Bactericidal and fungicidal compounds

Abstract

The present invention relates to compounds of formula (I) and compositions containing same, processes for producing the compounds and methods of using the compounds and compositions to prevent or control fungi in plants. JPEG2025533164000044.jpg115122

Description

[Technical Field]

[0001] The present invention relates to novel compounds having pesticidal activity. The present invention also relates to compositions containing the compounds, to processes for preparing the compounds, and to the use of the compounds or compositions in agriculture or horticulture to prevent or control phytopathogenic infestations of plants, harvested food crops, seeds, or non-living organisms. [Background technology]

[0002] Fungicides are widely used in agriculture to protect plants from damage caused by fungi. Fungicides can be of chemical or biological origin. Due to some adverse environmental effects of chemical fungicides, there is an increasing need for fungicides of biological, e.g., microbial, origin. Known microorganisms that produce antibiotics against fungi are actinomycetes, such as Streptomyces sp. A very well-known species is Streptomyces natalensis, which produces the antifungal compound natamycin, used in food and crop protection. U.S. Patent No. 5,356,624 discloses a Streptomyces rimosus strain found to be active against several wood-decomposing fungi. WO 2022 / 038180 discloses that novel Streptomyces species produce several known antifungal compounds, such as streptimidon, natamycin (pimaricin), or arbofungin. 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.

[0003] Another compound produced by Streptomyces sp., No. AC-69, is lipopeptin A, which is known to be active against several plant pathogenic fungi (Tsuda, Suzuki (1980), The Journal of Antibiotics Vol. 33, No. 2, pp. 247-248). Summary of the Invention

[0004] Due to the development of fungal resistance to fungicides, government regulations and societal pressures, there is a continuing need to search for new compounds with fungicidal and bactericidal activity of biological origin.

[0005] The present invention relates to a compound of formula (I) [ka] (Wherein R1=CH3 or C2H5) or a salt thereof.

[0006] Surprisingly, it has been found that the novel compounds according to the present invention have a surprising level of biological activity for preventing or controlling plant pathogenic microorganisms, such as fungi. Biological activity as used herein includes bactericidal and fungicidal activity.

[0007] In a second aspect, the present invention relates to a composition comprising a compound according to the invention and a microorganism capable of producing the compound according to the invention.

[0008] In a third aspect, the present invention relates to a process for the production of a compound or composition according to the invention, which process comprises culturing a microorganism in a suitable fermentation medium under conditions that allow the production of the compound.

[0009] In a fourth aspect, the present invention relates to a method for controlling or preventing infestation by phytopathogenic microorganisms in plants, wherein an effective amount of a compound according to the present invention or a salt thereof, or a composition according to the present invention as disclosed herein, is applied to the plant, to a part thereof or to its habitat.

[0010] According to a fifth aspect of the present invention, there is provided the use of a compound or composition according to the present invention as a pesticide, preferably as a bactericide or fungicide, which, according to this aspect of the present invention, excludes methods for the treatment of the human or animal body by surgery or therapy. [Brief explanation of the drawings]

[0011] [Figure 1] Optical absorption (UV-VIS) spectrum of compounds according to formula I(a), formula I(b) or lipopetin from 200 to 400 nm [Figure 2] LC-ESI-MS / MS spectrum of the precursor of formula I(a) 1204.6 m / z (M+H)+ illustrating fragment peaks consistent with the amino acids: aspartic acid, hydroxy-glutamine, serine, methyl-asparagine, and methyl-phenylalanine. [Figure 3] LC-ESI-MS / MS / MS spectrum of the precursor of formula I(a) at 294.2 m / z illustrating peaks consistent with the molecule C14H25-OH2-C4H5ON [Figure 4] High field region of the 1D 1H NMR spectrum of the compound according to formula I(a) in CD3OD at 600 MHz [Figure 5] Low field region of the 1D 1H NMR spectrum of the compound according to formula I(a) in CD3OD at 600 MHz [Figure 6] Graphical representation of lipopeptide gene clusters DETAILED DESCRIPTION OF THE INVENTION

[0012] The present invention relates to a compound of formula (I) [ka] (wherein R1 is CH3 or C2H5) or a salt thereof.

[0013] The compounds according to the invention comprise or are lipopeptides.

[0014] Thus, compounds according to the present invention include compounds according to formula I(a) and / or compounds according to formula I(b), or salts thereof.

[0015] Compounds according to formula I(a) have the structural formula [ka] or a salt thereof.

[0016] Compounds according to formula I(a) have the molecular formula C 55 H 85 N11O 19 and an exact mass of 1203.602 g. The compound according to Formula I(a) has a solubility in DMSO of greater than 10,000 ppm. The compound according to Formula I(a) comprises or is a lipopeptide.

[0017] Compounds according to formula I(b) have the structural formula [ka] or a salt thereof.

[0018] The compound of formula I(b) has the molecular formula C 56 H 87 N 11 O 19 and an exact mass of 1217.618 g. The compound according to formula I(a) has a solubility in DMSO of greater than 10,000 ppm. The compound according to formula I(b) comprises or is a lipopeptide.

[0019] In a preferred embodiment, the compound according to Formula I is an isolated compound. The term "isolated" in reference to a compound means that the compound has been isolated from its natural environment.

[0020] In one aspect, the present invention relates to a composition comprising a compound according to the present invention and a microorganism capable of producing the compound as disclosed herein, wherein the compound or a composition comprising a compound according to the present invention and a microorganism capable of producing the compound according to the present invention is applied to a plant or part thereof to treat or protect the plant from a disease caused by a plant pathogenic microorganism, such as a fungus, bacterium, or virus.

[0021] Surprisingly, it has now been found that compounds and / or compositions according to the invention have advantageous levels of biological activity for treating or protecting plants from diseases caused by the infestation of plant pathogenic microorganisms such as fungi, bacteria or viruses. Surprisingly, compounds and / or compositions according to the invention have advantageous fungicidal activity against a variety of plant pathogenic fungi.

[0022] Preferably, the compounds and / or compositions according to the invention have fungicidal or bactericidal activity, and are therefore preferably fungicidal or bactericidal agents.

[0023] The term "fungicidally active compound" or "fungicide" means a compound that controls, modifies, or prevents the growth of fungi. The term "fungicidally effective amount," when used, means the amount of such a compound or combination of such compounds that is capable of producing an effect on the growth of fungi. A controlling or modifying effect includes any deviation from the natural progression, such as killing, retardation, etc., and prevention includes the formation of a barrier or other defense in or on the plant to prevent fungal infection.

[0024] The compounds and / or compositions according to the invention may be produced in any suitable manner, preferably by culturing a microorganism in a suitable fermentation medium that allows the production of the compounds and / or compositions of the invention. The microorganism is preferably a Streptomyces sp.

[0025] Microorganisms capable of producing the compounds or compositions according to the invention include or are microorganisms comprising 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 according to SEQ ID NO: 47 to 91, preferably the amino acid sequence of SEQ ID NO: 67 and / or SEQ ID NO: 68.

[0026] Preferably, the microorganism 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 10, 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, 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 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, 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, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, 0, SEQ ID NO:51, SEQ ID NO:52, 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, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:74, SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85, SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90, or SEQ ID NO:91, preferably a nucleotide sequence encoding the amino acid sequence of SEQ ID NO:67 and / or SEQ ID NO:68, or an amino acid sequence having at least 80, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity thereto.

[0027] Microorganisms capable of producing compounds or compositions according to the invention include microorganisms comprising at least one nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity or 100% identity to the nucleotide sequences of SEQ ID NOs: 2 to 46, and preferably SEQ ID NO: 22 and / or SEQ ID NO: 23.

[0028] Preferably, the microorganism comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, 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, 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 of SEQ ID NO: 2, SEQ ID NO: nucleotide sequence of SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, 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, or a nucleotide sequence having at least 80, 85%, 90%, 95%, 96%, 97%, 98%, 99% identity thereto.

[0029] The microorganism capable of producing the compounds according to the present invention may be a naturally occurring microorganism or a recombinant microorganism. Recombinant microorganisms can be produced by methods known to those skilled in the art. Recombinant microorganisms can be produced by transforming a microorganism with at least one nucleotide sequence encoding at least one protein of the amino acid sequence according to SEQ ID NOs: 47 to 91, preferably the amino acid sequence of SEQ ID NOs: 67 and 68, preferably at least one of the nucleotide sequences of SEQ ID NOs: 2 to 47, preferably the nucleotide sequence of SEQ ID NO: 22 or 23, or at least one nucleotide sequence having at least 80% identity thereto, preferably at least 85%, preferably at least 90%, preferably at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, preferably at least 99% identity thereto.

