4] 2D Dept-edited 1H-13C HSQC NMR spectrum of compound I in DO at 600 MHz showing a negative (CH2) signal. [Figure 5] Optical absorption spectra (UV-VIS) from 200 to 400 nm of lipopeptides 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 lipopeptide of formula II(a), showing fragment peaks consistent with the amino acids: aspartic acid, hydroxy-glutamine, serine, methyl-asparagine, and methyl-phenylalanine. [Figure 7] LC-ESI-MS / MS / MS spectrum of the precursor of lipopeptide of formula II(a) at 294.2 m / z, showing peaks consistent with the molecule C14H25-OH2-C4H5ON [Figure 8] Upfield region of 1D 1H NMR spectra of lipopeptides according to formula II(a) in CD3OD at 600 MHz [Figure 9] Low field region of the 1D 1H NMR spectrum of the lipopeptide according to formula II(a) in CD3OD at 600 MHz [Figure 10] Optical absorption spectrum (UV-VIS) of polyene compounds from 200 to 500 nm [Figure 11] Graphical representation of lipopeptide gene clusters [Figure 12] Graphical representation of polyene gene clusters DETAILED DESCRIPTION OF THE INVENTION

[0139] Example 1 Example 1.1. Sources and Fermentation Fermentation of Streptomyces sp. Streptomyces species were ordered from the culture collections listed in Table 1. Streptomyces sp. Saigon 413 was deposited at the Westerdijk Institute under accession number CBS149411. This 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. Streptomyces species were cultured in Erlenmeyer flasks in a liquid medium consisting of (g / L) casein hydrolysate 10, glucose 40, KHPO 1.25, soytone 2, and tryptone 8, and incubated at 28°C for 4 days in a shaking incubator at 150 rpm with a 25 mm throw.

[0140] Large-scale fermentation For large-scale fermentation, standard procedures were applied to cultivate Streptomyces sp., such as Streptomyces chrestomyceticus NRRL3672 and Streptomyces chrestomyceticus CBS149411, to high cell densities using fed-batch fermentation. After harvesting, the broth was spray-dried or freeze-dried according to methods known to those skilled in the art. The final product (TGAI) after spray-drying or freeze-drying was 1 x 10 6 ~1×10 13 The cell count was CFU / g dry mass. TGAI: Technical grade active ingredient (unformulated product).

[0141] Example 1.2. Formulation For the preparation of different formulations of spray-dried or freeze-dried products (microorganisms), general formulation techniques were used as disclosed in Croda Crop Care, The Nouryon formulator toolbox and: Formulation of Microbial Biopesticides: Beneficial microorganisms, nematodes and seed treatments (412 p., 6 December 2012) eds. Burgess HD, Springer, ISBN 978-94-011-4926-6. For foliar treatment of different crop diseases (Example 3), whole fermentation broths of Streptomyces chrestomyceticus NRRL B-3672 and Streptomyces sp. Saigon 413, as well as Streptomyces Saigon 413 TGAI formulated as oil dispersions (OD) and non-aqueous dispersions (NAD), were used. 5 ~10 10 Spray-dried fermentation broth was tested for cfu / g TGAI. In some cases, the surfactant polyethylene glycol sorbitan monolaurate (Tween 20®) was used. OD40 wt% was prepared from 40% dry fermentation broth in vegetable oil containing an emulsifier such as a surfactant. UV protectants such as lauryl gallate can be added. 35% or 40% NAD formulations were prepared containing 35% or 40% w / w dry material, respectively, in alcohol, e.g., methyl-diproxitol, and a rheology modifier (organoclay AEROSIL KLUCEL G IND-cellulose 200).

[0142] In the seed treatment (Example 4), 10 7 ~10 10Spray-dried or freeze-dried S. chrestomyceticus CBS149411 (Streptomyces Saigon 413) TGAI at cfu / g TGAI was used as is or formulated as a free-flowing concentrate (FS) or oil dispersion (OD). FS300 is a free-flowing concentrate containing 30% w / w spray-dried TGAI in water, a surfactant such as a silicon-based antifoam, and a dispersant such as styrene / methacrylic acid. OD400 formulations contain 40% w / w spray-dried TGAI.

[0143] 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.

[0144] The species of Streptomyces sp. Saigon413 was identified by comparing the 16S rRNA sequence according to SEQ ID NO:1 with publicly available 16S rRNA sequences extracted using barrnap v0.9 with whole-genome sequence assemblies of genomes from Streptomyces species (based on the Genome Taxonomy Database GTDB (Parks, DH, et al. (2021). GTDB: Nucleic Acids Research, 50:D785-D794)). Based on this comparative analysis, Streptomyces sp. Saigon413 was identified as the species Streptomyces chrestomyceticus. The identity between the 16S rRNA sequences of S. sp. Saigon413 and S. chrestomyceticus NRRL-3672 was 99.87%, determined using Muscle v3.8.31 and the R package Seqinr v4.2-16.

[0145] Whole genome sequencing using genomic DNA from Streptomyces sp. Saigon413 was completed using both Pacific Biosciences and Illumina sequencing technologies. The genome was assembled using HFAP4 and purified using Illumina reads with Pilon. Genomic DNA was also extracted from Streptomyces rimosus CBS492.64, Streptomyces rimosus CBS570.66, Streptomyces rimosus CBS569.66, Streptomyces chrestomyceticus DSM41224, Streptomyces rimosus subsp. rimosus DSM40673, and Streptomyces rimosus subsp. rimosus DSM41057 using the method described in Kieser et al. (2000) Practical Streptomyces Genetics. Whole genome sequencing of these strains was completed using nanopore sequencing technology, and the genomes were assembled using Flye (Kolmogorov, M., et. al., (2019), Nature Biotechnology, 37, 540).

[0146] After assembling the genome from Streptomyces sp. Saigon413 and publicly available genomes, we calculated the average nucleotide identity (ANI) between Streptomyces sp. Saigon413 and closely related Streptomyces strains using fastANI (Jain, C., et al. (2018), Nature Communications, 9, 5114) (Table 1). The highest ANI for the Streptomyces sp. Saigon413 genome was 96.9% with the genome of S. chrestomyceticus NRRL B-3672.

[0147] Using 16S RNA sequence identity and ANI scores (%), strains CBS 596.66, CBS570.66, and DSM 41429 were also found to be Streptomyces chrestomyceticus strains, as indicated by the depository, and not Streptomyces rimosis strains or Streptomyces paromomycinus strains. Table 1 shows the percent identity of the whole genome and 16S RNA sequences of Streptomyces sp. Saigon 413 and Streptomyces chrestomyceticus NRRL B-3672.

[0148] [Table 1-1]

[0149] [Table 1-2]

[0150] [Table 1-3]

[0151] Example 1.4. Analysis of streptimidin, cyclothiazomycin C, and malonomycin The structure of cyclothiazomycin C is disclosed on page 3 of WO 2015191789, and can be extracted and analyzed according to the method disclosed in Wang et al. (2010) Appl. Environmental Microbiology, Vol. 76, No. 7, p. 2336. Malonomycin can be extracted and analyzed according to the method disclosed in Example I(B) of WO 2006 / 078939. Streptimidin can be extracted and isolated according to the method disclosed in Lee et al. J. of Antibiotics (2020) 73: 184-188 (including supplementary information).

[0152] Example 1.5. Purification of Oligosaccharide Compound I Whole broths of Streptomyces species listed in Table 1, such as Streptomyces chrestomyceticus CBS149411 (Streptomyces sp. Saigon413), were centrifuged to produce aqueous extracts and pellets. The aqueous extracts were freeze-dried. The material was resuspended in a minimum volume of water and fractionated with ethyl acetate to remove lipophilic components. The aqueous suspension was retained, freeze-dried, and resuspended in a minimum volume of water before being applied to an activated carbon column. The column was washed with water and eluted with water:acetone (50:50). Compound I was further purified by hydrophilic interaction liquid chromatography (HILIC) using mass-guided fractionation and ELSD detection. For example, a Waters XBridge Amide, 5 micron, 30 x 100 mm column was used, using a gradient of acetonitrile and 10 mM ammonium acetate.

[0153] Characterization of oligosaccharide compound I Compound I was measured in the clarified fermentation broth according to the method disclosed below.

[0154] Molecular composition and total molecular weight The molecular composition and total molecular weight are 53 H 90 N2O 44 and 1458.487 g, respectively, as determined using MS-MS and NMR spectroscopy as disclosed below.

[0155] solubility The solubility of oligosaccharide compound I in water at pH 7.01 was >10,000 ppm, and in DMSO it was >9772 ppm.

