Bactericidal and fungicidal compounds

Abstract

The present invention relates to a compound of formula (I) JPEG2025533160000027.jpg94160 and compositions containing same, processes for producing said compounds and methods of using said compounds and compositions for preventing or controlling (fungal) fungi in plants and in agriculture.

Description

[Technical Field]

[0001] The present invention relates to novel compounds having fungicidal and bactericidal activity, compositions containing the compounds, methods for preparing the compounds, and the use of the compounds or compositions for preventing or controlling fungal infestations in plants, harvested food crops, seeds, or non-living materials in agriculture or horticulture. [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. Biological fungicides can be, for example, of microbial origin or can be plant extracts. Known microorganisms that produce antibiotics against fungi are, for example, actinomycetes, such as Streptomyces sp. A very well-known species is Streptomyces natalensis, which produces natamycin, an antifungal compound used in food and crop protection. U.S. Patent No. 5,356,624 discloses a strain of Streptomyces rimosus that was found to be active against several wood-decaying fungi. WO 2022 / 038180 discloses novel Streptomyces species that produce several known antifungal compounds, such as streptimidon, natamycin (pimaricin), or albofungin. Extracts of these bacterial strains have been found to exhibit activity against well-known plant pests, such as Fusarium graminearum, Zymoseptoria tritici, and Puccinia striiformis.

[0003] The development of resistance to fungicides in fungi, government regulations, and strong societal demands have led to a continuing need to search for new compounds of biological origin with fungicidal and bactericidal activity. Summary of the Invention [Means for solving the problem]

[0004] The present invention is 53 H 90 N2O 44 and preferably has a molecular mass of 1458.487 g, and is further characterized by an NMR spectrum as set out in Tables 1 and 2, or a salt thereof.

[0005] The present invention further provides a compound of structural formula (I): [ka] or a salt thereof.

[0006] 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.

[0007] In a third aspect, the present invention relates to a process for producing a compound or composition according to the invention, comprising culturing a microorganism in a suitable fermentation medium under conditions that allow the production of the compound.

[0008] In a fourth aspect, the present invention relates to a method for controlling or preventing infestation of phytopathogenic microorganisms in plants, comprising applying to the plant, its part or its habitat an effective amount of a compound or composition according to the invention.

[0009] 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 a bactericide or fungicide, and / or as a priming agent, which use, according to this aspect of the invention, excludes surgical or therapeutic treatment of the human or animal body. [Brief explanation of the drawings]

[0010] [Figure 1] 1 is a one-dimensional 1H NMR spectrum of the compound of the present invention measured at 600 MHz in D2O. [Figure 2] 1 is a one-dimensional 13C NMR spectrum of the compound of the present invention measured at 600 MHz in D2O. [Figure 3] 1 is a two-dimensional 1H-13C HSQC NMR spectrum of the compound of the present invention measured in D2O at 600 MHz by the Dept method, showing a positive (CH) signal. [Figure 4] 1 is a two-dimensional 1H-13C HSQC NMR spectrum of the compound of the present invention measured at 600 MHz in D2O by the Dept method, showing a negative (CH2) signal. DETAILED DESCRIPTION OF THE INVENTION

[0011] According to the present invention, C 53 H 90 N2O 44 and further characterized by the NMR spectra set forth in Tables 1 and 2. 53 H 90 N2O 44 has a molecular mass of 1458.487 g. Preferably, the compounds according to the invention have a solubility at pH 7 of more than 10000 ppm and / or a solubility in DMSO of more than 9772 ppm.

[0012] Preferably, the compound according to the invention is an isolated compound. The term "isolated" in reference to a compound means that the compound has been isolated from its native environment. Preferably, the compound according to the invention is an oligosaccharide.

[0013] The compounds according to the present invention have the formula (I): [ka] The compound may be a compound having the structural formula: or a salt thereof.

[0014] 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 disclosed herein, wherein the 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 cure or protect the plant from a disease caused by a plant pathogenic microorganism, such as a fungus, bacterium, or virus.

[0015] It has now been surprisingly found that the compounds and / or compositions according to the invention have a level of biological activity advantageous for curing or protecting plants from diseases caused by infestations of plant pathogenic microorganisms, such as fungi, bacteria, or viruses. Surprisingly, the compounds and / or compositions according to the invention have advantageous bactericidal and / or fungicidal activity against various plant pathogenic fungi, such as Zymoseptoria tritici, Puccinia recondita f.sp. tritici, Magnaporthe grisea, Blumeria graminis f.sp. tritici, and / or Parastagonospora nodorum.

[0016] The composition according to the present invention is a non-natural composition.

[0017] Preferably, the compounds and / or compositions according to the invention have fungicidal or bactericidal activity, and therefore are preferably fungicidal or bactericidal agents.

[0018] As used herein, the term "fungicidally active compound" or "fungicide" refers to a compound that controls, modifies, or prevents the growth of fungi. When the term "fungicidally effective amount" is used, it refers to an amount of such a compound or combination of such compounds that is capable of affecting the growth of fungi. A control or modification effect includes any deviation from natural growth, such as killing, retardation, etc., and prevention includes the formation of a barrier or other defense in or on a plant where fungal infection is to be prevented.

[0019] Preferably, the microorganism capable of producing the compounds of the present invention, for example in the compositions disclosed herein, is a bacterium belonging to the genus Streptomyces, and preferably the bacterium is Streptomyces chrestomyceticus, Streptomyces rimosus, Streptomyces albofaciens, Streptomyces paromomycinus, or Streptomyces 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., preferably Streptomyces sp. Saigon413 deposited at the Westerdijk Institute under accession number CBS149411, which 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.

[0020] Preferably, the microorganism capable of producing the compound according to the invention comprises a genome sequence having at least 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity or 100% identity to the complete genome of Streptomyces sp. Saigon413 deposited at the Westerdijk Institute under accession number CBS149411.

[0021] As used herein, the terms "percent identity" and "percent identical" refer to the relatedness of two or more nucleotide or amino acid sequences and can be calculated by (i) optimally aligning and comparing two sequences over a comparison window, (ii) determining the number of positions where identical nucleic acid bases (in the case of nucleotide sequences) or amino acid residues (in the case of proteins) occur in both sequences to obtain the number of matched positions, (iii) dividing the number of matched positions by the total number of positions within 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 without specifying a specific comparison window, the percent identity is determined by dividing the number of matched positions over the aligned region by the full length of the reference sequence. Therefore, for purposes of the present invention, when two sequences (query and subject) are optimally aligned (allowing for gaps in the alignment), the "percent identity" with respect to a query sequence is equal to the number of identical positions between the two sequences divided by the total number of positions over the full length (or comparison window) of the query sequence, multiplied by 100 percent.

[0022] Also disclosed herein is a microorganism which is Streptomyces sp. Saigon413, deposited at the Westerdijk Institute under accession number CBS149411.

[0023] Surprisingly, it has been found that Streptomyces sp. Saigon413, deposited at the Westerdijk Institute under accession number CBS149411, has advantageous properties compared to Streptomyces species microorganisms known in the art.

