An antifungal composition comprising natamycin and at least one insecticide

Abstract

The present invention relates to a composition comprising natamycin and at least one insecticide that interferes with the nervous and / or muscular systems of insects and nematodes. The present invention further relates to the use of said composition comprising natamycin and at least one insecticide that interferes with the nervous and / or muscular systems of insects and nematodes, preferably for protecting plants or plant parts, for improving the growth and / or yield of agricultural plants, and for protecting the soil and / or growth substrate.

Description

Technical Field

[0001] The present invention relates to compositions for controlling fungal diseases of plants and plant parts and improving plant growth and yield.

Background Art

[0002] Plants can be attacked by various phytopathogenic fungi that cause significant losses to crops worldwide. In addition, the growth of fungi can cause loss of nutrients, formation of off-odors, and destruction of tissues, leading to a decrease in quality after processing. In many cases, fungal infections occur in the field and then, if conditions are favorable, the fungus grows during storage, resulting in post-harvest losses such as in grains, seeds, bulbs, seed tubers, fruits, and vegetables, and mold growth in processed foods such as breakfast cereals, juices, or cut fruits.

[0003] Phytopathogenic fungi in the soil, in the field (on crop parts such as seeds, bulbs, and seedlings), and after harvest (on, for example, grains, vegetables, and fruits) are generally controlled by fungicides, especially synthetic fungicides. However, many fungicides lose their activity over time with repeated use, and resistant bacteria develop. This phenomenon has occurred even with newly launched fungicides, for example, point mutations related to strobilurin fungicides have occurred in the relevant bacteria. More generally, resistance develops gradually, such as the expression of resistance of Zymoseptoria tritici to azole fungicides, so that the pathogen population gradually loses sensitivity. The development of resistance always leads to an increase in the number of treatments and the application of larger amounts and / or two or more fungicides.

[0004] Many of the currently commercially available fungicides are destructive to natural ecosystems, for example, by contaminating water sources or having adverse effects on non-target organisms. In addition to environmental pollution, human health issues, especially worker safety, have also become important problems. Furthermore, when consuming agricultural products, the fact that harmful fungicides remain at high levels exceeding the residue standards is a serious problem. Concerns among consumers and government regulatory authorities are increasing, leading to strengthened regulations in the EU, the United States, Japan, and many other countries.

[0005] Although many commercially available fungicides are available and widely used, it can be concluded that fungi occur in almost all crops and harvested agricultural products. Also, in the agricultural field, it can be concluded that there is a high need for environmentally friendly alternatives to the harmful synthetic fungicides currently in use.

[0006] For decades, the polyene macrolide antifungal agent natamycin has been mainly used to prevent mold growth in foods such as cheese and dry fermented sausages. Natamycin was first reported in 1957 and is produced by fermentation using bacteria of the genus Streptomyces (e.g., Streptomyces natalensis). Currently, this natural antimicrobial is widely used worldwide as a food additive.

[0007] Natamycin has a long history of being used safely, and more importantly, no resistant bacteria have been discovered in nature so far. Over the years, many documents have been published describing the potential use of natamycin in many agricultural applications. However, it has been observed that there has been little commercial use of natamycin in agriculture. This seems to be due to the fact that the price of natamycin, which always needs to be purified before use, is high for agricultural applications, especially for field use. SUMMARY OF THE INVENTION PROBLEMS TO BE SOLVED BY THE INVENTION

[0008] There is a need for improved efficacy of natamycin for fungal control and reduction of economic losses in agriculture, particularly for commercial use in agriculture.

Means for Solving the Problems

[0009] The present invention provides an antifungal composition comprising natamycin and at least one insecticide that interferes with the nervous system and / or muscle tissue of insects and nematodes. The at least one insecticide preferably belongs to subgroups 2, 3, 4, 6 and / or 28 described in the document "IRAC Mode of Action Classification Scheme" (June 2019; version 9.3).

[0010] However, a biopesticide called funnel spider peptide, or "SPEAR biopesticide", may be an exception. This peptide is thought to target nicotinic acetylcholine receptors and has recently been discovered to have a novel "neuro-muscular" mode of action. This funnel spider peptide has been inserted into a new class 32, which may be included as an insecticide that interferes with the nervous system and / or muscle system of insects and nematodes.

[0011] Insecticides that disrupt the action of the nerves or muscles of insects have been found to enhance the efficacy of natamycin against pathogenic fungi. This enhancement of the efficacy of natamycin has not been found when used in combination with insecticides having other modes of action such as growth regulation, microbial disruption of the insect midgut, energy metabolism (respiration), or unknown or non-specific modes of action (see "IRAC Mode of Action Classification Scheme" (June 2019; version 9.3)).

[0012] Polyene fungicides such as natamycin have been reported to interact with cell membranes, particularly fungal membrane sterols. Although the mechanism of action of natamycin has been reported to be different from that of other polyene fungicides, there are also reports that natamycin interacts with ergosterol, the major sterol in fungi, thereby regulating membrane fluidity and the function of membrane-bound enzymes (te Welscher et al., 2008. J Biol Chem 283: 6393-6401). Ergosterol is the most abundant sterol in the fungal cell membrane and regulates membrane permeability and fluidity (Douglas and Konopka, 2014. Annu Rev Microbiol 68: 377-393).

[0013] Without being bound by theory, the marked stimulation of the activity of natamycin by a group of insecticides that interfere with the nervous system and / or muscular system of insects and nematodes is thought to be based on the mechanism that in fungi or fungal spores, these insecticides affect the cell membrane and / or receptor, making the cell membrane and / or receptor more vulnerable to natamycin. In these vulnerable membranes, natamycin can effectively interfere with the ergosterol in the cell membrane. As a result, the generation of spores and hyphae by fungi is more effectively inhibited by natamycin.

[0014] It has been pointed out that "triazoles" such as propiconazole inhibit the synthesis of ergosterol. Therefore, since "triazole fungicides" do not directly interfere with the components of the cell membrane, it is less likely that triazole compounds exhibit a synergistic effect with the insecticides in subgroups 2, 3, 4, 6 and / or 28.

[0015] The antifungal composition according to the present invention preferably contains 1% to 98% (w / w) of natamycin (preferably 6% to 60% (w / w) of natamycin) and 1% to 99% (w / w) (preferably 5 to 50% (w / w)) of at least one of the above-mentioned insecticides. The ratio of natamycin to at least one of the above-mentioned insecticides in subgroups 2, 3, 4, 6 and / or 28 is preferably 1:1 to 1:5000 (w / w).

[0016] A preferred antifungal composition according to the present invention contains natamycin and at least one insecticide selected from fipronil, lambda-cyhalothrin, acetamiprid, clothianidin, imidacloprid, thiacloprid, thiamethoxam, abamectin and chlorantraniliprole. A more preferred antifungal composition according to the present invention contains at least two insecticides selected from fipronil, lambda-cyhalothrin, acetamiprid, clothianidin, imidacloprid, thiacloprid, thiamethoxam, abamectin and chlorantraniliprole.

[0017] The antifungal composition according to the present invention may further contain an agriculturally acceptable carrier.

[0018] The antifungal composition according to the present invention is preferably an aqueous or oily composition.

[0019] In one embodiment, the natamycin in the antifungal composition according to the present invention is produced by fermenting biomass by a fermenting organism.

[0020] The natamycin in the antifungal composition according to the present invention is preferably fractionated to an average particle diameter (volume particle diameter) of 0.5 to 3 μm, for example by grinding.

[0021] The antifungal composition according to the present invention preferably further contains an insoluble polyelectrolyte complex of a polyanion (such as a lignin compound) and a polycation (such as chitosan or poly-allylamine) in a relative amount of 1:2 to 60:1 (w / w).

[0022] The present invention further provides a method for protecting agricultural plants or plant parts, which includes preparing the antifungal composition according to the present invention and applying the composition to the agricultural plants or plant parts. Preferred plant parts are seeds, bulbs, fruits or vegetables.

[0023] The present invention further provides a method for improving the growth and / or yield of agricultural plants, which includes preparing the composition according to the present invention and bringing the plants into contact with the composition.

[0024] The present invention further provides a method for protecting soil and / or a growth substrate, which includes applying the composition according to the present invention to the soil and / or the growth substrate. A preferred growth substrate is a mushroom growth substrate.

[0025] In a preferred method according to the present invention, the antifungal composition is used undiluted or diluted up to 10 times with an aqueous solution or oil. 6 The antifungal composition may be used undiluted or diluted up to 100 times with an aqueous solution before applying the composition to the seeds. The antifungal composition may preferably be diluted 10 to 10 times in an aqueous solution or oil before being applied to plants, plant parts, soil and / or a growth substrate. 6 times.

[0026] The present invention further provides the use of the antifungal composition according to the present invention for protecting plants, plant parts, soil and / or a growth substrate from fungi.

Embodiments for Carrying Out the Invention

[0027] <Definition> As used herein, the term "suspension concentrate" means a suspension of solid particles in a liquid, which is intended to be diluted with water before use.

[0028] As used herein, the term "soluble liquid" means a solution in a liquid, which is intended to be diluted with water before use. The above liquid may be an aqueous liquid or a non-aqueous liquid, such as a petroleum solvent like xylene or kerosene.

[0029] As used herein, the term "suspo emulsion" means a suspension of solid particles in water, which is combined with an oil phase in the form of an emulsion and is intended to be diluted with water before use.

[0030] As used herein, the term "dispersion concentrate" means a dispersion of solid particles in a liquid, which is intended to be diluted with water before use.

[0031] As used herein, the term "water-dispersible granule" means a formulation in the form of granules that can be dispersed in water to form a dispersion such as a suspension or a solution.

[0032] As used herein, the term "wettable powder" means a powder formulation that is intended to be mixed with water or another liquid before use.

[0033] As used herein, the term "water slurriable powder" means a powder formulation that is intended to be made into a slurry in water before use.