[0156] MS-MS spectroscopy and liquid chromatography Spectra were recorded on a Thermo Scientific Orbitrap ID-X Tribrid mass spectrometer equipped with an OptaMax NG heated electrospray source (spray voltage: static, ion polarity (V): 3400 (positive ion mode) and 2400 (negative ion mode), sheath gas (Arb): 40, auxiliary gas (Arb): 5, sweep gas (Arb): 1, ion transfer tube temperature: 350°C, vaporizer temperature: 350°C). Scan parameters were as follows: Experiment 1: MS OT (Orbitrap resolution: 60,000, scan range (m / z): 200-2000, RF lens (%): 60, AGC target: standard, max injection time mode: auto, microscan: 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 resolving power: 30,000, RF lens (%): 60, polarity: negative). The mass spectrometer was connected to a Thermo Scientific Vanquish Flex UHPLC 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 5 μm 4.6 x 50 mm, PN 35005-054630. Temperature: 40 °C, DAD wavelength range: 250-260 nm, solvent gradient: Solvent A: HO with 0.1% formic acid, Solvent B: CHCN with 0.1% formic acid, Gradient: 0 min 1% B, 99% A; 4.00 min 50% B, 50% A; 4.25 min 100% B; 4.50 min 100% B; 4.95 min 1% B, 99% A; 6.00 min 1% B, 99% A, Flow rate: 1.0 mL / min, Injection volume: 2 µL, Total run time: 6.0 min.

[0157] NMR spectroscopy A standard Bruker pulse sequence was used to measure the 5 mm Bruker ( 1 H / 19 F) / 13 C / 15 NMR spectra were recorded on a Bruker AVIII 600 NMR spectrometer equipped with an N TCI cryoprobe and fitted with a Z gradient. Samples were dissolved in DO and spectra were recorded at 300°K. 1 2.225 ppm for H and 13 Acetone at 31.07 ppm was used as the standard for C. Figures 1 to 4 show the NMR spectra of the compounds of the present invention.

[0158] one bond 1 H- 13The C correlation spectrum contains peaks corresponding to one methyl (CH) group and 40 methine (CH) groups (listed in Table 1) and 9 methylene (CH) groups (listed in Table 2). 13 The C spectrum contains signals from three quaternary carbons at 104.7 ppm, 159.3 ppm, and 175.2 ppm (±0.1).

[0159] [Table 2-1]

[0160] [Table 2-2]

[0161] [Table 3]

[0162] Example 1.6. Lipopeptide Compound of Formula II Purification of lyopeptides according to Formula II (Formula II(a) and Formula II(b)) Mycelia from the fermentation broth of Streptomyces sp. listed in Table 1, e.g., Streptomyces sp. Saigon 413, were separated by centrifugation, and the supernatant was treated with butanol. The butanol was removed, and the extract was fractionated between water and ethyl acetate. Lipopeptides were purified from the ethyl acetate fraction by preparative reverse-phase (C18) HPLC. The lipopeptides were relatively easy to separate and eluted in the higher organic fractions in a gradient system using 0.1% formic acid and acetonitrile (0.1% formic acid). A 60% aqueous to 40% aqueous gradient using the above solvents allowed for the separation of the lipopeptide compounds according to Formula II(a) and II(b).

[0163] The compounds were detected by UV-VIS (Figure 5), mass spectrometry (Figures 6 and 7), and NMR spectroscopy (Figures 8 and 9).

[0164] Lipopeptin A Lipopeptin A was purchased from Fundacion MEDINA, Centro de Excelencia en Investigacion de Medicamentos Innovadores en Andalucia, Avda. del Conocimiento 34, Edificio Centro de Desarrollo Farmaceutico y Alimentario, Parque Tecnologico de Ciencias de la Salud, 18016 Granada (ESPANA).

[0165] Characterization of the lipopeptide of formula II Liquid chromatography and high-resolution mass spectrometry Spectra were recorded on a Thermo Scientific Orbitrap ID-X Tribrid mass spectrometer equipped with an OptaMax NG heated electrospray source (spray voltage: static, ion polarity (V): 3400 (positive ion mode) and 2400 (negative ion mode), sheath gas (Arb): 40, auxiliary gas (Arb): 5, sweep gas (Arb): 1, ion transfer tube temperature: 350°C, vaporizer temperature: 350°C). Scan parameters were as follows: Experiment 1: MS OT (Orbitrap resolution: 50,000, scan range (m / z): 200-2000, RF lens (%): 60, AGC target: standard, max injection time mode: auto, microscan: 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 resolving power: 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 resolving power: 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 resolving power: 30,000, RF lens (%): 60, polarity: positive). The mass spectrometer was interfaced to a Thermo Scientific Vanquish Flex UHPLC using a Vanquish Split Sampler FT, a Vanquish Binary Pump F, a Vanquish Column Compartment H, a Vanquish Diode Array Detector FG and a Vanquish Charged Aerosol Detector. Liquid chromatography conditions included: Waters ACQUITY UPLC C18 column 1.7 µm 3.0 x 50 mm, PN 186004660. Temperature: 40 °C. DAD wavelength range: 250-260 nm. Solvent gradient: Solvent A: HO with 0.1% formic acid, Solvent B: CHCN with 0.1% formic acid. Gradient: 0 min 10% B, 90% A; 4.00 min 90% B, 10% A; 4.25 min 90% B, 10% A; 4.50 min 10% B, 90% A; 5.00 min 10% B, 90% A. Flow rate: 1.0 mL / min. Injection volume: 2 µL. Total run time: 5.0 min. Purified fermentation broth as described above was injected. 6 and 7 show the LC-ESI-MS / MS / MS spectrum of the compound of formula II(a).

[0166] NMR spectroscopy A standard Bruker pulse sequence was used to measure the 5 mm Bruker ( 1 H / 19 F) / 13 C / 15NMR spectra were recorded on a Bruker AVIII 600 NMR spectrometer equipped with an N TCI cryoprobe and fitted with a Z gradient. Samples were dissolved in CD3OD and spectra were recorded at 300°K. 1 For H, the residual solvent signal at 3.31 ppm was used as the reference. Figures 8 and 9 each show the results of the analysis of the compound according to Formula II(a). 1 Covers half of the H NMR spectrum.

[0167] Molecular composition and mass The molecular compositions and masses of the lipopeptides according to Formula II(a) and Formula II(b) were determined using the liquid chromatography and high-resolution mass spectrometry results disclosed above. The lipopeptide compounds of Formula II(a) and Formula II(b) have the following compositions: Formula II(a) Lipopeptide 1204: molecular composition C55H85N11O19 and exact mass of 1203.602. Formula II(b) Lipopeptide 1218: molecular composition C56H87N11O19 and exact mass of 1217.618. Reference lipopeptin A has the following composition C54H84N10O19 and exact mass 1176.591421.

[0168] solubility The solubility of compounds of Formula II(a), Formula II(b) and Lipopeptin A was measured in water and DMSO.

[0169] [Table 4]

[0170] Identification of a biosynthetic gene cluster producing lipopeptide according to formula II in Streptomyces sp. Saigon 413 To identify genes involved in the production of lipopeptides according to Formula II, the assembled genome (see Example 1.4) was subjected to AntiSMASH (version 5.1.1, Blin et al., Nucleic Acids Res (2019) doi:10.1093 / nar / gkz310), a commonly used tool to aid in the identification of biosynthetic gene clusters responsible for the production of secondary metabolites.

[0171] The identification of lipopeptide compounds as belonging to the lipopeptide family (see above) and the AntiSMASH output allowed us to deduce that lipopeptide compounds are produced by nonribosomal peptide synthetases (NRPS gene clusters). The identification of the NRPS gene cluster involved in the biosynthesis of lipopeptide compounds was based on structural analysis of the compounds and the amino acids incorporated into the depsipeptide core of the lipopeptide compounds. Within Streptomyces sp. Saigon413, a unique NRPS biosynthetic gene cluster (Figure 11) was identified that allows the incorporation of amino acid precursors, including asparagine, aspartate, glutamate, phenylalanine, serine, and threonine, and was therefore associated with the production of lipopeptide compounds of Formula II(a) and II(b).

[0172] The NRPS biosynthetic gene cluster contains 45 coding sequences, including two coding sequences for NRPS genes, coding sequences for regulatory factors, and coding sequences responsible for the biosynthesis of the precursor incorporated into the lipopeptide of Formula II (Figure 11 and Table 4).

[0173] [Table 5-1]

[0174] [Table 5-2]

[0175] [Table 5-3]

[0176] Deletion and phenotypic analysis of the genomic region containing CDS_21 (SEQ ID NO: 22) and CDS_22 (SEQ ID NO: 23) from Streptomyces sp. Saigon413 To confirm that the identified biosynthetic gene cluster was associated with the production of [insert compound ID here], the region containing two nonribosomal peptide synthetase genes, encoded by ctg_7318 and ctg_7319 (SEQ ID NOs: 22 and 23), was deleted from Streptomyces sp. Saigon413. Plasmid pBCon2192 was used to generate Streptomyces sp. Saigon413Δ7318-7319. Plasmid pBCon2192 was prepared from pRAR017 and contained regions of homology on either side of the region to be deleted from the strain (facilitating primary and secondary crossovers). Plasmid pBCon2192 was transformed into E. coli ET12567 / pUZ8002 using standard electroporation techniques and then introduced into Streptomyces sp. Saigon413 by hyphal mating (T. Kieser et al., Practical Streptomyces Genetics, 2000, John Innes Foundation, Norwich). Thiostrepton-resistant colonies were patched onto ISP-4 agar supplemented with 40 μg / ml thiostrepton and 25 μg / ml nalidic acid. These patches were initially incubated at 28°C for 6 days to allow for plasmid replication. After 6 days at 28°C, the colonies were repatched onto ISP-4 agar supplemented with 40 μg / ml thiostrepton and incubated at 37°C for an additional 6 days for primary enrichment. After 6 days at 37°C, the resulting strains were transferred to ISP-4 solid agar medium without selection and incubated at 28°C for 15 days to undergo a second crossover.