[0024] 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 plant pests. The compounds of the present invention are distinguished by their excellent activity at low application rates, for example, 5 to 300 ppm, for example, 10 to 250 ppm, for example, 20 to 200 ppm, their good plant tolerance, and their environmental safety. They possess highly useful curative, preventive, and systemic properties, allowing them to be used for the protection of many cultivated plants. The compounds of the present invention can be used to suppress or eliminate pests that occur on plants or plant parts (fruits, flowers, leaves, stems, tubers, roots) of various crop plants, while simultaneously protecting later-developing plant parts.

[0025] The compounds according to the invention or compositions comprising the compounds according to the invention can be used as such 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 encapsulations.

[0026] Thus, in one embodiment, a composition comprising a compound according to the invention disclosed herein further comprises an adjuvant. Preferably, the adjuvant is an agriculturally acceptable adjuvant.

[0027] Suitable adjuvants are known in the art and include, for example, solvents, liquid carriers, solid carriers or extenders, surfactants, dispersants, emulsifiers, wetting agents, adjuvants, solubilizers, penetration enhancers, protective colloids, adhesives, thickeners, humectants, water repellents, attractants, feeding stimulants, compatibilizers, bactericides, antifreeze agents, antifoaming agents, colorants, tackifiers and binders.

[0028] 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 may be water or DMSO.

[0029] Suitable solid carriers include, for example, talc, titanium dioxide, pyroferritic clay, silica, attapulgite clay, kieselguhr, chalk, diatomaceous earth, lime, calcium carbonate, bentonite clay, fuller's earth, cottonseed hulls, wheat flour, soybean flour, pumice, wood flour, walnut shell flour, and lignin.

[0030] Examples of adjuvants include surfactants, crystallization inhibitors, viscosity modifiers, suspending agents, spray droplet regulators, pigments, antioxidants, foaming agents, antifoaming agents, light-blocking agents, compatibilizers, sequestering agents, neutralizing agents and buffers, corrosion inhibitors, dyes, odorants, spreading agents, penetration aids, micronutrients, softening agents, lubricants, and adhesives.

[0031] The compositions disclosed herein are preferably agriculturally acceptable compositions.

[0032] The compositions of the present invention may be in any conventional form, for example, in a twin dosage form. The compound may be used in the form of a dry seed treatment powder (DS), seed treatment emulsion (ES), seed treatment flowable (FS), seed treatment solution (LS), seed treatment water dispersible powder (WS), seed treatment capsule suspension (CF), seed treatment gel (GF), emulsifiable concentrate (EC), suspension concentrate (SC), suspoemulsion (SE), capsule suspension (CS), water dispersible granule (WG), emulsifiable granule (EG), water-in-oil emulsion (EO), oil-in-water emulsion (EW), microemulsion (ME), oil dispersion (OD), oil mixture flowable (OF), oil mixture liquid (OL), soluble concentrate (SL), ULV suspension (SU), ULV liquid (UL), technical concentrate (TK), dispersible concentrate (DC), wettable powder (WP) or in any technically feasible formulation in combination with an agriculturally acceptable adjuvant.

[0033] Such compositions can be prepared in a conventional manner, for example, by mixing the active ingredient with suitable inert ingredients (diluents, solvents, fillers, and optionally other ingredients, such as surfactants, biocides, antifreeze agents, spreading agents, thickeners, and compounds providing auxiliary effects). If long-lasting efficacy is desired, conventional sustained-release formulations can also be used. In particular, formulations applied in the form of sprays, such as water-dispersible concentrates (e.g., EC, SC, DC, OD, SE, EW, EO, etc.), wettable powders, and granules, may contain surfactants such as wetting agents and dispersants, as well as other compounds providing auxiliary effects, such as condensation products of formaldehyde with naphthalenesulfonates, alkylarylsulfonates, ligninsulfonates, fatty alkyl sulfates, and ethoxylated alkylphenols and ethoxylated fatty alcohols.

[0034] Compositions containing a compound according to the present invention typically contain 0.5 to 95 wt. % of the active ingredient, for example, 1 to 90 wt. %, for example, 2 to 80 wt. %, for example, 5 to 60 wt. The compound 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 comprises at least one additional active ingredient. An active ingredient, as defined herein, is an ingredient having fungicidal and / or insecticidal and / or herbicidal activity or plant growth regulator activity. The compound of the present invention or the compositions disclosed herein may be mixed with one or more additional ingredients having pesticidal activity known in the art, such as fungicides, insecticides, herbicides, microbicides, miticides, molluscicides, and / or plant growth regulators, if appropriate. The one or more additional ingredients having pesticidal activity may be of biological or chemical origin. The pesticides referred to herein using their common names are known, for example, from "The Pesticide Manual", 19th Ed., British Crop Protection Council 2021.

[0035] The use of an additional ingredient with pesticidal activity, e.g., bactericidal or fungicidal activity, can result in unexpected synergistic activity. Thus, a composition comprising a compound according to the present invention and an additional active ingredient, e.g., malonomycin, may exhibit synergistic activity. When the effect of the combination of active ingredients exceeds the sum of the effects of the individual ingredients, a synergistic effect occurs. The expected effect E of a given combination of active ingredients can be calculated according to the so-called COLBY formula as follows (COLBY, SR "Calculating synergistic and antagonistic responses of herbicide combination", Weeds, Vol. 15, pages 20-22; 1967): ppm = milligrams of active ingredient (=ai) per litre of spray mixture X = Effect of active ingredient (%) when active ingredient A is used in ppm Y = Effect of active ingredient (%) when q ppm of active ingredient (B) is used

[0036] According to COLBY, the (additive) effect expected from active ingredients A) + B) when used at p + q ppm of active ingredients is as follows:

number

[0037] If the observed effect (O) exceeds the expected effect (E), the combination is supra-additive, i.e., synergistic. Mathematically, synergy corresponds to a positive difference in (OE). If the activity is purely complementary additive (expected activity), the difference (OE) is zero. If the difference (OE) is negative, this indicates a decrease in activity compared to the expected activity.

[0038] Additional pesticidal and / or plant growth regulator ingredients can be used in the methods of the present invention in combination with the compositions of the present invention and applied simultaneously or sequentially with the compositions of the present invention. If applied simultaneously, these ingredients can be formulated together with the compositions of the present invention or mixed, for example, in a spray tank. As an alternative to direct mixing, these pesticidal ingredients can be used in separate fungicide / fungicide, insecticide, or herbicide applications as part of a program to control fungi, insects, or herbs over part or all of the growing season.

[0039] The at least one additional ingredient having pesticidal activity and / or being a plant growth regulator can be any suitable known bactericide, fungicide, insecticide, herbicide, and / or plant growth regulator. The at least one additional ingredient having pesticidal activity and / or being a plant growth regulator can be of chemical or biological origin, such as plant or microbial origin. The at least one additional ingredient having pesticidal activity in the compositions disclosed herein can be produced by a microorganism capable of producing a compound according to the present invention.

[0040] Additionally, the compositions of the present invention can 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 salicylates and the commercially available SAR inducer acibenzolar-S-methyl.