[0034] As used herein, the term "surfactant" means an ionic or non-ionic surfactant. Examples of surfactants include alkyl-terminated capped ethoxylated glycols, alkyl-terminated capped alkyl block alkoxylated glycols, dialkyl sulfosuccinates, phosphate esters, alkyl sulfonates, alkyl aryl sulfonates, tristerylphenol alkoxylates, natural or synthetic fatty acid alkoxylates, natural or synthetic fatty alcohol alkoxylates, alkoxylated alcohols (such as n-butyl alcohol polyglycol ether), block copolymers (such as ethylene oxide-propylene oxide block copolymers and ethylene oxide-butylene oxide block copolymers), or combinations thereof.

[0035] As used herein, the term "improvement in bioactivity" means an improvement in the healing, prevention, and / or sustained performance of the active ingredient.

[0036] As used herein, the term "plant part" refers to single cells, cell aggregates, and plant tissues (including tissue cultures). Examples of plant parts include, but are not limited to, pollen, ovary, leaf, embryo, root, root tip, anther, flower, fruit, shoot, scion, stock, seed, protoplast, and calli, etc., and preferably seeds.

[0037] <Antifungal composition> The present invention provides an antifungal composition comprising natamycin and at least one insecticide that interferes with the nervous and / or muscular systems of insects and nematodes. The at least one insecticide is preferably a subgroup 2 (GABA-gated chloride channel blocker), subgroup 3 (sodium channel modulator), subgroup 4 (competitive modulator of nicotinic acetylcholine receptors), subgroup 6 (allosteric modulator of glutamate-gated chloride channels) and / or subgroup 28 (ryanodine receptor modulator), and these subgroups are described in the document "IRAC Mode of Action Classification Scheme" (June 2019; version 9.3). It has surprisingly been found that the combination of one or more of the above insecticides with natamycin improves the biological activity of the natamycin, i.e., the bactericidal activity of natamycin.

[0038] A preferred insecticide is a GABA-gated chloride channel blocker. GABA-gated chloride channel blockers are thought to act on the nervous system of insects. Preferred GABA-gated chloride channel blockers are cyclodiene-based, organochlorine-based and / or phenylpyrazole-based. A preferred GABA-gated chloride channel blocker is fipronil ((±)-5-amino-1-(2,6-dichloro-α,α,α-trifluoro-p-tolyl)-4-trifluoromethylsulfinylpyrazole-3-carbonitrile).

[0039] Preferred insecticides are sodium channel modulators. Sodium channel modulators are thought to act on the insect nervous system. Preferred sodium channel modulators are pyrethroids and / or pyrethrins. A preferred sodium channel modulator is cyhalothrin ([cyano(3-phenoxyphenyl)methyl]-3-[(Z)-2-chloro-3,3,3-trifluoropropen-1-yl]-2,2-dimethylcyclopropane-1-carboxylate). Preferably, it is λ-cyhalothrin ([(R)-cyano(3-phenoxyphenyl)methyl](1S,3S)-3-[(Z)-2-chloro-3,3-trifluoropropen-1-yl]-2,2-dimethylcyclopropane-1-carboxylate).

[0040] Preferred insecticides are competitive modulators of nicotinic acetylcholine receptors. Competitive modulators of nicotinic acetylcholine receptors are thought to act on the insect nervous system. Preferred competitive modulators of nicotinic acetylcholine receptors are neonicotinoids. Preferred competitive modulators of nicotinic acetylcholine receptors are acetamiprid (N-[(6-chloropyridin-3-yl)methyl]-N’-cyano-N-methylethanamide), imidacloprid (N-[1-[(6-chloropyridin-3-yl)methyl]-4,5-dihydroimidazol-2-yl]nitramide, thiacloprid ([3-[(6-chloropyridin-3-yl)methyl]-1,3-thiazolidin-2-ylidene]cyanamide), and / or thiamethoxam (N-[3-[(2-chloro-1,3-thiazol-5-yl)methyl]-5-methyl-1,3,5-oxadiazinan-4-ylidene]nitramide).

[0041] Preferred insecticides are allosteric regulators of the glutamate-gated chloride channel. Allosteric regulators of the glutamate-gated chloride channel are thought to act on the insect nervous system. Preferred allosteric regulators of the glutamate-gated chloride channel are avermectin and / or milbemycin. A preferred allosteric regulator of the glutamate-gated chloride channel is abamectin ((1’R,2R,3S,4’S,6S,8’R,10’E,12’S,13’S,14’E,16’E,20’R,21’R,24’S)-2-butane-2-yl-21’,24’-dihydroxy-12’-[(2R,4S,5S,6S)-5-[(2S,4S,5S,6S)-5-hydroxy-4-methoxy-6-methyloxan-2-yl]oxy-4-methoxy-6-methyloxan-2-yl]oxy-3,11’,13’,22’-tetramethylspiro[2,3-dihydropyran-6,6’-3,7,19-trioxatetracyclo[15.6.1.14,8.020,24]pentacosa-10,14,16,22-tetraene]-2’-one; (1’R,2R,3S,4’S,6S,8’R,10’E,12’S,13’S,14’E,16’E,20’R,21’R,24’S)-21’,24’-dihydroxy-12’-[(2R,4S,5S,6S)-5-[(2S,4S,5S, 6S)-5-hydroxy-4-methoxy-6-methyloxan-2-yl]oxy-4-methoxy-6-methyloxan-2-yl]oxy-3,11’,13’,22’-tetramethyl-2-propan-2-ylspiro[2,3-dihydropyran-6,6’-3,7,19-trioxatetracyclo[15.6.1.14,8.020,24]penta-10,14,16,22-tetraene]-2'-one), emamectin benzoate([(2S,3S,4S,6S)-6-[(2S,3S,4S,6R)-6-[(1’R,2R,3S,4’S,6S,8’R,10’E,12’S,13’S,14’E,16’E,20’R,21’R,24’S)-2-[(2S)-butan-2-yl]-21’,24’-dihydroxy-3,11’,13’,22’-tetramethyl-2’-oxospiro[2,3-dihydropyran-6,6’-3,7,19-trioxatetracyclo[15. 6.1.14,8. 020,24]penta-10,14,16,22-tetraene]-12’-yl]oxy-4-methoxy-2-methyloxan-3-yl]oxy-4-methoxy-2-methyloxan-3-yl]-methylazanium; benzoate), lepimectin([(1R,4S,5’S,6R,6’R,8R,10E,12R,13S,14E,16E,20R,21R,24S)-6’-ethyl-21,24-dihydroxy-5’,11,13,22-tetramethyl-2-oxospiro[3,7,19-trioxatetracyclo[15.6.1.14,8.020,24]penta-10,14,16,22-tetraene-6,2’-oxane]-12-yl](2Z)-2-methoxyimino-2-phenylacetate), and / or milbemectin((1R,4S,5’S,6R,6’R,8R,10E,13R,14E,16E,20R,21R,24S)-21,24-dihydroxy-5’,6’,11,13,22-pentamethylspiro[3,7,19-trioxatetracyclo[15.6.1.14,8.020,24]penta-10,14,16,22-tetraene-6,2’-oxane]-2-one).

[0042] Preferred insecticides are ryanodine receptor regulators. Ryanodine receptor regulators are thought to act on the insect nervous system. Preferred ryanodine receptor regulators are diamides. A preferred diamide is chlorantraniliprole (5-bromo-N-[4-chloro-2-methyl-6-(methylcarbamoyl)phenyl]-2-(3-chloropyridin-2-yl)pyrazole-3-carboxamide).

[0043] The composition of the present invention preferably contains 1% to 98% (w / w) of natamycin, preferably 6% to 60% (w / w) of natamycin. The composition of the present invention preferably contains 1% to 99% (w / w), preferably 5 to 50% (w / w) of at least one of the above insecticides. The stock solution composition of the present invention preferably contains 1% to 98% (w / w) of natamycin, preferably 6% to 60% (w / w) of natamycin and 1% to 90% (w / w), preferably 5% to 50% (w / w) of at least one of the above insecticides.

[0044] A synergistic effect between the bactericidal activity of natamycin and insecticides of subgroups 2, 3, 4, 6 and / or 28 (refer to the IRAC mode of action classification system) is observed at a wide range of insecticide concentrations. This is thought to be because the insecticide affects the proteins in the cell membrane of the fungus, making this membrane more vulnerable to natamycin. As a result, the generation of spores and hyphae is more effectively inhibited by natamycin.

[0045] The composition is preferably characterized by the ratio (w / w) of natamycin:insecticide. It should be noted that it is difficult to determine the upper limit range of the at least one insecticide, since the insecticide itself begins to have bactericidal activity at a higher concentration. Experiments suggest that there may be no upper limit range for the insecticide. In the examples, it has been shown that a synergistic effect regarding the bactericidal activity of natamycin can be observed at a ratio of 1:2500, and even 1:7520 (w / w) (natamycin:insecticide). The preferred ratio of natamycin to the insecticides of subgroups 2, 3, 4, 6 and / or 28, preferably to at least one insecticide selected from fipronil, lambda-cyhalothrin, acetamiprid, clothianidin, imidacloprid, thiacloprid, thiamethoxam, abamectin and chlorantraniliprole is thus (natamycin:insecticide) 1:1 (w / w) to 1:10,000 (w / w), for example 1:2 to 1:250, for example 1:50 to 1:100. Preferred ratios include 1:2, 1:3, 1:4 and 1:5.

[0046] Regarding application to seeds and soil, it will be understood by those skilled in the art that the above ratio is preferably (natamycin:insecticide) 1:1 to 1:100 (w / w). This is because there is a need to treat the seeds with a large amount of insecticide so that the protection by the insecticide is extended to the embryo, preferably up to the seedling stage.