[0177] After 15 days of growth, strains were harvested in 20% glycerol. 100 μl of cell suspension was used to inoculate fresh plates, giving a maximum of 10 -10 Serial dilutions were performed. Then, 100 μl of 10 -8 ~10 -10 The resulting mixture was plated onto ISP-4 agar plates. The plates were incubated at 28°C until single colonies were observed.

[0178] Single colonies were double-patched onto nonselective and thiostrepton-selective ISP-4 agar plates, and susceptible patches (representing secondary recombinants) were then screened by PCR using gDNA isolated with the FastSpin kit for soil (MP Biomedicals) to identify the correct colonies.

[0179] To confirm that the region containing both SEQ ID NO:22 and SEQ ID NO:23 had been removed from the strain, a primer pair binding outside the deleted region was used. Sanger sequencing of the PCR product and alignment with the Streptomyces sp. Saigon413 genome confirmed the deletion of the genomic region containing SEQ ID NO:22 and SEQ ID NO:23. Further, whole-genome analysis using Illumina-PCR-free sequencing confirmed that no other modifications had been made to the genome.

[0180] Cultivation of Streptomyces sp. Saigon413Δ7318-7319 and analysis of extracts from this strain confirmed that the lipopeptide compound according to Formula II(a) was no longer produced by this strain and confirmed that SEQ ID NO:22 and SEQ ID NO:23 are essential for the production of the lipopeptide compound according to Formula II(a).

[0181] Example 1.7. Polyene Compounds Purification of polyene compounds A spray-dried sample from a culture of S. chrestomyceticus CBS149411 (Streptomyces sp. Saigon413) was washed with water. The solid residue was extracted twice with isopropanol to remove the isopropanol. The resulting solid was purified by preparative reverse-phase (C18) HPLC using an acetonitrile:water gradient. Further purification was achieved by preparative reverse-phase HPLC using a Zorbax C8 column eluted with an acetonitrile:water gradient. The compounds were detected by UV-VIS (Figure 10).

[0182] Characterization of polyene compounds Liquid chromatography and high-resolution mass spectrometry Spectra were recorded on a Thermo Scientific Orbitrap ID-X Tribrid mass spectrometer equipped with an OptaMax NG heated electrospray source (spray voltage: static, ion polarity (V): 3400 (positive ion mode) and 2400 (negative ion mode), sheath gas (Arb): 40, auxiliary gas (Arb): 5, sweep gas (Arb): 1, ion transfer tube temperature: 350°C, vaporizer temperature: 350°C). Scan parameters were as follows: Experiment 1: MS OT (Orbitrap resolution: 50,000, scan range (m / z): 200-2000, RF lens (%): 60, AGC target: standard, max injection time mode: auto, microscan: 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 resolving power: 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 resolving power: 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 resolving power: 30,000, RF lens (%): 60, polarity: positive). The mass spectrometer was interfaced to a Thermo Scientific Vanquish Flex UHPLC using a Vanquish Split Sampler FT, a Vanquish Binary Pump F, a Vanquish Column Compartment H, a Vanquish Diode Array Detector FG and a Vanquish Charged Aerosol Detector. Liquid chromatography conditions included: Waters ACQUITY UPLC C18 column 1.7 μm 3.0 × 50 mm, PN 186004660. Temperature: 40 °C. DAD wavelength range: 250–260 nm. Solvent gradient: Solvent A: HO with 0.1% formic acid, Solvent B: CHCN with 0.1% formic acid. Gradient: 0 min 10% B, 90% A; 4.00 min 90% B, 10% A; 4.25 min 90% B, 10% A; 4.50 min 10% B, 90% A; 5.00 min 10% B, 90% A. Flow rate: 1.0 mL / min. Injection volume: 2 μL. Total run time: 5.0 min. The clarified fermentation broth described in section 1.8.1 was injected.

[0183] The significant peaks observed were: Anion: C67H114NO25[MH] - Predicted: 1332.7685, Observed: 1332.7679 Cation: C67H114NO24[M-H2O+H] +Predicted: 1316.7725, Observed: 1316.7709 Cation: C67H115NO24 [M-H2O+2H] 2+ Predicted: 658.8899, Observed: 658.8895 Cation: C67H113NO23[M-2(H2O)+2H] 2+ Predicted: 649.8846, Observed: 649.8843 Cation: C67H111NO22[M-3(H2O)+2H] 2+ Predicted: 640.8793, Observed: 640.8790

[0184] Molecular composition and mass The molecular composition and mass of the polyene compound were determined using the results of liquid chromatography and high-resolution mass spectrometry disclosed above. The polyene compound had the molecular composition C 67 H 115 NO 25 and has an exact mass of 1333.7758.

[0185] solubility The solubility of the compounds was determined in DMSO.

[0186] [Table 6]

[0187] Identification of a biosynthetic gene cluster producing polyene compounds in Streptomyces sp. Saigon 413 To identify the genes involved in the production of the polyene compounds identified in 1.8.2 above, the assembled genome (see Example 1.4 above) was subjected to AntiSMASH (version 5.1.1, Blin et al., Nucleic Acids Res (2019) doi:10.1093 / nar / gkz310), a tool commonly used to assist in the identification of biosynthetic gene clusters responsible for the production of secondary metabolites.

[0188] The identification of polyene compounds (see above) and the AntiSMASH output allowed us to infer that the polyenes in these compounds are produced by modular type I polyketide synthase (PKS) gene clusters. The identification of modular type I PKS gene clusters responsible for the biosynthesis of polyene compounds was based on the analysis of biosynthetic gene clusters associated with the production of characteristic polyenes, such as filipin by Streptomyces filipinensis, amphotericin by Streptomyces nodosus, and thailin A and B by Actinokineospora bangkokensis 44EHW. Within Streptomyces sp. Saigon413, a single large modular type I PKS biosynthetic gene cluster (Figure 12) was identified and associated with the production of polyene compounds in this study. The modular type I PKS biosynthetic gene cluster contains 24 coding sequences, including eight coding sequences for modular type I PKS genes, coding sequences for regulatory factors, and coding sequences responsible for the biosynthesis of precursors incorporated into polyene compounds (Figure 12 and Table 5).

[0189] [Table 7]

[0190] Deletion and phenotypic analysis of the genomic region containing CDS_31 (SEQ ID NO: 67) and CDS_32 (SEQ ID NO: 68) from Streptomyces sp. Saigon413 To confirm that the identified biosynthetic gene cluster was associated with the production of polyene compounds, a region containing two polyketide synthase genes encoded by CDS_31 and CDS_32 (SEQ ID NO:67 and SEQ ID NO:68) was deleted from Streptomyces sp. Saigon413. Plasmid p073-031 was used to generate Streptomyces sp. Saigon413Δ941-942. Plasmid p073-031 was prepared from pRAR017 and contained regions of homology on either side of the region to be deleted from the strain (facilitating primary and secondary crossovers).

[0191] Plasmid p073-031 was transformed into E. coli ET12567 / pUZ8002 using standard electroporation techniques and then introduced into Streptomyces sp. Saigon413 by hyphal conjugation (T. Kieser et al., Practical Streptomyces Genetics, 2000, John Innes Foundation, Norwich). Thiostrepton-resistant colonies were patched onto ISP-4 agar supplemented with 40 μg / ml thiostrepton and 25 μg / ml nalidic acid. These patches were initially incubated at 28°C for 6 days to allow for plasmid replication. After 6 days at 28°C, strains were repatched onto ISP-4 agar supplemented with 40 μg / ml thiostrepton and incubated at 37°C for an additional 6 days for primary enrichment. After 6 days at 37°C, the resulting strains were transferred to ISP-4 solid agar medium without selection and incubated at 28°C for 15 days to undergo a second crossover.

[0192] After 15 days of growth, strains were harvested in 20% glycerol. 100 μl of cell suspension was used to inoculate fresh plates, giving a maximum of 10 -10 Serial dilutions were performed. Then, 100 μl of 10 -8 ~10 -10The resulting mixture was plated onto ISP-4 agar plates. The plates were incubated at 28°C until single colonies were observed.

[0193] Single colonies were double-patched onto nonselective and thiostrepton-selective ISP-4 agar plates, and susceptible patches (representing secondary recombinants) were then screened by PCR using gDNA isolated with the FastSpin kit for soil (MP Biomedicals).

[0194] To confirm that the region containing both SEQ ID NO:67 and SEQ ID NO:68 had been removed from the strain, a primer pair binding outside the deleted region was used. Sanger sequencing of the PCR product and alignment with the Streptomyces sp. Saigon413 genome confirmed the deletion of the genomic region containing SEQ ID NO:67 and SEQ ID NO:68. Further, whole-genome analysis using Illumina-PCR-free sequencing confirmed that no other modifications had been made to the genome.

