Fungicidal compounds

Abstract

The present invention relates to a compound of formula (I) and a composition containing the compound, a method for producing the compound and a method for preventing or controlling fungi in plants and agriculture using the compound and the composition. # imgabs0 #

Description

[0001] The present invention relates to novel compounds having fungicidal activity. The present invention also relates to compositions comprising the compounds, methods for preparing the compounds and the use of the compounds or the compositions in agriculture or horticulture for preventing or controlling fungal infestation of plants, harvested food crops, seeds or non-living materials. Background Art

[0002] Fungicides are widely used in agriculture to protect plants from damage caused by fungi. Fungicides can come from chemical or biological sources. Biofungicides can, for example, come from microbial sources or can be plant extracts. Known microorganisms that produce antifungal antibiotics are, for example, actinomycetes, such as Streptomyces species (Streptomyces sp.). A very well-known species is Natal Streptomyces (Streptomyces natalensis), which produces the antifungal compound natamycin for food and crop protection. In US 5,356,624, a strain of Streptomyces rimosus is disclosed, which is found to be active against several wood-degrading fungi. In WO 2022 / 038180, new Streptomyces species are disclosed, which produce several known antifungal compounds, such as streptoglutarimide, natamycin (pimaricin) or white fungi. Extracts of these bacterial strains were found to be active against well-known plant pests such as Fusarium graminearum, Zymoseptoria tritici and Puccinia striiformis.

[0003] The development of fungal resistance to fungicides, coupled with government regulation and societal pressure, has led to a continued search for new compounds with fungicidal activity from biological sources. Summary of the invention

[0004] The present invention relates to a compound comprising 53 H 90 N 2 O 44 The molecular formula is preferably 1458.487 g, the compound further characterized by the NMR spectra listed in Table 1 and Table 2, or a salt thereof.

[0005] The present invention further relates to a compound having structural formula (I):

[0006]

[0007] or a salt thereof.

[0008] In a second aspect, the present invention relates to a composition comprising a compound according to the invention and a microorganism capable of producing a compound according to the invention.

[0009] In a third aspect, the invention relates to a method for producing a compound or composition according to the invention, the method comprising culturing a microorganism in a suitable fermentation medium under conditions allowing the production of the compound.

[0010] In a fourth aspect, the present invention relates to a method for controlling or preventing infestation of plants by phytopathogenic microorganisms, wherein an effective amount of a compound or composition according to the present invention is applied to the plant, its part or the locus thereof.

[0011] 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 pesticidal agent, preferably a fungicide and / or as an initiator. According to this aspect of the present invention, the use does not include a method of treating the human or animal body by surgery or therapy. DETAILED DESCRIPTION

[0012] According to the present invention, there is provided a compound comprising 53 H 90 N 2 O 44 The compound is further characterized by the NMR spectra listed in Table 1 and Table 2, or a salt thereof. 53 H 90 N 2 O 44 The compound has a molecular weight of 1458.487 g. Preferably, the compound according to the invention has a solubility of more than >10'000 ppm at pH 7 and / or a solubility in DMSO of more than >9772 ppm.

[0013] Preferably, the compound according to the invention is an isolated compound. The wording "isolated" with respect to a compound means that the compound has been separated from its natural environment. Preferably, the compound according to the invention is an oligosaccharide.

[0014] The compound according to the present invention may be a compound having the structural formula (I):

[0015]

[0016] or a salt thereof.

[0017] In one aspect, the present invention relates to a composition comprising a compound according to the present invention and a microorganism capable of producing a compound disclosed herein. The composition comprising a compound according to the present invention and a microorganism capable of producing a compound according to the present invention is applied to a plant or part thereof to treat or protect the plant against diseases caused by plant pathogenic microorganisms (e.g., fungi, bacteria or viruses).

[0018] Surprisingly, it has now been found that the compounds and / or compositions according to the invention have a favourable level of biological activity for the treatment or protection of plants against diseases caused by infection with phytopathogenic microorganisms, such as fungi, bacteria or viruses. Surprisingly, the compounds and / or compositions according to the invention have a favourable fungicidal activity against various phytopathogenic fungi, such as Puccinia recondita f.sp.tritici, Magnaporthe grisea, Blumeria graminis f.sp.tritici and / or Parastagonospora nodorum.

[0019] The compositions according to the invention are non-naturally occurring compositions.

[0020] Preferably, the compounds and / or compositions according to the invention have fungicidal activity. Therefore, the compounds and / or compositions according to the invention are preferably fungicides.

[0021] As used herein, the term "compound having fungicidal activity" or "fungicide" means a compound that controls, modifies, or prevents the growth of fungi. The term "fungicidally effective amount" when used means an amount of such a compound or combination of such compounds that is capable of producing an effect on the growth of fungi. Controlling or modifying effects include all deviations from natural development, such as killing, retardation, etc., and prevention includes the formation of a barrier or other defense in or on a plant to prevent fungal infection.

[0022] Preferably, the microorganism capable of producing the compound according to the invention (e.g. in the composition disclosed herein) is a bacterium of the genus Streptomyces, preferably the bacterium is Streptomyces chrestomyceticus, Streptomyces chrestomyceticus or Streptomyces albofaciens, Streptomyces paromomycinus or Streptomyces monomicini. Preferably, the composition comprises Streptomyces species Saigon 413 deposited in the Fungal Culture Collection under the accession number CBS149411. Preferably, the microorganism is a Streptomyces species, preferably a Streptomyces species Saigon 413 deposited in the Fungal Culture Collection under the accession number CBS149411, wherein the Streptomyces species has a 16S RNA sequence that is 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% identical to SEQ ID NO:1.

[0023] Preferably, the microorganism capable of producing the compound according to the present invention comprises a genomic sequence that is at least 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identical or 100% identical to the whole genome of Streptomyces sp. Saigon 413 (deposited at the Fungal Culture Collection under the accession number CBS149411).

[0024] As used herein, the terms "percent identity" and "percent identical" refer to the relatedness of two or more nucleotide or amino acid sequences, which can be calculated by: (i) comparing two optimally aligned sequences over a comparison window, (ii) determining the number of positions at which the same nucleic acid base (for nucleotide sequences) or amino acid residue (for proteins) is present in the two sequences to produce the number of matched positions, (iii) dividing the number of matched positions by the total number of positions in the comparison window, and then (iv) multiplying the quotient by 100% to obtain the percent identity. If "percent identity" is calculated relative to a reference sequence without specifying a particular comparison window, the percent identity is determined by dividing the number of matched positions over the alignment region by the total length of the reference sequence. Therefore, for the purposes of the present invention, when two sequences (query sequence and subject sequence) are optimally aligned (allowing gaps in the alignment), the "percent identity" of the query sequence is equal to the number of identical positions between the two sequences divided by the total number of positions in the query sequence over its length (or comparison window), and then multiplied by 100%.

[0025] The present invention also discloses a microorganism, which is Streptomyces species Saigon 413 deposited in the Fungal Culture Collection Center with the deposit number CBS149411.

[0026] Surprisingly, it was found that the Streptomyces sp. Saigon 413 deposited with the Fungal Culture Collection under the accession number CBS 149411 has advantageous properties compared to the Streptomyces sp. known in the art.

[0027] Compound of the present invention or the composition comprising compound of the present invention can be used for agricultural sector and relevant use field, for example as the active component of controlling plant harmful organisms.Compound according to the present invention is characterised in that there is excellent activity under low application rate (for example 5 to 300ppm, for example 10 to 250ppm, for example 20 to 200ppm), plant tolerance is good and environmentally safe.It has very useful therapeutic, preventive and systemic characteristics and can be used for protecting many plants.Compound of the present invention can be used for suppressing or destroying harmful organisms occurring on the plant or plant part (fruit, flower, leaf, stem, tuber, root) of different plant crops, and simultaneously also protects those plant parts grown later.