[0047] The composition of the present invention is preferably fractionated, for example, by grinding using a bead mill such as Dynomill (registered trademark). The volume-based average particle diameter of natamycin is preferably 0.2 μm to 10 μm, more preferably 0.5 μm to 5 μm, and even more preferably 0.5 μm to 2 μm. The method for determining the volume-based average particle diameter of the composition according to the present invention is known to those skilled in the art. For example, Hukkanen and Braatz, 2003. Sensors and Actuators B 96: 451-459 discuss various methods that can be used to determine the average particle diameter of a composition, including forward light scattering and ultrasonic extinction. A preferred method is, for example, based on laser diffraction analysis using an Analysette 22-MicroTec plus laser particle size analyzer (Fritsch, Idar-Oberstein, Germany).

[0048] The antifungal composition according to the present invention may contain cellular matter. The natamycin in the composition according to the present invention is preferably produced by fermentation of biomass by a fermenting organism, and the cellular matter present in the composition is from the natamycin-producing fermenting organism. Examples of the natamycin-producing fermenting organism include Streptomyces natalensis and Streptomyces gilvosporeus.

[0049] The cellular matter preferably contains a compound that is the residue of a natamycin-producing bacterium or a compound excreted by a natamycin-producing bacterium. Examples of such compounds include compounds of the cell envelope containing the cell membrane and cell wall of a natamycin-producing bacterium. Such compounds include phospholipids such as phospholipids and glycolipids, and upon hydrolysis such as the addition of sodium hydroxide, they become fatty acids such as C16-C18 fatty acids.

[0050] Methods for producing natamycin by fermenting biomass with fermenting bacteria such as Streptomyces natalensis and Streptomyces gilvosporeus are known in the art. Methods for separating and purifying the produced natamycin from the bulk of the biomass are known in the art. For example, disruption of the biomass can result in lysis and destruction of all cells of the production organism. The broth containing the obtained natamycin may be filtered to obtain a filter cake, and then treated with alcohol (preferably methanol and / or ethanol) to disrupt the biomass and dissolve at least a portion of the natamycin. If necessary, the pH may be increased to solubilize the natamycin. Subsequent neutralization will cause at least a portion of the natamycin to precipitate.

[0051] The composition of the present invention is preferably an aqueous or non-aqueous (preferably oily) concentrated stock composition that may be diluted with a suitable diluent such as water or oil before use, or an aqueous or non-aqueous ready-to-use composition.

[0052] The composition of the present invention can be used for soil treatment or for preparing seed treatment materials such as seed dressing or seed coating, coating emulsions (e.g., for field fruits or plants), waxes applied to fruits (e.g., pineapples, oranges or apples), and oils sprayed onto field plants (e.g., bananas). The composition of the present invention also includes concentrated dry compositions (e.g., granules, powders and / or tablets) that can be used to prepare compositions for dipping, spraying or soaking agricultural products.

[0053] The antifungal agent composition of the present invention is preferably a suspension concentrate (SC), water-dispersible granule (WG), wettable powder (WP), suspo emulsion (oil-based) (SE), oil dispersion (OD), dispersion concentrate (DC), dry powder seed treatment composition (DS), water slurriable powder (WS), flowable seed treatment composition (FS), water-dispersible granule seed treatment composition (WG), suspo emulsion (SE), or soluble liquid (SL).

[0054] The antifungal composition of the present invention is a polyelectrolyte complex of a polyanion and a polycation as described in the published international patent application WO2013 / 133706 incorporated herein by reference, or other encapsulation techniques known in the art, for example, liposomes, lipid structures in which the composition of the present invention is encapsulated, or composed of, for example, empty cells of yeast.

[0055] The above polyelectrolyte complex is a complex of polyelectrolytes (a polyanion and a polycation) having opposite charges that form strong electrostatic bonds. The above polyelectrolyte complex is the above polyelectrolyte complex. The above polyelectrolyte complex is an insoluble complex. This complex alone has no antibacterial effect. The polyelectrolyte complex has adhesiveness and includes a polar part (charged part) and a non-polar part. The aromatic sites in the complex may have an affinity for antibacterial compounds such as natamycin. The combination with the adhesiveness of the polyelectrolyte complex enables the antibacterial compound to be optimally deposited and adhered to the soil for use in agriculture, horticulture, and mushroom cultivation.

[0056] The polyelectrolyte complex consists of polyanions such as lignin compounds (e.g., lignosulfonic acid, humic acid, chondroitin sulfate, and poly(acrylic acid)), and polycations such as chitosan, ε-poly(L)-lysine, and poly-allylamine. The relative amounts thereof are 1:2 to 60:1 (w / w), more preferably 1:1 to 50:1, still more preferably 2:1 to 30:1, for example, about 2:1, about 5:1, about 10:1, about 15:1, about 20:1, about 25:1, and about 30:1 (w / w). The relative amounts of the polyanion (preferably a lignin compound) and the polycation (preferably chitosan) in the polyelectrolyte complex are most preferably about 5:1 (w / w).

[0057] The polyelectrolyte complex is preferably present in the composition of the present invention at a concentration of 5 to 800 g / l, more preferably 50 to 500 g / l, and most preferably 75 to 250 g / l.

[0058] The antifungal composition according to the present invention may further contain one or more agriculturally acceptable carriers. The above-mentioned agriculturally acceptable carriers are preferably stabilizers, wetting agents, dispersants, antifreeze agents, antifoaming agents and / or thickeners, or contain them. By adding a small amount of one or more agriculturally acceptable carriers, parameters such as the stability and / or efficacy of the composition according to the present invention can be affected, preferably improved. Adding a small amount of one or more agriculturally acceptable carriers preferably enhances the stability, efficacy and / or rainfastness of the composition according to the present invention.

[0059] When a stabilizer is present, it is preferably selected from carboxylic acids such as citric acid and acetic acid, and / or dodecylbenzenesulfonic acid, dodecylbenzenesulfonic acid orthophosphate, and their suitable salts. The composition of the present invention may also contain two or more different stabilizers. The stabilizer is preferably present in an amount of 0% to a maximum of 10% (w / v), more preferably 0.01% to a maximum of 5% (w / v), still more preferably 0.02% to a maximum of 1% (w / v), and still more preferably about 0.05% (w / v).

[0060] The wetting agent is preferably selected from dioctyl succinate, polyoxyethylene / polypropylene, and tristearyl sulfonate / phosphate. The composition of the present invention may also contain two or more different wetting agents. The wetting agent is preferably present in an amount of from 0% to a maximum of 10% (w / v), more preferably from 0.01% to a maximum of 5% (w / v), more preferably from 0.02% to a maximum of 1% (w / v), and more preferably about 0.05% (w / v).

[0061] When a dispersant is present, it is preferably selected from Morwet® D425, lignin sulfonate, alkyl polysaccharide, styrene acrylic polymer, acrylic copolymer, and ethoxylated tristyrenephenol phosphate (e.g., polyethoxylated fosforic acid). The composition of the present invention may also contain two or more different dispersants. The dispersant is preferably present in an amount of from 0% to a maximum of 10% (w / v), more preferably from 0.01% to a maximum of 5% (w / v), more preferably from 0.02% to a maximum of 1% (w / v), and more preferably about 0.05% (w / v).

[0062] When a cryoprotectant is present, it is preferably selected from glycerin, ethylene glycol, hexylene glycol, and propylene glycol. The composition of the present invention may also contain two or more different cryoprotectants. The cryoprotectant is preferably present in an amount of from 0% to a maximum of 10% (w / v), more preferably from 0.01% to a maximum of 5% (w / v), more preferably from 0.02% to a maximum of 1% (w / v), and more preferably about 0.05% (w / v).

[0063] When present, the defoaming agent is preferably selected from polymethylsiloxane, polydimethylsiloxane, simethicone octanol, and silicone oil. The composition of the present invention may also contain two or more different anti-foaming agents. The defoaming agent is preferably present in an amount of from 0% to a maximum of 10% (w / v), more preferably from 0.05% to a maximum of 5% (w / v), more preferably from 0.1% to a maximum of 1% (w / v), and more preferably about 0.05% (w / v).

[0064] When a thickening agent is present, it is preferably selected from agar, alginic acid, alginate, carrageenan, gellan gum, xanthan gum, succinoglycan gum, guar gum, acetylated distarch adipate, acetylated oxidised starch, arabinogalactan, ethyl cellulose, methyl cellulose, locust bean gum, sodium octenyl succinate starch, and triethyl citrate. Also, the composition of the present invention may contain two or more different thickening agents. The thickening agent is preferably present in an amount of from 0% to a maximum of 10% (w / v), more preferably from 0.01% to a maximum of 5% (w / v), more preferably from 0.02% to a maximum of 1% (w / v), and more preferably about 0.05% (w / v).

[0065] The composition according to the present invention provides a stable aqueous suspension containing a high concentration of natamycin and at most about 30% (w / v) of the at least one insecticide described above, and the aqueous suspension has improved bactericidal activity compared to commercially available formulations of the above natamycin in the presence of a relatively low amount of adjuvant as an agriculturally acceptable carrier.

[0066] In some embodiments, the composition further comprises one or more physical stabilizers and / or additives such as a buffering agent, a souring agent, and an anti-drift agent, a pigment, a safety agent, and a preservative.

[0067] <Method of Use> The present invention further provides a method for protecting a crop plant or a crop plant part, comprising providing natamycin and at least one insecticide that interferes with the nervous system and / or muscular system of insects and nematodes, and applying the natamycin and the at least one insecticide to the crop plant or the crop plant part such that the crop plant or the crop plant part is contacted with a sufficient amount of the natamycin and the at least one insecticide.

[0068] A preferred method for protecting a crop plant or a crop plant part comprises providing a composition according to the present invention and applying the composition to the crop plant or the plant part such that the crop plant or the crop plant part is contacted with a sufficient amount of the composition.

[0069] Preferably, the above method is for protecting a plant or a plant part from fungi (preferably molds).