[0041] The compounds and / or compositions according to the present invention can induce plant resistance through a priming mechanism. Priming is a mechanism that leads plants to a physiological state that allows them to respond more quickly 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. 1, October 2014.

[0042] In one embodiment, the composition according to the invention further comprises cyclothiazomycin C (CtmC), streptimidon and / or malonomycin.

[0043] The structure of cyclothiazomycin C is disclosed on page 3 of WO2015191789 and can be prepared as disclosed in Example 4 of WO2015 / 191789.

[0044] Malonomicin (sometimes referred to as "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 has the formula II: [ka] It has the structural formula:

[0045] Malonomycin can be prepared as disclosed in WO 2006 / 078939, Example I. Malonomycin 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 Catalyses | VOL 1 | DECEMBER 2018 | 977-984).

[0046] Streptimidon has the formula III: [ka] is a known compound.

[0047] 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). In one embodiment, the active ingredients cyclothiazomycin C, streptimidone and / or malonomycin are produced by a microorganism capable of producing the compounds according to the invention as defined herein above.

[0048] The present invention also relates to a compound of the molecular formula C 53 H 90 N2O 44 The present invention also relates to a composition comprising a compound having the formula: 53 H 90 N2O 44 It has been found that a compound having a molecular formula according to the formula (I), preferably a composition comprising a compound according to formula (I) and malonomycin, can exhibit an unexpected synergistic fungicidal effect. It has been found that a composition comprising a compound according to the present invention and malonomycin exhibits a surprising synergistic fungicidal effect against, for example, Puccinia recondita.

[0049] Compositions comprising a compound of the invention and at least one additional active ingredient, such as malonomycin, are preferably mixed in a 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 particularly 5:1 to 1:5, in particular a ratio of 2:1 to 1:2, as well as a ratio of 4:1 to 2:1, in particular a ratio of 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 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 mixing ratio of the composition containing the mixture of the compound according to the present invention and malonomycin is 1000:1 to 1:1000, preferably 500:1 to 1:500, preferably 450:1 to 1:300, preferably 400:1 to 1:150, preferably 350:1 to 1:100 or 100:1 to 1:80. The mixture described above can be used in a method for controlling pests such as phytopathogenic microorganisms, which method comprises applying a composition containing the mixture described above to the pests or their environment, but excludes methods for surgical or therapeutic treatment of the human or animal body and diagnostic methods performed on the human or animal body.

[0050] Compositions comprising the compounds of the present invention described above and one or more additional ingredients that are pesticidal or plant growth regulators can be applied, for example, in the form of a single "ready-mixed" component, in a spray mixture consisting of a combination of separate formulations of a single active ingredient, such as a "tank mix," and in combination applications in which the single active ingredients are applied sequentially, i.e., one after the other, over a reasonably short period of time, such as a few hours, days, or weeks.

[0051] In one aspect, the present invention relates to a process for producing a compound or composition according to the invention, comprising culturing a microorganism in a suitable fermentation medium under conditions that allow the production of said compound or composition. The microorganism that is fermented in the process disclosed herein is a microorganism capable of producing a compound according to the invention as disclosed herein above.

[0052] Culturing microorganisms in a suitable fermentation medium to produce the compounds or compositions according to the present invention is known to those skilled in the art. The 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 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, peptides, or amino acids. The process for producing the compounds according to the present invention can be carried out by batch culture, fed-batch culture, or continuous culture.

[0053] In one embodiment, the process comprises producing a compound according to the present invention or a composition comprising a compound according to the present invention and a microorganism capable of producing the compound according to the present invention. The composition comprising a compound according to the present invention and a microorganism capable of producing the compound according to the present invention can be a fermentation broth. The microorganism capable of producing the compound according to the present invention in the process of the present invention can also produce further active ingredients as defined herein above, such as cyclothiazomycin C, streptimidon and / or malonomycin.

[0054] The process according to the present invention may further comprise a step of recovering the compound or composition according to the present invention. The compound according to the present invention may be recovered by a suitable method known in the art, for example, 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.

[0055] The process for producing a compound according to the invention may further comprise the step of formulating this compound into a suitable formulation or composition as defined herein above.

[0056] In a further aspect, the present invention relates to a method for controlling or preventing an infestation of plants, plant propagation material, habitats and / or harvested food crops by treating the plants, plant propagation material, habitats and / or harvested food crops and / or applying an effective amount of a compound according to the invention or a composition according to the invention to the plants, parts thereof, plant propagation material, habitats and / or harvested food crops.

[0057] In methods for controlling or preventing plant investment, applying an effective amount of a compound of the invention includes applying a compound of the invention at 5 g to 5 kg (active ingredient (ai) per hectare (ha)), preferably 10 g to 1 kg ai / ha, most preferably 20 g to 600 g ai / ha.

[0058] When the compounds or compositions of the invention are used for seed treatment, a rate of compound of the invention of 0.001 to 50 g per kg of seeds, preferably 0.01 to 10 g per kg of seeds, is generally sufficient.

[0059] Preferably, the compounds of the invention or compositions comprising compounds according to the invention are applied either prophylactically, ie before disease onset, or therapeutically, ie after disease onset.