[0195] Cultivation of Streptomyces sp. Saigon413Δ941-942 and analysis of extracts from this strain confirmed that polyene compounds were no longer produced by this strain, confirming that SEQ ID NO:67 and SEQ ID NO:68 are essential for the production of the polyene compounds disclosed herein.

[0196] Example 2. 2.1. Determination of the metabolites of malonomycin, CtmC, and streptimidon in the fermentation broth of several Streptomyces strains The presence of the following metabolites was measured in the fermentation broth of Streptomyces sp. Saigon 413 and other Streptomyces species: malonomycin, cyclothiazomycin C, and streptimidon, an oligosaccharide compound according to compound I, a lipopeptide of formula II, and a polyene compound. These metabolites were determined by isolation and purification according to the methods described in sections 1.4–1.7.

[0197] Table 6A shows that S. chrestomyceticus strains produced at least the metabolites malonomycin, CtmC, streptimidon, and at least the metabolites C in the fermentation broth of Streptomyces sp. Saigon 413 and several other Streptomyces strains. 53 H 90 N2O 44 Oligosaccharide compound I of formula II, lipopeptide according to formula II, and polyene C 67 H 115 NO 25 It is shown that it produces

[0198] [Table 8-1]

[0199] [Table 8-2]

[0200] 2.2. Effect of some Streptomyces strains against Zymoseptoria tritici and Puccinia recondite in wheat The strains listed in Table 6A were tested for their ability to control Zymoseptoria tritici and Puccinia recondite in wheat. Strains were fermented in Erlenmeyer flasks as described above. After fermentation, broth samples were stored frozen at -80°C until use in greenhouse testing. On the day of application, samples were thawed, mixed, and diluted with water supplemented with Tween 20 (0.025% v / v) ranging from 10% (submerged fermentation broth, v / v) to 0.3%. Spray volume was 400 liters / hectare, and application was 1 day before infection. Positive statements (Yes) reported in Table 6B indicate that the Streptomyces fermentation broth prevented or reduced disease severity on treated leaves at all of the dilution ratios tested.

[0201] For infection with Zymoseptoria tritici (EPPO code: SEPTTR), test plants of the wheat cultivar Riband were inoculated by spraying a spore suspension (1.5 μg / ml in water supplemented with 0.01% Tween 20) one day after application. After a 4-day incubation period at 22°C / 21°C (day / night) and 95% rh, the inoculated test plants were maintained in a greenhouse at 22°C / 21°C (day / night) and 70% rh. Efficacy was assessed visually once an appropriate level of disease appeared on untreated control plants (16–19 days after application).

[0202] For infection with Puccinia recondita (EPPO code: PUCCRE), wheat cultivar Arina test plants were inoculated by spraying a spore suspension (80,000 spores per ml in water supplemented with 0.1% Tween 20) one day after application. After a one-day incubation period at 20°C and 95% rh, the inoculated test plants were maintained in a greenhouse at 20°C and 60% rh. The percentage of leaf area covered by disease was visually assessed once an appropriate level of disease appeared on untreated control plants (9–12 days after infection).

[0203] conclusion The results in Table 6B show that fermentation broth from a Streptomyces chrestomyceticus strain is capable of controlling the growth of the fungal pathogens Puccinia recondita and Zymoseptoria tritici in wheat. Streptomyces sp. CBS492.64 is capable of controlling only Puccinia recondita, but not S. albofaciens or S. rimosis, either of the diseases tested.

[0204] [Table 9]

[0205] Example 3. Control of fungal infections by foliar treatment with S. chrestomyceticus (Streptomyces sp. Saigon 413) compositions 3.1 Treatment of Zymoseptoria tritici and Puccinia recondite infected wheat with S. chrestomyceticus fermentation broth Streptomyces isolate CBS149411 (deposited at the Westerdijk Fungal Diversity Institute, Uppsalalaan 8, 3584 CT, Utrecht, The Netherlands) and NRRL B-3672 (the type strain of Streptomyces chrestomycetius, deposited at the ARS Culture Collection (NRRL), 1815 N. University Street, Peoria, IL 61604, USA) were fermented in a 200 L fermenter using the same method as described above. After fermentation, the broth was stored frozen at -20°C until use in the assay. Wheat seedlings of cv. Riband (for Zymoseptoria tritici testing) or cv. Arina (for Puccinia recondita testing) were grown in a greenhouse until 14 days after sowing. One day before infection, the plants were treated with various dilutions of the fermentation end broth (diluted in water, supplemented with 0.025% Tween® 20), the spray volume used was 400 L / ha. In Table 3, the time points for disease assessment are given in days post infection (DAI). The effectiveness of the treatments is reported as the percentage reduction of symptoms compared to untreated checks.

[0206] For infection with Zymoseptoria tritici (EPPO code: SEPTTR), test plants were inoculated by spraying a spore suspension (1.5 μl spores / ml in water supplemented with 0.01% Tween 20) one day after application. After a 4-day incubation period at 22°C / 21°C (day / night) and 95% rh, the inoculated test plants were maintained in a greenhouse at 22°C / 21°C (day / night) and 70% rh. Efficacy was assessed directly after an appropriate level of disease appeared on untreated check plants (16–19 days after application).

[0207] For infection with Puccinia recondita (EPPO code: PUCCRE), test plants were inoculated by spraying with a spore suspension (80,000 spores per ml in water supplemented with 0.1% Tween 20) one day after application. After a one-day incubation period at 20°C and 95% rh, the inoculated test plants were maintained in a greenhouse at 20°C and 60% rh. The percentage of leaf area covered by disease was visually assessed once an appropriate level of disease appeared on untreated check plants (9–12 days after infection).

[0208] [Table 10]

[0209] Conclusion: The results in Table 7 show that both Streptomyces chrestomyceticus strains CBS149411 and NRRLB-3672 provided control of both Puccinia recondita and Zymoseptoria tritici on wheat seedlings when applied as broth at the end of fermentation. Further dilutions of the broth showed that the broth from S. chrestomyceticus CBS149411 was more potent than S. chrestomyceticus NRRL B-3672, showing higher levels of control against both brown rust and Septoria leaf blotch at a 0.16% dilution.

[0210] 3.2. Preventive and Curative Treatment of Fungal Infections by Zymoseptoria tritici and Puccinia recondita on Wheat with Streptomyces Saigon 413 and Solatenol Wheat seedlings of cv. Riband (for Zymoseptoria tritici tests) or cv. Arina (for Puccinia recondita tests) were grown in a greenhouse until 14 days after sowing. One day before infection (1-d preventative assay) or three days after infection (3-d curative assay), plants were treated with various dilutions of an OD40% formulation of Streptomyces Saigon 413, CBS 149411. The spray volume used was 200 L / ha. Some treatments included the addition of a tank-mix adjuvant (spreader and retention aid at typical application rates). The efficacy of the CBS 149411-derived formulation was compared to a commercially available reference fungicide (Elatus™ Plus containing SOLATENOL™ from Syngenta, Basel, Switzerland).

[0211] For infection with Zymoseptoria tritici (EPPO code: SEPTTR), test plants were inoculated by spraying a spore suspension (1.5 μg spores / ml in water supplemented with 0.01% Tween 20). After a 4-day incubation period at 22°C / 21°C (day / night) and 95% rh, the inoculated test plants were maintained in a greenhouse at 22°C / 21°C (day / night) and 70% rh. Efficacy was assessed directly after an appropriate level of disease appeared on untreated check plants (16-19 days after infection (DAI)). For infection with P. recondita (EPPO code: PUCCRE), test plants were inoculated by spraying with a spore suspension (80,000 spores per ml in water supplemented with 0.1% Tween 20). After a 1-day incubation period at 20°C and 95% rh, the inoculated test plants were maintained in a greenhouse at 20°C and 60% rh. The percentage leaf area covered by disease was assessed visually (9-12 days after infection (DAI)) once an appropriate level of disease appeared on untreated check plants.

[0212] [Table 11]

[0213] Conclusion: The results in Table 8 show that Streptomyces sp. Saigon 413, CBS 149411 (OD 40% formulation) provided control of both brown rust and Septoria leaf blight in wheat, similar to conventional fungicides such as SOLATENOL™. Effective disease control was obtained when S. chrestomyceticus CBS 149411 OD 40% was applied as a preventative or curative spray. The use of tank-mix adjuvants, alkyl polyglucosides and polyoxyethylene (6) C9-C11 alcohol, improved the efficacy of the OD 40% formulation against both brown rust and Septoria leaf blight in preventative and curative treatments.

[0214] 3.3 Treatment of wheat infected with stripe rust, Puccinia striiformis, by Streptomyces sp. Saigon 413 and CBS 149411 Wheat seedlings of the cultivar Loft were grown in a greenhouse until 14 days after sowing. One day before infection (1-d preventative assay) or four days after infection (4-d curative assay), plants were treated with a single dilution of the OD40% formulation. The spray volume used was 200 L / ha. A tank-mix adjuvant was added (spreader and retention aid at typical application rates). The efficacy of S. chrestomyceticus CBS was compared to a commercially available reference fungicide (Elatus™ Plus containing SOLATENOL™ from Syngenta, Basel, Switzerland).