[0028] The compound according to the invention or the composition comprising the compound according to the invention can be used as is or formulated with an adjuvant (preferably an agriculturally acceptable adjuvant) for use. Formulations known in the art are, for example, emulsifiable concentrates, coatable pastes, sprayable or dilutable solutions or suspensions, powders, dusts, granules and encapsulates.

[0029] Therefore, in one embodiment, the composition disclosed herein comprising the compound according to the present invention further comprises an adjuvant. Preferably, the adjuvant is an agriculturally acceptable adjuvant.

[0030] 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, wetting agents, repellents, attractants, feeding stimulants, extenders, bactericides, antifreeze agents, defoamers, colorants, tackifiers and adhesives.

[0031] Suitable solvents and liquid vehicles 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 vehicle known in the art. The solvent or liquid vehicle may be water or DMSO.

[0032] Suitable solid carriers include, for example, talc, titanium dioxide, pyrophyllite clay, silica, attapulgite clay, kieselguhr, chalk, diatomaxeous earth, lime, calcium carbonate, bentonite, Fuller's earth, cottonseed hulls, wheat flour, soybean flour, pumice, wood flour, walnut shell flour and lignin.

[0033] Adjuvants can be surfactants, crystallization inhibitors, viscosity regulators, suspending agents, spray droplet regulators, pigments, antioxidants, foaming agents, defoamers, opacifiers, compatibilizers, masking agents, neutralizing and buffering agents, corrosion inhibitors, dyes, flavor enhancers, spreading agents, penetration aids, micronutrients, emollients, lubricants and fixatives.

[0034] The compositions disclosed herein are preferably agriculturally acceptable compositions.

[0035] The composition of the present invention can be used in any conventional form, for example, in the form of a twin pack, a dry seed treatment powder (DS), a seed treatment emulsion (ES), a seed treatment flowable concentrate (FS), a seed treatment solution (LS), a seed treatment water dispersible powder (WS), a seed treatment capsule suspension (CF), a seed treatment gel (GF), an emulsion concentrate (EC), a suspension concentrate (SC), a suspoemulsion (SE), a capsule suspension (CS), a water dispersible granule (WG), an emulsifiable granule (EG), an oil-in-water emulsion (EO), an oil-in-water emulsion (EW), a microemulsion (ME), an oil dispersant (OD), an oil-miscible flowable concentrate (OF), an oil-soluble liquid (OL), a soluble concentrate (SL), an ultra low volume suspension (SU), an ultra low volume liquid (UL), a master batch (TK), a dispersible concentrate (DC), a wettable powder (WP) or any technically feasible formulation in combination with an agriculturally acceptable adjuvant.

[0036] Such compositions can be produced in a conventional manner, for example by mixing the active ingredient with appropriate formulation inerts (diluents, solvents, fillers and optionally other formulation ingredients, such as surfactants, biocides, antifreeze agents, adhesives, thickeners and compounds providing auxiliary effects). If it is desired to obtain a lasting effect, conventional slow-release formulations can also be used. In particular, formulations to be 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, can contain surfactants such as wetting agents and dispersants and other compounds providing auxiliary effects, such as condensation products of formaldehyde and naphthalene sulfonates, alkyl aryl sulfonates, lignin sulfonates, fatty alkyl sulfates, and ethoxylated alkylphenols and ethoxylated fatty alcohols.

[0037] The composition comprising the compound according to the present invention typically comprises 0.5w / w% to 95w / w% of active ingredient, such as 1w / w% to 90w / w%, such as 2w / w% to 80w / w%, such as 5w / w% to 60w / w%. The compound according to the present invention can be the only active ingredient in the composition disclosed herein. In one embodiment, the composition comprising the compound of the present invention further comprises at least one other active ingredient. The active ingredient defined herein is a composition with fungicidal and / or insecticidal and / or herbicidal activity or with activity as a plant growth regulator. The compound of the present invention or the composition disclosed herein can be mixed with one or more other ingredients (such as fungicides, insecticides, herbicides, bactericides, acaricides, nematicides) with pesticidal activity known in the art, and / or the other ingredients include plant growth regulators in appropriate cases. One or more other ingredients with pesticidal activity can be from biological or chemical sources. The pesticides mentioned herein using their common names are known, for example, from “The Pesticide Manual”, 19th edition, British Crop Protection Council 2021.

[0038] Additional ingredients with pesticidal activity (e.g., fungicidal activity) can promote unexpected synergistic activity. Therefore, compositions comprising compounds according to the invention and additional active ingredients (e.g., malonomicin) can show synergy. As long as the effect of the active ingredient combination is greater than the sum of the effects of the individual components, there is synergy. For a given active ingredient combination, the expected effect E is subject to the so-called COLBY formula and can be calculated as follows (COLBY, SR "Calculating synergistic and antagonistic responses of herbicide combination". Weeds [weeds], Vol. 15, pp. 20-22; 1967):

[0039] ppm = milligrams of active ingredient (=ai) per liter of spray mixture

[0040] X = effect of active ingredient A) in %, p ppm of active ingredient used

[0041] Y = action in % of active ingredient B), q ppm of active ingredient used.

[0042] According to COLBY, using p+q ppm of active ingredient, the expected (additive) effect of active ingredient A)+B) is:

[0043]

[0044] If the effect actually observed (O) is greater than the expected effect (E), then the effect of the combination is superadditive, i.e. there is a synergistic effect. In mathematical terms, synergism corresponds to a positive value of the difference (OE). In the case of a superposition of completely complementary activities (expected activity), the difference (OE) is zero. A negative value of the difference (OE) indicates a loss of activity compared to the expected activity.

[0045] Other ingredients with pesticidal activity and / or as plant growth regulators can be combined with the composition of the present invention and used in the method of the present invention and applied simultaneously or sequentially with the composition of the present invention. When applied simultaneously, these other ingredients can be formulated or mixed in, for example, a spray tank together with the composition of the present invention. As an alternative to directly mixing these other ingredients with pesticidal activity, these components can be used in separate fungicidal, insecticidal or weeding applications as a part of a fungus, insect or grass control program carried out in part or throughout the growing season.

[0046] At least one additional component having pesticidal activity and / or being a plant growth regulator can be any suitable known fungicide, insecticide, herbicide and / or plant growth regulator. At least one additional component having pesticidal activity and / or being a plant growth regulator can be from a chemical source or a biological source, for example from a plant or microbial source. At least one additional component having pesticidal activity in the composition disclosed herein can be produced by a microorganism capable of producing a compound according to the present invention.

[0047] In addition, the compositions of the invention may also be administered with one or more inducers of systemic acquired resistance ("SAR" inducers). SAR inducers are known and described, for example, in U.S. Pat. No. 6,919,298, and include, for example, salicylates and the commercial SAR inducer acibenzolar-S-methyl.

[0048] The compounds and / or compositions according to the invention may induce resistance in plants by a priming mechanism. Priming is a mechanism leading to a physiological state that enables a plant to respond more quickly and / or more robustly after exposure to a biotic or abiotic stress, as described 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.

[0049] In one embodiment, the composition according to the present invention further comprises cyclothiazolin C (CtmC), streptoglutarimide and / or malenomycin.

[0050] The structure of cyclothiazomycin C is disclosed on page 3 of WO 2015191789 and can be produced as disclosed in Example 4 of WO 2015 / 191789.

[0051] 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 and has the structural formula II.

[0052]

[0053] Malenomycin can be produced as disclosed in Example I of WO 2006 / 078939. Malenomycin can also be prepared according to the methods disclosed in Examples IA and B of EP 1860939 or according to Law et al., 2018 (Nature Catalysis | Vol. 1 | December 2018 | 977-984).