[0030] Preferably, the microorganism in the composition or process of the present invention as disclosed herein is a bacterium of the genus Streptomyces, and preferably the bacterium is Streptomyces chrestomyceticus, S. rimosus, S. paromomycinus, or S. monomicini. Preferably, the composition comprises Streptomyces sp. Saigon 413, deposited at the Westerdijk Institute under accession number CBS 149411. Preferably, the microorganism is a Streptomyces sp., for example Streptomyces sp. Saigon413 deposited at the Westerdijk Institute under accession number CBS149411, wherein the Streptomyces sp. has a 16S RNA sequence having at least 98%, preferably at least 98.2%, 98.4%, 98.6%, 98.8%, preferably at least 99%, 99.2%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, preferably at least 99.9% or 100% identity to SEQ ID NO:1.

[0031] Preferably, the microorganism in the composition or process according to the invention, such as Streptomyces chrestomyceticus, comprises a genome sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% identity or 100% identity to the complete genome of Streptomyces chrestomyceticus NRRL-3672 or to the complete genome of Streptomyces sp. Saigon413 deposited at the Westerdijk Institute under accession number CBS149411. In one embodiment, a composition or process according to the invention comprises Streptomyces chrestomyceticus, Streptomyces sp. Saigon 413, deposited at the Westerdijk Institute under accession number CBS149411.

[0032] As used herein, the terms "percent identity" and "percent identical" refer to the relatedness of two or more nucleotide or amino acid sequences, which may be calculated by (i) comparing two optimally aligned sequences over a window of comparison, (ii) determining the number of positions where the same nucleic acid base (for nucleotide sequences) or amino acid residue (for proteins) occurs in both sequences to obtain the number of matched positions, (iii) dividing the number of matched positions by the total number of positions in the window of comparison, and then (iv) multiplying this quotient by 100 percent to 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 region of alignment 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 being allowed), "percent identity" with respect to a 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) and then multiplied by 100 percent.

[0033] The present invention also relates to a microorganism which is Streptomyces sp. Saigon413 deposited at the Westerdijk Institute under accession number CBS149411.

[0034] Surprisingly, it has been found that Streptomyces sp. Saigon413, deposited at the Westerdijk Institute under accession number CBS149411, has advantageous properties compared to Streptomyces sp. known in the art.

[0035] The compounds of the present invention, or compositions containing the compounds of the present invention, can be used in the agricultural sector and related fields of use, for example, as active ingredients for controlling phytopathogenic microorganisms. The compounds of the present invention are distinguished by their excellent activity at low application rates, such as 2 to 250 ppm, e.g., 10 to 200 ppm, e.g., 20 to 100 ppm, while being well tolerated by plants and environmentally safe. They have very useful preventative properties and can be used to protect a large number of plants. The compounds of the present invention can be used to suppress or destroy phytopathogenic microorganisms occurring on plants or plant parts (fruits, flowers, leaves, stems, tubers, roots) or different crops of plants. The compounds can also protect later-developing plant parts.

[0036] The compounds according to the invention and / or compositions comprising the compounds according to the invention can be used as is or can be formulated with adjuvants, preferably agriculturally acceptable adjuvants. Formulations known in the art are, for example, emulsifiable concentrates, coatable pastes, sprayable or dilutable solutions or suspensions, powders, dusts, granules and encapsulants.

[0037] Thus, in one embodiment, a composition comprising a compound according to the present invention as disclosed herein further comprises an adjuvant. Preferably, the adjuvant is an agriculturally acceptable adjuvant.

[0038] 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.

[0039] 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 (dimethyl sulfoxide).

[0040] Suitable solid carriers include, for example, talc, titanium dioxide, pyrophillide 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.

[0041] Adjuvants can be surfactants, crystallization inhibitors, viscosity modifiers, suspending agents, spray droplet conditioners, pigments, antioxidants, foaming agents, antifoaming agents, light-blocking agents, compatibilizers, sequestering agents, neutralizing and buffering agents, corrosion inhibitors, dyes, colorants, spreading agents, penetration aids, micronutrients, emollients, lubricants, and adhesives.

[0042] Compositions as disclosed herein are preferably agriculturally acceptable compositions.

[0043] Compositions containing the compounds according to the present invention as disclosed herein typically contain 0.5 to 95% w / w, such as 1 to 90% w / w, such as 2 to 80% w / w, such as 5 to 60% w / w, of the active ingredient. The compounds according to the present invention may be the only active ingredient in the compositions disclosed herein. In one embodiment, a composition containing a compound of the present invention further contains at least one additional active ingredient. The active ingredient, as defined herein, has fungicidal and / or insecticidal and / or herbicidal activity, or activity as a plant growth regulator. The compounds or compositions of the present invention may be mixed with one or more additional ingredients having pesticidal activity, such as fungicides, insecticides, herbicides, bactericides, miticides, nematicides, and / or plant growth regulators, as appropriate. 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.

[0044] Additional ingredients with pesticidal, e.g. fungicidal or fungicidal, activity may result in unexpected synergistic activity. Thus, a composition comprising a lipopeptide compound according to formula I and an additional active ingredient, e.g., malonomycin, may exhibit a synergistic effect. A synergistic effect occurs whenever the effect of the active ingredient combination is greater than the sum of the effects of the individual components. The expected effect E for a given active ingredient combination can be calculated according to the so-called COLBY formula (COLBY, SR "Calculating synergistic and antagonistic responses of herbicide combinations", Weeds, Vol. 15, pages 20-22; 1967): ppm = milligrams of active ingredient (=ai) per litre of spray mixture X = % effect of active ingredient A) using p ppm of active ingredient Y = % effect of active ingredient B) using q ppm of active ingredient.

[0045] According to COLBY, the expected (additive) effect of active ingredients A) + B) using p + q ppm of active ingredients is:

number

[0046] If the actual observed effect (O) is greater than the expected effect (E), the effect of the combination is superadditive, i.e., a synergistic effect exists. In mathematical terms, synergy corresponds to a positive value for the difference (OE). In the case of a purely complementary addition of activities (expected activity), the difference (OE) is zero. A negative value for the difference (OE) indicates a loss of activity compared to the expected activity.

[0047] In addition to the actual synergistic action with regard to fungicidal and bactericidal activity, the compositions according to the invention may also have other unexpected advantageous properties, such as better degradability, improved toxicological and / or ecotoxicological behavior, or increased seedling emergence, crop yield, more developed root systems, increased tillering, increased plant height, larger leaf blades, fewer dead basal leaves, stronger tillers, greener leaf color, less fertilizer needed, fewer seeds needed, more productive tillers, earlier flowering, earlier grain maturity, less plant lodging (lodging), increased shoot growth, improved plant vigor, and earlier germination.

[0048] Additional ingredients having pesticidal activity and / or plant growth regulators can be combined with the compositions of the present invention and used in the methods of the present invention, and applied simultaneously or sequentially with the compositions of the present invention. If applied simultaneously, these additional ingredients can be formulated together with the compositions of the present invention or mixed, for example, in a spray tank. As an alternative to directly mixing these additional pesticidal ingredients, the ingredients can be used in separate fungicidal, insecticidal, or herbicidal applications as part of a program of fungal, insect, or weed control that is extended over part or all of the growing season.

[0049] The at least one additional component having pesticidal activity and / or being a plant growth regulator may be any suitable known bactericide / fungicide, insecticide, herbicide and / or plant growth regulator. The at least one additional component having pesticidal activity and / or plant growth regulator may be of chemical or biological origin, for example, plant or microbial origin. The at least one additional component having pesticidal activity in the composition as disclosed herein may be produced by a microorganism capable of producing a compound according to formula (I) according to the present invention as disclosed hereinabove.

[0050] Additionally, the compositions of the present invention may also 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.

[0051] The compounds and / or compositions according to the present invention may induce plant resistance through a priming mechanism, which leads to a physiological state that allows plants to respond faster and / or more robustly after exposure to biotic or abiotic 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. October 1, 2014.

[0052] In one embodiment, the composition according to the invention further comprises cyclothiazomycin C, streptimidon and / or malonomycin.