[0215] Puccinia striiformis (EPPO code: PUCCST) was inoculated onto wheat seedlings by spraying a spore suspension (80,000 spores per ml in water supplemented with 0.1% Tween 20). After a 48-day incubation period at 11°C and 95% rh, the inoculated test plants were maintained in a greenhouse at 17°C and 60% rh. The percentage of leaf area covered by disease was visually assessed (16–25 days after infection (DAI)) once appropriate levels of disease appeared on untreated control plants.

[0216] [Table 12]

[0217] Conclusion: The results in Table 9 show that S. chrestomyceticus CBS149411 formulated at OD 40% provided control of yellow rust in wheat similar to conventional fungicides such as SOLATENOL™ in both preventative and curative treatments.

[0218] Example 3.4. Treatment of Black Sigatoka (Mycosphaerella fijensis) on Bananas with Streptomyces sp. Saigon 413 Spray-dried powder of S. chrestomyceticus (CBS149411) was applied to banana plants in combination with spray oil (Orchex® 796) and emulsifier (Emulsogen M). Banana plants (variety Dwarf Cavendish, grown in a greenhouse for 6 weeks from commercially available in vitro explants) were treated twice with this triple mixture, 8 days and 1 day before infection. The efficacy of CBS149411 powder was compared with treatment with spray oil and emulsifier alone at the same spray timing. The fungicide MIRAVIS® Bold (together with the fungicide ADEPIDYN® from Syngenta, Basel, Switzerland), a suspension concentrate (SC200), in the same mixture with spray oil and emulsifier, was used as a positive control in the experiment and was sprayed once 1 day before infection. Orchex® 796 is an agricultural spray oil (a product of Calmette Specialty Products Partners, Indianapolis, USA). Emulsogen M is an emulsifier for mineral oils (a product of Clariant international, Muttenz, Switzerland) that contains oleyl alcohol polyglycol ether.

[0219] Mycosphaerella fijiensis (EPPO code: MYCOFI) was inoculated onto banana plants by spraying a spore suspension (100,000 spores per ml in water supplemented with 0.1% Tween 20). After 48 h of incubation under a hood to maintain high relative humidity (RH), the inoculated test plants were maintained at 24°C and 90% RH in a greenhouse until symptoms were assessed. The percentage of leaf area covered by disease was visually assessed after the appropriate level of disease appeared on untreated control plants (7–10 weeks after infection, expressed as days since infection (DAI)).

[0220] conclusion The results in Table 10 show that treating banana plants with S. chrestomyceticus CBS149411 powder in combination with spray oil and emulsifier provided control of black Sigatoka on bananas.

[0221] [Table 13]

[0222] Example 3.5. Treatment of fungal diseases of rice with Streptomyces Saigon 413 OD40% or NAD40% formulations of Streptomyces Saigon413, CBS149411 combined with 0.05% Tween® 20 were sprayed on rice plants at a spray volume of 400 L / ha 1-2 days before infection.

[0223] For infection with Pyricularia oryzae (EPPO code: PYRIOR), 14-day-old rice cultivars of the cultivar Koshihikari were grown in pots. The day after spraying, plants were inoculated with a spore suspension (approximately 85,000 spores per milliliter) in water supplemented with 0.04% Tween 20 using a paintbrush. Plants were maintained at 23°C, 14 hours of light, and high relative humidity (>80%) until evaluation. The percentage of leaf area covered by disease was visually assessed once an appropriate level of disease appeared on untreated control plants (approximately 8 days after infection (DAI)).

[0224] For infection with Rhizoctonia solani (EPPO code: RHIZSO), 14-day-old rice cultivars of the cultivar Koshihikari were grown in pots. The day after spraying the product, the plants were inoculated with a mycelium suspension in water supplemented with 0.1% Tween 20 using a paintbrush (mycelium was sedimented in a blender and filtered through a sieve). The plants were maintained at 23°C, 14 hours of light, and high relative humidity (>80%) until evaluation. The percentage of leaf area covered by disease was visually assessed once an appropriate level of disease appeared on untreated control plants (approximately 7 days after infection (DAI)).

[0225] For infection with Xanthomonas oryzae pv. oryzae (EPPO code: XANTOR), 3-week-old rice cultivars (cv. Koshihikari) were treated with formulated test compounds in a spray chamber. Two days after application, rice plants were infected by cutting at the upper leaf level with scissors pre-soaked in the bacterial suspension. After a 14-day incubation period at 23°C and high relative humidity (>80%), visual assessment of disease level was performed.

[0226] The products Tilt® (containing the fungicide propiconazole from Syngenta, Basel, Switzerland) and Amistar® (containing the fungicide azoxystrobin from Syngenta, Basel, Switzerland) were used as positive controls in the experiment.

[0227] Conclusion: The results in Table 11 show that Streptomyces chrestomyceticus CBS149411 formulated as either OD40% or NAD40% provided control of rice blast disease (PYRIOR), sheath blight (RHIZSO), and bacterial blight (XANTOR) in rice.

[0228] [Table 14]

[0229] Example 3.6. Treatment of powdery mildew (Blumeria graminis f.sp. tritici) in wheat with Streptomyces sp. Saigon 413 and SOLATENOL™ Streptomyces chrestomyceticus CBS149411 TGAI powder was applied to wheat plants in combination with a tank-mix adjuvant (methylcellulose (3%) in water) to improve retention and spread. The spray volume used was 200 L / ha. The efficacy of the CBS149411-derived formulation was compared to a commercially available reference fungicide (Elatus™ Plus containing SOLATENOL™ from Syngenta, Basel, Switzerland).

[0230] Blumeria graminis f.sp. tritici (EPPO code: ERYSGT) was inoculated by dusting 14-day-old wheat seedlings (cv. Arina) with spores from well-sporulated infected plants. The inoculated test plants were maintained in a greenhouse at 20°C and 60% rh. The percentage leaf area covered by disease was visually assessed once an appropriate level of disease appeared on untreated check plants (approximately 7 days after infection (DAI)).

[0231] conclusion The results in Table 12 show that S. chrestomyceticus CBS149411 TGAI in combination with a tank-mix adjuvant provided control of powdery mildew (Blumeria graminis f.sp. tritici) in wheat.

[0232] [Table 15]

[0233] Example 3.7. Treatment of Fungal Infections in Tomato, Cucumber, and Groundnut with Streptomyces sp. Saigon 413 and SOLATENOL™ An OD40% formulation of S. chrestomyceticus CBS149411 was sprayed on tomato, cucumber, and groundnut plants one day before infection at a spray volume of 400 L / ha in combination with 0.025% Tween® 20. Elatus™ Plus containing SOLATENOL™ (Syngenta, Basel, Switzerland) was used as the reference fungicide.

[0234] Alternaria solani (EPPO code: ALTESO) Four-week-old tomato plants of the cultivar Roter Gnom were sprayed in a spray chamber with the formulated test compound diluted in water. Two days after application, the test plants were inoculated by spraying with a spore suspension. The inoculated test plants were incubated in a greenhouse at 22 / 18°C (day / night) and 95% rh, and the percentage leaf area covered by disease was assessed when an appropriate level of disease appeared on untreated check plants (5-7 days after application).

[0235] Colletotrichum orbiculare (EPPO code: COLLLA) One-week-old cucumbers cv. Wisconsin were treated with formulated test compounds in a spray chamber. One day after application, plants were inoculated by spraying a spore suspension (1 x 105 spores / ml) onto the test plants. After a one-day incubation period at 25°C and 100% rh, the plants were kept in a greenhouse at 22°C and 70% rh for 6 days. Disease incidence was assessed 7 days after inoculation (DAI).

[0236] Mycosphaerella arachidis (EPPO code: MYCOAR) Three-week-old peanut plants of the cultivar Florunner were treated with the formulated test compounds in a spray chamber. One day after application, the plants were inoculated by spraying a spore suspension (3 x 105 spores / ml) onto the test plants. After a 4-day incubation period at 24°C and 95% rh, the plants were kept in a greenhouse at 26°C and 70% rh for 7 days. Disease incidence was assessed 11 days after inoculation (DAI). Rh: relative humidity

[0237] conclusion Table 13 shows that the OD 40% formulation of S. chrestomyceticus CBS149411 provided control of early blight of tomato (Alternaria solani), anthracnose of cucumber (Colletotrichum orbiculare), and early leaf spot of groundnut (Mycosphaerella arachidis).

[0238] [Table 16]

[0239] Example 3.8. Treatment of Fungal Infections in Barley and Wheat with S. chrestomyceticus CBS 149411 (Streptomyces sp. Saigon 413) and Chemical Fungicides An OD40% and NAD40% formulation of S. chrestomyceticus CBS149411 was sprayed on barley and wheat at a spray volume of 200 L / ha one day before infection. Elatus™ Plus containing SOLATENOL™ (Syngenta, Basel, Switzerland) was used as the reference fungicide.

[0240] Ramularia collo-Cygni (EPPO code: RAMUCC) Twelve-day-old wheat plants of the Venture cultivar were treated with the formulated test compounds in a spray chamber. One day after application, a spore suspension (3 x 10 5 Plants were inoculated by spraying 17 days after inoculation (17 spores / ml) of the fungus. After a 2-day incubation period under a hood, the plants are kept in a greenhouse at 21°C / 19°C (day / night) and 80% relative humidity for 15 days. Disease incidence was assessed 17 days after inoculation (DAI).