[0054] Streptoglutarimide is a known compound having formula III

[0055]

[0056] Streptomycin glutarimide 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, aglutarimide antibiotic from Streptomyces rimosus paromomycinus. Eur. J. Org. Chem. (20), 3459-3462 (2000). In one embodiment, the active ingredients cyclothiazomycin C, streptamidone and / or malenomycin are produced by a microorganism capable of producing a compound according to the invention as defined above.

[0057] The present invention also relates to a composition comprising the compound of the present invention having the molecular formula C 53 H 90 N 2 O 44 Surprisingly, it was found that the compound according to the present invention has a 53 H 90 N 2 O 44 The composition of the compound of the molecular formula, preferably the compound according to formula (I) and malenomycin can exhibit unexpected synergistic fungicidal effects. For example, it was found that the composition comprising the compound according to the present invention and malenomycin has a surprising synergistic fungicidal effect on Puccinia reclusa.

[0058] The mixing ratio of the composition comprising the compound of the invention and at least one additional active ingredient (e.g. malenomycin) is preferably 100:1 to 1:6000, especially 50:1 to 1:50, more especially 20:1 to 1:20, even more especially 10:1 to 1:10, very especially 5:1 and 1:5, particularly preferably 2:1 to 1:2, and 4:1 to 2:1 ratio is likewise preferred, especially 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. Those mixing ratios are by weight. The mixing ratio of the composition comprising a mixture of a compound according to the present invention and malenomycin 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 as described above can be used in a method for controlling harmful organisms (e.g., plant pathogenic microorganisms), which method comprises applying a composition comprising a mixture as described above to the harmful organism or its environment, except for methods for treating the human or animal body by surgery or therapy and diagnostic methods performed on the human or animal body.

[0059] Compositions comprising a compound of the invention and one or more additional ingredients having pesticidal activity or plant growth regulators as described above can be applied, for example, in a single "ready-to-use" form, in a combined spray mixture consisting of separate formulations of these single active ingredient components (e.g., a "tank mix"), and in combination with these single active ingredients when applied in a sequential manner (i.e., one after another within a suitably short period of time, e.g., hours, days or weeks).

[0060] In one aspect, the present invention relates to a method for producing a compound or composition according to the present invention, the method comprising culturing a microorganism in a suitable fermentation medium under conditions that allow the production of the compound or composition. The microorganism fermented in the method disclosed herein is a microorganism capable of producing a compound according to the present invention, as disclosed above.

[0061] Cultivating the microorganism for producing the compound according to the present invention or composition in a suitable fermentation medium is well known to those skilled in the art. Microorganism can ferment under aerobic or anaerobic conditions. Microorganisms belonging to Streptomyces species are typically cultivated under aerobic conditions. Suitable fermentation medium comprises nutrients, such as suitable carbon source (such as sugar cane or beet molasses, polysaccharides, flour, starch, sugar or glucose) and suitable nitrogen source (such as casein hydrolysate, tryptone, ammonium sulfate, ammonia, yeast extract, peptone or urea peptide or amino acid). The method for producing the compound according to the present invention can be carried out with batch culture, fed-batch culture or continuous culture.

[0062] In one embodiment, the method comprises producing a compound or comprising a compound according to the present invention and a composition that can produce a microorganism of a compound according to the present invention. The composition comprising a compound according to the present invention and a microorganism that can produce a compound according to the present invention can be a fermentation broth. The microorganism that can produce a compound according to the present invention in the method of the present invention can produce other active ingredients as defined above, for example cyclothiazomycin C, streptoglutarimide and / or malenomycin.

[0063] The method according to the present invention may further include the step of reclaiming the compound or composition according to the present invention. The compound according to the present invention may be reclaimed by suitable method known in the art (e.g., via crystallization or chromatography, e.g., HPLC). Reclaiming the compound according to the present invention may further include the step of purifying the compound.

[0064] The method for producing a compound according to the invention may further comprise the step of formulating the compound into a suitable formulation or composition as defined above.

[0065] In a further aspect, the present invention relates to a method for controlling or preventing infestation of plants, plant propagation materials, loci and / or harvested food crops by treating plants, plant propagation materials, loci and / or harvested food crops, wherein an effective amount of a compound according to the invention or a composition according to the invention is applied to the plant, its part, the plant propagation material, the loci and / or the harvested food crops.

[0066] Application of an effective amount of a compound of the invention in a method for controlling or preventing plant infestations comprises applying 5 g to 5 kg of the compound of the invention (active ingredient (ai)) per hectare (ha), preferably 10 g to 1 kg ai / ha, most preferably 20 g to 600 g ai / ha.

[0067] When the compounds or compositions according to the invention are used to treat seeds, a rate of 0.001 to 50 g of compound according to the invention per kg of seeds, preferably 0.01 to 10 g per kg of seeds, is generally sufficient.

[0068] Suitably, the compounds of the invention or compositions comprising the compounds according to the invention are administered prophylactically (meaning before the disease develops) or therapeutically (meaning after the disease develops).

[0069] Phytopathogenic microorganisms affected by the compounds of the invention are fungi and fungal vectors of diseases as well as phytopathogenic bacteria and viruses. The phytopathogenic microorganisms in the method according to the invention include the following fungi and fungal vectors of diseases as well as phytopathogenic bacteria:

[0070] Absidia corymbifera, Albugo candida, Alternaria spp. (including A. solani), Aphanomyces spp., Ascochyta spp., Aspergillus spp. (including A. flavus, A. fumigatus, A. nidulans, A. niger, A. terrus), Aureobasidium spp. (including A. pullulans), Bacillus subtilis, Blastomyces dermatitidis, Blumeria graminis, Blumeria graminis wheat-specific type, Blumeriella jaapii), Botryosphaeria spp. (including B. dothidea, B. obtusa), Botrytis spp. (including Botrytis cinerea), Bremialactucae, Cadophora gregata, Candida spp. (including C. albicans, C. glabrata, C. krusei, C. lusitaniae, C. parapsilosis, C. tropicalis), Cephaloascus fragrans, Ceratocystis spp., Cercospora spp.) include C.arachidicola, C.beticola, C.kikuchii, C.sojina), Cercosporidium personatum, Cladosporium spp., Clarireedia homoeocarpa, Clavibacter spp., Claviceps purpurea, Coccidioidesimmitis, Cochliobolus spp., Colletotrichum spp. including C. dematium, C. lindemuthianum, C. musae, C. orbiculare, C. truncatum, Corynespora cassicola, Cryptococcus neoformans, Diaporthe spp., Dickeya zeae), Didymella sp., Drechslera spp., Elsinoe spp., Epidermophyton spp., Eremothecium gossypiim, Erwinia spp. (including E. amylovora and E. carotovora), Erysiphe spp. (including E. cichoracearum and E. necator), Eutypa lata, Fusarium spp. spp.) (including Fusarium graminearum, Fusarium graminearum, Fusarium langsethiae, Fusarium moniliforme, Fusarium oxysporum, Fusarium poae, Fusarium proliferatum, Fusarium pseudograminearum, Fusarium sacchari, Fusarium sambucinum, Fusarium subglutinans, Fusarium solani, Fusarium sporotrichioides, Fusarium trilineata, Fusarium spp.tricinctum), F. virguliforme), Gaeumannomyces graminis, Gibberella spp. (including G. avenacea, G. fujikuroi, G. intricans, G. moniliformis, G. zeae), Gloeodes pomigena, Gloeosporium musarum, Glomerella cingulate, Golovinomyces cichoracearum, Gymnosporangium juniperi-virginianae, Guignardia bidwellii, Helminthosporium spp.), Hemileia spp., Histoplasma spp. (including H. capsulatum), Hyaloperonospora parasitica, Kabatiella zeae, Laetisaria fuciformis, Leptographium lundbergii, Leveillula taurica, Lophodermium seditiosum, Microdochium majus, Microdochium snow mold, Microsporum spp., Monilinia spp. (including M. fructicola), Monographella spp.) (including M. nivalis), Mucor spp., Mycosphaerella spp. (including M. arachidis, M. fijiensis, M. graminicola, M.pomi), Nakataea oryzae, Neopseudocercosporella spp., Oculimacula spp., Oncobasidium theobromaeon, Ophiostoma spp., Pantoea stewartia, Paracoccidioides spp., Parastagonospora nodorum, Pectobacterium spp., Penicillium spp. (including P. digitatum, P. italicum), Petriellidium spp., Peronosclerospora spp.) (including P. maydis, P. philippinensis and P. sorghi), Peronospora spp. (including P. destroyer), Phaeosphaeria nodorum, Phakopsora pachyrhizi, Phellinus igniarus, Phialophora spp., Phlyctema vagabunda, Phoma spp., Phomopsis viticola, Phyllactoderma pomigena, Phyllosticta spp., Physoderma maydis, Phytophthora spp. spp.) (including P. capsica, P. infestans), Plasmodiophora brassicae, Plasmopara spp. (including P. halstedii, P. viticola), Pleodorus spp., Pleospora spp., Podosphaera spp. (including P.leucotricha), Polymyxa graminis, Polymyxa betae, Pseudocercosporafijiensis, Pseudocercosporella herpotrichoides, Pseudomonas spp. (including P. syringae), Pseudoperonospora spp. (including P. cubensis, P. humuli), Pseudopeziza tracheiphila, Pseudopyrenochaeta lycopersici, Puccinia spp. spp.) (including P. hordei, P. cryptica, P. stripe, P. triticina), Pyrenopeziza spp., Pyrenophora spp., Pyricularia spp. (including Pyricularia oryzae), Pythium spp. (including P. ultimum), Ralstonia solanacearum, Ramularia spp., Rathayibacter spp., Remotididymella destructiva, Rhizoctonia spp., Rhizomucor pusillus, Rhizopus arrhizus, Rhynchosporium spp.), Robbsiaandropogonis, Sarocladium oryzae, Scedosporium spp. (including S. apiospermum and S. prolificans), Schizothyrium pomi, Sclerophthora macrospora, Sclerotinia spp. (including S.sclerotiorum), Sclerotium spp, Septoria spp (including S. nodorum, S. tritici), Setosphaeria turcica, Sphaerotheca macularis, Sphaerotheca fusca, Sphaerotheca fuliginea, Spiroplasma kunkelii, Sporothorix spp, Stagonospora nodorum, Stagonosporopsis cucurbitacearum, Stemphylium spp, Stenocarpella macrospora, Stereum hirsutum), Streptomyces species, Thanatephorus cucumeris, Thielaviopsis basicola, Tilletia spp., Tranzschelia discolor, Trichoderma spp. (including T. harzianum, T. pseudokoningii, T. viride), Trichophyton spp., Typhula spp., Uncinula necator, Urocystis spp., Uromyces spp., Ustilago spp., Venturia spp. (including V. inaequalis), Verticillium spp. spp), Wilson's sore throat (Wilsonomyces carpophilus) or Xanthomonas spp (including Xanthomonas oryzae and Xanthomonas campestris), Xylella spp, Psoralea corylifolia. .

[0071] Surprisingly it has been found that the phytopathogenic microorganisms affected by the compounds and / or compositions according to the invention are fungi, for example belonging to Puccinia tritici, Puccinia recondita, Pyricularia grisea or Bletilla striella graminearum.

[0072] Any suitable plant, plant propagation material, locus or food crop may be treated according to the method of the invention as defined herein.

[0073] As used herein, the term "locus" means a place where or on which plants grow, or where seeds of cultivated plants are sown, or where seeds are to be placed in the soil. It includes soil, seeds, and seedlings, together with established vegetation.

[0074] The term "plant" refers to all tangible parts of a plant, including seeds, seedlings, young trees, roots, tubers, stems, stalks, leaves, and fruits. Germinated plants and young plants that will be transplanted after germination or after emergence can also be mentioned. These young plants can be protected before transplanting by fully or partially processing through dipping.

[0075] The term "plant propagation material" is understood to mean reproductive parts of plants, such as seeds, which can be used for the multiplication of plants, and vegetative material, such as cuttings or tubers (such as potatoes), roots, fruits, bulbs, rhizomes or parts of plants.

[0076] The term plant relates to "useful plants" or "crops". The expressions "useful plants" and "crops" are used interchangeably herein. "Useful plants" and "crops" include perennial and annual crops, such as berry plants, for example blackberries, blueberries, cranberries, raspberries and strawberries; cereals, for example barley, maize (corn), millet, oats, rice, rye, sorghum, triticale and wheat; fiber plants, for example cotton, flax, hemp, jute and sisal; field crops, for example sugar beets and fodder beets, coffee beans, hops, mustard, rapeseed (canola), poppies, sugar cane, sunflower, tea and tobacco; fruit trees, for example apples, apricots, avocados, bananas, cherries, citrus, nectarines, peaches, pears and plums; grasses, for example Bermuda grass, bluegrass, bentgrass, centipede grass, fescue, ryegrass, St. Augustine's The term "plant" also includes woody crops, such as pine or pine, and woody plants.

[0077] The term "useful plants" is to be understood as also including useful plants that have been rendered tolerant to herbicides such as bromoxynil or classes of herbicides such as, for example, HPPD inhibitors, ALS inhibitors, for example primisulfuron, prosulfuron and trifloxysulfuron, EPSPS (5-enol-pyruvate-shikimate-3-phosphate-synthetase) inhibitors, GS (glutamine synthetase) inhibitors or PPO (protoporphyrinogen oxidase) inhibitors as a result of conventional methods of breeding or genetic engineering.

[0078] The term "useful plants" is to be understood as also including useful plants which have been transformed by the use of recombinant DNA techniques in such a way that they are able to synthesize one or more selectively acting toxins, as are known, for example, from toxigenic bacteria, in particular those of the genus Bacillus.

[0079] Preferably, the plant, plant propagation material or food crop is rice, wheat, corn, soybean or banana.The wording soya herein includes soybean.

[0080] Controlling or preventing in the methods of the present invention means that the infestation by phytopathogenic microorganisms, especially fungi, is reduced to a level that demonstrates an improvement.

[0081] The preferred method of controlling or preventing crop plants from being infected by plant pathogenic microorganisms (especially fungi) is to apply the compound and / or composition according to the present invention to the leaves. The frequency of application and the rate of application will depend on the risk of infection by the corresponding pathogen or insect. Alternatively, the compound and / or composition according to the present invention can penetrate the plant through the root (systemic effect) by soaking the plant in a liquid formulation or by applying the compound in a solid form, for example, in a granular form to the soil (soil application). In rice crops, such granules can be applied to the paddy field of irrigation. The compound according to the present invention can also be applied to seeds (coating) by impregnating seeds or tubers with a liquid formulation of a fungicide or coating it with a solid formulation.

[0082] The compounds of the present invention can also be used as seed dressings for treating plant propagation materials (e.g., seeds, such as fruits, tubers or cereals, or plant cuttings) to prevent fungal infections and to prevent plant pathogenic fungi that occur in the soil. Propagation materials can be treated with compounds and / or compositions according to the present invention before planting: for example, seed dressing can be used before sowing. Such methods for treating plant propagation materials, and plant propagation materials treated in this way are disclosed herein.