[0053] Cyclothiazomicin C is a known compound, the structure of which is disclosed on page 3 of WO 2015191789, and which can be produced as disclosed in Example 4 of WO 2015 / 191789.

[0054] Malonomicin (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 of formula II. [ka]

[0055] Maronomycin can be produced as disclosed in WO 2006 / 078939, Example I. Maronomycin can also be prepared according to the methods disclosed in EP 1 860 939, Examples IA and B, or according to Law et al., 2018 (Nature Catalysis | VOL 1 | DECEMBER 2018 | 977-984).

[0056] Streptimidon is a known compound of formula III. [ka]

[0057] Streptimidone 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).

[0058] The present invention also relates to a composition comprising a lipopeptide compound according to formula (I), preferably according to formula I(a), and maronomycin. Surprisingly, it has been found that a composition comprising a lipopeptide compound according to formula (I) and maronomycin can exhibit an unexpected synergistic fungicidal and fungicidal effect. The surprising synergistic fungicidal and fungicidal effect of a composition comprising a lipopeptide compound according to formula (I), preferably according to formula I(a), and maronomycin has been found against, for example, Zymoseptoria tritici, Fusarium culmorum, Microdochium nivale, Botrytis cinerea, Puccinia recondita, and Pyricularia oryzae.

[0059] In one embodiment, the active ingredients cyclothiazomycin C, streptimidon and / or malonomycin are produced by a microorganism capable of producing the compound according to formula (I) according to the invention as defined herein above.

[0060] Compositions comprising a mixture of the compound of the invention with at least one additional active ingredient are preferably in a mixing ratio of 100:1 to 1:6000, in particular 50:1 to 1:50, more in particular 20:1 to 1:20, even more in particular 10:1 to 1:10, very in particular 5:1 to 1:5, with ratios of 2:1 to 1:2 being particularly preferred, and ratios 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. 1:2, 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. The composition comprising a mixture of a lipopeptide compound according to formula (I) and malonomycin comprises a ratio of the lipopeptide compound according to formula (I) to malonomycin of 2000:1 to 1:2000, preferably 1000:1 to 1:1000, for example 800:1 to 1:800, for example 600:1 to 1:600, or 500:1 to 1:500, 400:1 to 1:400, 300:1 to 1:300, or 300:1 to 1:200. A mixture as described above can be used in a method for controlling pests, which method comprises applying a composition comprising a mixture as described above to the pests or their environment, except for methods for treating the human or animal body by surgery or therapy or diagnostic methods performed on the human or animal body.

[0061] Compositions comprising a mixture of a compound of the present invention with one or more active ingredients as described above, such as malonomycin, can be applied, for example, in a single "ready-to-use" form, in a combined spray mixture made up of separate formulations of the single active ingredient components, such as a "tank mix," and in a sequential manner, i.e., in combination with the single active ingredients when applied one after the other with a reasonably short period of time between them, such as a few hours or days.

[0062] In one aspect, the present invention relates to a process for the production of a compound or composition according to the invention, the process comprising culturing a microorganism in a suitable fermentation medium under conditions that allow the production of the compound. The microorganism that is fermented in the process disclosed herein is a microorganism that is capable of producing a compound according to the invention as defined herein above.

[0063] Microorganisms capable of producing the compounds according to the invention are, for example, bacteria of the genus Streptomyces, as disclosed herein above.

[0064] Culturing microorganisms in a suitable fermentation medium to produce the compounds according to the present invention is known to those skilled in the art. Microorganisms can be fermented under aerobic or anaerobic conditions. Microorganisms belonging to the genus Streptomyces are typically cultured under aerobic conditions. A suitable fermentation medium contains nutrients such as a suitable carbon source, such as sugarcane molasses or sugarbeet molasses, polysaccharides, flour, starch, sugar, or glucose, and a suitable nitrogen source, such as casein hydrolysate, tryptone, ammonium sulfate, ammonia, yeast extract, peptone or urea peptide, or amino acids. The production process of the compounds according to the present invention can be carried out in batch, fed-batch, or continuous culture.

[0065] In one embodiment, the process further comprises producing a composition comprising a compound according to the invention as defined herein. The microorganism capable of producing the compound or composition according to the invention is defined herein above. The microorganism capable of producing the compound or composition according to the invention may be capable of producing further active ingredients as defined herein above, such as cyclothiazomycin C, streptimidinone and / or malonomycin.

[0066] The process according to the present invention may further comprise a step of recovering the compound according to the present invention or a salt thereof. The compound according to the present invention may be recovered by a suitable method known in the art, for example, by crystallization or chromatography, for example, HPLC. The recovery of the compound according to the present invention may further comprise a step of purifying the compound.

[0067] The process for the preparation of the compounds according to the invention may further comprise the step of formulating the compounds into a suitable formulation or composition as defined herein above.

[0068] In a further aspect, the present invention relates to a method for controlling or preventing infestation of plants, plant propagation material and / or harvested food crops by phytopathogenic microorganisms by treating the plants, plant propagation material and / or harvested food crops, wherein an effective amount of a compound or composition according to the present invention is applied to the plant, part thereof or its habitat, plant propagation material and / or harvested food crops.

[0069] In methods for controlling or preventing plant beetles, applying an effective amount of a compound or composition of the invention includes applying 0.01 to 5 kg per hectare (ha) (active ingredient (ai) per hectare (ha), preferably 0.015 g to 500 g ai / ha, preferably 0.020 g to 100 g ai / ha, preferably 0.025 g to 50 g ai / ha, preferably 0.030 g to 5 g ai / ha, preferably 0.035 g to 500 mg ai / ha).

[0070] When the compounds of the invention or compositions of the invention are used to treat seeds, a rate of 0.0001 to 10 g of compound of the invention per kg of seeds, for example 0.0002 to 0.1 g per kg of seeds, for example 0.0005 to 0.001 g per kg of seeds, is generally sufficient.

[0071] Suitably, the compounds or compositions of the invention are applied either prophylactically, meaning before the onset of disease, or therapeutically, meaning after the onset of disease.

[0072] The phytopathogenic microorganisms affected by the compounds of the invention are disease fungi and fungal vectors as well as phytopathogenic bacteria and viruses. The phytopathogenic microorganisms in the method according to the invention include the following disease fungi and fungal vectors as well as 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. niger, and A. terrus; Aureobasidium spp. including A. pullulans; Bacillus subtilis; Blastomyces dermatitidis Botryosphaeria spp. including B. dermatidis, Blumeria graminis, Blumeriella jaapii, B. dothidea, and B. obtusa; Botrytis spp. including B. cinerea; Bremia lactucae, Cadophora gregata, C. albicans, C. glabrata, C. krusei, C. lusitaniae, C. parapsilosis, and C. tropicalis; Cercospora spp., including 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.Fusarium species, including F. pseudograminearum, F. sacchari, F. sambucinum, F. subglutinans, F. solani, F. sporotrichioides, F. tricinctum, and F. virguliforme, Gaeumannomyces graminis, Gibberella spp. including G. graminis, G. avenacea, G. fujikuroi, G. intricans, G. moniliformis, G. zeae, Gloeodes pomigena, Gloeosporium musarum, Glomerella cingulate, Golovinomyces cichoracearum, Gymnosporangium juniperi-virginianae, Guignardia bidwellii, bidwellii, Gymnosporangium juniperi-virginianae, 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, Microsporum spp., Monilinia spp. including M. fructicola, and Monographella spp. 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, and Ophiostoma spp. Pantoea stewartia, Paracoccidioides spp., Parastagonospora nodorum, Pectobacterim spp., Penicillium spp. including P. digitatum, P. italicum, Petriellidium spp., Peronosclerospora spp. including P. maydis, P. philippinensis and P. sorghi, Peronosclerospora spp. including P. destructor, Peronospora spp. including P. destructor, spp., Phaeosphaeria nodorum, soybean rust (Phakopsora pachyrhizi), Phellinus igniarus, Phialophora spp., Phlyctema vagabunda, Phoma spp., Phomopsis viticola, Phyllachora pomigena, Phyllosticta spp., Physoderma maydis, P. capsica, P. infestans, and other Phytophthora spp. Plasmopara spp., including Plasmodiophora brassicae, P. halstedii, and P. viticola; Podosphaera spp., including Plenodomus spp., Pleospora spp., and 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, Pseudopeziza tracheiphila, Pseudopyrenochaeta Puccinia spp. including P. lycopersici, P. hordei, P. recondita, P. striiformis, and P. triticina; Pyrenopeziza spp., Pyrenophora spp., Pyricularia spp. including P. oryzae; Pythium spp. including P. ultimum; Ralstonia solanacearum; Ramularia spp.; Rathayibacter spp. spp, Remotididymella destructiva, Rhizoctonia spp, Rhizomucor pusillus, Rhizopus arrhizus, Rhynchosporium spp, Robbsia andropogonis, Sarocladium oryzae, S. apiospermum and S. prolificans.prolificans, Scedosporium spp., Schizothyrium pomi, Sclerophthora macrospora, Sclerotinia spp., including S. sclerotiorum, Sclerotium spp., Sclerotium spp., S. nodorum, Septoria spp., including S. tritici, Setosphaeria turcica, Sphaerotheca macularis, Sphaerotheca fusca, Sphaerotheca fuliginea, fuliginea), Spiroplasma kunkelii, Sporothorix spp., Stagonospora nodorum, Stagonosporopsis cucurbitacearum, Stemphylium spp., Stenocarpella macrospora, Stereum hirsutum, Streptomyces spp., Thanatephorus cucumeris, Thielaviopsis basicola, Tilletia spp., Tranzschelia discolor), T. harzianum, T. pseudokoningii, T. virideTrichoderma spp. including Trichoderma viride, Trichophyton spp., Typhula spp., Uncinula necator, Urocystis spp., Uromyces spp., Ustilago spp., Venturia spp. including V. inaequalis, Verticillium spp., Wilsononomyces carpophilus, or Xanthomonas spp. including X. oryzae and X. campestris, Xylella spp. spp) and Zymoseptoria tritici.