[0241] Parastagonospora nodorum (EPPO code: LEPTNO) Thirteen-day-old wheat plants of the variety Riband were treated with the formulated test compounds in a spray chamber. One day after application, a spore suspension (4 x 10 5 Plants were inoculated by spraying with 100 spores (100 spores / ml). After a 2-day incubation period under high relative humidity (>95%), plants are kept in a greenhouse at 21°C / 19°C (day / night) and 70% relative humidity for 5 days. Disease incidence was assessed 7 days after inoculation (DAI).

[0242] Rhynchosporium secalis (EPPO code: RHYNSE) Fourteen-day-old barley plants (cv. Pasadena) were treated with the formulated test compounds in a spray chamber. One day after application, a spore suspension (4 x 10 5 Plants were inoculated by spraying with 100 spores (0.001 spores / ml). After a 2-day incubation period under high relative humidity (>95%), plants are kept in a greenhouse at 21°C / 19°C (day / night) and 70% relative humidity for 13 days. Disease incidence was assessed 15 days after inoculation (DAI).

[0243] conclusion The results in Table 14 show that OD 40% or NAD 40% formulations of S. chrestomyceticus CBS148411 provided control of Ramularia early leaf spot in barley (Ramularia collo-Cygni), glume spot in wheat (Parastagonospora nodorum), or scalded disease in barley (Rhynchosporium secalis).

[0244] [Table 17]

[0245] Example 3.9. Treatment of Wheat Eyespot (Oculimacula yallundae) with S. chrestomyceticus CBS 149411 (Streptomyces sp. Saigon 413) and Cyprodinil A 40% NAD formulation of S. chrestomyceticus CBS149411 was sprayed onto the plants one day before infection at a spray volume of 200 L / ha. Unix™ WG750 (Syngenta, Switzerland) containing cyprodinil was used as the reference fungicide.

[0246] For infection with Oculimacula yallundae (EPPO code: PSDCHE) Eleven-day-old wheat plants of the variety Arina were treated with the formulated test compounds in a spray chamber. One day after application, a spore suspension (6 x 10 5Wheat plants were inoculated by spraying 100 spores (0.001 spores / ml) at 10°C / 11°C (day / night) on the stalks, specifically targeting the base of the plant. After a 2-day incubation period under a hood, the plants were kept in a greenhouse at 11°C / 10°C (day / night) and 80% relative humidity. Disease incidence was assessed when symptoms were fully developed on untreated checks (approximately 7-12 weeks after inoculation).

[0247] conclusion The results in Table 15 show that S. chrestomyceticus CBS149411 formulated as NAD 40% provided an effective means of controlling eyespot (Oculimacula yallundae) of wheat.

[0248] [Table 18]

[0249] Example 3.10. Off-site treatment of wheat fungal diseases with Streptomyces chrestomyceticus CBS149411 A 40% NAD formulation of S. chrestomyceticus 149411 was applied onto leaf sections one day before infection at a rate of approximately 400 L / ha. The efficacy of the CBS149411-derived formulation was compared to a commercially available reference fungicide (Elatus™ Plus containing SOLATENOL™ from Syngenta, Switzerland).

[0250] Wheat seedlings of either cv. Riband (for Zymoseptoria tritici testing) or cv. Arina (for Puccinia recondita testing) were grown in a greenhouse until 14 days after sowing. At this time, such seedlings typically have a fully emerged first leaf (designated L1), a fully emerged second leaf (L2), and a partially emerged, developing third leaf (L3). Using a permanent pen, two marks were applied to the second leaf, generating three sections of approximately equal size: the base, middle, and tip of the leaf. The compounds to be tested were diluted in water at the indicated concentrations (in ppm). The diluted compounds were applied to the central portion of leaf 2 using a conventional cotton stick. The cotton stick was dipped in the diluted compound and rubbed several times across the adaxial leaf surface between the two marks. One day later, intact plants were inoculated with a fungal spore suspension using a paintbrush. The spore suspension was applied until runoff. For infection with Zymoseptoria tritici (EPPO code: SEPTTR), test plants were inoculated one day after application by spraying with a spore suspension (1.5 μl spores / ml in water supplemented with 0.01% Tween 20). After a 4-day incubation period at 22°C / 21°C (day / night) and 95% rh, the inoculated test plants were maintained in a greenhouse at 22°C / 21°C (day / night) and 70% rh. Efficacy was assessed directly after the appropriate level of disease appeared on untreated control plants (16–19 days after application (DAI)).

[0251] For infection with Puccinia recondita (EPPO code: PUCCRE), test plants were inoculated by spraying with a spore suspension (80,000 spores per ml in water supplemented with 0.1% Tween 20) one day after application. After a one-day incubation period at 20°C and 95% rh, the inoculated test plants were maintained in a greenhouse at 20°C and 60% rh. The percentage of leaf area covered by disease was visually assessed (9–12 days after infection (DAI)) once an appropriate level of disease appeared on untreated check plants.

[0252] conclusion The results in Table 16 show that S. chrestomyceticus 149411 can protect treated leaf areas of wheat plants from infection by Septoria leaf blight (Zymoseptoria tritici) and brown rust (Puccinia recondita). The level of protection achieved by S. chrestomyceticus CBS 149411 was comparable to the effectiveness of conventional fungicides (e.g., SOLATENOL™). As with SOLATENOL™, the untreated apical leaf portion of leaf 2 was similarly well protected from both diseases. Surprisingly, the base of leaf 2 was also highly protected (>88%) from disease when the assay rate of 2500 ppm was used. Similarly, systemic leaf 3 was also protected by applying the product onto leaf 2. The protection of the basal parts of leaf 2 and pedicle 3 was found to be rate dependent.

[0253] [Table 19-1]

[0254] [Table 19-2]

[0255] Example 3.11. Treatment of Fusarium head disease in wheat with S. chrestomyceticus CBS 149411 (Streptomyces sp. Saigon 413) A 35% NAD formulation of S. chrestomyceticus CBS149411 was applied to leaf segments one day before infection in the presence of a tank-mix adjuvant at a spray volume of approximately 220 L / ha. Miravis™ Plus (Syngenta, Switzerland) containing ADEPIDYN™ was used as the reference fungicide.

[0256] Fusarium avenaceum (EPPO code: GIBBAV) Flowering wheat cultivar Monsun was treated with the formulated test compounds in a spray chamber. One day after application, a spore suspension (1 x 10 5 Plants were inoculated by spraying with 100 spores (100 spores / ml). After a 2-day incubation period with misting water at high relative humidity (>95%) and in the absence of light, the plants were kept in a greenhouse at 21°C / 19°C (day / night) and 70% relative humidity for 12 days. Disease incidence in panicles was assessed 14 days after inoculation. The inoculum consisted of a triple mixture of three different isolates (ratio approximately 1:1:1), each of which was confirmed to produce a different mycotoxin typical of this species (Enniatin A, Enniatin A1, Enniatin B, Enniatin B1, and moniliformin).

[0257] Fusarium graminearum (EPPO code: GIBBZE) Flowering wheat plants of variety Monsun were treated with the formulated test compounds in a spray chamber. One day after application, a spore suspension (1 x 10 5Plants were inoculated by spraying with 100 spores (100 spores / ml). After a 2-day incubation period with misting water at high relative humidity (>95%) and in the absence of light, the plants were kept in a greenhouse for 5 days at 21°C / 19°C (day / night) and 70% relative humidity. Disease incidence in the panicles was assessed 7 days after inoculation. The inoculum consisted of a triple mixture of three different isolates (ratio approximately 1:1:1), each of which was confirmed to produce a different mycotoxin typical of this species (DON = deoxynivalenol, 3- or 15-acetyldeoxynivalenol, zearalenone).

[0258] conclusion The results in Table 17 show that S. chrestomyceticus CBS 149411 formulated as NAD 35% provided an effective means of controlling Fusarium head disease (Fusarium graminearum) or eyespot disease (Oculimacula yallundae or Fusarium avenaceum) in wheat. Fusarium species with different chemotypes (dnitridole (DON)-producing and enniatin-producing) were controlled.

[0259] [Table 20]

[0260] Example 4. Treatment of seeds with Streptomyces chrestomyceticus CBS149411 Semi-Field Platform Methodology Used in Examples 4.1A and 4.1B The semi-field platform consisted of a water pool system containing pots in which the soil temperature was controlled. A cooling unit allowed for the water temperature to be controlled independently of external weather conditions according to the requirements for disease establishment. Plastic pots (60 L capacity) were filled with 45 L of freshly sieved soil.

[0261] Seed treatment Before sowing, the seeds were treated with a slurry containing 5 g and 10 g of test compound per kg of corn and wheat seeds, respectively, using a stirring mixer according to standard procedures known to those skilled in the art. The treated seeds were dried in open bottles at room temperature for 24 hours, and then stored in paper bags at room temperature until sowing.

