Fungicidal mixture containing cyclothiazomycin C and malonomycin.

Abstract

Compositions comprising cyclothiazomycin C and malonomycin, pesticide compositions comprising this mixture, and methods and uses of the compositions in controlling or preventing fungal infestation of plants or other substrates.

Description

[Technical Field]

[0001] The present invention relates to a mixture comprising two known compounds, cyclothiazomycin C and malonomycin, and its use, in particular for controlling fungi in agriculture or horticulture. The present invention also relates to a fungicidal composition, in particular an agrochemical fungicidal composition, comprising this mixture, and to a process for preparing this composition. [Background technology]

[0002] Cyclothiazomicin C has the formula I; [ka] is a known compound.

[0003] The structure of cyclothiazomycin C is disclosed on page 3 of WO2015191789. The disclosure also provides examples of the antibacterial activity of cyclothiazomycin C in Table 6 on page 31. WO2015191789 clearly states after Table 6 on page 31 that "the greatest inhibitory activity was observed against Bacillus. The inventors also decided to evaluate whether cyclothiazomycin C exhibited growth inhibitory activity against various fungal strains, but no such activity was observed."

[0004] Malonomicin (sometimes spelled "malonomycin") is a compound of formula (II) [ka] {[(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.

[0005] Maronomycin is a natural compound prepared by fermentation. U.S. Patent No. 3,536,811 discloses maronomycin for use as an antibiotic and protozoan growth inhibitor. European Patent No. 1,860,939 describes maronomycin as an agricultural fungicide.

[0006] Surprisingly, it has now been found that mixtures comprising cyclothiazomycin C and malonomycin can exhibit an unexpected synergistic fungicidal effect. Summary of the Invention [Means for solving the problem]

[0007] According to a first aspect of the present invention there is provided a mixture comprising cyclothiazomycin C and malonomycin.

[0008] According to a second aspect of the present invention, there is provided an agrochemical composition comprising a mixture comprising a fungicidally effective amount of cyclothiazomycin C and malonomycin. Such agrochemical composition may further comprise an agrochemically acceptable diluent or carrier.

[0009] Reference herein to an agrochemical composition includes chemical compositions and biological compositions, preferably non-naturally occurring biological compositions, e.g., biologically produced compositions, e.g., microbiologically produced compositions, or non-naturally occurring compositions.

[0010] The composition according to the present invention is an isolated composition. An isolated composition refers to a non-naturally occurring composition.

[0011] According to a third aspect of the present invention, there is provided a method for controlling or preventing fungal infestation of plants, wherein a fungicidally effective amount of an agrochemical composition comprising a mixture comprising cyclothiazomycin C and malonomycin is applied to a plant, its part or its habitat.

[0012] According to a fourth aspect of the present invention there is provided the use of a mixture comprising cyclothiazomycin C and malonomycin as a fungicide. According to this particular aspect of the invention, the use may not include methods involving the treatment of the human or animal body by surgery or therapy.

[0013] Cyclothiazomicin C is available as disclosed in WO 2015191789. In particular, it is produced by NRRL strain WC-3908 and can be isolated as described in paragraph

[0178] of WO 2015191789. Strain WC-3908 is publicly available from the ARS Culture Collection (NRRL), 1815 N University Street, Peoria, IL 61604.

[0014] Maronomycin can be prepared according to the method disclosed in Examples IA and B of EP 1860939 or according to Law et al, 2018 (Nature Catalysis | VOL 1 | DECEMBER 2018 | 977-984). DETAILED DESCRIPTION OF THE INVENTION

[0015] Mixtures containing cyclothiazomycin C and malonomycin can be used as active ingredients in agricultural and related fields of use, for example, to control fungal plant pests or in non-biological materials to control decay fungi or fungi potentially harmful to humans. Mixtures containing cyclothiazomycin C and malonomycin have surprising efficacy at low application rates and are well tolerated by plants. They have very useful curative and preventive properties and can be used to protect a wide variety of cultivated plants. Mixtures containing cyclothiazomycin C and malonomycin also have surprising synergistic effects. They can be used to suppress or eradicate fungi that appear on plants or plant parts (fruits, flowers, leaves, stems, tubers, roots) of different crops of useful plants, while simultaneously protecting those parts of the plant that grow later.

[0016] The present invention further relates to a method for controlling or preventing infestation of plants or plant propagation material and / or harvested food crops that are susceptible to fungal attack by treating the plants or plant propagation material and / or harvested food crops, wherein a mixture comprising a fungicidally effective amount of cyclothiazomycin C and malonomycin is applied to the plants, their parts or their habitat.

[0017] A mixture containing cyclothiazomycin C and malonomycin can also be used more broadly as a fungicide. As used herein, the term "fungicide" refers to a compound that controls, modifies, or prevents the growth of fungi. The term "fungicidally effective amount," when used, refers to the amount of such a compound or combination of such compounds that is capable of producing an effect on fungal growth. A controlling or modifying effect includes any deviation from natural occurrence, such as killing, delaying, etc., and prevention includes the formation of a barrier or other defense in the plant to prevent fungal infection.