[0073] Phytopathogenic microorganisms which have been found to be surprisingly affected by the compounds and / or compositions according to the invention are fungi, such as fungi belonging to the genus Blumeria, Botrytis sp., Cercospora sp., Fusarium sp., Glomerella, Microdochium Mycosphaerella, Zymoseptoria sp., Parastagonospora sp., Puccinia sp., Phaeosphaeria sp., Pyrenophora, Pyricularia sp., Sclerotinia sp., Zymoseptoria, preferably Blumeria graminis. Blumeria graminis f.sp. tritici, Botrytis cinerea, Cercospora arachidicola, Fusarium culmorum, Glomerella lagenarium, Microdochium nivale, Mycosphaerella arachidis, Parastagonospora nodorum, Puccinia recondite, Puccinia recondita f.sp. tritici, Phaeosphaeria nodorum nodorum) and Pyrenophora.

[0074] Phytopathogenic microorganisms which have been found to be surprisingly affected by the compounds and / or compositions according to the invention are fungi, for example fungi belonging to the genus Botrytis, Glomerella, Mycosphaerella, Puccinia, Phaeosphaeria, Pyrenophora or Zymoseptoria, preferably Botrytis cinerea, Glomerella lagenarium, Mycosphaerella arachidis, Puccinia recondita f.sp. tritici, Phaeosphaeria nodorum, It is a fungus belonging to the genus Pyrenophora nodorum, Pyrenophora teres, or Zymoseptoria tritici.

[0075] Control or prevention means reducing the infestation by phytopathogenic microorganisms, especially fungi, to a level such that an improvement is demonstrated.

[0076] A preferred method for controlling or preventing infestation of crop plants by phytopathogenic microorganisms, particularly fungi, or insects involves the application of a compound or composition according to the present invention, which is a foliar application. The frequency and rate of application will depend on the risk of infestation by the corresponding pathogen or insect. However, the compound or composition according to the present invention can also be introduced into the plant through the roots via the soil (systemic action) by flooding the plant habitat with a liquid formulation or by applying the compound in solid form to the soil (soil application), e.g., in granular form. In rice crops, such granules can be applied to flooded rice fields. The compound or composition according to the present invention can also be applied to seeds by impregnating the seeds or tubers with a liquid formulation of the fungicide or coating them with a solid formulation (coating).

[0077] For protection against fungal infection and phytopathogenic fungi occurring in the soil, the compounds or compositions according to the present invention can also be used as dressings for treating plant propagation material, for example, seeds such as fruit, tubers, or grains, or plant cuttings. The propagation material can be treated with the compounds and / or compositions according to the present invention before planting: for example, seeds can be dressed before sowing. The compounds and / or compositions according to the present invention can also be applied to grains (coatings) either by impregnating the seeds in a liquid formulation or by coating the seeds with a solid formulation. The compositions can also be applied to the planting site when the propagation material is planted, for example, in the sowing furrow during sowing. Disclosed herein are such methods for treating plant propagation material, and the plant propagation material thus treated.

[0078] The term "habitat" as used herein means the field in which plants are growing or where seeds of cultivated plants are sown or where seeds will be sown in the soil. It includes the soil, seeds, and seedlings, as well as established vegetation.

[0079] The term "plant" refers to any physical part of a plant, including seeds, seedlings, saplings, roots, tubers, stems, stalks, leaves, and fruits. It may also include germinated plants and young plants that are to be transplanted after germination or emergence from the soil. These young plants can be protected before transplanting by complete or partial treatment by immersion.

[0080] The term "plant propagation material" is understood to mean plant material such as reproductive parts of plants, such as seeds, cuttings or tubers (e.g. potatoes), roots, fruits, bulbs, rhizomes or plant parts, which can be used for their propagation.

[0081] The term plants includes "useful plants" or "crops". The expressions "useful plants" and "crops" are used interchangeably herein. "Useful plants" and "crops" include 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; agricultural crops such as sugar and fodder beet, coffee, hops, mustard, oilseed rape (canola), poppy, sugarcane, sunflower, tea and tobacco; fruits such as apples, apricots, avocados, bananas, cherries, citrus fruits, nectarines, peaches, pears and plums, grasses such as bermudagrass, strawberry grass, bentgrass, centipedegrass, fescue, ryegrass, St. Augustine grass and zoysiagrass; basil, borage, chives, coriander, lavender, lovage, mint, oregano, parsley, rosemary , herbs such as sage and thyme; legumes such as beans, lentils, peas and soybeans; nuts such as almonds, cashews, groundnuts, hazelnuts, peanuts, pecans, pistachios and walnuts; palms such as oil palms; ornamental plants such as flowers, shrubs and trees, other trees such as cocoa, coconut, olives and rubber; vegetables such as asparagus, eggplant, broccoli, cabbage, carrots, cucumber, garlic, lettuce, marrow, melon, okra, onion, pepper, potato, pumpkin, rhubarb, spinach and tomato; and perennial and annual crops such as vines, e.g., grapes. The term "plant" also includes timber crops such as pine trees or woody plants.

[0082] The term "useful plants" should also be understood to include useful plants which have been made tolerant to herbicides such as bromoxynil, or to several 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, etc.) or PPO (protoporphyrinogen-oxidase) inhibitors, either by conventional methods of breeding or as a result of genetic engineering.

[0083] The term "useful plants" should also be understood to include useful plants which have been transformed by the use of recombinant DNA techniques so that they are able to synthesize one or more selectively acting toxins, such as are known, for example, from toxin-producing bacteria, in particular from bacteria of the genus Bacillus.

[0084] Any suitable plant, plant propagation material or food 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 potato, tomato, grape, canola / rapeseed / colza, cucurbit, groundnut, wheat, barley, maize, rice, banana or soybean, preferably the plant is wheat or barley.

[0085] In another aspect, the present invention relates to the use of a compound or composition according to the present invention as a pesticide, preferably as a bactericide / fungicide, and / or as a priming agent. The relevant features of the compounds and compositions according to the present invention are as disclosed hereinabove. Accordingly, the present invention relates to the use of a compound and / or composition according to the present invention as a bactericide / fungicide. [Example]

[0086] Example 1. Sources and Extraction of Compounds of the Invention 1.1. Fermentation of Streptomyces sp. Streptomyces species were ordered from the strain collections disclosed in Table 1. Streptomyces sp. Saigon 413 was isolated in Vietnam before 1961. Streptomyces sp. Saigon 413 was deposited at the Westerdijk Institute under accession number CBS149411. The deposit was made by Syngenta Ltd., Jealott's Hill Research International Centre, Bracknell, Berkshire, RG42 6EY, UK, under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure.