[0083] In another aspect, the present invention relates to the use of a compound according to the invention or a composition comprising a compound according to the invention as a pesticidal agent, preferably as a fungicide and / or as an initiator. The features associated with the compound according to the invention and the composition comprising the compound according to the invention are as disclosed above. Therefore, the present invention relates to a method for using a compound according to the invention as a pesticidal agent, preferably as a fungicide and / or as an initiator. BRIEF DESCRIPTION OF THE DRAWINGS

[0084] Figure 1 The compound of the present invention is in D 2 1D at 600MHz in O 1 H NMR spectrum

[0085] Figure 2 The compound of the present invention is in D 2 1D at 600MHz in O 13 C NMR spectrum

[0086] Figure 3 The compound of the present invention is in D 2 2D Dept edited 1H-13C HSQC NMR spectrum at 600 MHz in O showing positive (CH) signal

[0087] Figure 4 The compound of the present invention is in D 22D Dept edited 1H-13C HSQC NMR spectrum at 600 MHz in O showed negative (CH 2 )Signal

[0088] Examples

[0089] Example 1. Source and extraction of compounds of the present invention

[0090] 1.1. Fermentation of Streptomyces species

[0091] Streptomyces species were ordered from the culture collections disclosed in Table 3. Streptomyces species Saigon 413 was isolated in Vietnam before 1961. Streptomyces species Saigon 413 was deposited with the Fungal Culture Collection under the accession number CBS 149411. The deposit was made by Syngenta Ltd. at Jealott's Hill Research International Centre, Bracknell, Berkshire RG42 6EY, United Kingdom under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure.

[0092] Streptomyces species were cultured in Erlenmeyer flasks containing a liquid medium consisting of (g / l) casein hydrolysate 10, glucose 40, K2HPO4 1.25, soytone 2, tryptone 8, and incubated at 28°C in an incubator with shaking at 150 rpm and an amplitude of 25 mm for 4 days.

[0093] 1.2.16S rDNA Isolation and Species Identification

[0094] Genomic DNA was isolated from Streptomyces species Saigon 413 using the method described in Kutchma et al. (1998) Biotechniques 24:452-457. The 16S rRNA gene was amplified using universal 16S primers and sequenced using Sanger sequencing. The 16S rRNA is shown in SEQ ID NO: 1.

[0095] The species of strain Streptomyces species Saigon413 was identified by comparing the 16S rRNA sequence according to SEQ ID NO: 1 with the publicly available 16S rRNA sequence extracted using the whole genome sequence assembly of the Streptomyces species genome (based on the genome classification database GTDB (Parks, DH, et al. (2021). GTDB: Nucleic Acids Research [Nucleic Acids Research], 50: D785-D794)), using barrnap v0.9. Based on this comparative analysis, Streptomyces species Saigon413 was identified as a Streptomyces species. The sequence identity between the 16S rRNA sequence of Streptomyces species Saigon413 and the publicly available Streptomyces crown mycelium NRRL-3672 determined using Muscle v3.8.31 and the R software package Seqinrv4.2-16 was 99.87%.

[0096] Whole genome sequencing was completed using genomic DNA from Streptomyces species Saigon 413 using Pacific Biosciences and Illumina sequencing technologies. The genome was assembled using HFAP4 and perfected using 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 rimosus 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).

[0097] After assembling the genome of Streptomyces sp. Saigon 413 and publicly available genomes, the average nucleotide identity (ANI) between Streptomyces sp. Saigon 413 and closely related Streptomyces strains was calculated using fastANI (Jain, C., et al. (2018), Nature Communications, 9, 5114) (Table 3). The highest percentage identity (ANI) of the genome of Streptomyces sp. Saigon 413 with the publicly available genome of Streptomyces coronatus NRRL B-3672 was 96.9%.

[0098] Using 16S RNA sequence identity and ANI scores (%), strains CBS 596.66, CBS 570.66, and DSM 41429 were also found to be Streptomyces coronatus strains, rather than Streptomyces rimosus or Streptomyces paromomycinus strains as indicated by the depository institute.

[0099] 1.3. Purification of the compounds of the present invention

[0100] The whole broth was centrifuged to produce an aqueous extract and a precipitate. The aqueous extract was freeze-dried. The material was resuspended in a minimum volume of water and partitioned with ethyl acetate to remove lipophilic components. The aqueous suspension was retained and freeze-dried, and then resuspended in a minimum volume of water and applied to an activated carbon column.

[0101] The column was washed with water and eluted with water:acetone (50:50).

[0102] The compounds of the present invention were further purified by hydrophilic interaction liquid chromatography (HILIC) using mass-guided fractionation and an ELSD detector. For example, Waters XBridge Amide (5 µm, 30x100 mm) was used with a gradient of acetonitrile and 10 mM ammonium acetate.

[0103] Example 2. Characterization of the compounds of the present invention.

[0104] The compounds of the present invention in the purified fermentation broth were assayed according to the methods disclosed below.

[0105] The results in Table 3 show that several Streptomyces species are capable of producing the compounds of the present invention.

[0106] 2.1. Molecular composition and total molecular weight

[0107] The molecular composition and total molecular weight are C 53 H 90 N 2 O 44and 1458.487 g, determined using MS-MS and NMR spectroscopy as disclosed in paragraphs 2.3 and 2.4.

[0108] Solubility

[0109] The solubility of the compounds of the invention was determined.

[0110] The solubility of the compounds of the invention in water (pH 7.01) is >10'000 ppm and in DMSO is >9772 ppm.

[0111] 2.3. MS-MS spectroscopy and liquid chromatography

[0112] Spectra were recorded on an Orbitrap ID-XTribrid mass spectrometer from Thermo Scientific equipped with an OptaMax NG heated electrospray source (spray voltage: static, polar ion (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;

[0113] Experiment 1: MS OT (Orbitrap resolution: 60,000, Scan range (m / z): 200 to 2000, RF lens (%): 60, AGC target: standard, Max injection time mode: automatic, Micro scan: 1, Data type: profile, Polarity: both),

[0114] Experiment 2: tMS2 OT CID (MSn level (n): 2, separation window (m / z): 1.6, activation type: CID, CID collision energy (%): 30, detector type: Orbitrap, Orbitrap resolution: 30,000, RF lens (%): 60, polarity: negative). The mass spectrometer was connected to a Vanquish Flex UHPLC from Thermo Fisher Scientific, using a Vanquish split sampler FT, Vanquish binary pump F, Vanquish column chamber H, Vanquish diode array detector FG, and Vanquish charged aerosol detector. Liquid chromatography conditions included: Thermo Scientific Hypercarb TM Porous graphitic carbon pillars 5 μm 4.6x50 mm, PN35005-054630. Temperature: 40 °C, DAD wavelength range: 250 to 260 nm, solvent gradient: Solvent A: HO with 0.1% formic acid 2 O, solvent B: CH with 0.1% formic acid3 CN, 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 uL, total run time: 6.0 min.

[0115] NMR spectroscopy

[0116] NMR spectra were recorded on a Bruker AVIII 600 NMR spectrometer equipped with a 5 mm Bruker ( 1 H / 19 F) / 13 C / 15 N TCI cryoprobe. Dissolve the sample in D 2 O, the spectra were recorded at 300°K, and for 1 H, referenced to acetone at 2.225 ppm, for 13 C, referenced to acetone at 31.07 ppm. Figures 1 to 4 The NMR spectra of the compounds of the present invention are shown.

[0117] single bond 1 H- 13 C-related spectra include the methyl groups (CH 3 ) and 40 methine (CH) groups (listed in Table 1) and 9 methylene (CH 2 ) groups (listed in Table 2).

[0118] In addition, 1D 13 The C spectrum includes signals from three quaternary carbons at 104.7, 159.3 and 175.2 ppm (±0.1).

[0119] Table 1. Single bonds of compounds according to the invention 1 H- 13 Methyl and methine signals in C correlation spectra and 1D 1 Proton multiplicity information resolved from the H spectrum.