[0018] For protection against fungal infections and phytopathogenic fungi occurring in the soil, a mixture containing cyclothiazomycin C and malonomycin may also be used as a dressing for treating plant propagation material, such as seeds, such as fruits, tubers, or grains, or plant cuttings. The propagation material can be treated with a composition containing cyclothiazomycin C and malonomycin before planting: for example, seeds can be dressed before sowing. Cyclothiazomycin C and malonomycin can also be applied to seeds (coatings) by impregnating the seeds in a liquid formulation or by coating the seeds with a solid formulation. The composition can also be applied to the planting site, for example, in the sowing furrow during sowing, when the propagation material is planted. The present invention also relates to a method for treating such plant propagation material, and to plant propagation material treated in this way.

[0019] Furthermore, mixtures containing cyclothiazomycin C and malonomycin can be used to control fungi, for example in the protection of technical materials and wood-related technical products, food preservation, pharmaceutical applications, veterinary applications and related fields of hygiene.

[0020] Additionally, the present invention can be used to protect non-living materials such as timber, wallboard, wallpaper and paint from fungal attack.

[0021] Examples of important fungi that need to be controlled in agriculture and other sectors are: Botrytis cinerea, Cercospora kikuchii, Cercospora sojina, Cochliobolus sativus, Colletotrichum lindemuthianum, Colletotrichum orbiculare, Corynespora cassiicola, Fusarium avenaceum, Fusarium culmorum, Fusarium langsethiae, Fusarium poae, Fusarium sporotrichioides sporotrichioides, Fusarium tricinctum, Fusarium virguliforme, Gibberella avenacea, Gibberella fujikuroi, Gibberella zeae, Microdochium majus, Monographella nivalis, Mycosphaerella arachidis, Phakopsora pachyrhizi, Puccinia triticinae triticina (=P. recondita), wheat yellow spot fungus (Pyrenophora tritici-repentis), Ramularia collo-cygni, Rhynchosporium secalis, Septoria glycines, Tilletia tritici, Ustilago segetum var.Tritici (Ustilago segetum var. Tritici), apple scab fungus (Venturia inaequalis), and Zymoseptoria tritici.

[0022] Preferred examples of fungi are Gibberella species, including Gibberella zeae (also known as Fusarium graminearum), Fusarium species, including Fusarium virguliforme (also known as Fusarium solani f.sp. glycines, or soybean sudden death syndrome), Microdochium species, such as Monographella nivalis (also known as Microdichium nivale or cereal head blight), and Zymoseptoria tritici (Septoria tritici, wheat leaf blight (also known as Mycosphaerella graminicola or Septoria leaf blight), in particular Zymoseptoria tritici, Microdichium nivale (=Monographella nivalis) and Gibberella zeae (=Fusarium graminearum).

[0023] Fusarium avenaceum, Fusarium culmorum, Fusarium langsethiae, Fusarium poae, Fusarium sporotrichioides, Fusarium tricinctum, Fusarium virguliforme, Fusarium verticillioides, Fusarium subglutinans, Zymoseptoria tritici, Puccinia triticina Particularly preferred are P. triticina (= P. recondita), Mycosphaerella fijiensis, Puccinia striiformis, Magnaporthe grisea and Rhizoctonia solani.

[0024] Target crops and / or useful plants to be protected are typically berry plants, such as blackberries, blueberries, cranberries, raspberries and strawberries; cereals, such as barley, maize (corn), millet, oats, rice, rye, sorghum, triticale and wheat; fiber plants, such as cotton, flax, hemp, jute and sisal; field crops, such as sugar and fodder 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; pasture grasses, such as bermudagrass, bluegrass, bentgrass, centipedegrass, fescue, rye, lawn grass and wild grass; Includes herbs such as basil, borage, chives, coriander, lavender, lovage, mint, 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, radish, melon, okra, onion, pepper, potato, pumpkin, rhubarb, spinach and tomato; and perennial and annual crops such as vines.

[0025] The term "crop" or "useful plant" should also be understood to include useful plants that have been rendered tolerant to herbicides such as bromoxynil or to certain classes of herbicides (e.g., HPPD inhibitors, ALS inhibitors such as primisulfuron, prosulfuron and trifloxysulfuron, EPSPS (5-enol-pyroyl-shikimate-3-phosphate-synthase) inhibitors, GS (glutamine synthetase) inhibitors or PPO (protoporphyrinogen-oxidase) inhibitors) by conventional methods of breeding (mutagenesis). Crops that have been rendered tolerant to imidazolinones, such as imazamox, by conventional methods of breeding (mutagenesis) include, for example, Clearfield® summer rape (canola). Crops that have been genetically engineered to be tolerant to herbicides or classes of herbicides include, for example, glyphosate- and glufosinate-tolerant corn varieties commercially available under the trade names RoundupReady®, Herculex I®, and LibertyLink®.

[0026] It should be understood that the term "crop" or "useful plant" also includes 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 Bacillus bacteria.

[0027] Mixtures containing cyclothiazomycin C and malonomycin may also be used, for example, on turf, ornamental plants such as flowers, shrubs, broadleaf trees, or evergreen trees such as conifers, as well as for trunk injections, pest control, and the like.

[0028] Preferred crops on which mixtures comprising cyclothiazomycin C and malonomycin can be used include bananas, cereals and pulses such as peanuts or soybeans, and grains such as wheat, barley, rice or corn.