[0070] As used herein, the terms "plant" and "crop" both mean plants, trees, or fungi cultivated for food, clothing, animal feed, biofuel, medicine, or other uses.

[0071] Preferably, the above plant parts are leaves, stems, seeds, bulbs, flower heads, seed tubers, roots, tubers, fruits and / or vegetables, and most preferably seeds, bulbs, fruits or vegetables.

[0072] The present invention further provides a method for improving the growth and / or yield of a crop plant, comprising providing natamycin and at least one insecticide that interferes with the nervous system and / or muscular system of insects and nematodes, and contacting the plant with the natamycin and the at least one insecticide that interferes with the nervous system and / or muscular system of insects and nematodes.

[0073] A preferred method for improving the growth and / or yield of a crop plant comprises providing a composition according to the present invention and contacting the plant with the composition.

[0074] A composition (preferably the composition of the present invention) comprising natamycin and at least one insecticide that interferes with the nervous system and / or muscular system of insects and nematodes can be applied in many different ways. For example, the above composition(s) can be applied as follows: (1) Optionally using a carrier such as wax or oil, spraying on plants in a field or greenhouse; (2) Soaking seeds, bulbs, or seed tubers; (3) Adding to plant parts (such as seeds or root systems) through, for example, soil; (4) Adding to plant parts (such as seeds, seed tubers, or bulbs) through seed coating or seed dressing; (5) Adding to the soil or growth substrate where seeds are planted or germinate, and / or where plants or mushrooms grow; (6) Adding to the applied water or watering system in, for example, a greenhouse or field; (7) Treating harvested plant parts such as bulbs, seeds, grains, soybeans, flowers, fruits, vegetables, or plants by, for example, dipping or spraying.

[0075] The composition of the present invention can be applied without dilution or after dilution. Usually, the composition of the present invention will be applied via an aqueous or oily diluent, via a dressing, coating, or wax. The composition according to the present invention is preferably undiluted or diluted. For seed treatment, the composition according to the present invention is preferably diluted up to 10 times or 100 times. For other uses in the method of the present invention, the composition according to the present invention is preferably diluted up to 10 times to 10 6 times in an aqueous solution or in oil. Since different treatments may be required depending on the use, it will be readily understood that the required amount of the composition of the present invention will vary for each use. However, generally, the amount of the ready-to-use composition such as an immersion or spray suspension required to treat a product (such as a growth substrate, soil, seeds, bulbs, field plants, or harvested fruits) will be 10 - 100,000 ppm of natamycin, more preferably 30 - 50,000 ppm of natamycin, and most preferably 50 - 5,000 ppm of natamycin.

[0076] The final amount of natamycin in soil or growth medium, on plants, or on harvested plant parts can be expressed in different ways. As a first example, for instance, the amount of natamycin on seeds, which is applied via, for example, seed dressing or seed coating, is 0.01 to 20.0 g of natamycin per kg of seeds, more preferably 0.05 to 5.0 g of natamycin per kg of seeds, and most preferably 0.1 to 2.0 g of natamycin per kg of seeds.

[0077] As a second example, the compositions of the present invention for dipping or spraying products such as flower bulbs, seed tubers, onions, apples, pears, bananas, and pineapples will generally contain 0.01 g / 1 to 100 g / l, preferably 0.03 g / l to 50 g / l, and most preferably 0.05 g / l to 5 g / l of natamycin.

[0078] After treating products such as flower bulbs, seed tubers, onions, apples, pears, bananas, and pineapples, the amount of natamycin on the products will typically be 0.01 to 20.0 mg / dm 2 and preferably 0.1 to 10.0 mg / dm 2 of it.

[0079] When treating a growth substrate such as a mushroom growth substrate, in a single spraying treatment, 0.01 to 5.0 g of natamycin per m 2 of the growth substrate is added, more preferably 0.02 to 1.0 g of natamycin per m 2 of the growth substrate.

[0080] When treating soil (for example, where vegetables or foliage plants are cultivated), 0.01 to 5.0 g of natamycin is applied per m 2 of the soil (preferably mixed into the surface layer of the soil), more preferably 0.02 to 1.0 g of natamycin per m 2From 0.1 to 1.0 g of natamycin is applied. When spraying on field crops, typical dosages are from 1 to 5000 g of natamycin per hectare, more preferably from 50 to 2000 g of natamycin per hectare. However, for crops such as bananas, the preferred dosage of natamycin is from 5 to 500 g per hectare, more preferably from 10 to 100 g per hectare.

[0081] Natamycin and at least one insecticide (preferably the composition of the present invention) that interferes with the nervous system and / or muscular system of insects and nematodes can be added to the growth medium, soil, plant or plant part at any suitable time using any suitable method, for example, before, during or after planting (e.g., of seeds, bulbs, seed tubers, cut flowers or young grass); during growth, after harvest or during storage in a field of fruits, vegetables, fruits or flower bulbs.

[0082] One aspect of the present invention provides the use of at least one insecticide that interferes with the nervous system and / or muscular system of insects and nematodes to enhance the biological activity of natamycin. By the above use according to the present invention, it may be possible to reduce the application frequency of the above natamycin and / or increase the biological activity of the above natamycin.

[0083] One aspect of the present invention provides the application of at least one insecticide that interferes with the nervous system and / or muscular system of insects and nematodes to enhance the biological activity of natamycin present in a plant, plant part, or soil. The above insecticide can increase the biological activity of the above natamycin. The above at least one insecticide preferably belongs to subgroup 2 (GABA-gated chloride channel blocker), subgroup 3 (sodium channel modulator), subgroup 4 (competitive modulator of nicotinic acetylcholine receptors), subgroup 6 (allosteric modulator of glutamate-gated chloride channels), and / or subgroup 28 (ryanodine receptor modulator) described in the document "IRAC Mode of Action Classification Scheme" (June 2019; version 9.3).

[0084] The terms "reduction in application frequency" and "increase in biological activity" may refer to an application frequency reduced by 10% or more, preferably 30% or more, when compared to the application frequency of natamycin not containing at least one insecticide.

[0085] The reduced application frequency may refer to an application frequency of 5 mg (a.i.) to 2.5 kg a.i. / ha of active ingredient per hectare, preferably 1 g a.i. / ha to 2 kg a.i. / ha, for example 100 to 750 g a.i. / ha (frequencies of 600 g a.i. / ha, 500 g a.i. / ha, 400 g a.i. / ha, 300 g a.i. / ha, 200 g a.i. / ha, and 100 g a.i. / ha are included).

[0086] The antifungal composition according to the present invention is suitable for controlling pests encountered in horticulture, agriculture, and forestry. The antifungal composition is active against both normally sensitive and resistant pest species and at all or individual stages of development. Prior to use, it is preferable to dissolve or disperse the composition containing the antifungal composition according to the present invention in water or dilute it with water to prepare an aqueous composition containing 0.001 to 10 w / v% of bioactive natamycin. Optionally, an agriculturally acceptable carrier such as an adhesive is added to the diluted aqueous composition.

[0087] Before contacting the composition according to the present invention with a plant, plant part, or soil, it is preferably diluted 2 to 5000 times, preferably about 200 times, with an aqueous solvent (preferably water) so as to contain 0.0001 to 10% (w / v) of natamycin.

[0088] To control agricultural pests, the present invention provides the use of natamycin and an insecticide (preferably, a composition according to the present invention comprising natamycin and at least one insecticide) that interferes with the nervous system and / or muscular system of insects and nematodes for protecting a plant or a part of a plant from pathogens. To achieve this effect, the above plant or part of a plant, or soil, is contacted with the above natamycin and an insecticide (preferably, the above composition comprising a diluted aqueous composition as described above herein) that interferes with the nervous system and / or muscular system of insects and nematodes. The above natamycin and an insecticide (preferably the above composition) that interferes with the nervous system and / or muscular system of insects and nematodes are used, for example, to control powdery mildew and Clostridium botulinum infection of food / feed crops (including tree fruits, vegetable crops, field crops, grapes, ornamental plants, and sod farms). Further, for example, it is also used for controlling scab such as common sacab, apple scab, and black scab on potatoes, pear scab, and powdery scab, brown rot of peaches, leaf spots of currants and gooseberries, Fusarium diseases, peanut leaf spots, and mildew of roses. Other usage methods include protecting greenhouse flowers, home gardens, and lawns of houses. Further, the above natamycin and an insecticide (preferably, the above composition comprising a diluted aqueous composition) that interferes with the nervous system and / or muscular system of insects and nematodes may be contacted with separated seeds, fruits, nuts, vegetables, and / or flowers.

[0089] The present invention further provides a method for protecting a plant or a plant part from a pathogen, the method comprising contacting the plant or the plant part with natamycin and an insecticide that interferes with the nervous system and / or muscular system of insects and nematodes (preferably, an aqueous dilution composition according to the present invention comprising natamycin and an insecticide that interferes with the nervous system and muscular system (preferably, an insecticide belonging to subgroups 2, 3, 4, 6 and / or 28 described in the "IRAC Mode of Action Classification Scheme" (June 2019; version 9.3))).

[0090] The present invention further provides a method for preventing, reducing and / or eliminating the presence of a pathogen in a plant or a part of a plant, the method comprising contacting the plant or the part of the plant with natamycin and an insecticide that interferes with the nervous system and / or muscular system of insects and nematodes (preferably, an aqueous composition according to the present invention).

[0091] For the above uses and methods, natamycin and an insecticide that interferes with the nervous system and / or muscular system of insects and nematodes (preferably, the above composition comprising a diluted aqueous composition) are preferably sprayed on the plant or a part thereof. Spraying applications, including the use of automated spraying systems, are known to reduce labor costs and be cost-effective. Methods and devices well known to those skilled in the art can be used therefor. Natamycin and an insecticide that interferes with the nervous system and / or muscular system of insects and nematodes (preferably, the above composition comprising a diluted aqueous composition) can be sprayed regularly when the risk of infection is high. When the risk of infection is low, the spraying interval may be longer, as is known to those skilled in the art.