[0060] The phytopathogenic microorganisms affected by the compounds of the present invention are fungi and disease-carrying fungi as well as phytopathogenic bacteria and viruses. The phytopathogenic microorganisms in the method according to the present invention include the following fungi and disease-carrying fungi and phytopathogenic bacteria: Alternaria spp. such as Absidia corymbifera, Albugo candida, and Alternaria solani; Aphanomyces spp.; Ascochyta spp.; Aspergillus spp. such as Aspergillus flavus, Aspergillus fumigatus, Aspergillus nidulans, Aspergillus niger, and Aspergillus terrus; and Aureobasidium spp. such as Aureobasidium pullulans. Botryosphaeria spp. such as Bacillus subtilis, Blastomyces dermatitidis, Blumeria graminis, Blumeria graminis f.sp.tritici, Blumeria jaapii, Botryosphaeria dothidea, and Botryosphaeria obtusa, Botrytis spp. such as Botrytis cinerea, Bremia lactucae, and Cadophora gregata. Candida spp., such as Candida gregata, Candida albicans, Candida glabrata, Candida krusei, Candida lusitaniae, Candida parapsilosis, and Candida tropicalis.), Cephaloascus fragrans, Ceratocystis spp., Cercospora spp. such as Cercospora arachidicola, Cercospora beticola, Cercospora kikuchii, and Cercospora sojina, Cercosporidium personatum, Cladosporium spp., Clarireedia homoeocarpa, Clavibacter spp., and Claviceps purpurea. Colletotrichum spp. such as C. purpurea, Coccidioides immitis, Cochliobolus spp., Colletotrichum spp. such as C. dematium, C. lindemuthianum, C. musae, C. orbiculare, and C. truncatum, Corynespora cassiicola, Cryptococcus neoformans, Diaporthe spp., Dickeya zeae, zeae, Didymella spp., Drechslera spp., Elsinoe spp., Epidermophyton spp., Erwinia spp. such as Eremothecium gossypiim, Erwinia amylovora, and Erwinia carotovora.Erysiphe spp. such as E. cichoracearum, E. necator, Eutypa lata, Fusarium culmorum, Fusarium graminearum, Fusarium langsethiae, Fusarium moniliforme, Fusarium oxysporum, Fusarium poae, Fusarium proliferatum, and Fusarium pseudograminearum. Fusarium microorganisms such as Fusarium nearum, Fusarium sacchari, Fusarium sambucinum, Fusarium subglutinans, Fusarium solani, Fusarium sporotrichioides, Fusarium tricinctum, and Fusarium virguliforme. Gibberella spp., such as Gaeumannomyces graminis, G. avenacea, G. fujikuroi, G. intricans, G. moniliformis, and G. zeae.), Gloeodes pomigena, Gloeosporium musarum, Glomerella cingulata, Golovinomyces cichoracearum, Gymnosporangium juniperi-virginianae, Guignardia bidwellii, Gymnosporangium juniperi-virginianae, Helminthosporium spp., Hemileia spp., Histoplasma species such as Histoplasma capsulatum, spp.), Hyaloperonospora parasitica,. Kabatiella zeae, Laetisaria fuciformis, Leptographium lundbergii, Leveillula taurica, Lophodermium seditiosum, Microdochium majus, Microdochium nivale, Microsporum spp., Monilinia spp. such as M. fructicola, Monographella spp. such as M. nivalis, Mucor spp. Mycosphaerella spp., such as Mycosphaerella arachidis, Mycosphaerella fijiensis, Mycosphaerella graminicola, and Mycosphaerella pomi, Nakataea oryzae, Neopseudocercosporella spp., Oculimacula spp., Oncobasidium theobromaeon, Ophiostoma spp., Pantoea stewartia, and Paracoccidioides spp. Penicillium spp., such as Parastagonospora nodorum, Pectobacterium spp., Penicillium digitatum, and Penicillium italicum.), Petriellidium spp., Peronosclerospora spp. such as Peronosclerospora maydis, Peronosclerospora philippinensis, and Peronosclerospora sorghi, Peronosclerospora spp. such as Peronosclerospora destructor, Phaeosphaeria nodorum, Phakopsora pachyrhizi, Phellinus igniarus, Phialophora Phytophthora spp. such as Phlyctema vagabunda, Phoma spp., Phomopsis viticola, Phyllachora pomigena, Phyllosticta spp., Physoderma maydis, Phytophthora spp. such as P. capsica, Phytophthora infestans, and Plasmodiophora brassicae, Plasmopara spp. such as P. halstedii and P. viticola, Podosphaera spp., such as Plenodomus spp., Pleospora spp., and Podosphaera spp., including P. leucotricha.Pseudomonas spp. such as Polymyxa graminis, Polymyxa betae, Pseudocercospora fijiensis, Pseudocercosporella herpotrichoides, and Pseudomonas syringae, Pseudoperonospora spp. such as Pseudoperonospora cubensis and Pseudoperonospora humuli, Pseudoperonospora tracheiphila, and Pseudopyrenochaeta Puccinia spp. such as Puccinia lycopersici, Puccinia hordei, Puccinia recondita, Puccinia striiformis, and Puccinia triticina, Pyrenopeziza spp., Pyrenophora spp., Pyricularia spp. such as Pyricularia oryzae, Pythium spp. such as Pythium ultimum, Ralstonia solanacearum, Ramularia spp., and Rathayibacter spp.), Remotididymella destructiva, Rhizoctonia spp.), Rhizomucor pusillus, Rhizopus arrhizus, Rhynchosporium spp., Scedosporium spp. such as Robbsia andropogonis, Sarocladium oryzae, Scedosporium spp. such as S. apiospermum and S. prolificans, Sclerotinia spp. such as Schizothyrium pomi, Sclerophthora macrospora, and Sclerotinia sclerotiorum, spp.), Sclerotium spp., Septoria spp. such as Septoria nodorum, Septoria tritici, Setosphaeria turcica, Sphaerotheca macularis, Sphaerotheca fusca (Sphaerotheca fuliginea), Spiroplasma kunkelii, Sporothorix spp., Stagonospora nodorum, Stagonosporopsis cucurbitaceum cucurbitacearum), Stemphylium spp.), Stenocarpella macrospora, Stereum hirsutum, Streptomyces spp., Thanatephorus cucumeris, Thielaviopsis basicola, Tilletia spp., Tranzschelia discolor, Trichoderma spp. such as T. harzianum, T. pseudokoningii, and T. viride, Trichoderma spp., Trichophyton spp., and Typhula spp. spp.), Uncinula necator, Urocystis spp., Uromyces spp., Ustilago spp., Venturia spp. such as V. inaequalis, Verticillium spp., Xanthomonas spp. such as Wilsononomyces carpophilus or Xanthomonas oryzae and Xanthomonas campestris, Xylella spp., Zymoseptoria tritici.

[0061] The phytopathogenic microorganisms which have surprisingly been found to be affected by the compounds and / or compositions according to the invention are fungi, for example those belonging to the genera Zymoseptoria tritici, Puccinia recondita f.sp. tritici, Magnaporthe grisea or Blumeria graminis f.sp. tritici.

[0062] Any suitable plant, plant propagation material, habitat or food crop can be treated in the method according to the invention as defined herein.

[0063] As used herein, the term "habitat" means a field in or on which plants are growing, or a field in which seeds of cultivated plants are sown, or a field in whose soil seeds will be sown. It includes the soil, seeds and seedlings, and established vegetation.

[0064] The term "plant" refers to all physical parts of a plant, including seeds, seedlings, saplings, roots, tubers, trunks, stems, foliage, 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 also be protected by treating them in whole or in part by dipping before transplanting.

[0065] The term "plant propagation material" is understood to refer to reproductive parts of plants such as seeds and cuttings or vegetative bodies such as tubers (e.g. potatoes), roots, fruits, bulbs, rhizomes or plant parts that can be used for the propagation of plants.

[0066] 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; field crops such as sugar beet and fodder beet, 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; grasses such as dactylon, strawberry vine, bean grass, Japanese laurel wort, fescue, rye, St. Augustine grass, and lawn grass; aromatic herbs such as perennial and annual crops such as basil, borage, chives, coriander, lavender, lovage, peppermint, oregano, parsley, rosemary, 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 palm; ornamental plants such as flowers, shrubs and trees; other trees such as cocoa, coconut, olive and rubber; vegetables such as asparagus, eggplant, broccoli, cabbage, carrots, cucumber, garlic, lettuce, squash, melon, okra, onion, pepper, potato, pumpkin, rhubarb, spinach and tomato; and vines such as grapes. The term "plants" also includes woody crops or woody plants such as pines.

[0067] The term "useful plants" should also be understood to include useful plants that have been conferred resistance to herbicides or herbicide classes such as bromoxynil as a result of conventional breeding or genetic engineering methods (e.g., HPPD inhibitors, ALS inhibitors such as primisulfuron, prosulfuron and trifloxysulfuron, EPSPS (5-enol-pyrovyl-shikimate-3-phosphate synthase) inhibitors, GS (glutamine synthetase) inhibitors or PPO (protoporphyrinogen oxidase) inhibitors, etc.).