[0029] As used herein, the term "habitat" means the field in which the plant is growing or in which the seeds of the plant to be cultivated have been sown or will be sown in the soil. It includes the soil, the seeds and seedlings, and the established vegetation.

[0030] The term "plant" refers to all physical parts of a plant, including seeds, seedlings, saplings, roots, tubers, stems, stalks, leaves and fruits.

[0031] The term "plant propagation material" is understood to refer to reproductive parts, such as seeds, of plants and vegetative parts, such as cuttings or tubers, which can be used for their propagation, for example potatoes. Examples include seeds (in the strict sense), roots, fruits, tubers, bulbs, rhizomes and parts of plants. Mention may also be made of germinated plants and seedlings that are to be transplanted after germination or emergence from the soil. These seedlings can be protected by a complete or partial immersion treatment before transplanting. Preferably, "plant propagation material" is understood to refer to seeds.

[0032] The mixtures containing cyclothiazomycin C and malonomycin can be used in their original form or, preferably, with adjuvants conventionally used in the formulation art. For this purpose, they can be formulated in a known manner into emulsifiable concentrates, coatable pastes, ready-to-spread or dilutable solutions or suspensions, diluted emulsions, wettable powders, soluble powders, dusts, granules, or even encapsulated in polymeric materials. The application method, such as spraying, atomizing, dusting, scattering, coating, or pouring, as well as the type of composition, are selected depending on the intended purpose and the circumstances. The compositions can also contain further adjuvants, such as stabilizers, antifoaming agents, viscosity modifiers, binders, or tackifiers, as well as fertilizers, sources of trace elements, or other ingredients for special effects.

[0033] Furthermore, when cyclothiazomycin C is obtained from a microorganism, it can be isolated from the microorganism as described in WO2015191789. Alternatively, significant amounts of cyclothiazomycin C may be present in the medium in which the microorganism is grown, in which case the medium or liquid medium can be used together with malonomycin to formulate the fungicidal composition. Maronomycin can also be produced by microorganisms such as those disclosed in Examples IA and IB of EP1860939. As a further alternative, the microorganism can produce both cyclothiazomycin C and malonomycin, in which case the microorganism itself can be used to formulate the composition. Thus, a process for producing cyclothiazomycin C and malonomycin is disclosed, comprising fermenting a microorganism in a suitable fermentation medium under conditions that result in the production of cyclothiazomycin C and malonomycin. In such cases, the microorganism can be formulated as viable cells that actively produce cyclothiazomycin C and malonomycin, or can be inactivated, for example, by heat treatment. The microorganisms can be concentrated, if desired, by centrifugation or other conventional techniques.

[0034] Suitable carriers and adjuvants, for example for agricultural applications, can be solid or liquid and are substances useful in formulation technology, such as natural or regenerated inorganic substances, solvents, dispersants, wetting agents, tackifiers, thickeners, binders or fertilizers. Such carriers are described, for example, in WO 97 / 33890.

[0035] Suspension concentrates are formulations in which finely divided solid particles of the active compound are suspended in a liquid. Such formulations contain anti-settling and dispersing agents and may further contain wetting agents, as well as anti-foaming and crystal growth inhibitors to enhance activity. When used, these concentrates are diluted in water and typically applied as a spray to the area to be treated. The amount of active ingredient can range from 0.5% to 95% of the concentrate.

[0036] Wettable powders are in the form of finely divided particles that disperse readily in water or other liquid carriers. The particles contain the active ingredient held in a solid matrix. Typical solid matrices include fuller's earth, kaolin clay, silica, and other readily wet organic or inorganic solids. Wettable powders usually contain 5% to 95% of the active ingredient and small amounts of wetting agents, dispersing agents, or emulsifying agents.

[0037] Emulsifiable concentrates are homogeneous liquid compositions that are dispersible in water or other liquids and may consist solely of the active compound and a liquid or solid emulsifier, or may also contain a liquid carrier such as xylene, high-boiling aromatic naphtha, isophorone, and other non-volatile organic solvents. When used, these concentrates are dispersed in water or other liquid and typically applied as a spray to the area to be treated. The amount of active ingredient can range from 0.5% to 95% of the concentrate.

[0038] Granular formulations include both extrudates and relatively coarse particles, and are usually applied undiluted to the area where treatment is required. Typical carriers for granular formulations include sand, fuller's earth, attapulgite clay, bentonite clay, montmorillonite clay, vermiculite, perlite, calcium carbonate, brick, pumice, pyrophyllite, kaolin, dolomite, gypsum, wood flour, ground corn cobs, ground peanut shells, sugar, sodium chloride, sodium sulfate, sodium silicate, sodium borate, magnesia, mica, iron oxide, zinc oxide, titanium oxide, antimony oxide, cryolite, gypsum, diatomaceous earth, calcium sulfate, and other organic or inorganic materials that can absorb or be coated with the active compound. Granular formulations usually contain 5% to 25% active ingredient, which may include surfactants or vegetable oils such as high-boiling aromatic naphtha, kerosene, and other petroleum fractions; and / or adhesives such as dextrin, glue, or synthetic resins.

[0039] Dusts are free-flowing mixtures of the active ingredient and finely divided solids such as talc, clays, flours and other organic and inorganic solids which act as dispersants and carriers.