[0092] Other methods suitable for contacting a plant or a part thereof with natamycin and an insecticide (preferably the composition of the present invention) that interferes with the nervous system and / or muscular system of insects and nematodes are also part of the present invention. These include, but are not limited to, dipping, watering, soaking, introduction into a dump tank, vaporization, atomization, fumigation, painting, brushing, spraying, spreading, foaming, spraying, packaging, coating (e.g., by means of wax or static electricity), etc. Further, natamycin and an insecticide (preferably the composition of the present invention, including a diluted aqueous composition) that interferes with the nervous system and / or muscular system of insects and nematodes may be injected into the soil.

[0093] For example, a plant and a part thereof may be coated with natamycin and an insecticide (preferably a diluted aqueous composition containing natamycin and at least one insecticide according to the present invention) that interferes with the nervous system and / or muscular system of insects and nematodes, and a plant or a part thereof may be protected from pathogens and / or the presence of pathogens in the plant or a part of the plant strip may be prevented, reduced and / or eliminated by dipping the plant or a part thereof in natamycin and an insecticide (preferably the diluted aqueous composition according to the present invention) that interferes with the nervous system and / or muscular system of insects and nematodes.

[0094] A preferred part of a plant coated with natamycin and an insecticide (preferably the composition according to the present invention, or a dilution thereof) that interferes with the nervous system and / or muscular system of insects and nematodes is the seed. A more preferred part of a plant coated with natamycin and an insecticide (preferably the composition according to the present invention, or a dilution thereof) that interferes with the nervous system and / or muscular system of insects and nematodes is the fruit. Preferably, they are post-harvest fruits such as citrus fruits like oranges, mandarins, limes, etc., pome fruits like apples, pears, etc., stone fruits like almonds, apricots, cherries, damsons, nectarines, tomatoes, watermelons, etc., and tropical fruits like bananas, mangoes, lychees, tangerines, etc. Preferred fruits are citrus fruits such as oranges and / or tropical fruits such as bananas.

[0095] The present invention further provides a method for controlling diseases caused by pathogenic fungi on plants or their propagation materials, the method comprising contacting the plant or its propagation materials with natamycin and an insecticide that interferes with the nervous system and / or muscular system of insects and nematodes (preferably the composition according to the present invention comprising an aqueous dilution composition).

[0096] The present invention also provides a method for controlling pests by contacting (i) pests or their locus, (ii) plants or their locus or propagation materials, (iii) soil, and / or (iv) a location where pest infestation is to be prevented, with natamycin and an insecticide that interferes with the nervous system and / or muscular system of insects and nematodes (preferably the composition of the present invention comprising natamycin and at least one insecticide).

[0097] The present invention also provides an improved method for pest control, comprising applying natamycin and an insecticide that interferes with the nervous system and / or muscular system of insects and nematodes (preferably the composition described herein) to plants or soil.

[0098] The present invention also provides a method for prolonging the control effect of natamycin on plants, plant parts or soil, comprising applying natamycin and an insecticide that interferes with the nervous system and / or muscular system of insects and nematodes (preferably the composition of the present invention or its diluent) to the plants, plant parts or soil.

[0099] In some embodiments, the target is a plant, a plant part, soil or a growth substrate. In some embodiments, the target is a fungus.

[0100] The present invention also provides a method for controlling pests by prevention, treatment or continuous treatment of plant diseases caused by pathogenic fungi, comprising contacting a plant, its locus or its propagation materials with an effective amount of natamycin and an insecticide that interferes with the nervous system and / or muscular system of insects and nematodes (preferably the composition according to the present invention comprising natamycin and at least one insecticide).

[0101] The composition according to the invention comprising natamycin and at least one insecticide may be applied to healthy or diseased plants. The composition described can be used on various plants including, but not limited to, crops, seeds, bulbs, propagation material, or ornamental species.

[0102] The present invention provides a method for controlling diseases caused by pathogenic fungi on plants or their propagation materials, the method comprising contacting the plant, its locus or its propagation material with natamycin and an insecticide that interferes with the nervous system and / or muscular system of insects and nematodes (preferably, a composition according to the invention comprising natamycin and at least one insecticide).

[0103] In some embodiments, the fungus is one of Mycosphaerella graminicola; anamorph: Septoria tritici (Leaf Blotch of Wheat), Puccinia triticina (Brown Rust of Wheat), Puccinia striiformis f. sp. tritici (Stripe Rust of Wheat), Venturia inaequalis (scab of apple), Ustilago maydis (Blister Smut of Maize), Uncinula necator (Powdery Mildew of Grape), Rhynchosporium secalis (Barley scald), Magnaporthe grisea (Blast of Rice), Phakopsora pachyrhizi (Rust of Soybean), Leptosphaeria nodorum (Glume Blotch of Wheat), Blumeria graminis f. sp. tritici (Powdery Mildew of Wheat), Blumeria graminis f. sp. hordei (Powdery Mildew of Barley), Erysiphe cichoracearum (Powdery Mildew of Cucurbits), Glomerella lagenarium (Anthracnose of Cucurbits), Cercospora beticola (Leaf Spot of Beet), Alternaria solani (Early Blight of Tomato), and Pyrenophora teres (Net Blotch of Barley).

[0104] In some embodiments, natamycin and an insecticide that interferes with the nervous system and / or muscular system of insects and nematodes (preferably, the composition according to the present invention comprising natamycin and at least one insecticide) are applied at a frequency effective for controlling pests. In some embodiments, natamycin and an insecticide that interferes with the nervous system and / or muscular system of insects and nematodes (preferably, the composition according to the present invention comprising natamycin and at least one insecticide) are applied at a frequency effective for preventing the invasion of pests. In some embodiments, natamycin and an insecticide that interferes with the nervous system and / or muscular system of insects and nematodes (preferably, the composition according to the present invention comprising natamycin and at least one insecticide) are applied at a frequency effective for curing the invasion of pests.

[0105] In some embodiments, the method of the present invention is effective for preventing the invasion of pests. In some embodiments, the method of the present invention is effective for curing the spread of pests. In some embodiments, the method is effective for increasing the insecticidal activity of natamycin. In some embodiments, the method is effective for extending the insecticidal effect of natamycin.

[0106] In some embodiments, the method of the present invention is effective for reducing the half-maximal effective concentration (EC50) of natamycin. In some embodiments, the method is effective for reducing the EC50 by 10% or more. In some embodiments, the method is effective for reducing the EC50 by 25% or more. In some embodiments, the method is effective for reducing the EC50 by 35% or more. In some embodiments, the method is effective for reducing the EC50 by 50% or more.

[0107] In some embodiments, the method of the present invention is effective in reducing the LC50 of natamycin. In some embodiments, the method is effective in reducing the LC50 by 10% or more. In some embodiments, the method is effective in reducing the LC50 by 25% or more. In some embodiments, the method is effective in reducing the LC50 by 50% or more. In some embodiments, the method is effective in reducing the LC50 by 75% or more. In some embodiments, the method is effective in reducing the LC50 by 90% or more.

[0108] In some embodiments, the method of the present invention is effective in reducing the LC90 of natamycin. In some embodiments, the method is effective in reducing the LC90 by 10% or more. In some embodiments, the method is effective in reducing the LC90 by 25% or more. In some embodiments, the method is effective in reducing the LC90 by 50% or more. In some embodiments, the method is effective in reducing the LC90 by 75% or more. In some embodiments, the method is effective in reducing the LC90 by 90% or more.

[0109] In some embodiments, the method of the present invention further comprises applying at least one additional pesticide to the pest, plant part, plant, its locus, or its propagation material. The additional pesticide may be admixed in the tank or applied sequentially with natamycin and an insecticide that interferes with the nervous system and / or muscular system of insects and nematodes, and is preferably applied to the plant, plant part, soil or growing substrate together with a composition comprising natamycin and at least one insecticide.

[0110] The present invention will be illustrated by the following examples, which are not intended to be limiting.

[0111] <Example> <Overview> [Natamycin formulation used in the experiment] Natamycin was tested on apples at concentrations of 0 ppm (control), 50 ppm, 100 ppm, and 200 ppm. Natamycin was formulated as shown in Table 1.

[0112]

Table 1

[0113] [Insecticide formulations used in the experiment] The insecticide active ingredients thiamethoxam (Merck; 37924), imidacloprid (Merck; 37894), abamectin (Merck; 31732), and chlorpyrifos (Merck; 45395) were formulated in the compositions shown in Table 2.

[0114]

Table 2-1

[0115]

Table 2-2

[0116]

Table 2-3

[0117]

Table 2-4

[0118] As the active ingredients, fipronil, lambda-cyhalothrin, alpha-cypermethrin, acetamiprid, emamectin benzoate, chlorantraniliprole, flubendiamide, fenoxycarb, Bacillus thuringiensis, chlorfenapyr, and azadirachtin were commercially available products (see Table 3).

[0119] [Dosage rates of the insecticides applied in the examples] In the examples, natamycin was combined with the labeled dosages of standard insecticides. The amounts of the insecticide active ingredients used in the examples are shown in Table 3.

[0120]

Table 3

[0121] [Measurement of synergistic effect] There are also examples where the stimulation of the antifungal activity of natamycin by insecticides has been found to be synergistic. The Colby formula (Colby, 1967. Weeds 15: 20 - 22) calculates the expected value (E in %) of the antifungal activity of a combination containing two or more active ingredients:

Equation

[0122] <Example 1: Efficacy test of insecticides against the antifungal activity of natamycin in artificially infected plant tissues [Materials and methods] Apple cv Elstar is from organic cultivation and has obtained SKAL certification. SKAL is a semi-governmental and semi-private organization in the Netherlands and manages organic cultivation in the Netherlands.