[0068] 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.

[0069] Preferably, the plant, plant propagation material or food crop is rice, wheat, corn, soybean or banana. References herein to soya include soybean.

[0070] Controlling or preventing in the method of the present invention means reducing the infestation of phytopathogenic microorganisms, especially fungi, to a level at which an improvement is observed.

[0071] A preferred method for controlling or preventing infestations of phytopathogenic microorganisms, particularly fungi, in crop plants is foliar application of the compounds and / or compositions according to the present invention. The frequency and rate of application depend on the risk of damage by the corresponding pathogen or insect. Alternatively, the compounds and / or compositions according to the present invention can be introduced into the plants through the soil via the roots (systemic application) by drenching the plant habitat with a liquid formulation or by applying a solid form of the compound to the soil, for example in granular form (soil application). In rice cultivation, such granules can be applied to flooded rice fields. The compounds according to the present invention can also be applied to seeds (coating) by either immersing the seeds or tubers in a liquid formulation of the fungicide or coating them with a solid formulation.

[0072] The compounds of the present invention can also be used as dressings to treat plant propagation material, such as seeds of fruits, tubers, or grains, or plant cuttings, to protect them from fungal infection as well as from plant pathogenic fungi occurring in the soil. The propagation material can be treated with the compounds and / or compositions according to the present invention before planting, for example, by dressing seeds before sowing. This specification also discloses such methods for treating plant propagation material and the plant propagation material thus treated.

[0073] In another aspect, the present invention also relates to the use of a compound according to the present invention or a composition comprising a compound according to the present invention as a pesticide, preferably as a bactericide or fungicide, and / or as a priming agent. The characteristics of the compound according to the present invention and the composition comprising a compound according to the present invention are as disclosed hereinabove. The present invention therefore relates to a method for using a compound according to the present invention as a pesticide, preferably as a bactericide or fungicide, and / or as a priming agent. [Example]

[0074] Example 1. Sources and Extraction of Compounds of the Invention 1.1. Streptomyces sp. fermentation Streptomyces species were obtained from the culture collections disclosed in Table 3. Streptomyces sp. Saigon 413 was isolated in Vietnam before 1961. 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.

[0075] This Streptomyces species microorganism was cultured in an Erlenmeyer flask using a liquid medium consisting of 10 parts casein hydrolysate, 40 parts glucose, 1.25 parts KHPO, 2 parts soytone, and 8 parts tryptone (all (g / l)). The culture was incubated at 28°C in an incubator with shaking at 150 rpm and an amplitude of 25 mm for 4 days.

[0076] 1.2.16S rDNA isolation and species identification Genomic DNA was isolated from Streptomyces sp. Saigon413 using the method described by Kutchma et al. (1998) Biotechniques 24:452-457. The 16S rRNA gene was amplified using 16S universal primers and sequenced using Sanger sequencing. The 16S rRNA is shown in SEQ ID NO:1.

[0077] The species of Streptomyces sp. Saigon413 was identified using barrnap v0.9 by comparing the 16S rRNA sequence (SEQ ID NO:1) with publicly available 16S rRNA sequences extracted using whole-genome sequence assemblies of Streptomyces species (based on the Genome Taxonomy Database GTDB (Parks, D.H., 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 of the 16S rRNA sequences between Streptomyces sp. Saigon413 and the publicly available Streptomyces chrestomyceticus NRRL-3672 was 99.87%, as determined using Muscle v3.8.31 and the R package Seqinr v4.2-16.

[0078] Whole-genome sequencing of Streptomyces sp. Saigon413 was completed using genomic DNA from both Pacific Biosciences and Illumina sequencing technologies. The genome was assembled using HFAP4 and corrected 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 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).

[0079] After assembling the genome from Streptomyces sp. Saigon413 and publicly available genomes, the average nucleotide identity (ANI) between Streptomyces sp. Saigon413 and closely related Streptomyces strains was calculated using fastANI (Jain, C., et al. (2018), Nature Communications, 9, 5114) (Table 3). The genome of Streptomyces sp. Saigon413 had the highest ANI to the publicly available genome of Streptomyces chrestomyceticus NRRL B-3672, at 96.9%.

[0080] Using the 16S RNA sequence identity and ANI score (%), it was also determined that strains CBS 596.66, CBS570.66, and DSM 41429 were Streptomyceticus strains, and not Streptomyces rimosis or Streptomyces paromomycinus strains as indicated by the depository.

[0081] 1.3. Purification of the Compound of the Present Invention The whole culture broth was centrifuged to obtain an aqueous extract and pellet. The aqueous extract was freeze-dried. This material was resuspended in a minimum amount of water and partitioned with ethyl acetate to remove lipophilic components. The aqueous suspension was retained, freeze-dried, resuspended in a minimum amount of water, and then applied to an activated charcoal column.

[0082] The column was washed with water and eluted with water:acetone (50:50).

[0083] Compounds of the present invention were further purified by hydrophilic interaction liquid chromatography (HILIC) using mass-guided fractionation and ELSD detection, e.g., a Waters XBridge Amide, 5 micron, 30 x 100 mm, with a gradient of acetonitrile and 10 mM ammonium acetate.

[0084] Example 2. Characterization of compounds of the present invention The compounds of the present invention were identified in fermentation broths purified according to the methods disclosed below.

[0085] The results in Table 3 show that several species of Streptomyces produce the compounds according to the present invention.

[0086] 2.1. Molecular composition and overall molecular mass The molecular composition and total molecular mass are C 53 H 90 N2O 44 and 1458.487 g, which was determined using MS-MS and NMR spectroscopy as disclosed in paragraphs 2.3 and 2.4.

[0087] 2.2. Solubility The solubility of the compounds of the present invention was determined.

[0088] The compounds of the present invention have a solubility in water at pH 7.01 of greater than 10,000 ppm and a solubility in DMSO of greater than 9,772 ppm.

[0089] 2.3. MS-MS analysis and liquid chromatography Spectra were recorded on a Thermo Scientific Orbitrap ID-X Tribrid mass spectrometer equipped with an OptaMax NG heated electrospray ion source (spray voltage: static, ionization voltage (Polarity Ion) (V): 3400 (positive ion mode) and 2400 (negative ion mode), sheath gas (Arb): 40, auxiliary gas (Arb): 5, sweep gas (Arb): 1, ion inlet temperature: 350°C, vaporizer temperature: 350°C). Scanning parameters were as follows: Experiment 1: MS OT (Orbitrap resolution: 60,000, scan range (m / z): 200-2000, RF lens (%): 60, AGC target: Standard, Maximum 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 method: CID, CID collision energy (%): 30, detector type: Orbitrap, Orbitrap resolution: 30,000, RF lens (%): 60, polarity: negative). The mass spectrometer was connected to a Vanquish Flex UHPLC (Thermo Scientific) 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 the following: Thermo Scientific Hypercarb™ Porous Graphitic Carbon column, 5 μm, 4.6 × 50 mm, product number 35005-054630; temperature: 40 °C; DAD wavelength range: 250–260 nm; solvent gradient: Solvent A: 0.1% formic acid in HO; Solvent B: 0.1% formic acid in CHCN; 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.