[0040] Microcapsules are typically droplets or granules of an active ingredient enclosed in an inert, porous shell that allows the encapsulated material to be released into the environment at a controlled rate. The encapsulated droplets are typically 1 to 50 microns in diameter. The encapsulated liquid typically constitutes 50 to 95% of the capsule's weight and may contain solvent in addition to the active compound. Encapsulated granules are generally porous granules with a porous membrane that seals the pore openings of the granule, retaining the active species in liquid form within the pores of the granule. Granules typically range in diameter from 1 millimeter to 1 centimeter, preferably 1 to 2 millimeters. Granules are formed by extrusion, agglomeration, or prilling, or are naturally occurring. Examples of such materials are vermiculite, calcined clay, kaolin, attapulgite clay, sawdust, and granular carbon. Shell or membrane materials include natural and synthetic rubbers, cellulosic materials, styrene-butadiene copolymers, polyacrylonitriles, polyacrylates, polyesters, polyamides, polyureas, polyurethanes, and starch xanthates.

[0041] Other useful formulations for pesticide applications include simple solutions of the active ingredient in a solvent in which it is completely soluble at the desired concentration, such as water, acetone, alkylated naphthalenes, xylene, and other organic solvents. Pressurized sprayers can also be used, in which the active ingredient is dispersed in finely divided form as a result of evaporation of the low-boiling dispersant solvent carrier.

[0042] Suitable agricultural adjuvants and carriers useful in formulating the compositions of the present invention in the formulation types described above are well known to those skilled in the art.

[0043] Liquid carriers that can be used include, for example, water, vegetable oil, toluene, xylene, petroleum naphtha, crop oil, acetone, methyl ethyl ketone, cyclohexanone, acetic anhydride, acetonitrile, acetophenone, amyl acetate, 2-butanone, chlorobenzene, cyclohexane, cyclohexanol, alkyl acetate, diacetone alcohol, 1,2-dichloropropane, diethanolamine, p-diethylbenzene, diethylene glycol, diethylene glycol abietate, diethylene glycol butyl ether, diethylene glycol ethyl ether, diethylene glycol methyl ether, N,N-dimethylformamide, dimethyl sulfoxide, 1,4-dioxane, dipropylene glycol, dipropylene glycol methyl ether, dipropylene glycol dibenzoate, diproxitol, alkylpyrrolidinone, ethyl acetate, 2-ethylhexanol, ethylene carbonate, 1,1,1-Trichloroethane, 2-heptanone, alpha-pinene, d-limonene, ethylene glycol, ethylene glycol butyl ether, ethylene glycol methyl ether, gamma-butyrolactone, glycerol, glycerol diacetate, glycerol monoacetate, glycerol triacetate, hexadecane, hexylene glycol, isoamyl acetate, isobornyl acetate, isooctane, isophorone, isopropyl benzene, isopropyl myristate, lactic acid, laurylamine, mesityl oxide, methoxy-propanol, methyl isoamyl ketone, methyl isobutyl ketone, methyl laurate, methyl octanoate, methyl oleate, methylene chloride, m-xylene, n-hexane, n-octyl Examples of suitable carriers include methylamine, octadecanoic acid, octylamine acetate, oleic acid, oleylamine, o-xylene, phenol, polyethylene glycol (PEG 400), propionic acid, propylene glycol, propylene glycol monomethyl ether, p-xylene, toluene, triethyl phosphate, triethylene glycol, xylene sulfonic acid, paraffin, mineral oil, trichloroethylene, perchloroethylene, ethyl acetate, amyl acetate, butyl acetate, methanol, ethanol, isopropanol, and higher molecular weight alcohols such as amyl alcohol, tetrahydrofurfuryl alcohol, hexanol, octanol, ethylene glycol, propylene glycol, glycerin, and N-methyl-2-pyrrolidinone. Water is generally the carrier of choice for dilution of concentrates.

[0044] Suitable solid carriers include, for example, talc, titanium dioxide, pyrophyllite clay, silica, attapulgite clay, kieselguhr, chalk, diatomaceous earth, lime, calcium carbonate, bentonite clay, fuller's earth, cottonseed hulls, wheat flour, soy flour, pumice, wood flour, walnut hulls, and lignin.

[0045] A wide variety of surfactants are advantageously employed in both the liquid and solid compositions, particularly those designed to be diluted with a carrier before application. These agents, when used, will usually comprise from 0.01% to 15% by weight of the formulation. They may be anionic, cationic, nonionic, or polymeric in character and may be used as emulsifying agents, wetting agents, suspending agents, or for other purposes. Typical surfactants include Tween 20, salts of alkyl sulfates, such as diethanolammonium lauryl sulfate; alkylarylsulfonate salts, such as calcium dodecylbenzenesulfonate; alkylphenol-alkylene oxide adducts, such as nonylphenol-C.sub.18 ethoxylate; alcohol-alkylene oxide adducts, such as tridecyl alcohol-C.sub.16 ethoxylate; soaps, such as sodium stearate; alkylnaphthalenesulfonate salts, such as sodium dibutylnaphthalenesulfonate; dialkyl esters of sulfosuccinate salts, such as sodium di(2-ethylhexyl)sulfosuccinate; sorbitol esters, such as sorbitol oleate; quaternary amines, such as lauryltrimethylammonium chloride; polyethylene glycol esters of fatty acids, such as polyethylene glycol stearate; block copolymers of ethylene oxide and propylene oxide; and salts of mono- and dialkylphosphate esters.