[0123] As a spore suspension of Botrytis cinerea, one containing 10 5 spores / ml is used.

[0124] Using a cork borer with a diameter of 0.6 cm and a maximum depth of 0.5 cm, damage the peel, and make 2 wounds per apple fruit. Apply 30 μl of a fresh spore suspension of B. cinerea (about 10 5 spores / ml) to each wound with a pipette. Then, air-dry the spore suspension for 3 hours. As shown in Tables 4 and 5 below, apply 50 μl of natamycin and / or insecticide to each wound with a pipette.

[0125] All fruits are stored at room temperature (20 °C). The wounds are inspected daily and recorded after 10 days.

[0126] All treatments are performed on 6 apples, each with 2 wounds, and 12 wounds are made in one treatment. The recorded antifungal activity is the decrease (%) in the average surface area of the rot observed in 12 wounds compared to the surface area of the rot of the untreated control.

[0127] [Results] The results are shown in Tables 4 and 5. Each letter indicates the difference in the infection rate (surface area), with A having the highest infection area, and B, C, and D having decreasing infection areas (B is significantly lower than A, C is significantly lower than B, and D is significantly lower than C). From these results, it is clear that the insecticide itself does not have a bactericidal effect. The insecticide in Table 4, which acts through the insect's nervous or muscular system, stimulates the bactericidal effect of natamycin on apple plant tissue, while the insecticide in Table 5, which does not act through the insect's nervous or muscular system, does not have this effect.

[0128]

Table 4

[0129]

Table 5

[0130] <Example 2: Test on the Bactericidal Effect of Natamycin with and without Insecticide on Pathogenic Fungi in Seeds of Artificially Infected Soil> [Materials and Methods] Seeds are coated with the natamycin formulation described in Table 1 in the presence or absence of the insecticide composition described in Table 2. Soybean seeds are placed in artificially infected soil (see the method below).

[0131] Soil infection is obtained by culturing soil using dead sorghum seeds that have been infected. For this purpose, 100 g of sorghum seeds and 100 ml of water are placed in a 500 ml bottle and autoclaved twice (121 °C, 15 minutes, 15 psi). The dead sorghum seeds are infected with Fusarium graminearum segments (circular segments with a height of 5 mm on agar, completely grown with fresh Fusarium graminearum hyphae cut out from a petri dish using a cork borer with a diameter of 6 mm). Ten agar pieces of F. graminearum are placed in a 500 ml bottle containing 100 ml of water with autoclaved (dead) sorghum seeds and cultured at 25 °C for 2 weeks (16 hours of daylight, 8 hours of darkness). To prepare the infected soil, soil (Lentse potgrond type 821201030, purchased from Horticoop) and sand (river sand from van Leusden in Wageningen) are put into plug trays (the tray dimensions are 52 cm × 30 cm, with 40 round cells (modiform; Leusden, the Netherlands) with a diameter of 5 cm and a depth of 4.5 cm) in a 1:1 mixture.

[0132] Three infected sorghum seeds that serve as carriers of F. graminearum are sown into each plug containing the sandy soil mixture. The trays are left in the greenhouse (16 hours of sunlight, day temperature 20 °C, night temperature 18 °C, humidity 60%) for 1 week.

[0133] Thereafter, soybean seeds are washed with 70% ethanol and thoroughly rinsed with water.

[0134] Natamycin (0.06 g, 0.12 g, or 0.24 g of active ingredient per 1 kg of seeds) and the indicated insecticide (dose refer to Table 3) are suspended or dissolved in 30 ml of water to prepare the user solution. Next, 250 g of seeds are put into a rotary coater, and the user solution is applied to the seeds. The seeds are rotated for 45 seconds with each solution. The solution is evenly distributed to the seeds through the rotating disk of the rotary coater. Thereafter, the seeds are taken out from the rotary coater and placed in a dryer at a temperature of 25 °C for 15 minutes.

[0135] Seeds were sown at a depth of 2 cm in the cells (plugs) of a plug tray containing a mixture of soil and sand, with 20 seeds per treatment and 1 seed per cell. On the 14th day after sowing, the emergence and quality evaluation of the seedlings were carried out.

[0136] [Results] Tables 6 and 7 show the effects of the combined and non-combined use of an insecticide having activity on the nervous or muscular system of insects (Table 6), or an insecticide having activity on other systems of insects (Table 7) and natamycin on the growth quality of soybean seeds. Fourteen days after sowing, the proportions of healthy seedlings (compared to the growth of soybean seeds in non-infected soil), abnormal seedlings (e.g., stunted growth, small discolored plants, deformed leaves) and dead seeds were determined. The results in Tables 6 and 7 represent the proportion of healthy seedlings.

[0137] [Table 6]

[0138] [Table 7]

[0139] [Results] In Tables 6 and 7, each individual letter indicates the difference in the proportion of healthy seedlings, where A has the lowest proportion of healthy seedlings, and B, C, and D indicate an increasing proportion of healthy seedlings (B is significantly higher than A, C is significantly higher than B, and D is significantly higher than C).

[0140] From these results, it is clear that the insecticides themselves have no bactericidal power, but the insecticide in Table 6 with a mode of action via the nervous or muscular system of insects stimulates the bactericidal action of natamycin on seeds, and the insecticide in Table 7 with a mode of action not via the nervous or muscular system of insects does not inhibit its action.

[0141] [Example 3: Effects of Natamycin and Insecticides Alone or in Combination against Fungal Attack on Maize Seeds] [Materials and Methods] The dosage of the insecticide is as shown in Table 3. The treatment method of the corn seeds is the same as the method described for the soybean seeds in Example 2. Fourteen days after sowing, the percentages of healthy seedlings (equivalent to the growth of corn seeds in non-infected soil), abnormal seedlings (e.g., stunted growth, small discolored plants, deformed leaves), and dead seeds are determined. The results in Table 8 show the percentage of healthy seedlings.

[0142] Table 8 shows the effect on the growth quality of corn seeds when an insecticide having activity on the nervous or muscular system of insects is used in combination with natamycin.

[0143] [Table 8]

[0144] From the results in Table 8, it can be seen that the insecticide acting through the nerve or muscle tissue itself has no bactericidal power, but stimulates the bactericidal action of natamycin on corn seeds, and healthier seedlings can be obtained.

[0145] Table 9 shows the effect on the growth quality of corn seeds treated with a combination of an insecticide having no activity on the nervous or muscular system of insects and natamycin.

[0146] [Table 9]

[0147] From the results in Table 9, it can be seen that the insecticide having no mechanism of action through the nervous or muscular system itself has no bactericidal activity and does not stimulate the bactericidal action of natamycin on seeds.

[0148] [Example 4: Effects of Natamycin and Abamectin (acting on the nervous system of insects) Alone and in Combination on the Attack of Various Pathogenic Fungi on Corn Seeds] [Materials and Methods] The method is as described for the seed experiment in Example 2. Fusarium graminearum was replaced with the pathogenic fungi described in Table 10. The natamycin and insecticide compositions are shown in Tables 1 and 2.

[0149]

Table 10

[0150] [Results] From the results in Table 10, it can be seen that abamectin, an insecticide with a mechanism of action via the nervous or muscular system, has no bactericidal effect on its own, but stimulates the bactericidal effect of natamycin against different fungal pathogens of seeds, resulting in healthier seedlings.

[0151] <Example 5: Effects of Natamycin and Imidacloprid (Acting on the Insect Nervous System), Alone or in Combination, on Different Types of Seeds> [Materials and Methods] Except for using 75 g of sugar beet seeds, 75 g of turfgrass seeds, and 25 g of tomato seeds, different types of seeds were planted in artificially infected soil (Fusarium culmorum) in the same manner as described in Example 2. The percentage of healthy seedlings was evaluated after 14 days of cultivation.

[0152] [Results] From the results in Table 11, it can be seen that imidacloprid has no bactericidal effect on its own, but stimulates the bactericidal effect of natamycin on seeds of different plants, resulting in healthier seedlings.

[0153]

Table 11

[0154] <Example 6: Influence of Fipronil on the Efficacy of Natamycin Against Fusarium culmorum> [Materials and Methods for Examples 6 to 19] In a 100 ml Duran bottle, 3.9 g of potato dextrose agar (PDA) from Carl-Roth (Carl-Roth GmbH + Co. KG, Karlsruhe, Germany) and 100 ml of deionized water were mixed, and the agar medium was prepared by autoclaving the Duran bottle at 120 °C for 15 minutes. After autoclaving, the solution was placed in an oven at 48 °C for about 2 hours to cool. Thereafter, the doses of natamycin and / or insecticide specified in Table 12 were carefully mixed into the semi-liquid PDA solution. The medium in the Duran bottle was dispensed onto 5 Petri dishes (90 × 15 mm) at 20 ml per dish using a 25 ml serum pipette (ROTILABO®; Carl-Roth). Each natamycin treatment and / or insecticide treatment was carried out in 5 replicates.

[0155] A suspension of fungal spores was prepared by immersing a sufficiently grown Petri dish in sterile water. The fungus was scraped off and filtered through Miracloth (pore size: 22 - 25 μm) (Merck KGaA, Darmstadt, Germany; catalog number: 475855). The number of spores was counted using a hemocytometer, and the fungal suspension was adjusted to 10 6 cells / ml. Thereafter, 5 μl of the prepared spore suspension was pipetted onto the center of the agar plate. The plates were incubated at 25 °C. The growth of the fungus was measured using a caliper at different time points.

[0156] In this example, the synergistic effect with natamycin was verified using BASF's product REGENT® containing fipronil, which is an active ingredient belonging to IRAC group 2. The insecticide concentration "N" used (see below) was 0.94 g of the insecticide product per 100 ml of PDA agar. 0.5N is 0.47 g. The concentration of natamycin in the natamycin-containing incubation was 1 ppm. The ratio of natamycin:fipronil was 1:7520 (w / w) and 1:3760 (w / w). After culturing for 7 days in a 25 °C incubator, the efficacy was evaluated. The calculation of the synergistic effect was performed using the Colby equation.