[0120]

[0121]

[0122] Table 2. Single bonds of compounds according to the invention 1 H- 13 C correlation spectra of methylene signals and 1D 1Proton multiplicity information resolved from the H spectrum.

[0123]

[0124]

[0125] Table 3. Identification of compounds of the invention in different Streptomyces species and ANI (%) and 16S RNA identity % relative to Streptomyces species Saigon 413

[0126]

[0127]

[0128] CBS Fungal Collection or Westerdijk Fungal Diversity Institute: Uppsala Boulevard 8, Utrecht, The Netherlands (3584 CT)

[0129] DSMZ German Collection of Microorganisms and Cell Cultures: Inhofenstrasse 7B, 38124 Braunschweig, Germany.

[0130] ARS ARS Culture Collection (NRRL), 1815 North University Street, Peoria, IL (61604), USA

[0131] Example 3. Activity of the compounds according to the invention on plants

[0132] 3.1. Leaf disc or leaf segment testing in orifice plates

[0133] Leaf discs or leaf segments of different plant species are cut from plants grown in a greenhouse. The cut leaf discs or leaf segments are placed on water agar in a multiwell plate (24-well format). The leaf discs are sprayed with the test solution before (preventive) or after (curative) inoculation. The compound to be tested is prepared as an aqueous solution (up to 10 mg / ml) which is diluted to the appropriate concentration with 0.025% Tween20 just before spraying. The inoculated leaf discs or leaf segments are incubated under defined conditions (temperature, relative humidity, light, etc.) according to the corresponding test system. Depending on the disease system, a single assessment of the disease level is performed 3-9 days after inoculation. The disease control percentage relative to the untreated control leaf discs or leaf segments is then calculated.

[0134] Puccinia recursa wheat specific type / wheat / leaf discs (preventive) (brown rust)

[0135] Wheat leaf segments cv. Kanzler were placed on agar in multiwell plates (24-well format) and sprayed with formulated test compounds diluted in water. One day after application, the leaf disks were inoculated with a spore suspension of the fungus. The inoculated leaf segments were incubated in a climate chamber at 19° C. and 75% rh under a lighting regime of 12 h light / 12 h dark, and the activity of the compound was assessed as the percentage of disease control compared to untreated leaves when appropriate levels of disease damage appeared on untreated control leaf segments (7-9 days after application).

[0136] Puccinia recursa wheat-specific type / wheat / leaf discs (curative) (brown rust)

[0137] Wheat leaf segments cv. Kanzler were placed on agar in a multiwell plate (24-well format). The segments were inoculated with a spore suspension of the fungus. The plates were stored in the dark at 19°C and 75% rh. The formulated test compound diluted with water was applied 1 day after inoculation. The leaf segments were incubated at 19°C and 75% rh in a climate chamber under a lighting scheme of 12h light / 12h dark, and the activity of the compound was assessed as the percentage of disease control compared to the untreated when the appropriate level of disease damage appeared on the untreated control leaf segments (6-8 days after application).

[0138] Rice blast fungus (Pyricularia oryzae) / Rice / Leaf disc (preventive) (Rice blast)

[0139] Rice leaf segments cv. Ballila were placed on agar in multiwell plates (24-well format) and sprayed with the formulated test compound diluted in water. 2 days after application, the segments were inoculated with a spore suspension of the fungus. The inoculated segments were incubated in a climate chamber at 22° C. and 80% rh under a lighting regime of 24 h dark followed by 12 h light / 12 h dark, and the activity of the compound was assessed as the percentage of disease control compared to the untreated when the appropriate level of disease damage appeared on the untreated control leaf segments (5-7 days after application).

[0140] Table 4. Reduction (%) in fungal development of Puccinia reconditatis wheat-specific (preventive), Puccinia reconditatis wheat-specific (curative) and Magnaporthe oryzae (preventive) on leaf disks or segments in the presence of several concentrations of compounds of the invention.

[0141]

[0142] The results in Table 4 show that compounds of the invention reduced fungal growth of several fungal species starting at a concentration of 22 ppm in both leaf disk and leaf segment tests. Compounds of the invention provided both preventative (compound application followed by pathogen infection) and curative (pathogen infection followed by compound application) activity to control the rust pathogen Puccinia reconditatis.

[0143] 3.2. Wheat seedling leaf smear assay

[0144] Seedlings of wheat varieties Riband (for wheat leaf blight test) or varieties Arina (for hidden stem rust test) were grown in a greenhouse until 14 days after sowing. At this time, such seedlings usually have a first leaf (designated as L1) that has fully emerged, a second leaf (L2) that has fully emerged, and a third leaf (L3) that has partially emerged and is in the process of development. Two points were applied on the second leaf using a permanent pen to form three roughly equal-sized sections: Section A (leaf base), Section B (leaf middle), and Section C (leaf tip). A stock solution of a compound of formula (I) was produced in water, and a stock solution of benzovinflumazole was produced in DMSO at a concentration of 10,000 ppm. The stock solution was then further diluted in water supplemented with Tween20 to a final concentration of 200 ppm or higher for the test compound (containing 0.05% Tween 20 and 2% DMSO) (only for compounds with DMSO stock solution). The diluted compound was applied to the middle section of L2 using a regular cotton swab; the swab was soaked in the diluted compound and rubbed several times on the adaxial surface of the leaf between the two marks. One day later, the fungal spore suspension was inoculated onto the entire plant using a paintbrush. The spore suspension was applied until it was almost runny.

[0145] For infection with Psoralea corylifolia (preventive), the test plants were inoculated one day after application by spraying with a spore suspension (1.5 million spores / ml in water supplemented with 0.01% Tween 20). After a 4-day incubation period at 22°C / 21°C (day / night) and 95% rh, the inoculated test plants were kept in a greenhouse at 22°C / 21°C (day / night) and 70% rh. Efficacy was evaluated directly when appropriate levels of disease appeared on untreated control plants (16-19 days after application).

[0146] For infection with Puccinia recondita (preventive): One day after application, the test plants are inoculated by spraying with a spore suspension (80,000 spores / ml of a spore suspension in water supplemented with Tween 20 (0.1%)). After a 1-day incubation period at 20°C and 95% rh, the inoculated test plants are kept in a greenhouse at 20°C and 60% rh. When an appropriate level of disease appears on untreated control plants (9-12 days after infection), the percentage of leaf area covered by disease is assessed visually.

[0147] The three segments of the leaf were evaluated separately.

[0148] Table 5. Reduction in fungal growth of Puccinia recondita tritici (preventive) and Psoralea corylifolia (preventive) on wheat leaves in the presence of compounds of the invention and benzovindiflupyr in leaf smear assay (%)

[0149]

[0150]

[0151] nt.: Not tested

[0152] The leaf smear test showed that the compounds of the invention have anti-fungal activity against the growth of Puccinia recondita and Psoralea tritici. The compounds of the invention provide control over the treated area (mid-segment) as well as above the treated area (leaf tip). The activity identified in plants was independent of the surfactant Tween 20 or the solvent DMSO (also present at low concentrations when the compounds were tested). The shift of the efficacy to the apical segment is similar to the SDHI fungicide benzovindiflupyr (which is known to exhibit such a shift).

[0153] 3.3. Powdery mildew determination in wheat

[0154] One-week-old wheat leaves were used for the experiment. 5 cm long segments were cut from the leaf tip under water and placed in a cuvette containing 100 μL of the test solution. After 1.5 h, the leaves were removed and left to stand on a paper towel for 1 h. The leaves were then placed on a water agar plate (1%) and stored in a light box (illumination for 8 h at 18 ° C). The day before the harvested leaf segments were inoculated, wheat plants infected with powdery mildew (B. graminis wheat-specific type) were gently shaken to remove older conidia, thereby forming fresh conidia. For inoculation, the plate containing the leaf segments was spread flat on the ground, and then the infection hood was placed on top. The wheat plants infected with powdery mildew were placed in the hood through the outlet on the hood and gently shaken to evenly distribute the spores. After 48 h of inoculation, the leaves were transferred to a reaction tube containing 10 mL of 80% ethanol. Ten days after decolorization of the leaf tissue, fungal spores were stained with 10% (v / v) ink, 25% (v / v) acetic acid in ddH2O. The ratio of successful penetration to papillae formation was evaluated at one hundred interaction sites between the spores and the plant cells using an optical microscope. The test was repeated four times, and the values ​​shown in Table 4 are the average of four tests.