[0046] Other adjuvants commonly used in agricultural compositions include crystallization inhibitors, viscosity modifiers, suspending agents, spray droplet modifiers, pigments, antioxidants, foaming agents, antifoaming agents, light-blocking agents, compatibilizers, defoamers, sequestering agents, neutralizing agents and buffers, corrosion inhibitors, dyes, odorants, spreading agents, penetration aids, micronutrients, emollients, lubricants and adhesives.

[0047] In addition, other agrochemically active ingredients or compositions may be combined with the compositions of the present invention and used in the methods of the present invention, and applied simultaneously or sequentially with the compositions of the present invention. When applied sequentially, the compositions of the present invention may be applied to plants during a different growth phase than the other agrochemically active ingredients. When applied simultaneously, these additional active ingredients may be formulated together with the compositions of the present invention or mixed, for example, in a spray tank. These additional agrochemically active ingredients may be fungicides, herbicides, insecticides, bactericides, acaricides, nematicides, growth stimulants, systemic acquired resistance compounds, and / or plant growth regulators.

[0048] Pesticides are referred to herein using their common names and are known, for example, from "The Pesticide Manual", 19th Ed., British Crop Protection Council 2021.

[0049] The mixtures according to the invention can be mixed with one or more pesticides known in the art.

[0050] The mixture comprising cyclothiazomycin C and malonomycin can be used in the form of agrochemical compositions, and can be applied simultaneously or sequentially to crop areas or treated plants with additional compounds.These additional compounds can be, for example, fertilizers or trace element sources or other preparations that affect plant growth.They can also be selective or non-selective herbicides, as well as insecticides, fungicides, bactericides, nematicides, molluscicides, or mixtures of several of these preparations, and if necessary, also contain additional carriers, surfactants, or application-promoting adjuvants commonly used in the formulation field.

[0051] The mixture containing cyclothiazomycin C and malonomycin can be used in the form of a (fungicidal) composition for the control or protection against plant pathogenic microorganisms, which contains as active ingredients cyclothiazomycin C and malonomycin and at least one of the above-mentioned adjuvants.

[0052] Thus, the present invention provides a composition, preferably a fungicidal composition, comprising cyclothiazomycin C, malonomycin, an agriculturally acceptable carrier, and optionally an adjuvant. An agriculturally acceptable carrier is, for example, a carrier suitable for agricultural use. Agricultural carriers are well known in the art. Preferably, the composition may contain, in addition to cyclothiazomycin C and malonomycin, at least one or more pesticidally active compounds, such as additional fungicidal active ingredients.

[0053] The compositions according to the invention may also comprise further solid or liquid auxiliaries, such as stabilizers, for example non-epoxidized or epoxidized vegetable oils (for example epoxidized palm oil, rapeseed oil or soybean oil), antifoaming agents, for example silicone oils, preservatives, viscosity regulators, binders and / or tackifiers, fertilizers or other active ingredients for achieving specific effects, for example fungicides, bactericides / fungicides, nematicides, plant activators, molluscicides or herbicides.

[0054] The compositions according to the invention are prepared in a manner known per se in the absence of auxiliaries, for example by grinding, sieving and / or compressing the solid active ingredient, and in the presence of at least one auxiliary, for example by intimately mixing the active ingredient with one or more auxiliaries and / or grinding. These methods for preparing the compositions and the use of compound (I) for preparing these compositions are also the subject of the present invention.

[0055] Another aspect of the present invention relates to the use of a composition comprising cyclothiazomycin C and malonomycin, or a fungicidal or insecticidal mixture comprising cyclothiazomycin C and malonomycin, in admixture with other fungicides or insecticides as mentioned above, for the control or prevention of infestation by insects or by phytopathogenic microorganisms, preferably fungal organisms, on plants, such as useful plants, e.g. crop plants, their propagation material, e.g. seeds, harvested crops, e.g. harvested food crops, or non-living material.

[0056] A further aspect of the present invention relates to a method for controlling or preventing infestation by insects, or by plant pathogenic or spoilage microorganisms, or by organisms potentially harmful to humans, in particular fungal organisms, on plants, such as useful plants, e.g. crop plants, their propagation material, e.g. seeds, harvested crops, e.g. harvested food crops, or non-living material, which method comprises applying a mixture comprising cyclothiazomycin C and malonomycin as active ingredients to the plant, plant parts or their habitat, their propagation material, or any part of the non-living material.

[0057] Control or prevention means the reduction of infestation by plant pathogenic or spoilage microorganisms or by organisms potentially harmful to humans, especially fungal organisms, to the extent that an improvement can be demonstrated.

[0058] Preferred methods for controlling or preventing infestation of crop plants by phytopathogenic microorganisms, particularly fungal organisms, or insects include foliar application of a compound of formula (I) and a compound of formula (II), or an agrochemical composition containing at least one of the aforementioned compounds. The frequency and amount of application will depend on the risk of infestation by the corresponding pathogen. However, the compositions of the present invention can also be applied to plants through the roots via the soil by drenching the plant habitat with a liquid formulation or by applying the compound to the soil in solid form, for example, in granular form (soil application). In rice crops, such granules can be applied to flooded rice fields. The mixtures of the present invention can also be applied to seeds by impregnating the seeds or tubers with a liquid formulation of the fungicide or by coating them with a solid formulation (coating).