[0090] 2.4.NMR spectroscopy NMR spectra were recorded on a 5 mm Bruker ( 1 H / 19 F) / 13 C / 15 Spectra were recorded using a Bruker AVIII 600 NMR spectrometer equipped with an N TCI cryoprobe using standard Bruker pulse sequences. Samples were dissolved in DO and spectra were recorded at 300°K. 1 For H, 2.225 ppm of acetone; 13In the case of C, 31.07 ppm of acetone was used as the reference. Figures 1 to 4 show the NMR spectra of the compounds of the present invention.

[0091] 1 H- 13 The C single-bond correlation spectrum contains peaks corresponding to one methyl (CH3) group and 40 methine (CH) groups (listed in Table 1) and nine methylene (CH2) groups (listed in Table 2).

[0092] In addition, 1D 13 The C spectrum contains three signals from quaternary carbons at 104.7, 159.3, and 175.2 ppm (±0.1).

[0093] [Table 1]

[0094] [Table 2]

[0095] [Table 3]

[0096] CBS Westerdijk Fungal Diversity Institute, Uppsalalaan 8, 3584 CT, Utrecht, The Netherlands DSMZ German Collection of Microorganisms and Cell Cultures: Inhoffenstrasse 7B, 38124 Braunschweig, Germany. ARS ARS Culture Collection (NRRL),1815 N.University Street,Peoria,IL 61604,USA

[0097] Example 3. Plant activity of compounds according to the present invention 3.1. Test using leaf discs or leaf segments in well plates Leaf discs or segments of various plant species were excised from greenhouse-grown plants. The excised leaf discs or segments were placed on plain agar medium in a multiwell plate (24-well format). The test solution was sprayed onto the leaf discs before (preventive) or after (therapeutic) inoculation. The test compounds were prepared as aqueous solutions (up to 10 mg / ml) and diluted to the appropriate concentration with 0.025% Tween 20 immediately before spraying. The inoculated leaf discs or segments were incubated under the specified conditions (temperature, relative humidity, light, etc.) according to each test method. Depending on the pathogen response system, the severity of disease was evaluated once, 3 to 9 days after inoculation. The disease control rate was then calculated compared to untreated control leaf discs or segments.

[0098] Puccinia recondita f.sp.tritici / Wheat / Leaf disc (preventive) (leaf rust) Wheat leaf segments of the Kanzler variety were placed on agar medium in a multiwell plate (24-well format) and sprayed with formulated test compounds diluted in water. One day after application, the leaf discs were inoculated with a fungal spore suspension. The inoculated leaf segments were incubated in a constant temperature room at 19°C and 75% RH under a 12-hour light / 12-hour dark photoperiod. When the disease damage on untreated control leaf segments reached an appropriate level (7-9 days after application), the activity of the compounds was evaluated as the disease control rate compared to untreated leaves.

[0099] Puccinia recondita f.sp.tritici / wheat / leaf disc (therapeutic) (leaf rust) Leaf segments of wheat cultivar Kanzler were placed on agar medium in multiwell plates (24-well format). The leaf segments were inoculated with a fungal spore suspension. The plates were stored in the dark at 19°C and 75% RH. The formulated test compounds were diluted with water and applied one day after inoculation. The leaf segments were incubated in a constant temperature room at 19°C and 75% RH under a 12-hour light / 12-hour dark photoperiod. When the disease damage on untreated control leaf segments reached an appropriate level (6-8 days after application), the activity of the compounds was evaluated as the disease control rate compared to the untreated control.

[0100] Magnaporthe grisea (Pyricularia oryzae) / Rice / Leaf disc (preventive) (Rice blast) Leaf segments of Ballila rice were placed on agar medium in a 24-well multiwell plate and sprayed with formulated test compounds diluted in water. Two days after application, the leaf segments were inoculated with a fungal spore suspension. The inoculated leaf segments were incubated in a constant temperature room at 22°C and 80% RH under a 24-hour dark followed by a 12-hour light / 12-hour dark photoperiod. When the disease damage on untreated control leaf segments reached an appropriate level (5-7 days after application), the activity of the compounds was evaluated as the disease control rate compared to the untreated control.

[0101] [Table 4]

[0102] The results in Table 4 show that the compounds of the present invention inhibit fungal growth of several fungal species at concentrations of 22 ppm and above in leaf disc and leaf segment tests. The compounds of the present invention provide both preventative (application of compound followed by pathogen infection) and curative (pathogen infection followed by compound application) activity for the control of the rust pathogen Puccinia recondita.

[0103] 3.2. Leaf painting assay on wheat seedlings Wheat seedlings of either variety Riband (for Zymoseptoria tritici testing) or variety Arina (for Puccinia recondite testing) were grown in the greenhouse for 14 days after sowing. At this time, such seedlings typically have a fully expanded first leaf (designated L1), a fully expanded second leaf (L2), and a partially expanded third leaf (L3). Two dots were made on the second leaf using a permanent marker to separate it into three approximately equal-sized segments: segment A (base), segment B (middle), and segment C (tip). Stock solutions of the compound of formula (I) in water and benzovindiflupyr in DMSO were prepared at a concentration of 10,000 ppm. The stock solution was then further diluted with water containing Tween 20 to a final concentration of ≥200 ppm test compound, 0.05% Tween 20, and 2% DMSO (only for compounds using DMSO stock solutions). The diluted compound was applied to the center of the L2 using a conventional cotton swab. The cotton swab soaked in the diluted compound was rubbed several times between two marks on the adaxial surface of the leaf. One day later, the entire plant was inoculated with a fungal spore suspension using a paintbrush. The spore suspension was applied until runoff.

[0104] For Zymoseptoria tritici (preventative infection), test plants were inoculated by spraying a spore suspension (1.5 million spores per ml of water containing 0.01% Tween 20) one day after application. After 4 days of incubation 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 when the disease appeared on untreated control plants to the appropriate extent (16–19 days after application).

[0105] For Puccinia recondita infection (preventive): One day after application, test plants were inoculated by spraying with a spore suspension (80,000 spores per ml of water containing 0.1% Tween 20). After one day of incubation at 20°C and 95% RH, the inoculated test plants were kept in a greenhouse at 20°C (day / night) and 60% RH. When the disease on untreated control plants reached the appropriate severity (9-12 days after infection), the percentage of leaf area covered by disease was visually assessed.

[0106] The three sections of the leaf were evaluated individually.

[0107] [Table 5]

[0108] Foliar application studies have shown that compounds of the present invention are active against the growth of the fungi Puccinia recondite and Zymoseptoria tritici. The compounds of the present invention demonstrated control not only in the treated area (center section) but also above the treated area (tip). The activity observed in plants was independent of the surfactant Tween 20 or the solvent DMSO, which were also present at low concentrations when the compounds were tested. The translocation of activity to acropetal growth sites was similar to that of benzovindiflupyr, an SDHI fungicide / fungicide well known to exhibit such translocation.