[0087] 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 in a 150 rpm shaking incubator with a 25 mm throw for 4 days.

[0088] The presence of compounds according to Formula I, such as compounds according to Formula I(a) and Formula I(b), in the fermentation broth was determined by isolation and purification according to the methods specified in Section 1.5 and disclosed in Section 2.

[0089] The results in Table 1 show that compounds according to Formula I(a) were present in the fermentation broths of various Streptomyces species.

[0090] 1.2.16S rDNA isolation, whole genome sequencing 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.

[0091] Strain Streptomyces sp. Saigon413 was identified by comparing the 16S rRNA sequence according to SEQ ID NO: 1 using barrnap v0.9 with publicly available 16S rRNA sequences extracted using 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 Streptomyces chrestomyceticus. The sequence identity between the 16S rRNA sequences of S. sp. Saigon413 and the publicly available S. chrestomyceticus NRRL-3672 was 99.87%, determined using Muscle v3.8.31 and the R package Seqinr v4.2-16.

[0092] Additionally, 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 polished with Pilon using Illumina reads. Genomic DNA was also extracted from Streptomyces rimosus CBS 492.64, Streptomyces rimosus CBS 570.66, Streptomyces rimosus CBS 569.66, Streptomyces chrestomyceticus DSM 41224, Streptomyces rimosus subsp. rimosus DSM 40673, and Streptomyces rimosus subsp. rimosus DSM 41057 using the method described by Kieser et al. (2000) in Practical Streptomyces Genetics. Whole genome sequencing of these strains was completed using Nanopore sequencing technology, and the genomes were assembled in Flye (Kolmogorov, M., et. al. (2019), Nature Biotechnology, 37, 540).

[0093] After assembly of the genome from Streptomyces sp. Saigon413 and publicly available genomes, the average nucleotide identity (ANI) was calculated 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 publicly available genome of S. chrestomyceticus NRRL 3672.

[0094] Using 16S RNA sequence identity and ANI scores (%), strains CBS 596.66, CBS570.66, and DSM 41429 were determined to be Streptomyces chrestomyceticus strains, and not Streptomyces rimosis or Streptomyces paromomycinus strains, as indicated by the depository.

[0095] Table 1 shows the percentage identity of the whole genome and 16S RNA sequences of several Streptomyces species to those of Streptomyces sp. Saigon413.

[0096] [Table 1-1]

[0097] [Table 1-2]

[0098] 1.3. Identification of the biosynthetic gene cluster producing the lipopeptide according to Formula I in Streptomyces sp. Saigon 413 To identify genes involved in the production of lipopeptides according to Formula I, the assembled genome (see Example 1.2) was scanned with AntiSMASH (version 5.1.1, Blin et al., Nucleic Acids Res (2019) doi:10.1093 / nar / gkz310), a tool commonly used to aid in the identification of biosynthetic gene clusters involved in the production of secondary metabolites.

[0099] The identification of lipopeptide compounds as a family of lipopeptides (see 1 above and the anti-SMASH 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 species 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 the lipopeptide compounds of Formulas I(a) and I(b).

[0100] The NRPS biosynthetic gene cluster contains 45 coding sequences, including two coding sequences for NRPS genes, coding sequences for regulatory factors, and coding sequences involved in the biosynthesis of the precursor incorporated into the lipopeptide of Formula I (Figure 6 and Table 4).

[0101] [Table 2-1]

[0102] [Table 2-2]

[0103] 1.4. 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. Saigon 413 To confirm that the identified biosynthetic gene cluster was associated with the production of [insert compound ID here], a 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 pBCCon2192 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).

[0104] Plasmid pBCon2192 was transformed into E. coli ET12567 / pUZ8002 using standard electroporation techniques and then introduced into Streptomyces species Saigon413 by mycelial 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 to force primary integration. After 6 days at 37°C, the resulting strains were transferred onto ISP-4 solid agar medium without selection and incubated at 28°C for 15 days to allow a second crossover.

[0105] After 15 days of growth, strains were harvested in 20% glycerol. 100 μl of cell suspension was used to inoculate fresh plates as well as 10 -10 Serial dilutions were performed up to 10 -8 ~10 -10 100 μl of the culture was then plated onto ISP-4 agar plates. The plates were incubated at 28° C. until single colonies were observed.

[0106] 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.

[0107] To confirm that both SEQ ID NO:22 and the region containing 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 to the Streptomyces sp. Saigon413 genome confirmed the deletion of the genomic region containing SEQ ID NO:22 and SEQ ID NO:23. In addition, whole-genome analysis using Illumina-PCR-free sequencing confirmed that no other modifications had been made to the genome.

[0108] Cultivation of Streptomyces sp. Saigon413Δ7318-7319 and analysis of extracts from the strain confirmed that the lipopeptide compound according to Formula I(a) was no longer produced by the strain, and confirmed that SEQ ID NO:22 and SEQ ID NO:23 were essential for the production of the lipopeptide compound according to Formula I(a).

[0109] 1.5. Purification of the Compounds According to the Invention Mycelia from fermentation broth derived from the Streptomyces strains disclosed in Table 1 were separated by centrifugation, and the supernatant was treated with butanol. The butanol was removed, and the extract was partitioned between water and ethyl acetate. Lipopeptides were purified from the ethyl acetate fraction by preparative reverse-phase (C18) HPLC. Lipopeptides are relatively nonpolar and elute 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 compounds according to Formula I(a) and Formula I(b).

[0110] Similarly, the fermentation broth of Streptomyces sp. Saigon413 was treated.

[0111] Stock solutions for compounds of Formula I(a) and Formula I(b) were made in DMSO (up to 10 mg / ml) which were further diluted with water plus 0.025% Tween 20 to produce suitable working concentrations for biological efficacy assays. Compounds were detected by UV-VIS (Figure 1), mass spectrometry (Figures 2 and 3), and NMR spectroscopy (Figures 4 and 5).

[0112] 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).

[0113] Example 2. Characterization of compounds of formula I 2.1. 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 resolution: 30,000, RF lens (%): 60, polarity: positive), Experiment 3: tMS2 OT HCD (MSn level (n): 2, isolation window (m / z): 1.0, activation type: HCD, HCD collision energy (%): 30, detector type: Orbitrap, Orbitrap resolution: 30,000, RF lens (%): 60, polarity: positive), experiment 4: tMS3 OT HCD (MSn level (n): 3, isolation window (m / z): 1.6, activation type: HCD, HCD collision energy (%): 30, MS2 isolation window (m / z): 2, MS2 activation type: HCD, MS2 HCD collision energy (%): 30, detector type: Orbitrap, Orbitrap resolution: 30,000, RF lens (%): 60, polarity: positive). The mass spectrometer was 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 were as follows: 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. 2 and 3 show the LC-ESI-MS / MS / MS spectrum of the compound of formula I(a). Compounds according to Formula I(a) and Formula I(b) were identified in the fermentation broth of Streptomyces sp. Saigon 413, deposited under accession number CBS 149411.

[0114] 2.2.NMR spectroscopy NMR spectra were recorded on a 5 mm Bruker ( 1 H / 19 F) / 13 C / 15 Spectra were recorded on a Bruker AVIII 600 NMR spectrometer equipped with an N TCI cryoprobe. Samples were dissolved in CD3OD and spectra were recorded at 300°K. 1 The residual solvent signal at 3.31 ppm for H was used as the reference. Figures 4 and 5 each cover half of the 1H NMR spectrum of a compound according to Formula 1(a).

[0115] 2.3. Molecular Composition and Mass The molecular composition and mass of the compounds according to Formula I(a) and Formula I(b) were determined using the results of liquid chromatography and high-resolution mass spectrometry as disclosed in 2.1. The compounds of Formula I(a) and I(b) have the following compositions: Formula I(a) Lipopeptide 1204: molecular composition C55H85N11O19 and exact mass of 1203.602. Formula I(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.