[0059] Formulations, e.g., compositions, containing the mixtures of the present invention and, if desired, solid or liquid adjuvants or monomers encapsulating the mixtures of the present invention, can typically be prepared in a known manner by intimately mixing and / or grinding the compounds together with extenders, e.g., solvents, solid carriers, and, optionally, surface-active compounds (surfactants).

[0060] The weight / molar ratio of cyclothiazamicin C to malonomycin is preferably 10:1 to 1:500, more preferably 1:1 to 1:300, and most preferably 1:10 to 1:100.

[0061] The preferred application rate is generally 0.1 g to 6 kg of active ingredient (ai; the total weight of cyclothiazamicin C and maronomycin) per hectare (ha), preferably 0.1 g to 1 kg ai / ha, and most preferably 10 g to 800 g ai / ha.

[0062] When the combinations according to the invention are used for seed treatment, a rate of from 0.001 to 100 g of active ingredient per kg of seed, preferably from 0.01 to 10 g per kg of seed, is generally sufficient.

[0063] Preferably, the composition comprising cyclothiazomycin C and malonomycin according to the invention is applied either prophylactically, meaning before the onset of disease, or curatively, meaning after the onset of disease.

[0064] Certain mixtures of cyclothiazamycin C and malonomycin may exhibit synergistic effects. This occurs whenever the effect of the combination of active ingredients is greater than the sum of the effects of the individual components. 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 combinations". Weeds, Vol. 15, pages 20-22; 1967): ppm = milligrams of active ingredient (=ai) per liter of spray mixture X = % effect of active ingredient A) using p ppm of active ingredient Y = % effect of active ingredient B) using q ppm of active ingredient.

[0065] According to COLBY, the expected (additive) effect of active ingredients A) + B) using p+q ppm of active ingredients is as follows:

number

[0066] If the actual observed effect (O) is greater than the expected effect (E), the effect of the combination is superadditive, i.e., a synergistic effect is present. In mathematical terms, synergy corresponds to a positive value for the difference (OE). If the activities are purely complementary and additive (expected activity), the difference (OE) is zero. A negative value for the difference (OE) indicates a loss of activity compared to the expected activity.

[0067] However, in addition to the actual synergistic action with respect to fungicidal activity, the composition according to the present invention may also have other unexpected advantageous properties.Examples of such advantageous properties that may be mentioned include: more advantageous decomposition; improved toxicological and / or ecotoxicological behavior; or improved useful plant properties, including emergence, crop yield, more developed root system, increased tillers, increased plant height, larger leaf blades, fewer dead basal leaves, stronger tillers, greener leaf color, less fertilizer required, less seed required, more productive tillers, earlier flowering, earlier grain maturation, less plant lodging (lodging), increased shoot growth, improved plant vigor, and earlier germination.

[0068] The compositions of the present invention may be in any conventional form, such as, for example, two-part systems, dry seed treatment powders (DS), seed treatment emulsions (ES), seed treatment flowable concentrates (FS), seed treatment solutions (LS), seed treatment water dispersible powders (WS), seed treatment capsule suspensions (CF), seed treatment gels (GF), emulsion concentrates (EC), suspension concentrates (SC), suspoemulsions (SE), capsule suspensions (CS), water dispersible granules (WG), emulsifiable granules (WG), They may be employed in the form of granules (EG), water-in-oil emulsions (EO), oil-in-water emulsions (EW), microemulsions (ME), oil dispersions (OD), oil-miscible flowables (OF), oil-miscible liquids (OL), soluble concentrates (SL), ultra-low volume suspensions (SU), ultra-low volume liquids (UL), technical concentrates (TK), dispersible concentrates (DC), wettable powders (WP), or any technically feasible formulation in combination with agriculturally acceptable adjuvants.

[0069] Such compositions can be prepared in a conventional manner, for example, by mixing the active ingredient with suitable formulation inerts (diluents, solvents, fillers, and optionally other formulation ingredients such as surfactants, biocides, antifreeze agents, spreading agents, thickeners, and compounds providing auxiliary effects). Conventional sustained-release formulations can also be employed when long-lasting effectiveness is intended. In particular, formulations applied in spray form, 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, 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.

[0070] The seed dressing formulation is applied to seeds in a manner known per se, using the combination of the present invention and a diluent in a suitable seed dressing formulation form, such as an aqueous suspension or a dry powder form with good adhesion to the seeds. Such seed dressing formulations are known in the art. The seed dressing formulation may contain a single active ingredient or a combination of active ingredients in encapsulated form, for example, as slow-release capsules or microcapsules.