[0155] The compound INA (2,6-dichloro-isonicotinic acid, CAS: 5398-44-7) is a synthetic salicylic acid analogue and is included as a reference for elicitor activity (Krauss et al., 1992, Plant Journal 2, 655-60).

[0156] The test sample AEF1 is a sample enriched in the compound of formula I. This sample corresponds to the fraction most enriched in the compound of formula I eluted from the activated carbon column.

[0157] Table 6. Haustoria formation in wheat leaves treated with different compounds 48 h after infection with B. graminis. The reported values ​​are haustoria frequencies relative to the mean control values. Different letters indicate significant differences (P value < 0.05)

[0158]

[0159] The results in Table 6 demonstrate that wheat pre-treated with a compound of the invention or a fraction enriched with a compound of the invention results in a reduction in haustoria formation by the powdery mildew fungus Blumeria graminicola. This effect was observed at concentrations of 25 μM or higher. The enriched fraction and the purified compound had similar effects at the highest tested rate.

[0160] 3.4. Reactive oxygen species (ROS) burst assay in wheat leaf discs

[0161] 200 μL of the test solution (analyte) or the corresponding control (water) of the appropriate concentration were pipetted into a white 96-well plate (Nunc, Langenselbold, Germany). A 5 mm leaf disk of a 2-week-old wheat plant was obtained using a tissue punch and then floated on the test solution. The plate was stored at RT for 24 h. The next day, 50 μL ddH2O was used to replace the solution, and the leaf disk was placed in the dark at RT for at least one hour for regeneration. At the same time, a freshly prepared appropriate master mixture with or without the exciter flg22 (see below) was prepared in a black 5 mL reaction tube. After regeneration, 50 μL of the corresponding master mixture was added to the hole containing the leaf disk. Subsequently, luminescence was recorded with a plate reader (BMG Labtech; Ortenberg, Germany) for 40 minutes. The compound INA (2,6-dichloro-isonicotinic acid, CAS: 5398-44-7) is a synthetic salicylic acid analogue and was included as a reference for elicitor activity (Kauss et al., 1992).

[0162] Master mix - flg22: 4.98 mL ddH2O, 10 μL HRP (10 mg / mL), 10 μL L-012 (20 mM)

[0163] Master mix + flg22: 4.979 mL ddH2O, 10 μL HRP (10 mg / mL), 10 μL L-012 (20 mM), 1 μL flg22 (10 μM)

[0164] Abbreviations: flg22 (22 amino acid flagellin peptide, Eurogentec catalog number AS-62633); HRP (horseradish peroxidase), L-012 sodium salt (CAS number: 143556-24-5) was used.

[0165] Test sample AEF1 is a sample enriched with the compound of formula (I). This sample corresponds to the fraction with the highest enrichment of the compound of formula I eluted from the activated carbon column.

[0166] Table 7. Fold changes in peak ROS production. Values ​​represent the ratio of peak values ​​observed in the treated groups to the control group, using the maximum value measured in each condition over a 40-minute time course. Values ​​indicated are the average of two replicates.

[0167]

[0168] The results in Table 7 show that wheat pre-treated with fractions enriched with compounds of the invention (at concentrations ranging from 1 ppm to 100 ppm) increased ROS production induced by peptide flg22 by 2-fold or more. This response was similar to or stronger than that observed with treatment with INA (2,6-dichloro-isonicotinic acid), a well-known initiator (Krauss et al., 1992, Plant Journal 2: 655-60).

[0169] Example 4. Fungicidal activity of a mixture of an oligosaccharide compound according to the invention and malenomycin in a leaf disc assay

[0170] Add 0.025% in water A stock solution of the oligosaccharide compound according to the invention was produced in 20. Malenomycin was produced according to Law et al., 2018 (Nature Catalysis | Vol. 1 | December 2018 | 977-984). 0.025% A stock solution of malenomycin was generated in 20.

[0171] To test the efficacy of a mixture of a compound of formula (I) in combination with malenomycin in controlling fungal pathogens in a leaf disc assay, an assay for two 24-well plates was designed.

[0172] Table 8 and Table 9: Overview of 24-well test plates (1) and (2) (including the concentration of the compound sprayed in each well). The upper numbers in the cells indicate the concentration of the oligosaccharide compound of the present invention (ppm), and the lower numbers indicate the concentration of malenomycin (ppm). (1) is listed as a dilution series of the compound of formula (I), and (2-D) is a dilution series of malenomycin. Well 2-D-(1) represents an untreated control. This design was applied to a test involving the use of preventive and curative spray times for Puccinia recursa (EPPO code: PUCCRE).

[0173] Table 8. 24-well plate (1)

[0174]

[0175] Table 9. 24-well plate (2)

[0176]

[0177] Table 10. The mixing ratios of the compounds sprayed in the 24-well plate assay are summarized in Tables 8 and 9. The numbers represent the ratio of Compound 1: Compound 2. Compound 1 is malenomycin and Compound 2 is an oligosaccharide compound of the present invention. The plate designs of the two 24-well plates span a wide range from 64:1 to 1:270.

[0178]

[0179] Table 11 and Table 12: Overview of 24-well test plates (3) and test plates (4) (including the concentration of the compound in each well). The upper numbers in the cells indicate the concentration of the oligosaccharide compound of the present invention (ppm), and the lower numbers indicate the concentration of malenomycin (ppm). (1) is listed as a dilution series of the compound having formula (I), and (4-D) is a dilution series of malenomycin. Well 4-D-(1) represents an untreated control. This design was applied to tests including the wheat-specific type of B. graminis (EPPO code: ERYSGT) using a preventive spraying time and the test of Pseudomonas erythematosus (EPPO code: LEPTNO) using a preventive spraying time.

[0180] Table 11. 24-well plates (3)

[0181]

[0182] Table 12. 24-well plates (4)

[0183]

[0184] Table 13. Mixing ratios of compounds sprayed in 24-well plate assays are summarized in Tables 11 and 12. Numbers represent the ratio of Compound 1: Compound 2. Compound 1 is malenomycin and Compound 2 is a compound of formula (I) according to the present invention. The plate designs of two 24-well plates span a wide range from 64:1 to 1:81.

[0185]

[0186] A first set of master plates were prepared with 1-fold concentrated spray solutions of the stock solutions of the compounds of formula (I) diluted in water according to the concentrations in Tables 8, 9, 11 and 12, respectively. Each well contained 0.025% 20. Accordingly, a second set of master plates was prepared with a 1x concentrated malenomycin stock solution diluted in water. Each well of the second set contained 0.025% 20. Leaf segments placed on agar in a 24-well plate were sprayed with 8 ul of a solution containing a compound of formula (I) from the mother plate, the leaf segments were allowed to dry, and 2 hours later, they were sprayed with 8 ul of a solution containing malenomycin from the mother plate. After the second spray drying, the leaf segments were infected with fungal spores to obtain a preventive application time. Alternatively, leaf segments that had been infected one day before spraying the compound were used to obtain a therapeutic spray time. In addition, several plates were made in which the leaves were sprayed with fungal spores in the absence of the test compound (containing only 20) were sprayed twice as an untreated control sample. The percentage leaf coverage of disease symptoms was assessed for each leaf segment. The percentage reduction in leaf coverage relative to the untreated control was calculated. The efficacy of the mixture was tested in duplicate against different fungal species. The reported efficacy value is the average of the two replicate results.