[0109] 3.3. Powdery mildew test on wheat One-week-old wheat leaves were used for the experiment. A 5 cm section from the leaf tip was excised underwater and placed in a cuvette containing 100 μL of test solution. After 1.5 hours, the leaves were removed and placed on a paper towel for 1 hour. The leaves were then placed on plain agar plates (1%) and stored in a light room (18°C, 8 hours of light). One day before inoculation of the excised leaf segments, wheat plants infected with powdery mildew (Blumeria graminis f.sp. tritici) were gently shaken to remove old conidia and allow new conidia to be produced. To inoculate the excised leaf segments, the plates containing the leaf segments were placed on the ground and covered with an infection hood. A powdery mildew-infected wheat plant was inserted through the hood's outlet and gently shaken to distribute the spores evenly. Forty-eight hours after inoculation, the leaves were transferred to reaction tubes containing 10 mL of 80% ethanol. Ten days after the leaf tissue had discolored, fungal spores were stained with ddH2O containing 10% (v / v) ink and 25% (v / v) acetic acid. The percentage of successful penetration (papilla formation) was assessed at 100 spore-plant cell interaction sites using a light microscope. This test was repeated four times. The values ​​shown in Table 4 are the average of the four experiments.

[0110] The compound INA (2,6-dichloro-isonicotinic acid, CAS: 5398-44-7) is a synthetic salicylic acid analogue and was included as a reference substance for priming activity (Krauss et al., 1992, Plant Journal 2, 655-60).

[0111] Test sample AEF1 is a sample enriched in the compound of formula I. It corresponds to the fraction eluted from the activated carbon column that is enriched in the compound of formula I.

[0112] [Table 6]

[0113] The results in Table 6 demonstrate that wheat pretreated with a compound of the present invention or a compound-enriched fraction of the present invention reduced haustoria formation by the powdery mildew fungus Blumeria graminis f.sp. tritici. This effect was observed at concentrations of 25 μM and above. The compound-enriched fraction and the purified compound were equally effective at the highest ratios tested.

[0114] 3.4. Reactive oxygen species (ROS) explosion test 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 floated on the test solution. The plates were stored at room temperature for 24 h. The next day, the solution was replaced with 50 μL of ddH2O, and the leaf discs were left to regenerate for at least 1 h in the dark at room temperature. Meanwhile, the appropriate master mix, with or without the elicitor flg22 (see below), was freshly prepared in a 5 mL black reaction tube. After regeneration, 50 μL of the master mix corresponding to the well containing the leaf disc was added. Luminescence was then recorded for 40 min using 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 reference substance for priming activity (Kauss et al., 1992). Master Mix-flg22: 4.98 mL ddHO, 10 μL HRP (10 mg / mL), 10 μL L-012 (20 mM) Master mix + flg22: 4.979 mL ddH2O, 10 μL HRP (10 mg / mL), 10 μL L-012 (20 mM), 1 μL flg22 (10 μM) Abbreviations: flg22 (flagellin peptide consisting of 22 amino acids, Eurogentec catalogue number AS-62633); HRP (horseradish peroxidase), L-012 sodium salt (CAS#: 143556-24-5) was used.

[0115] Test sample AEF1 is a sample enriched in the compound of formula (I), which corresponds to the fraction eluted from the activated carbon column that contains the compound of formula I most.

[0116] [Table 7]

[0117] The results in Table 7 show that wheat plants pretreated with the compound-enriched fractions at concentrations ranging from 1 ppm to 100 ppm exhibited a more than two-fold increase in ROS production induced by peptide flg22, a response comparable to or stronger than that observed by treatment with the well-known priming agent INA (2,6-dichloro-isonicotinic acid) (Krauss et al., 1992, Plant Journal 2:655-60).

[0118] Example 4. Bactericidal and fungicidal activity of a mixture of the oligosaccharide compound according to the present invention and malonomycin in the leaf disc test Stock solutions for oligosaccharide compounds according to the invention were prepared in water with the addition of 0.025% Tween® 20. Malonomycin was prepared according to Law et al., 2018 (Nature Catalysis | VOL 1 | DECEMBER 2018 | 977-984). Stock solutions of malonomycin were prepared in water with the addition of 0.025% Tween® 20.

[0119] A test to determine the efficacy of mixtures of compounds of formula (I) in combination with malonomycin for control of pathogenic fungi in a leaf disc test was designed in two 24-well plates.

[0120] Tables 8 and 9: Overview of 24-well test plate (1) and test plate (2), including the compound application concentration per well. The upper number in the cell indicates the concentration (ppm) of the oligosaccharide compound of the present invention, and the lower number indicates the concentration (ppm) of malonomycin. The vertical column (1) contains the dilution series of the compound of formula (I), and the horizontal column (2-D) contains the dilution series of malonomycin. Well 2-D-(1) represents the untreated control. This design was applied to a test involving Puccinia recondita (EPPO code: PUCCRE), with application at preventive and therapeutic time points.

[0121] [Table 8]

[0122] [Table 9]

[0123] [Table 10]

[0124] Tables 11 and 12: Overview of 24-well test plate (3) and test plate (4), including the compound concentrations per well. The upper number in the cell indicates the concentration (ppm) of the oligosaccharide compound of the present invention, and the lower number indicates the concentration (ppm) of malonomycin. Column (1) contains a dilution series of the compound of formula (I), and column (4-D) contains a dilution series of malonomycin. Well 4-D-(1) represents an untreated control. This design was applied to a test involving Blumeria graminis f.sp. tritici (EPPO code: ERYSGT) sprayed preventively and a test involving Parastagonospora nodorum (EPPO code: LEPTNO) sprayed preventively.

[0125] [Table 11]

[0126] [Table 12]

[0127] [Table 13]

[0128] A first set of master plates is prepared by diluting a 1x stock spray solution of the compound of formula (I) with water to the concentrations shown in Tables 8, 9, 11, and 12, respectively. Each well contains 0.025% Tween® 20. Accordingly, a second set of master plates is prepared by diluting a 1x stock of malonomycin with water. Each well of the second set contains 0.025% Tween® 20. Leaf sections placed on agar medium in a 24-well plate are sprayed with 8 μl of solution from the master plate containing the compound of formula (I), allowed to dry, and then sprayed with 8 μl of solution from the master plate containing malonomycin after 2 hours. After the second spray has dried, the leaf sections are infected with fungal spores to obtrain the preventive application time. Alternatively, leaf sections infected one day before spraying with the compound are used. This represents the therapeutic application time point. In addition, several plates were prepared in which leaf segments were sprayed at 2x in the absence of test compound (Tween® 20 only) to serve as untreated controls. For each leaf segment, the leaf area covered with disease symptoms was assessed. The loss of leaf coverage relative to the untreated control was calculated. The efficacy of the mixtures was tested in duplicate against different fungal species. The reported efficacy values ​​are the average of two replicates.