[0116] 2.4. Solubility The solubility of compounds of formula I(a), formula I(b) and lipopeptin A was measured in water and DMSO:

[0117] [Table 3]

[0118] Example 3. Plant activity of compounds according to the present invention 3.1. Fungicidal activity in liquid culture assays Fungal mycelial fragments or conidial suspensions, either freshly prepared from fungal liquid cultures or cryogenically stored, were mixed directly into the nutrient broth. Stock solutions of Formula I(a) and Formula I(b) compounds were prepared in DMSO (up to 10 mg / ml) and diluted with water plus 0.025% Tween® 20 to produce 10x concentrated samples, and 10 μl of this solution was pipetted into a 96-well microtiter plate. The nutrient broth containing the fungal spores / mycelial fragments was then added to obtain the final concentration of the test compound. The test plates were incubated in the dark at 24°C and 96% rh. Inhibition of fungal growth was measured photometrically after 2 to 7 days, depending on the pathogen response system, and percent antifungal activity was calculated relative to an untreated control. The effects of the test compounds (Formula I(a), I(b) and Lipopeptin A) were tested against the following fungi under the conditions outlined above and specifically herein below:

[0119] Botryotinia fuckeliana (Botrytis cinerea) / Liquid culture (gray mold) Fungal conidia from cryogenic storage were mixed directly into nutrient broth (Vogels broth) and growth inhibition was measured photometrically 3-4 days after application.

[0120] Glomerella lagenarium (Colletotrichum lagenarium) / Liquid culture (Anthracnose) Fungal conidia from cryogenic storage were mixed directly into nutrient broth (PDB potato dextrose broth). Growth inhibition was measured photometrically 3-4 days after application.

[0121] Mycosphaerella arachidis (Cercospora arachidicola) / Liquid culture (Early spot disease) Fungal conidia from cryogenic storage were mixed directly into nutrient broth (PDB potato dextrose broth). Growth inhibition was measured photometrically 4-5 days after application.

[0122] Zymoseptoria tritici (Septoria tritici) / Liquid Culture (Septoria leaf spot) Fungal conidia from cryogenic storage were mixed directly into nutrient broth (PDB potato dextrose broth). Growth inhibition was measured photometrically 4-5 days after application.

[0123] The results are shown in Tables 2 and 3.

[0124] [Table 4]

[0125] [Table 5]

[0126] The results in Tables 2 and 3 show that the lipopeptide compounds of Formula I(a) and Formula I(b) were active against several plant pathogenic fungi, providing up to 100% control. The compounds of Formula I(a) and Formula I(b) had similar efficacy, while lipopeptin A was less active.

[0127] 3.2 Leaf disc or leaf segment test in well plates Leaf discs or leaf segments of various plant species were excised from plants grown in a greenhouse. The excised leaf discs or leaf segments were placed on plain agar medium in multiwell plates (24-well format). Before inoculation with fungal spores, the leaf discs were sprayed with a test solution. The compounds to be tested were prepared as DMSO solutions (up to 10 mg / ml) and diluted to the appropriate concentration with water plus 0.025% Tween® 20 immediately before spraying. The inoculated leaf discs or leaf segments were incubated under specified conditions (temperature, relative humidity, light, etc.) depending on the respective test system. A single assessment of disease level was performed by visual evaluation 3 to 9 days after inoculation, depending on the pathogen response system as shown below. The percent disease control was then calculated compared to untreated control leaf discs or leaf segments.

[0128] Puccinia recondita f.sp. tritici / wheat / leaf rust (leaf rust) Wheat cv. Kanzler leaf segments were placed on agar in multiwell plates (24-well format) and sprayed with the formulated test compounds diluted in water. The leaf segments were inoculated with a fungal spore suspension one day after application. The inoculated leaf segments were incubated in a climate cabinet at 19°C and 75% rh under a 12-h light / 12-h dark light regime, and the activity of the compounds was evaluated as percent disease control compared to untreated control segments when appropriate levels of disease appeared on untreated control segments (7-9 days after application).

[0129] Phaeosphaeria nodorum (Septoria nodorum) / Wheat / Leaf blight (Wheat leaf blight) Wheat cv. Kanzler leaf segments were placed on agar in multiwell plates (24-well format) and sprayed with the formulated test compounds diluted in water. The leaves were inoculated with a fungal spore suspension two days after application. The inoculated test leaf segments were incubated in a climate cabinet at 20°C and 75% rh under a 12-h light / 12-h dark light regime, and the activity of the compounds was evaluated as percent disease control compared to untreated control segments when an appropriate level of disease appeared on the untreated control segments (5-7 days after application).

[0130] Pyrenophora teres / Barley / Leaf spot preventative (net blotch) Barley leaf segments (cv. Hasso) were placed on agar in multiwell plates (24-well format) and sprayed with the formulated test compounds diluted in water. The leaf segments were inoculated with a fungal spore suspension two days after application. The inoculated leaf segments were incubated in a climate cabinet at 20°C and 65% rh under a 12-h light / 12-h dark light regime, and the activity of the compounds was evaluated as disease control compared to untreated control segments when appropriate levels of disease appeared on untreated control segments (5-7 days after application).

[0131] [Table 6]

[0132] [Table 7]

[0133] The results in Tables 4 and 5 show that compounds according to Formula I(a) or I(b) were active in the assays, providing up to a 95% reduction in fungal growth in several fungal species. Compounds of Formula I(a) and Formula I(b) have similar efficacy, while lipopeptin A was found to be less active.

[0134] 3.3. Leaf smear assay on wheat seedlings Wheat seedlings of cv. Riband (for Zymoseptoria tritici tests) or cv. Arina (for Puccinia recondite tests) 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 dots were made on the second leaf to generate three sections of approximately equal size: section A (base), section B (middle), and section C (top of the leaf). Stock solutions of test compounds were prepared at a concentration of 10,000 ppm in DMSO. The stock solution was then further diluted with water supplemented with Tween® 20 to a final concentration of ≥200 ppm test compound, 0.05% Tween® 20, and 2% DMSO. The diluted compound was applied to the central section of the L2 using a conventional cotton stick; the cotton stick was dipped in the diluted compound and rubbed several times on the adaxial leaf surface between the two marks. One day later, the intact plants were inoculated with a fungal spore suspension using a paintbrush and applying the spore suspension until runoff.

[0135] For infection with Zymoseptoria tritici: Test plants were inoculated by spraying a spore suspension (1.5 μg / ml in water supplemented with 0.01% Tween® 20) on them 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 kept in a greenhouse at 22°C / 21°C (day / night) and 70% rh. Efficacy was assessed by visual evaluation as soon as an appropriate level of disease appeared on untreated control plants (usually >80% disease coverage 16-19 days after inoculation).

[0136] For infection with Puccinia recondita: Test plants were inoculated by spraying them 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 kept in a greenhouse at 20°C and 60% rh. The percentage of leaves covered by disease was assessed by visual evaluation when an appropriate level of disease appeared on untreated control plants (usually 50-80% disease coverage 9-12 days after infection). Three sections of the leaf were evaluated separately.

[0137] [Table 8]

[0138] The results in Table 6 show that compounds of Formula I(a) and Formula I(b) were active in the assay, providing up to 70% control of Puccinia recondite and Zymoseptoria tritici on the treated area (center section). Compounds of Formula I(a) and Formula I(b) have similar efficacy.

[0139] 3.4. Reactive oxygen species (ROS) burst assay using wheat leaf discs 200 μL of the appropriate concentration of test solution (analyte) or the respective control (water) was pipetted into a white 96-well plate (Nunc, Langenselbold, Germany). 5 mm leaf discs were obtained from 2-week-old wheat plants using a tissue punch and then floated on the test solution. The plate was stored at room temperature (RT) for 24 h. The next day, the solution was replaced with 50 μL of ddH2O, and the leaf discs were left for at least 1 h at room temperature in the dark for regeneration. Meanwhile, the appropriate master mix was freshly prepared in a black 5 mL reaction tube with or without the elicitor flg22 (see below). After regeneration, 50 μL of the corresponding master mix was added to the leaf disc-containing wells. Luminescence was then recorded for 40 min on a plate reader (BMG Labtech; Ortenberg, Germany). The compound INA (2,6-dichloro-isonicotinic acid, CAS: 5398-44-7) is a synthetic salicylic acid analogue and was included as a benchmark for priming activity (Kauss et al., 1992). Mastermix-flg22:4.98mL ddH2O, 10μL HRP (10mg / mL), 10μL L-012 (20mM) Mastermix+flg22:4.979mL ddH2O, 10μL HRP (10mg / mL), 10μL L-012 (20mM), 1μL flg22 (10μM) Abbreviations: flg22 (22 amino acid flagellin peptide. Eurogentec Cat. No. AS-62633); HRP (horseradish peroxidase), L-012 sodium salt (CAS#: 143556-24-5) was used.