[0071] Generally, the formulation contains 0.01 to 90% by weight of the active agent, 0 to 20% by weight of an agriculturally acceptable surfactant, and 10 to 99.99% by weight of a solid or liquid inert compounding agent and adjuvants, including cyclothiazomycin C and malonomycin, optionally together with other active agents, particularly microbicides or preservatives. Concentrate forms of the composition generally contain about 2 to 80% by weight, preferably about 5 to 70% by weight, of the active agent. The application form of the formulation can contain, for example, 0.01 to 20% by weight, preferably 0.01 to 5% by weight, of the active agent. Commercially available products are preferably formulated as concentrates, but end users usually use diluted formulations. [Example]

[0072] The following examples illustrate the invention. Temperatures are given in degrees Celsius (°C), rh means relative humidity, and ppm means parts per million by weight. Colby, supra, see COLBY, SR, "Calculating synergistic and antagonistic responses of herbicide combinations." Weeds, Vol. 15, pages 20-22; 1967.

[0073] 1. Fungicidal activity of mixtures with malonomycin in liquid culture assays method: Cyclothiazomicin C was prepared as disclosed in WO 2015191789. A stock solution of cyclothiazomycin C was prepared in DMSO (up to 10 mg / ml). Malonomycin was prepared according to Law et al., 2018 (Nature Catalyses | VOL 1 | DECEMBER 2018 | 977-984). A stock solution of malonomycin was prepared in water with 0.025% Tween® 20.

[0074] An assay to test the effectiveness of a mixture of cyclothiazomycin C in combination with malonomycin for control of fungal pathogens in liquid culture assays was designed for 96-well plates.

[0075] [Table 1]

[0076] The 96-well plate design allows for the comparison of the disease control of the mixture to that of each single compound at the same rate. By comparing the estimated efficacy from the mixture to the efficacy calculated from the same mixture according to Colby, it is possible to describe whether the mixture is additive (efficacy is similar to the Colby calculation), synergistic (efficacy is better than the Colby calculation), or antagonistic (efficacy is worse than the Colby calculation). The same 96-well plate design also allows for the evaluation of whether the two compounds can be mixed at different use rates and mix ratios. The plate design outlined in Table 1 provides the following mix ratios, as shown in Table 2:

[0077] [Table 2]

[0078] A master plate was prepared containing 10x concentrated solutions of compound stock solutions diluted in water with 0.025% Tween 20. The concentrations of DMSO (from cyclothiazomycin C stock) and Tween 20 were kept constant in all cells of the master plate. 10 μl was transferred from the master plate to a 96-well test plate. Nutrient broth containing fungal spores / mycelium fragments was then added to the test plate to obtain a 1x final concentration of the test compound (as outlined in Table 1). The test plate was incubated in the dark at 24°C and 96% rh. Inhibition of fungal growth was determined photometrically after approximately 3 days, and the percentage of fungal growth reduction relative to untreated controls was calculated. The effectiveness of the mixtures was tested on various fungal species:

[0079] Zymoseptoria tritici (EPPO code: SEPTTR) Cryopreserved fungal conidia were mixed directly into nutrient broth (PDB: potato dextrose broth). Test plates were incubated at 24°C and growth inhibition was determined photometrically after 72 hours.

[0080] Fusarium culmorum (EPPO code: FUSACU) Cryopreserved fungal conidia were mixed directly into nutrient broth (PDB: potato dextrose broth). Test plates were incubated at 24°C and growth inhibition was determined photometrically after 72 hours.

[0081] Microdochium nivale (EPPO code: MONGNI) Cryopreserved fungal conidia were mixed directly into nutrient broth (PDB: potato dextrose broth). Test plates were incubated at 24°C and growth inhibition was determined photometrically after 72 hours.

[0082] Botrytis cinerea (EPPO code: BOTRCI) Cryopreserved fungal conidia were mixed directly into nutrient liquid medium (Vogel's minimal medium). Test plates were incubated at 24°C and growth inhibition was determined photometrically after 72 hours.

[0083] result:

[0084] [Table 3]

[0085] [Table 4]

[0086] [Table 5]

[0087] [Table 6]

[0088] [Table 7]

[0089] [Table 8]

[0090] [Table 9]

[0091] [Table 10]

[0092] Conclusion: Cyclothiazomycin C and malonomycin can be mixed to obtain complete or partial control of various fungal pathogens. The mixing ratio of the two compounds can vary widely within the mixture, yet still result in over 50% control of fungal growth. Surprisingly, the efficacy of many mixtures was superior to that predicted based on calculations from Colby, indicating that mixtures of cyclothiazomycin C and malonomycin have a synergistic effect on the control of fungal pathogens. This surprising synergy was observed in tests to control Zymoseptoria tritici, Fusarium culmorum, Microdochium nivale, and Botrytis cinerea.

[0093] 2. Fungicidal activity of a mixture of cyclothiazomycin C and malonomycin in the leaf disc assay method: Cyclothiazomicin C was prepared as disclosed in WO 2015191789. A stock solution of cyclothiazomycin C was prepared in DMSO (up to 10 mg / ml). Maronomycin was prepared according to Law et al., 2018 (Nature Catalyses | VOL 1 | DECEMBER 2018 | 977-984). A stock solution of maronomycin was prepared in water with 0.025% Tween 20.

[0094] The assay to test the effectiveness of a mixture of cyclothiazomycin C in combination with malonomycin for control of fungal pathogens in a leaf disc assay was designed for two plates.