[0129] Preventive time-point spraying against Puccinia recondita (EPPO code: PUCCRE). Wheat (Kanzler) leaf segments are placed on agar medium in a 24-well multi-well plate and sprayed with the test solution (8 μl per well). After drying, the leaf discs are inoculated with a fungal spore suspension. After appropriate incubation, the activity of the compounds is evaluated as preventive fungicidal and fungicidal activity 8 dpi (days post-inoculation).

[0130] Therapeutic application for Puccinia recondita (EPPO code: PUCCRE). Wheat (Kanzler variety) leaf segments are placed on agar medium in a 24-well multi-well plate. The leaf discs are then inoculated with a fungal spore suspension. One day after inoculation, the test solution is sprayed (8 μl per well). After appropriate incubation, the activity of the compounds is evaluated 8 days post-inoculation (dpi) as therapeutic fungicidal and fungicidal activity.

[0131] Preventive spraying against Blumeria graminis f.sp.tritici (EPPO code: ERYSGT). Wheat (Kanzler) leaf segments are placed on agar medium in a 24-well multi-well plate and sprayed with the test solution (8 μl per well). After drying, the leaf discs are inoculated with a fungal spore suspension. After appropriate incubation, the activity of the compounds is evaluated as preventive fungicidal and fungicidal activity 7 dpi (days post-inoculation).

[0132] Parastagonospora nodorum (EPPO code: LEPTNO) Wheat (Kanzler) leaf segments are placed on agar medium in a 24-well multi-well plate and sprayed with the test solution (8 μl per well). After drying, the leaf discs are inoculated with a fungal spore suspension. After appropriate incubation, the activity of the compounds is evaluated as preventive fungicidal and fungicidal activity 4 dpi (days post-inoculation).

[0133] result The results of fungicidal experiments using mixtures of the compounds according to the invention described above with malonomycin are shown in Tables 14 to 19.

[0134] [Table 14]

[0135] For each test condition in Table 14 where efficacy was 50% or greater (effective mixture), the respective mixture ratios could be assigned from Table 10. It was found that effective mixtures for the control (preventive) of Puccinia recondita were between 64:1 and 1:90.

[0136] [Table 15]

[0137] [Table 16]

[0138] For each test condition in Table 16 where efficacy was 50% or greater (effective mixtures), the respective mixture ratios could be assigned from Table 10. Mixtures effective for the control (curative) of Puccinia recondita were found to be between 64:1 and 1:270.

[0139] [Table 17]

[0140] [Table 18]

[0141] For each test condition in Table 18 where efficacy was 50% or greater (effective mixture), the respective mixture ratios could be assigned from Table 11. It was found that effective mixtures for the control (preventive) of Blumeria graminis f.sp. tritici were between 64:1 and 1:4.

[0142] [Table 19]

[0143] For each test condition in Table 19 where efficacy was 50% or greater (effective mixture), the respective mixture ratios could be assigned from Table 11. It was found that effective mixtures for the control (preventive) of Parastagonospora nodorum were between 64:1 and 1:9.

[0144] conclusion Mixtures of the oligosaccharide compounds of the present invention and malonomycin control a variety of pathogenic fungi. Even when the mixture ratio of the two compounds varied widely, foliar application still resulted in greater than 50% control of fungal growth. Examples of ratios ranging from 64:1 to 1:270 for several fungal species are provided, but examples of ratios between the stated ratios (ratio of malonomycin compound to oligosaccharide compound of the present invention) are also included. Surprisingly, some mixtures exhibited efficacy that exceeded that predicted based on calculations from Colby, suggesting that foliar application of mixtures of the oligosaccharide compounds of the present invention and malonomycin exhibits a synergistic effect in controlling pathogenic fungi. This surprising synergistic effect was confirmed in tests controlling Puccinia recondita.

Claims

1. C 53 H 90 N 2 O 44 and further characterized by an NMR spectrum set forth in Table 1 and Table 2, or a salt thereof.

2. Structural formula (I): 【Chemical 1】 Formula (I) 2. The compound or salt thereof, optionally according to claim 1, having the formula:

3. 3. A composition comprising a compound according to claim 1 or 2 and a microorganism capable of producing the compound according to claim 1 or 2.

4. 10. A composition comprising a compound according to claim 1 or 2 or a composition according to claim 3 or 4, further comprising an adjuvant.

5. The compound according to claim 1 or 2 or the composition according to any one of claims 3 to 5, wherein the compound or the composition has bactericidal and fungicidal activity.

6. The composition according to any one of claims 3 to 5, further comprising at least one additional ingredient with pesticidal activity and / or at least one plant growth regulator.

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

8. 8. A process for producing a compound according to claim 1 or 2 or a composition according to any one of claims 3 to 7, comprising culturing a microorganism in a suitable fermentation medium under conditions that allow the production of said compound.

9. The microorganism is a Streptomyces sp., preferably Streptomyces chrestromyceticus, Streptomyces rimosus, Streptomyces paromomycinus or Streptomyces albofaciens, preferably a Streptomyces sp. having a 16S RNA sequence having at least 98% identity to SEQ ID NO: 1, and preferably the microorganism is Streptomyces sp. deposited at the Westerdijk Institute under accession number CBS149411. The composition according to any one of claims 3 to 7 or the process according to claim 8, wherein the hydroxybenzoate is hydroxybenzoate (sp.) Saigon 413.

10. 10. A method for controlling or preventing infestation of phytopathogenic microorganisms in plants, plant propagation material, habitats and / or harvested food crops by treating said plants, plant propagation material, habitats and / or harvested food crops, comprising applying to said plants, parts thereof, said plant propagation material, habitats and / or harvested food crops an effective amount of a compound according to claim 1 or 2 or a composition according to any one of claims 3 to 7 or 9.

11. 11. The method of claim 10, wherein the effective amount comprises 5 g to 5 kg per hectare of the compound of claim 1 or 2.

12. 11. The method of claim 10, wherein the plant propagation material is a seed and the effective amount comprises 0.001 to 50 g of the compound of claim 1 or 2 per kg of seed.

13. The plant pathogenic microorganism is a fungus, preferably a fungus belonging to the genus Zymoseptoria, Puccinia, Magnaporthe, Blumeria or Parastagonospora, preferably Zymoseptoria tritici, Puccinia recondite, Puccinia recondita forma specialis tritici, Magnaporthe grisea, ...ita forma specialis tritici, Magnaporthe grisea, Zymoseptoria tritici, Puccinia recondita forma specialis tritici, Magnaporthe grisea, Zymoseptoria tritici, Puccinia recondita forma specialis tritici, Magnaporthe grisea, Zymoseptoria tritici, Puccinia recondita forma specialis tritici, Magnaporthe grisea, Zymoseptoria The method according to any one of claims 10 to 12, wherein the fungus is a fungus belonging to Blumeria graminis f.sp.tritici, Blumeria graminis f.sp.tritici, or Parastagonospora nodorum.

14. The method according to any one of claims 10 to 13, wherein the plant is rice, wheat, corn, soybean or banana.

15. Use of a compound according to claim 1 or 2 or a composition according to any one of claims 3 to 7 as a pesticide, preferably as a bactericide / fungicide and / or as a priming agent.

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