[0262] Example 4.1A. Efficacy of Streptomyces chrestomyceticus Saigon 413 against soilborne Fusarium graminearum in maize under semi-field conditions In this semi-field experiment, the topsoil layer was mixed with a substrate pre-inoculated with the test fungus, namely Fusarium graminearum. A total of 100 maize seeds of the cultivar Arma were sown per pot. A complete randomized block design was used with four replications (pots) per treatment. The pots were placed in aquaria containing cold water (10°C). The water temperature was maintained at approximately 10°C for the first three weeks, then at 15–20°C for the next two weeks.

[0263] The following treatments were applied to corn seeds using the standard procedures disclosed above: 200g TGAI of Streptomyces chrestomyceticus Saigon 413 spray-dried broth per 100kg of seeds Freeze-dried supernatant, per 100 kg of seeds, 200 g of freeze-dried supernatant, Streptomyces chrestomyceticus Saigon 413 TGAI Standard (reference treatment) fludioxonil (product: Celest, FS025) at 2.5 g ai per 100 kg of seeds

[0264] The study included two control treatments: an infected control and a non-infected control. The activity (%) of each seed treatment was calculated based on the % emergence (number of plants) of plants at final standing compared to infected controls.

[0265] result: Disease pressure in the experiment was high: final standing number was reduced by 72% in the infected control treatment. The reference treatment provided 97% activity. The spray-dried broth provided 84% activity. The freeze-dried supernatant provided 69% activity.

[0266] Example 4.1B. Effect of seed-applied Streptomyces chrestomyceticus Saigon 413 on soil-borne infection by Fusarium graminearum of corn and Fusarium culmorum of wheat under semi-field conditions. In this semi-field experiment, the topsoil layer was mixed with a substrate pre-inoculated with the test fungus, namely Fusarium graminearum (K-6102) or Fusarium culmorum. A total of 100 maize seeds (cv. Andromeda) or 100 wheat seeds (CV. Taifun) were sown per pot. A complete randomized block design was used with four replicates (pots) per treatment. The pots were placed in aquaria containing cold water (12°C).

[0267] Using the standard procedures disclosed above, the following treatments were applied to corn and wheat seeds: Wheat seeds were treated with a free-flowing concentrate (FS300) of Streptomyces chrestomyceticus Saigon 413 at 200 g TGAI per 100 kg of seeds. Corn seeds were treated with an oil dispersion of Streptomyces chrestomyceticus Saigon 413 at OD400 at 200 g TGAI per 100 kg of seeds. o Standard (reference treatment), fludioxonil (product: Celest, FS025) o 2.5g ai per 100kg of seed for experiments with maize o 5g ai per 100 kg of seed for experiments with wheat

[0268] Two control treatments were included in the study: an infected control and an uninfected control. The % activity of each seed treatment was calculated based on the % emergence of plants at the final plant rest compared to the infected control.

[0269] result: Disease pressure in the experiments was moderate; final plant survival in the infected control treatments was reduced by 19% in wheat and 34% in corn. The reference treatments provided 100% activity in the corn experiment and 93% activity in the wheat experiment. In the wheat experiment with Fusarium culmorum, Streptomyces Saigon 413 provided 92% activity at 200 g TGAI. In the corn experiment with Fusarium graminearum, Streptomyces Saigon 413 provided 99% activity at 200 g TGAI. Results indicate that both the oil dispersion formulation as a flowable concentrate formulation provided greater than 90% activity.

[0270] Example 4.1. Effect of soil Fusarium graminearum on corn seed under controlled conditions in a greenhouse when applied to Streptomyces sp. Saigon 413. Corn seed variety Andromeda was treated according to standard procedures known to those skilled in the art using a stirring mixer with a slurry volume of 5 g per kg of seed. The treated seeds were allowed to dry in open bottles at room temperature for 24 hours. Four replicates (soil trays) containing 25 seeds each were used. Prior to sowing, the soil was incubated with fungal spores of the test pathogen, Fusarium graminearum, at 18°C ​​for 7 days.

[0271] The following seed-applied treatments were compared: Streptomyces chrestomyceticus Saigon 413 TGAI was formulated as FS300 at 200g TGAI per 100 kg of seeds. Standard (reference treatment) fludioxonil (product: Celest, FS025) at 2.5 g ai per 100 kg of seeds

[0272] The study included two control treatments: an infected control and a non-infected control. The percent activity of each seed treatment was calculated based on the percent emergence of plants at final standing compared to infected controls.

[0273] result: Disease pressure in the experiment was moderately high; final plant volume in the infected control treatment was reduced by 45%. The reference treatment (fludioxonil) at 2.5 g ai provided 96% activity. Streptomyces chrestomyceticus Saigon 413 TGAI provided 82% activity at 200 g TGAI.

[0274] Example 4.2. Efficacy of seed-applied Streptomyces chrestomyceticus Saigon 413 against soil-borne infection by Fusarium virguliforme in soybean under controlled conditions in a greenhouse Soybean seeds (cultivar Toliman) were treated using the standard procedure disclosed above. Five replicates (pots) containing five seeds each were used. The soil was mixed with a pre-inoculated substrate of the test fungus, namely, Fusarium virguliforme. The following seed-applied treatments were compared: Streptomyces chrestomyceticus Saigon 413 TGAI was formulated as FS300 at 200g TGAI per 100 kg of seeds. Standard (reference treatment) ADEPIDYN® (active ingredient: pydiflumetofen; FS500) at 40 g ai per 100 kg of seeds

[0275] Two control treatments were included in the test: an infected control and an uninfected control. Calculation of % activity of the seed treatments was based on a disease index derived from the severity of leaf symptoms compared to the infected control.

[0276] result: Disease pressure in the experiment was high, i.e., disease incidence was 80%. The reference treatment (ADEPIDYN®) provided 89% activity at 40 g ai. Streptomyces chrestomyceticus Saigon 413 TGAI provided 58% activity at 200 g ai.

[0277] Example 4.3. Effect of seed-applied Streptomyces chrestomyceticus Saigon 413 on soil-borne infection by Aphanomyces cochlioides (oomycete) in sugar beet under controlled conditions in a greenhouse Sugar beet seeds (cultivar Jagger) were treated according to standard procedures known in the art, as disclosed above. Five replicates (pots) containing 25 seeds each were used. Seven days after sowing, a mycelium suspension of the plant pathogen Aphanomyces cochlioides strain K-9164 was added to each pot.

[0278] The following seed-applied treatments were compared: Streptomyces Saigon 413 TGAI formulated as FS300 at TGAI g ai per million seeds Standardized TGAI g ai per million seeds (reference treatment), hymexazole (product Tachigaren, WS070)

[0279] Two control treatments were included in the test: an infected control and an uninfected control. The activity (%) of each seed treatment was calculated based on the reduction of symptoms (count of dead plants) compared to the infected control.

[0280] result: Disease pressure in the experiment was moderately high: 56% dead plants in the infected control. The reference treatment (hymexazol) provided 74% activity. Streptomyces Saigon 413 TGAI provided 53% activity.

[0281] Example 4.4. Efficacy of seed-applied Streptomyces chrestomyceticus Saigon 413 against soil infection by Gaeumannomyces graminis in wheat under controlled conditions in a greenhouse Wheat seeds of the cultivar Arina were treated according to the standard procedure disclosed above. Four replicates (pots) containing 15 seeds each were used. At the time of sowing, the soil substrate was mixed with the fungal inoculum.

[0282] The following seed-applied treatments were tested: S. chrestomyceticus (Streptomyces Saigon 413) TGAI as FS300 (FS: Seed Treatment Fluid Concentrate) at 100g ai per 100kg of seeds

[0283] The study included two control treatments: an infected control and a non-infected control. The activity (%) of the seed treatment was calculated based on root health (severity of disease symptoms) compared to the control treatment.

[0284] result: Disease pressure in the experiment was high: 76% unhealthy roots in infected controls. Streptomyces Saigon 413 TGAI provided 25% activity (efficacy). The results of Examples 4.1 to 4.4 are summarized in Table 18.

[0285] conclusion The results of Experiments 4.1 to 4.4 shown in Table 18 indicate that Streptomyces chrestomyceticus CBS149411 was effective in controlling diseases caused by several fungal (Fusarium and Gaeumannomyces) and oomycete infections on seeds.

[0286] [Table 21]

[0287] Example 5. Activity of Streptomyces chrestomyceticus Saigon 413 against a broad species spectrum of Fusarium using in vitro bioassays Various Fusarium spp. (see Table 19) were tested in an in vitro bioassay using Petri dishes (9 cm diameter) containing Luria broth agar growth medium and spray-dried Streptomyces chrestomyceticus Saigon 413 (formulated as OD400) at different concentrations. Three replicates (Petri dishes) were included for one treatment concentration and one control. Agar plugs were taken from Fusarium growth colonies using a cork borer (φ6 mm), and one plug was placed in the center of each agar plate. The fungi were incubated in the dark at 22°C. After incubation, mycelial growth (diameter) was measured for efficacy and median effective concentration, or EC-50, calculation.

[0288] [Table 22]

[0289] Example 6. Effect of Streptomyces chrestomyceticus Saigon 413 TGAI against Botrytis cinerea and Sclerotinia sclerotiorum in a microprofiling screen Botrytis cinerea (gray mold): Fungal conidia stored at low temperatures were mixed directly into nutrient broth (Vogel's minimal medium). A DMSO solution of the test compound was placed in a 96-well microtiter plate, and the nutrient broth containing the fungal spores was added. The test plates were incubated at 24°C, and growth inhibition was determined photometrically after 72 hours.