[0157] [Results]

Table 12

[0158] [Example 7: Influence of λ-cyhalothrin on the natamycin efficacy against Fusarium culmorum> [Materials and methods] Refer to Example 6. In this example, the Syngenta product KARATE ZEON® containing λ-cyhalothrin, an active ingredient belonging to IRAC group 3, was used.

[0159] The concentration of the insecticide "N" used (see below) was 20 μl of the insecticide formulation per 100 ml of PDA agar. 0.5N is 10 μl per 100 ml of PDA agar.

[0160] The concentration of natamycin in the natamycin-containing culture was 1 ppm. The ratio of natamycin:λ-cyhalothrin was 1:50 (w / w) and 1:25 (w / w). The efficacy was evaluated after culturing for 7 days in an oven at 25 °C.

[0161] [Results]

Table 13

[0162] [Example 8: Influence of λ-cyhalothrin on the efficacy of natamycin against Alternaria solani> [Materials and methods] Refer to Example 6. In this example, the Syngenta product KARATE ZEON® containing λ-cyhalothrin, an active ingredient belonging to IRAC group 3, was used.

[0163] The concentration of the insecticide "N" used (see below) was 500 μl of an insecticide solution diluted 25-fold with water per 100 ml of PDA agar. For 2N, the diluted solution in water was 1000 μl per 100 ml of PDA agar. For 5N, 100 μl of the stock solution of KARATE ZEON was used per 100 ml of PDA agar.

[0164] The natamycin concentration in the natamycin-containing culture was 0.5 ppm. The ratio of natamycin: λ-cyhalothrin was 1:100 (w / w), 1:200 (w / w), and 1:500 (w / w). After incubation in an oven at 25 °C for 4 days, the efficacy was evaluated.

[0165] [Results] [Table 14]

[0166] [Example 9: Effect of λ-cyhalothrin on the efficacy of natamycin against Sclerotinia sclerotiorum> [Materials and methods] Refer to Example 6. In this example, the Syngenta product KARATE ZEON® containing λ-cyhalothrin, an active ingredient belonging to IRAC group 3, was used.

[0167] The concentration of the insecticide "N" used (see below) was 500 μl of an insecticide solution diluted 25-fold with water per 100 ml of PDA agar. For 2N, the diluted solution in water was 1000 μl per 100 ml of PDA agar medium. For 5N, 100 μl of the stock solution was added to 100 ml of PDA agar.

[0168] The natamycin concentration in the natamycin-containing culture was 0.5 ppm. The ratio of natamycin: λ-cyhalothrin was 1:100 (w / w), 1:200 (w / w), and 1:500 (w / w). After incubation in an oven at 25 °C for 4 days, the efficacy was evaluated.

[0169] [Results] [Table 15]

[0170] [Example 10: Influence of λ-cyhalothrin on the efficacy of natamycin against Botrytis cinerea] [Materials and methods] Refer to Example 6. In this example, Syngenta's product KARATE ZEON® containing λ-cyhalothrin, an active ingredient belonging to IRAC group 3, was used.

[0171] For the insecticide concentration "N" used (see below), 20 μl of the insecticide product was used per 100 ml of PDA agar. 0.5N was 10 μl of the product per 100 ml of PDA agar. 0.25N was 5 μl of the stock solution of KARATE ZEON per 100 ml of PDA agar, and 0.125N was 2.5 μl of the insecticide per 100 ml of PDA agar.

[0172] The concentration of natamycin in the natamycin-containing culture was 1 ppm. The ratio of natamycin:λ-cyhalothrin was 1:12.5 (w / w), 1:25 (w / w), and 1:50 (w / w). After culturing for 6 days in an oven at 25 °C, the efficacy was evaluated.

[0173] [Results] [Table 16]

[0174] [Example 11: Influence of imidacloprid on the efficacy of natamycin against Fusarium graminerum] [Materials and methods] Refer to Example 6. In this example, imidacloprid, an insecticidal active ingredient belonging to IRAC group 4, was formulated together with additional compounds as shown in Table 17.

[0175]

Table 17

[0176] The concentration of the insecticide "N" used (see below) was 5 μl of the insecticide formulation per 100 ml of PDA agar. 0.5N is 2.5 μl of the formulation per 100 ml of PDA agar. The concentration of natamycin in the natamycin-containing culture was 1 ppm. The ratio of natamycin:imidacloprid was 1:20 (w / w) and 1:10 (w / w). After culturing in an oven at 25 °C for 8 days and 12 days, the efficacy was evaluated.

[0177] [Results]

Table 18

[0178]

Table 19

[0179] <Example 12: Influence of thiamethoxam on the efficacy of natamycin against Fusarium culmorum> [Materials and methods] Refer to Example 6. In this example, thiamethoxam, an active ingredient belonging to IRAC group 4, was mixed with water to make a 1000 ppm solution. The concentration of the insecticide "N" used (see below) was 5 ml of the insecticide solution per 100 ml of PDA agar. 2N is 10 ml of the thiamethoxam solution per 100 ml of PDA agar.

[0180] The natamycin concentration in the natamycin-containing culture was 0.5 ppm. The ratio of natamycin:thiamethoxam was 1:100 (w / w) and 1:200 (w / w). After culturing in an oven at 25 °C for 4 days, the efficacy was evaluated.

[0181] [Results]

Table 20

[0182] <Example 13: Influence of abamectin on the efficacy of natamycin against Fusarium graminerum> [Materials and methods] Refer to Example 6. In this example, the Syngenta product VERTIMEC® containing abamectin, an active ingredient belonging to IRAC group 6, was used.

[0183] For the pesticide concentration "N" used (see below), 75 μl of the pesticide product was used per 100 ml of PDA agar. For 0.5N, 37.5 μl of the product was used per 100 ml of PDA agar.

[0184] The natamycin concentration in the natamycin-containing culture was 1 ppm. The ratios of natamycin:abamectin were 1:13.5 (w / w) and 1:6.75 (w / w). After culturing at 25 °C on a stove for 8 and 12 days, the efficacy was evaluated.

[0185] [Results]

Table 21

[0186]

Table 22

[0187] <Example 14: Influence of abamectin on the efficacy of natamycin against Fusarium graminerum> [Materials and methods] Refer to Example 6. In this example, the Syngenta product VERTIMEC® containing abamectin, an active ingredient belonging to IRAC group 6, was used. The concentration of the insecticide "N" used (see below) was 75 μl of the insecticide product per 100 ml of PDA agar. 0.5N is 37.5 μl of the product per 100 ml of PDA agar. The concentration of natamycin in the natamycin-containing culture was 1 ppm. The ratio of natamycin:abamectin was 1:13.5 (w / w) and 1:6.75 (w / w). After culturing in a 25°C incubator for 7 days, the efficacy was evaluated.

[0188] [Results]

Table 23

[0189] <Example 15: Influence of Chlorantraniliprole on the Efficacy of Natamycin Against Botrytis cinerea> [Materials and Methods] Refer to Example 6. In this example, the Dupont product CORAGEN® containing chlorantraniliprole, an active ingredient belonging to IRAC group 28, was used.

[0190] The concentration of the insecticide "N" used (see below) was 125 μl of the insecticide formulation per 100 ml of PDA agar. Per 100 ml of PDA agar, a concentration of 0.25N contained 31.25 μl of collagen, 0.5N contained 62.5 μl of collagen, 2N contained 250 μl of collagen, 5N contained 625 μl of collagen, and 10N contained 1250 μl of the collagen product.

[0191] The concentration of natamycin in the natamycin-containing culture was 1 ppm. The ratio of natamycin:chlorantraniliprole was 1:62.5 (w / w), 1:125 (w / w), 1:250 (w / w), 1:500 (w / w), 1:1250 (w / w), and 1:2500 (w / w). After culturing on a stove at 25°C for 4 and 5 days, the efficacy was evaluated.

[0192]

Table 24

[0193]

Table 25

[0194] <Example 16: Influence of Hydroprene on the Efficacy of Natamycin Against Botrytis cinerea> [Materials and Methods] Refer to Example 6. In this example, the product GENTROL® IGR (Zoecon; Syngenta), which contains hydroprene, an active ingredient belonging to IRAC Group 7, was used.

[0195] The pesticide concentration "N" used (see below) was prepared by adding 300 μl of a solution prepared by dissolving 2.49 ml of Gentrol in 50 ml of water to 50 ml of PDA agar. For 0.5N: 150 μl of the pesticide solution was used per 100 ml of PDA agar, and for 0.25N: 75 μl of the pesticide solution was used.

[0196] The concentration of natamycin in the natamycin-containing culture was 1 ppm. The ratio of natamycin:hydroprene was 1:3.4 (w / w), 1:6.75 (w / w), and 1:13.5 (w / w). After culturing in a stove at 25°C for 4 days, the efficacy was evaluated.

[0197] [Results]

Table 26

[0198] <Example 17: Influence of Hydroprene on the Efficacy of Natamycin against Fusarium culmorum> [Materials and Methods] Refer to Example 6. In this example, the product GENTROL® IGR (Zoecon; Syngenta), which contains hydroprene as the active ingredient, was used.

[0199] The pesticide concentration “N” used (see below) was prepared by adding 300 μl of a solution obtained by dissolving 2.49 ml of Gentrol in 50 ml of water to 50 ml of PDA agar. For 0.5N, 150 μl of the pesticide solution and for 0.25N, 75 μl of the pesticide solution were used.

[0200] The natamycin concentration in the natamycin-containing culture was 1 ppm. The ratios of natamycin:hydroprene were 1:3.4 (w / w), 1:6.75 (w / w), and 1:13.5 (w / w). After culturing at 25 °C on a stove for 5 days, the efficacy was evaluated.