[0187] For Puccinia recondita (EPPO code: PUCCRE), use preventive spraying schedule.

[0188] Wheat (cultivar Kanzler) leaf segments were placed on agar in a multiwell plate (24-well format) and sprayed with the test solution (8 ul / well). After drying, the leaf disks were inoculated with a spore suspension of the fungus. After appropriate incubation, the activity of the compounds 8 days after inoculation (8 dpi) was evaluated as preventive fungicidal activity.

[0189] For Puccinia recondita (EPPO code: PUCCRE), use curative spraying schedules.

[0190] Wheat (cultivar Kanzler) leaf segments were placed on agar in a multiwell plate (24-well format). These leaf disks were then inoculated with a spore suspension of the fungus. One day after inoculation, the test solution (8ul / well) was sprayed. After appropriate incubation, the activity of the compound after 8 days of inoculation (8dpi) was evaluated as a therapeutic fungicidal activity.

[0191] For Blumeria graminearum, wheat-specific type (EPPO code: ERYSGT), adopt preventive spraying time.

[0192] Wheat (cultivar Kanzler) leaf segments were placed on agar in a multiwell plate (24-well format) and sprayed with the test solution (8 ul / well). After drying, the leaf disks were inoculated with fungal spores. After appropriate incubation, the compound activity was evaluated 7 days after inoculation (7 dpi) as preventive fungicidal activity.

[0193] Ascopolysporus lemborius (EPPO code: LEPTNO)

[0194] Wheat (cultivar Kanzler) leaf segments were placed on agar in a multiwell plate (24-well format) and sprayed with the test solution (8 ul / well). After drying, the leaf disks were inoculated with a spore suspension of the fungus. After appropriate incubation, the activity of the compounds 4 days after inoculation (4 dpi) was evaluated as preventive fungicidal activity.

[0195] result

[0196] The results of fungus control experiments conducted with mixtures of compounds according to the invention and malenomycin as described above are shown in Tables 14 to 19.

[0197] Table 14. Control of Puccinia recondita (preventive) by single compounds and mixtures. Plate designs including compounds of formula (I) and malenomycin concentrations are shown in Tables 8 and 9. Values ​​indicate control of fungal growth (% reduction in symptoms on leaf segments compared to untreated controls).

[0198]

[0199] For each test condition in Table 14 where the efficacy was 50% or greater (effective mix), the corresponding mix ratio can be assigned according to Table 10. Effective mix ratios for controlling Puccinia reconnaissance (preventative) were found to be 64:1 to 1:90.

[0200] Table 15. Comparison of measured values ​​for disease control of Puccinia recondita (preventive) (as reported in Table 14) with values ​​calculated for the same mixture using Colby's formula. The numbers reported in the table represent the difference between the measured efficacy (in %) minus the calculated efficacy (in %). Values ​​near 0 (zero) indicate additive activity, while positive values ​​indicate synergistic activity.

[0201]

[0202] Table 16. Control of Puccinia recondita (curative) by single compounds and mixtures. The plate design including the compound of formula (I) and malenomycin concentrations are shown in Table 8 and Table 8. The values ​​indicate control of fungal growth (% reduction of symptoms on leaf segments compared to untreated controls).

[0203]

[0204] For each test condition in Table 16 with an efficacy of 50% or greater (effective mix), the corresponding mix ratio can be assigned according to Table 10. Effective mix ratios for controlling Puccinia recondita (curative) were found to be 64:1 to 1:270.

[0205] Table 17. Comparison of measured values ​​for disease control of Puccinia recondita (curative) (as reported in Table 16) with values ​​calculated for the same mixture using Colby's formula. The numbers reported in the table represent the difference between the measured efficacy (in %) minus the calculated efficacy (in %). Values ​​near 0 (zero) indicate additive activity, while positive values ​​indicate synergistic activity.

[0206]

[0207] Table 18. Control of Blumeria graminearum (preventive) by single compounds and mixtures. The plate design including the compound of formula (I) and the concentration of malenomycin is shown in Table 11 and Table 12. The values ​​indicate the control of fungal growth (% reduction of symptoms on leaf segments compared to untreated controls).

[0208]

[0209] For each test condition in Table 18 with an efficacy of 50% or more (effective mix), the corresponding mix ratio can be specified according to Table 11. The effective mix ratio for controlling B. graminearum wheat-specific type (preventive) was found to be 64:1 to 1:4.

[0210] Table 19. Control of Ascochyta nodosum by single compounds and mixtures (preventive). Plate designs including compound of formula (I) and malenomycin concentrations are shown in Tables 11 and 12. Values ​​indicate control of fungal growth (% reduction of symptoms on leaf segments compared to untreated controls).

[0211]

[0212] For each test condition in Table 19 with an efficacy of 50% or greater (effective mix), the corresponding mix ratio can be assigned according to Table 11. The effective mix ratios for controlling Ascochyta nodosum (preventative) were found to be 64:1 to 1:9.

[0213] in conclusion

[0214] The mixture of oligosaccharide compounds and malenomycin according to the present invention controls a variety of fungal pathogens. When spraying the mixture on leaves, a variety of mixing ratios of the two compounds in the mixture can produce 50% or higher control on fungal growth. For several fungal species, examples of ratios ranging from 64:1 to 1:270 are shown, including examples with ratios between the ratios (compound malenomycin: ratio of oligosaccharide compounds of the present invention). Surprisingly, the efficacy of several mixtures is better than the efficacy predicted based on Colby calculations, indicating that the mixture of oligosaccharide compounds of the present invention and malenomycin has a synergistic effect on the control of fungal pathogens when sprayed on leaves. This surprising synergistic effect was observed in the test of controlling hidden stem rust.

[0215]

Claims

1. A compound comprising 53 H 90 N2O 44 The compound is further characterized by the NMR spectra listed in Table 1 and Table 2, or a salt thereof.

2. A compound, which is optionally a compound according to claim 1, having structural formula (I): or a salt thereof.

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

4. A composition comprising the compound according to claim 1 or 2 or the 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 fungicidal activity.

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

7. The composition according to claim 6, wherein the additional ingredients comprise cyclothiazolinyl, streptoglutarimide and / or malenomycin, preferably malenomycin.

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

9. The composition according to any one of claims 3 to 7, or the method according to claim 8, wherein the microorganism is a Streptomyces species, preferably Streptomyces coronamicinus, Streptomyces fissureus, Streptomyces paromomycinus or Streptomyces albus, preferably a Streptomyces species having a 16S RNA sequence that is at least 98% identical to SEQ ID NO: 1, preferably wherein the microorganism is a Streptomyces species deposited with the Fungal Culture Collection Saigon 413 under the accession number CBS149411.

10. A method for controlling or preventing infection of plants, plant propagation materials, places and / or food crops in the harvest by treating the plants, plant propagation materials, places and / or food crops in the harvest, wherein 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 is applied to the plants, parts thereof, the plant propagation materials, places thereof, and / or food crops in the harvest.

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

12. 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 method according to any one of claims 10 to 12, wherein the plant pathogenic microorganism is a fungus, preferably the fungus belongs to the genus Puccinia, Puccinia, Magnaporthe oryzae, Bleucophylla or Ascophyllosporium, preferably a fungus belonging to Puccinia tritici, Puccinia cryptica, Puccinia cryptica wheat-specific type, Magnaporthe oryzae, Bleucophylla wheat-specific type or Ascophyllosporium glumeboc.

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 any one of claims 1 or 2 or a composition according to any one of claims 3 to 7 as a pesticide, preferably as a fungicide and / or as an initiator.

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