[0290] Sclerotinia sclerotiorum (cotton rot, white mold, etc.): Fungal mycelial fragments prepared from fresh liquid cultures were mixed directly into nutrient broth (PDB potato dextrose broth). DMSO solutions of the test compounds were placed in microtiter plates (96-well format), and nutrient broth containing fungal spores was added. Test plates were incubated at 24°C, and growth inhibition was measured photometrically at 620 nm after 72 hours.

[0291] result: Botrytis cinerea: 100% efficacy at 37 ppm (mg / L) Sclerotinia sclerotiorum: 100% efficacy at 333 ppm (mg / L)

Claims

1. An isolated microbial strain containing a genome sequence that is at least 99.8% identical to the entire genome of Streptomyces sp. Saigon 413, deposited at the Westerdijk Institute under receipt number CBS149411.

2. The isolated microbial strain according to claim 1, wherein the strain comprises a nucleotide sequence having at least 99.9% identity with SEQ ID NO:

1.

3. The microbial strain according to claim 1, wherein the microbial strain is Streptomyces chrestomyceticus, preferably a species of the genus Streptomyces (Streptomyces sp.) Saigon 413 deposited with the Westerdigk Institute under receipt number CBS149411.

4. C 53 H 90 N 2 O 44 The molecular formula includes the following, further characterized by the NMR spectra listed in Tables 2 and 3, preferably structural formula I: 【Chemistry 1】 A strain of the genus Streptomyces characterized by, and at least one compound selected from the group consisting of malonomicin, cyclothiazomycin C, and streptimidone, an oligosaccharide compound by compound I, formula II: 【Chemistry 2】 A lipopeptide or a salt thereof according to (wherein R1 is CH 3 or C 2 H 5 ), as well as a polyene compound characterized by the molecular formula of C 67 H 115 NO 25 A composition comprising a polyene compound characterized by the molecular formula, wherein said polyene compound is further characterized by a light absorption spectrum as shown in Figure 10.

5. The composition according to claim 4, further comprising an auxiliary agent.

6. The composition according to claim 4, wherein the strain of the genus Streptomyces contains a genome sequence having at least 91% identity with the whole genome of Streptomyces chrestomyceticus NRRL B-3672, or at least 91% identity with the whole genome of a species of the genus Streptomyces, Saigon 413, deposited at the Westerdijk Institute under acceptance number CBS149411.

7. The composition according to claim 4, wherein the strain of Streptomyces comprises a nucleotide sequence having at least 99% identity with SEQ ID NO:

1.

8. The composition according to claim 4, wherein the composition is a fermentation broth, preferably a spray-dried fermentation broth or a freeze-dried fermentation broth, or a compound thereof.

9. The above composition contains 10 per 1g of dry weight. 2 ~10 12 The composition according to claim 4, comprising the number of cells of a strain of the genus Streptomyces or a microbial strain of cfu.

10. The isolated microbial strain according to claim 1, or the composition according to claim 4, wherein the microbial strain or the Streptomyces strain comprises at least one nucleotide sequence encoding a protein having at least 80% identity with the amino acid sequence described in SEQ ID NOs. 71 to 115, preferably SEQ ID NOs. 91 and / or SEQ ID NOs. 92, preferably at least one nucleotide sequence from among the nucleotide sequences described in SEQ ID NOs. 2 to 46, preferably at least 80% identity with the at least one nucleotide sequence described in SEQ ID NOs. 22 or 23.

11. The isolated microbial strain or the Streptomyces strain according to claim 1, or the composition according to claim 4, wherein the isolated microbial strain or the Streptomyces strain comprises at least one nucleotide sequence encoding a protein having at least 80% of the amino acid sequence described in SEQ ID NOs: 116 to 139, preferably the amino acid sequence described in SEQ ID NOs: 136 or 137, preferably at least one of the nucleotide sequences described in SEQ ID NOs: 47 to 70, preferably at least one nucleotide sequence having at least 80% identity with at least one nucleotide sequence described in SEQ ID NOs: 67 or 68.

12. A process for producing the microbial strain described in claim 1 or the composition described in claim 4, comprising culturing the microbial strain or the Streptomyces strain in a suitable fermentation medium under suitable fermentation conditions, and optionally comprising a step of recovering the microbial strain or composition.

13. The aforementioned microbial strain or the aforementioned strain of the genus Streptomyces is C 53 H 90 N 2 O 44 The molecular formula includes the following, and is further characterized by the NMR spectra listed in Tables 2 and 3, preferably compound I having structural formula I: 【Transformation 3】 Further characterized by malonomicin and at least one compound selected from the group consisting of cyclothiazomycin C and streptimon, an oligosaccharide compound by compound I, formula II: 【Chemistry 4】 A lipopeptide, or a salt thereof (wherein R is used in the formula) 1 However, CH 3 or C 2 H 5 (is), and C 67 H 115 NO 25 The process according to claim 12, wherein the polyene compound is characterized by the molecular formula, and the polyene is further characterized by the light absorption spectrum as shown in Figure 1.

14. A method for controlling or preventing infestation of plants, plant propagation materials and / or harvested food crops by plant pathogenic microorganisms, comprising applying an effective amount of Streptomyces chrestomyceticus, the microbial strain described in claim 1, or the composition described in claim 4 to the plants, a part thereof, or its habitat, or to the plant propagation materials and / or harvested food crops by treating the plants, plant propagation materials and / or harvested food crops.

15. The aforementioned plant pathogenic microorganisms are fungi, preferably Zymoseptoria, Puccinia, Mycorspherella, Pyricularia, Rhizoctonia, Xanthomonas, Blumeria, and Alternaria. (Alternaria), Colletotrichum, Ramularia, Parastagonospora, Rhynchosporium, Oculimacula, Fusarium, Gaeumannnomyces Fungi belonging to the genera Botrytis or Sclerotinia, preferably Zymoseptoria tritici, Puccinia recondita, Puccinia striiformis, Mycosphaerella fijiensis, Mycosphaerella arachidis, Pyricularia oryzae, Rhizoctonia solanii Xanthomonas oryzae pv. oryzae, Blumeria graminis f. sp. tritici, Alternaria solani, Colletotrichum lagenarium, Ramularia collo-cygni, Parastagonospora nodorum, Rhynchosporium cecalis secalis, OculimaculaFungi belonging to the species F. yallandae, Fusarium avenaceum, F. graminearum, F. culmorum, F. virguliforme, F. subglutinans, F. pseudogramaminearum, F. verticillioides, F. fujikuroi, F. subglutinans, F. oxysporum, for example, F. A species of the genus *Oxysporum* specialized to *Cubense* (F. oxysporum f. sp. cubense), a species of the genus *Oxysporum* specialized to *Melonis* (F. oxysporum f. sp. Melonis), a species of the genus *Oxysporum* specialized to *Vasinfectum* (F. oxysporum f. sp. vasinfectum), F. The method according to claim 14, wherein the plant pathogenic microorganism is a species of the genus Oxysporum f.sp. Lycopesici, Gaeumannomyces graminis, Botrytis cinerea, or Sclerotinia sclerotiorum, or the plant pathogenic microorganism is an oomycete belonging to Aphanomyces, preferably Aphanomyces cochlioides.

16. The effective amount is 2 x 10 per hectare. 2 ~5x10 17 The method according to claim 14, comprising colony-forming units (cfu), or 0.1 g to 10 kg of Streptomyces chrestomyceticus, or the isolated microorganism described in claim 1, or the composition described in claim 4.

17. The plant propagation material is seeds, and the effective amount is 2 x 10 per 1 kg of seeds. 2 ~5x10 15 The method according to claim 14, comprising (cfu), or 0.0001 g to 100 g of Streptomyces chrestomyceticus, or the isolated microbial strain described in claim 1, or the composition described in claim 4.

18. The method according to claim 14, wherein the plant comprises wheat, barley, rice, corn, soybean, sugar beet banana, tomato, cucumber, and / or ground nuts.

19. A plant or plant propagation material treated with the microbial strain described in claim 1, or the composition described in claim 4.

20. Use of the microbial strain described in claim 1, S. chrestomyceticus, or the composition described in claim 4, preferably as a pesticide and fungicide.

21. C 53 H 90 N 2 O 44 A compound comprising the molecular formula, further characterized by the NMR spectra listed in Tables 2 and 3, preferably characterized by structural formula I, at least one compound selected from the group consisting of malonomicin, cyclothiazomycin C, and streptimidone, an oligosaccharide compound of compound I, a lipopeptide of formula II, or a salt thereof (wherein R1 is CH 3 or C 2 H 5 (is), and C 67 H 115 NO 25 Use of a microbial strain according to claim 1, or a strain of the genus Streptomyces having at least 91% identity with the whole genome of Streptomyces chrestomyceticus NRRL B-3672, or at least 91% identity with the whole genome of Streptomyces sp. Saigon 413 deposited at the Westerdijk Institute under receipt number CBS149411.

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