[0140] [Table 9]

[0141] The results in Table 7 show that wheat pretreatment with a compound of Formula I(a) or Formula I(b) at a concentration of 100 ppm increased ROS production induced by peptide flg22 by a factor of more than 2.7, a response similar to or stronger than that observed from treatment with INA (2,6-dichloro-isonicotinic acid), a well-known priming agent (Krauss et al., 1992: Dichloroisonicotinic and salicylic acid, inducers of systemic acquired resistance, enhance fungal elicitor responses in parsley cells, Plant Journal 2:655-60).

[0142] Example 4. Fungicidal activity of mixtures of lipopeptides according to formula I(a) and malonomycin in liquid culture assays

[0143] method Stock solutions for compounds of Formula I(a) were prepared in DMSO (up to 10 mg / ml). Maronomycin was prepared according to Law et al., 2018 (Nature Catalysis | VOL 1 | DECEMBER 2018 | 977-984). Stock solutions of maronomycin were prepared in water plus 0.025% Tween® 20. An assay to test the effectiveness of mixtures of lipopeptide compounds of Formula I(a) in combination with maronomycin for controlling fungal pathogens in liquid culture assays was designed for 96-well plates as outlined in Table 1.

[0144] [Table 10]

[0145] The 96-well plate design allows for the disease control of the mixture to be compared to that of each single compound at the same ratio. Comparison of the calculated efficacy of the same mixture according to Colby with the estimated efficacy from the mixture allows for the statement of whether the mixture is additive (efficacy is similar to the Colby calculation), synergistic (efficacy is better than the Colby calculation), or antagonistic (efficacy is inferior to the Colby calculation). The same 96-well plate design also allows for the evaluation of whether the two compounds can be mixed at different use rates and mixing ratios. The plate design outlined in Table 1 would provide the following mixing ratios, as shown in Table 2:

[0146] [Table 11]

[0147] A master plate was prepared with 10x concentrated solutions of compound stock solutions diluted in water plus 0.025% Tween® 20. The concentrations of DMSO (from the stock of the compound of Formula I(a)) and Tween® 20 were kept constant in all cells of the master plate. 10 μL was transferred from the master plate to a 96-well test plate. Nutrient broth containing fungal spores / mycelial fragments was then added to the test plate to obtain a 1x final concentration for the test compound (as outlined in Table 8). The test plate was incubated in the dark at 24°C and 96% rh. Inhibition of fungal growth was determined photometrically after approximately 3 days, and the percent fungal growth reduction relative to the untreated control was calculated.

[0148] The effectiveness of the mixture was tested on different fungal species:

[0149] Zymoseptoria tritici (EPPO code: SEPTTR) Fungal conidia from cryogenic storage were mixed directly into nutrient broth (PDB: potato dextrose broth). Test plates were incubated at 24°C and growth inhibition was measured photometrically after 72 hours.

[0150] Fusarium culmorum (EPPO code: FUSACU) Fungal conidia from cryogenic storage were mixed directly into nutrient broth (PDB: potato dextrose broth). Test plates were incubated at 24°C and growth inhibition was measured photometrically after 72 hours.

[0151] Microdochium nivale (EPPO code: MONGNI) Fungal conidia from cryogenic storage were mixed directly into nutrient broth (PDB: potato dextrose broth). Test plates were incubated at 24°C and growth inhibition was measured photometrically after 72 hours.

[0152] Botrytis cinerea (EPPO code: BOTRCI) Fungal conidia from cold storage were mixed directly into nutrient broth (Vogel's minimal medium). Test plates were incubated at 24°C and growth inhibition was measured photometrically after 72 hours.

[0153] Pyricularia oryzae (EPPO code: PYRIOR) Fungal conidia from cryogenic storage were mixed directly into nutrient broth (PDB: potato dextrose broth). Test plates were incubated at 24°C and growth inhibition was measured photometrically after 72 hours.

[0154] Cercospora arachidicola (EPPO code: MYCOAR) Fungal conidia from cryogenic storage were mixed directly into nutrient broth (PDB: potato dextrose broth). Test plates were incubated at 24°C and growth inhibition was measured photometrically after 5-6 days.

[0155] Sclerotinia sclerotiorum (EPPO code: SCLESC) Fungal mycelial fragments prepared from fresh liquid cultures were mixed directly into nutrient broth (PDB: potato dextrose broth). Test plates were incubated at 24°C and growth inhibition was measured photometrically after 72 hours.

[0156] result

[0157] [Table 12]

[0158] For each of the assay conditions in Table 10 with an efficacy of 50% or greater (hereafter referred to as the effective mixture), the respective mixture ratio can be assigned from Table 9. Effective mixtures for the control of Zymoseptoria tritici were found at 641:1 to 1:100.

[0159] [Table 13]

[0160] [Table 14]

[0161] For each of the assay conditions in Table 12 with 50% or greater efficacy (effective mixtures), the respective mixture ratios can be assigned from Table 9. Effective mixtures for control of Fusarium culmorum were found at ratios between 641:1 and 20:1.

[0162] [Table 15]

[0163] [Table 16]

[0164] For each of the assay conditions in Table 14 with 50% or greater effectiveness (effective mixtures), the respective mixture ratios can be assigned from Table 9. Effective mixtures for the control of Microdochium nivale were found at 641:1 to 1:3.

[0165] [Table 17]

[0166] [Table 18]

[0167] For each of the assay conditions in Table 16 with an efficacy of 50% or greater (effective mixtures), the respective mixture ratios can be assigned from Table 9. Effective mixtures for the control of Botrytis cinerea were found at 641:1 to 1:6.

[0168] [Table 19]

[0169] [Table 20]

[0170] For each of the assay conditions in Table 18 with 50% or greater efficacy (effective mixtures), the respective mixture ratios can be assigned from Table 9. Effective mixtures for the control of Pyricularia oryzae were found at 641:1 to 1:3.

[0171] [Table 21]

[0172] [Table 22]

[0173] For each of the assay conditions in Table 20 with an efficacy of 50% or greater (effective mixtures), the respective mixture ratios can be assigned from Table 9. Effective mixtures for the control of Cercospora arachidicola were found at 641:1 to 1:100.

[0174] [Table 23]

[0175] For each of the assay conditions in Table 21 with 50% or greater efficacy (effective mixtures), the respective mixture ratios can be assigned from Table 9. Effective mixtures for control of Sclerotinia sclerotiorum ranged from 641:1 to 1:2.

[0176] conclusion Mixtures of lipopeptide compounds of Formula I(a) and malonomycin control a variety of fungal pathogens. A wide variety of mixture ratios of the two compounds, ranging from 641:1 to 1:100, produced greater than 50% control of fungal growth for several fungal species, including examples with ratios between the mentioned ratios (compound malonomycin:compound (a) of Formula I). ​​Surprisingly, the efficacy of many mixtures was better than predicted based on calculations from Colby, demonstrating that mixtures of lipopeptide compounds of Formula I(a) and malonomycin have a synergistic effect on fungal pathogen control. This surprising synergy was observed in trials to control Zymoseptoria tritici, Fusarium culmorum, Microdochium nivale, Botrytis cinerea, and Pyricularia oryzae.

[0177] Example 5. Fungicidal activity of mixtures of lipopeptide compounds according to formula I(a) and malonomycin in the leaf disc assay A stock solution of the lipopeptide compound of formula I(a) was prepared in DMSO (up to 10 mg / ml). Maronomycin was prepared according to Law et al., 2018 (Nature Catalysis | VOL 1 | DECEMBER 2018 | 977-984). A stock solution of maronomycin was prepared in water plus 0.025% Tween® 20. The assay to test the efficacy of mixtures of compounds of formula I(a) in combination with malonomycin for control of fungal pathogens in a leaf disc assay was designed for two 24-well plates.