[0201] [Results]

Table 27

[0202] <Example 18: Influence of Hydroprene on the Efficacy of Natamycin against Fusarium graminerum> [Materials and Methods] Refer to Example 6. In this example, the product GENTROL® IGR (Zoecon; Syngenta), which contains hydroprene as the active ingredient belonging to IRAC group 7, was used.

[0203] The concentration of the insecticide "N" used (see below) was prepared by adding 300 μl of a solution obtained by dissolving 2.49 ml of Gentrol in 50 ml of water to 50 ml of PDA agar. For 0.5N: 150 μl of the insecticide solution was added to 100 ml of PDA agar, and for 0.25N: 75 μl of the insecticide solution was used.

[0204] The natamycin concentration in the natamycin-containing culture was 1 ppm. The ratio of natamycin:hydroprene was 1:3.4 (w / w), 1:6.75 (w / w), and 1:13.5 (w / w). After culturing at 25°C on a stove for 5 days, the efficacy was evaluated.

[0205] [Results] [Table 28]

[0206] [Example 19: Influence of Bacillus thuringiensis on the Efficacy of Natamycin against Fusarium graminearum> [Materials and Methods] Refer to Example 6. In this example, Product XENTARI® (Bayer) containing Bacillus thuringiensis (15,000 IU / mg (IU = International Unit)), which is an active ingredient belonging to IRAC group 11, was used.

[0207] The concentration of the insecticide "N" used (see below) was prepared by adding 31.25 mg of the XENTARI product to 100 ml of PDA agar. For 0.5N: 15.63 mg of the XENTARI product was added to 100 ml of PDA agar.

[0208] The natamycin concentration in the natamycin-containing culture was 1 ppm. The ratio of natamycin:Bacillus thuringiensis was 1:156.25 and 1:312.5 (w / w based on the XENTARI product standard). After culturing at 25°C on a stove for 8 days, the efficacy was evaluated.

[0209] [Results] TIFF0007699576000033.tif48131

[0210] <Example 20: Influence of Chlorfenapyr on the Efficacy of Natamycin against Fusarium culmorum> [Materials and Methods] Refer to Example 6. In this example, the product SPECTRE® (ADAMA), which contains chlorfenapyr, an active ingredient belonging to IRAC group 13, was used.

[0211] The pesticide concentration "N" used (see below) was obtained by adding 1 ml of the Spectre product to 100 ml of PDA agar. For 100 ml of PDA agar, 0.5N was obtained by adding 0.5 ml of Spectre, and 0.25N was obtained by adding 0.25 ml of Spectre.

[0212] The natamycin concentration in the natamycin-containing culture was 1 ppm. The ratios of natamycin:chlorfenapyr were 1:536 (w / w), 1:1072 (w / w), and 1:2145 (w / w). After culturing for 7 days in an oven at 25 °C, the efficacy was evaluated.

[0213] [Results]

Table 30

[0214] <Example 21: Influence of Azadirachtin on the Efficacy of Natamycin against Fusarium graminerum> [Materials and Methods] Refer to Example 6. In this example, the product AZATIN® (Certis), which contains azadirachtin, an active ingredient belonging to an as yet unknown IRAC group (formulated to have a concentration of 217 g / L azadirachtin (resulting in a concentration of 26 g / L azadirachtin-A)), was used.

[0215] The concentration of the pesticide "N" used (see below) was obtained by adding 31 μl of the azatin formulation to 100 ml of PDA agar. 0.5N: 37.5 μl of azatin was added to 100 ml of PDA agar.

[0216] The concentration of natamycin in the natamycin-containing culture was 1 ppm. The ratio of natamycin to azadirachtin was 1:9.75 (w / w) and 1:19.5 (w / w) based on azadirachtin-A. After culturing for 8 days in a 25 °C incubator, the efficacy was evaluated.

[0217] [Results] TIFF0007699576000035.tif44129

[0218] <Example 22: Influence of emamectin benzoate on the efficacy of natamycin against Fusarium graminearum> [Materials and methods] Refer to Example 6. In this example, the product OPTIGARD® Cockroach gel Bait, (Syngenta), which contains emamectin benzoate, an active ingredient belonging to IRAC group 6, was used. The concentration of the pesticide "N" used (see below) was 250 mg of the pesticide product per 100 ml of PDA agar. 0.5N is 125 mg of the product per 100 ml of PDA agar. The concentration of natamycin in the natamycin-containing culture was 0.25 ppm. The ratio of natamycin to emamectin benzoate was 1:10 and 1:5 (w / w). After culturing for 3 days in a 25 °C incubator, the efficacy was evaluated.

[0219] [Results]

Table 32

Claims

1. An antifungal composition used for protecting crop plants or plant parts, improving the growth and / or yield of crop plants, protecting soil and / or growth substrates, or protecting plants, plant parts, soil and / or growth substrates from fungi, comprising natamycin and at least one insecticide that interferes with the nervous system and / or muscular system of insects and nematodes, wherein the at least one insecticide is selected from emamectin benzoate, alpha-cypermethrin, fipronil, lambda-cyhalothrin, acetamiprid, clothianidin, imidacloprid, thiacloprid, thiamethoxam, abamectin, and chlorantraniliprole, and the volume-based average particle diameter of the natamycin is 0.2 μm to 10 μm. The above antifungal composition.

2. The antifungal composition according to claim 1, comprising 1% to 98% (w / w) of natamycin and 1% to 99% (w / w) of the at least one insecticide.

3. The antifungal composition according to claim 1, wherein the ratio of natamycin to the at least one insecticide is 1:1 to 1:5000 (w / w).

4. The antifungal composition according to any one of claims 1 to 3, further comprising an agriculturally acceptable carrier.

5. The antifungal composition according to any one of claims 1 to 4, which is an aqueous composition or an oily composition.

6. The antifungal composition according to any one of claims 1 to 5, wherein the natamycin is ground to an average particle diameter of 0.5 μm to 3 μm.

7. The antifungal composition according to any one of claims 1 to 6, further containing an insoluble polyelectrolyte complex of polyanion and polycation with a relative amount of 1:2 to 60:1 (w / w). Claims 8. Preparing an antifungal composition comprising natamycin and at least one insecticide that interferes with the nervous system and / or muscular system of insects and nematodes, wherein the at least one insecticide is selected from emamectin benzoate, alpha-cypermethrin, fipronil, lambda-cyhalothrin, acetamiprid, clothianidin, imidacloprid, thiacloprid, thiamethoxam, abamectin, and chlorantraniliprole, the volume-based average particle diameter of the natamycin is 0.2 μm to 10 μm, and applying the composition to agricultural plants or plant parts, A method for protecting agricultural plants or plant parts, comprising: Claims 9. The method according to claim 8, wherein the plant part is a seed, bulb, fruit or vegetable. Claims 10. Preparing an antifungal composition comprising natamycin and at least one insecticide that interferes with the nervous system and / or muscular system of insects and nematodes, wherein the at least one insecticide is selected from emamectin benzoate, alpha-cypermethrin, fipronil, lambda-cyhalothrin, acetamiprid, clothianidin, imidacloprid, thiacloprid, thiamethoxam, abamectin, and chlorantraniliprole, the volume-based average particle diameter of the natamycin is 0.2 μm to 10 μm, and bringing the composition into contact with the plant, A method for improving the growth and / or yield of agricultural plants, comprising: Claims 11. A method for protecting soil and / or a growth substrate, comprising applying to the soil and / or growth substrate an antifungal composition comprising natamycin and at least one insecticide that interferes with the nervous system and / or muscular system of insects and nematodes, wherein the at least one insecticide is selected from emamectin benzoate, alpha-cypermethrin, fipronil, lambda-cyhalothrin, acetamiprid, clothianidin, imidacloprid, thiacloprid, thiamethoxam, abamectin, and chlorantraniliprole, wherein the volume-based average particle diameter of the natamycin is from 0.2 µm to 10 µm, said method.

12. The method according to claim 11, wherein the growth substrate is a mushroom growth substrate.

13. The method according to any one of claims 9 to 12, wherein the composition is undiluted or diluted with an aqueous solution or oil before providing the composition to a plant, plant part, soil and / or growth substrate.

14. Use of an antifungal composition comprising natamycin and at least one insecticide that interferes with the nervous system and / or muscular system of insects and nematodes for the protection of plants, plant parts, soil and / or growth substrate from fungi, wherein the at least one insecticide is selected from emamectin benzoate, alpha-cypermethrin, fipronil, lambda-cyhalothrin, acetamiprid, clothianidin, imidacloprid, thiacloprid, thiamethoxam, abamectin, and chlorantraniliprole, The volume-based average particle diameter of the natamycin is 0.2 μm to 10 μm for the use described above. **Claim 15**: The method according to any one of claims 8 to 13 or the use according to claim 14, comprising 1% to 98% (w / w) of natamycin and 1% to 99% (w / w) of the at least one insecticide. **Claim 16**: The method according to any one of claims 8 to 13 or the use according to claim 14, wherein the ratio of natamycin to the at least one insecticide is 1:1 to 1:5000 (w / w). **Claim 17**: The method according to any one of claims 8 to 13, claim 15, and claim 16, or the use according to any one of claims 14 to 16, further comprising an agriculturally acceptable carrier. **Claim 18**: The method according to any one of claims 8 to 13 and claims 15 to 17, or the use according to any one of claims 14 to 17, which is an aqueous composition or an oily composition. **Claim 19**: The method according to any one of claims 8 to 13 and claims 15 to 18, or the use according to any one of claims 14 to 18, wherein the natamycin is ground to an average particle diameter of 0.5 μm to 3 μm. **Claim 20**: The method according to any one of claims 8 to 13 and claims 15 to 19, or the use according to any one of claims 14 to 19, further containing an insoluble polyelectrolyte complex of polyanion and polycation with a relative amount of 1:2 to 60:1 (w / w).

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