[0095] Tables 11 and 12: Overview of Test Plate (1) and Test Plate (2), including the concentration of compounds sprayed per well. The upper number in the cell indicates the concentration of cyclothiazomycin C (ppm), and the lower number indicates the concentration of malonomycin (ppm). Column (1) holds the dilution series of cyclothiazomycin C, and row (2-D) holds the dilution series of malonomycin. Well 2-D-(1) represents the untreated test. This design was applied to a test involving Puccinia recondita (EPPO code: PUCCRE) at preventive and curative spray timings.

[0096] [Table 11]

[0097] [Table 12]

[0098] [Table 13]

[0099] A first set of master plates was prepared containing a 1x concentrated spray solution of cyclothiazomycin C stock diluted in water at the concentrations shown in Table 11 or Table 12, respectively. Each well contained 2% DMSO and 0.025% Tween 20. A second set of master plates was therefore prepared containing a 1x concentrated malonomycin stock diluted in water. Each well of the second set contained 0.025% Tween 20. Leaf sections placed on agar in the plates were sprayed with 8 μl of the solution from the master plate containing cyclothiazomycin C, allowed to dry, and then sprayed 2 hours later with 8 μl of the solution from the master plate containing malonomycin. After the second spray had dried, the leaf sections were infected with fungal spores to provide a preventative application timing. Alternatively, leaf sections infected one day before spraying the compound were used to provide a curative spray timing. Additionally, several plates were prepared in which leaf sections were sprayed twice in the absence of test compound (containing only DMSO and Tween 20) to represent untreated test samples. Percent leaf coverage of disease symptoms was assessed for each leaf section. Reduction in percent leaf coverage relative to the untreated test was calculated. The efficacy of mixtures was tested in duplicate. Reported efficacy values ​​are the average of the two replicates.

[0100] Preventive spray timing for Puccinia recondita (EPPO code: PUCCRE). Wheat (Kanzler variety) leaf segments are placed on agar in multiwell plates and sprayed with the test solution (8 ul per well). After drying, the leaf segments are inoculated with a fungal spore suspension. After appropriate incubation, the activity of the compounds is evaluated 8 dpi (days post-inoculation) as preventative fungicidal activity.

[0101] Puccinia recondita (EPPO code: PUCCRE) with therapeutic spray timing. Wheat (Kanzler variety) leaf segments are placed on agar in multi-well plates. The leaf segments are then inoculated with a fungal spore suspension. One day after inoculation, the test solution is sprayed (8 ul per well). After appropriate incubation, the activity of the compound is evaluated 8 dpi (days after inoculation) as curative fungicidal activity.

[0102] result:

[0103] [Table 14]

[0104] [Table 15]

[0105] [Table 16]

[0106] [Table 17]

[0107] Conclusion: Cyclothiazomicin C and malonomycin can be mixed to obtain complete or partial control of fungal pathogens. The mixing ratio of the two compounds can vary widely in the mixture, but still results in greater than 50% control of fungal growth when sprayed on leaves. Surprisingly, the efficacy of many mixtures was superior to that predicted based on calculations from Colby, indicating that mixtures of cyclothiazomycin C and malonomycin have a synergistic effect on the control of fungal pathogens when sprayed on leaves. This surprising synergy was observed in tests to control Puccinia recondita.

Claims

1. A composition comprising cyclothiazomycin C and malonomycin.

2. 2. The composition of claim 1, wherein the ratio of cyclothiazomycin C to malonomycin is from 10:1 to 1:500 by weight.

3. 3. The composition of claim 2, wherein the ratio of cyclothiazomycin C to malonomycin is from 1:1 to 1:300 by weight.

4. A pesticide composition comprising a fungicidally effective amount of a mixture comprising cyclothiazomycin C and malonomycin.

5. 5. The pesticide composition of claim 4, further comprising an agriculturally acceptable diluent or carrier.

6. The pesticide composition according to claim 4 or 5, comprising the composition according to any one of claims 1 to 3.

7. 10. A method for controlling or preventing fungal infestation of plants, wherein a fungicidally effective amount of a composition according to any one of claims 1 to 6 is applied to the plant, its parts or its habitat.

8. 8. The method of claim 7, wherein the composition is applied to plants at a rate of 0.1 g to 6 kg of the combined weight of cyclothiazomicin C and malonomycin per hectare (ha).

9. 8. The method of claim 7, wherein the composition is applied to seeds at a rate of 0.001 to 100 g of the combined weight of cyclothiazamicin C and malonomycin per kg of seeds.

10. The method according to any one of claims 7 to 9, wherein the plant is selected from the group consisting of cereals and pulses.

11. The fungus may be selected from the group consisting of Fusarium avenaceum, Fusarium culmorum, Fusarium langsethiae, Fusarium poae, Fusarium sporotrichioides, Fusarium tricinctum, Fusarium virguliforme, Fusarium verticillioides, Fusarium subglutinans, and the like.

11. The method according to any one of claims 7 to 10, wherein the selected fungus is selected from the group consisting of Puccinia tritici, Puccinia triticina (= P. recondita), Mycosphaerella fijiensis, Puccinia striiformis, Magnaporthe grisea and Rhizoctonia solani.

12. Use of a composition comprising cyclothiazomycin C and malonomycin according to any one of claims 1 to 3 as an antifungal or fungicidal agent.

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