[0178] [Table 24]

[0179] [Table 25]

[0180] [Table 26]

[0181] [Table 27]

[0182] A first set of master plates was prepared with a 1x concentrated spray solution of the lipopeptide compound of Formula I(a) stock diluted in water at the concentrations shown in Table 22, Table 23, Table 25, or Table 26, respectively. Each well contained 2% DMSO and 0.025% Tween 20. A second set of master plates was prepared accordingly with a 1x concentrated malonomycin stock diluted in water. Each well of the second set contained 0.025% Tween 20. Leaf sections placed on agar in a 24-well plate were sprayed with 8 μl of solution from the master plate containing the lipopeptide compound of Formula I(a), allowed to dry, and then sprayed with 8 μl of solution from the master plate containing malonomycin two hours later. After the second spray dried, the leaf sections were infected with fungal spores to obtain a preventative application timing. Alternatively, leaf sections infected one day before spraying with the compound were used to obtain a therapeutic spray timing. In addition, several plates were generated in which leaf sections were sprayed 2x in the absence of test compounds (with only DMSO and Tween 20) to represent untreated controls. For each leaf section, the percent leaf coverage of disease symptoms was assessed. The percent leaf coverage reduction relative to the untreated control was calculated. The efficacy of the mixtures was tested in duplicate on different fungal species. The reported efficacy values ​​are the average of the two replicates.

[0183] Puccinia recondita (EPPO code: PUCCRE) for preventive spraying. Wheat (variety Kanzler) leaf segments are placed on agar in multi-well plates (24-well format) and sprayed with the test solution (8 μl per well). After drying, the leaf segments are inoculated with a fungal spore suspension. After appropriate incubation, the activity of the compounds is evaluated 8 dpi (days after inoculation) as preventive fungicidal activity.

[0184] Puccinia recondita (EPPO code: PUCCRE) at the time of therapeutic spraying. Wheat (variety Kanzler) leaf segments are placed on agar in a multi-well plate (24-well format). The leaf segments are then inoculated with a fungal spore suspension. One day after inoculation, the test solution is sprayed (8 ul / well). After appropriate incubation, the activity of the compound is evaluated 8 dpi (days after inoculation) as therapeutic fungicidal activity.

[0185] Blumeria graminis f.sp. tritici (EPPO code: ERYSGT) for preventive spraying Wheat (variety Kanzler) leaf segments are placed on agar in multi-well plates (24-well format) and sprayed with the test solution (8 ul per well). After drying, the leaf segments are inoculated with fungal spores. After appropriate incubation, the activity of the compounds is evaluated 7 dpi (days after inoculation) as preventive fungicidal activity.

[0186] Parastagonospora nodorum (EPPO code: LEPTNO) Wheat (variety Kanzler) leaf segments are placed on agar in multi-well plates (24-well format) and sprayed with the test solution (8 ul per well). After drying, the leaf segments are inoculated with fungal spores. After appropriate incubation, the activity of the compounds is evaluated 4 dpi (days after inoculation) as preventive fungicidal activity.

[0187] result

[0188] [Table 28]

[0189] For each of the assay conditions in Table 28 with an efficacy of 50% or greater (effective mixtures), the respective mixture ratios can be assigned from Table 24. Effective mixtures for the control (preventative) of Puccinia recondita were found at 64:1 to 1:270.

[0190] [Table 29]

[0191] For each of the assay conditions in Table 29 with an efficacy of 50% or greater (effective mixtures), the respective mixture ratios can be assigned from Table 24. Effective mixtures for the control (curative) of Puccinia recondita were found at 64:1 to 1:270.

[0192] [Table 30]

[0193] [Table 31]

[0194] For each of the assay conditions in Table 31 with an efficacy of 50% or greater (effective mixtures), the respective mixture ratios can be assigned from Table 27. Effective mixtures for the control (preventative) of Blumeria graminis f.sp. tritici were found at 192:1 to 1:9.

[0195] [Table 32]

[0196] For each of the assay conditions in Table 32 with 50% or greater efficacy (effective mixtures), the respective mixture ratios can be assigned from Table 27. Effective mixtures for the control (preventative) of Parastagonospora nodorum were found at 192:1 to 1:3.

[0197] Conclusion: Compounds of Formula I(a) can be mixed with malonomycin to obtain complete or partial control of various fungal pathogens. The mixture ratio of the two compounds can vary widely within the mixture and still produce greater than 50% control of fungal growth when sprayed on leaves. For several fungal species, examples are provided for ratios ranging from 192:1 to 1:270, including many examples with ratios between the stated ratios (compound malonomycin:lipopeptide compound of Formula I(a)). Surprisingly, the efficacy of many mixtures was better than predicted based on calculations from Colby, indicating that mixtures of compounds of Formula I(a) and malonomycin have a synergistic effect on fungal pathogen control when sprayed on leaves. This surprising synergistic effect was observed in tests to control Puccinia recondita.

Claims

1. Formula (I) 【Chemical 1】 (Wherein R1 is CH 3 or C 2 H 5 is) A compound or a salt thereof according to

2. A composition comprising a compound of claim 1 and a microorganism capable of producing the compound of claim 1.

3. 10. A composition comprising the compound of claim 1 or the composition of claim 2, further comprising an adjuvant.

4. 4. The compound of claim 1 or the composition of claim 2 or 3, wherein the compound has bactericidal and fungicidal activity.

5. The composition according to any one of claims 2 to 4, further comprising at least one additional ingredient with pesticidal activity and / or at least one plant growth regulator.

6. 6. The composition of claim 5, wherein the additional components include cyclothiazomycin C, streptimidon and / or malonomycin, preferably malonomycin.

7. 10. A process for the production of a compound according to claim 1 or a composition according to any one of claims 2 to 6, comprising culturing a microorganism in a suitable fermentation medium under conditions that allow the production of said compound or said composition.

8. 8. The composition according to any one of claims 2 to 6 or the process according to claim 7, wherein the microorganism comprises at least one nucleotide sequence encoding a protein having at least 80% identity with at least one amino acid sequence according to SEQ ID NOs: 47 to 91, preferably SEQ ID NO: 67 or SEQ ID NO: 68, and preferably said at least one nucleotide sequence has at least 80% identity with at least one of the nucleotide sequences according to SEQ ID NOs: 2 to 46, preferably SEQ ID NO: 22 or SEQ ID NO:

23.

9. 9. The composition of any one of claims 2 to 6 or 8, or the process of any one of claims 7 or 8, wherein the microorganism is Streptomyces sp., preferably Streptomyces chretomyceticus, preferably a Streptomyces comprising a 16S RNA sequence having at least 98% identity to SEQ ID NO: 1, preferably Streptomyces sp. Saigon 413 deposited at the Westerdijk Institute under accession number CBS 149411.

10. A microorganism which is Streptomyces sp. Saigon 413 deposited at the Westerdijk Institute under accession number CBS149411.

11. 10. A method for controlling or preventing infestation of plants, plant propagation material and / or harvested food crops by plant pathogenic microorganisms by treating said plants, plant propagation material and / or harvested food crops, which comprises applying an effective amount of a compound according to claim 1 or a composition according to any one of claims 2 to 6, 8 or 9 to said plants, parts thereof or their habitats, said plant propagation material and / or harvested food crops.

12. 12. The method of claim 11, wherein the effective amount comprises 0.001 g to 5 kg of the compound of formula (I) per hectare.

13. 12. The method of claim 11, wherein the plant propagation material is a seed and the effective amount comprises 0.0001 to 50 g of the compound of formula (I) per kg of seed.

14. The phytopathogenic microorganisms are fungi, preferably Blumeria, Botrytis sp., Cercospora sp., Fusarium sp., Glomerella, Microdochium, Mycosphaerella, Zymoseptoria sp., Parastagonospora sp., Puccinia sp., Phaeosphaeria sp. sp.), Pyrenophora, Pyricularia sp., Sclerotinia sp., Zymoseptoria, preferably Blumeria graminis f. sp. Blumeria graminis f.sp. tritici, Botrytis cinerea, Cercospora arachidicola, Fusarium culmorum, Glomerella lagenarium, Microdochium nivale, Mycosphaerella arachidis, Parastagonospora nodorum, Puccinia recondite, The method according to any one of claims 11 to 13, wherein the fungus is a fungus belonging to the group consisting of Puccinia recondita f.sp. tritici, Puccinia recondita f.sp. tritici, Phaeosphaeria nodorum, Pyrenophora teres, Pyricularia oryzae, Sclerotinia sclerotiorum, or Zymoseptoria tritici.

15. 15. The method of any one of claims 11 to 14, wherein the plants include potato, tomato, grape, canola, cucurbit, groundnut, wheat, and / or barley, corn, rice, soybean, banana.

16. Use of a compound according to claim 1 or a composition according to any one of claims 2 to 6, 8 or 9 as a pesticide, preferably as a bactericide-fungicide and / or as a priming agent.

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