SPRAYABLE ANTIFUNGAL COMPOSITION FOR USE IN AGRICULTURE - ASSOCIATED PROCESS AND USE

Abstract

The present invention relates to a sprayable antifungal composition for use in agriculture. Characteristically, according to the invention, it contains a vehicle and as active ingredients at least one compound chosen from resveratrol, smilagenin and hecogenin and at least one trace element chosen from copper, iron, potassium, magnesium, molybdenum and manganese ions and in particular a mixture of these 6 ions.

Description

Title of the invention: SPRAYABLE ANTIFUNGAL COMPOSITION FOR USE IN AGRICULTURE - ASSOCIATED METHOD AND USE Technical field

[0001] The present invention relates to an antifungal composition usable in agriculture which comprises as active agents compounds of natural origin. Prior art

[0002] Pesticides are a major source of environmental pollution because they accumulate in soils and diffuse into groundwater. A large quantity of pesticides is used to treat specific crops such as vines, fruit trees, vegetables or other plants grown in intensive agriculture. This is the case for industrial crops such as potatoes and cereals such as wheat, for example.

[0003] A good approach to reducing the amount of plant protection products is to target the plant diseases for which the largest amounts of plant protection products are used.

[0004] In order to reduce pollution caused by synthetic pesticides, the effects of which on living organisms, particularly humans, are sometimes little known, it is recommended to use plant protection products that contain active ingredients of natural origin. The use of products derived from plants poses a problem of reproducibility of the compositions. Indeed, the composition of plants varies greatly depending on where they grow, the season and the stage at which they are harvested for use. Using a given plant to make an extract or an infusion does not make it possible to regularly obtain a product that contains the active ingredients at their effective dose.

[0005] Technical problem to be solved

[0006] A technical problem that the present invention seeks to solve is to provide an antifungal composition containing active ingredients of natural origin which can be easily produced according to reproducible and standardized processes to result in a composition whose active ingredient contents are identified, standardized and whose agronomic results are constant.

[0007] Another technical problem that the present invention seeks to solve is to provide an antifungal composition that is harmless to animals and in particular to mammals including humans.

[0008] Another technical problem that the present invention aims to solve is to provide an antifungal composition that is not phytotoxic.

[0009] Another technical problem which the present invention seeks to solve is to provide an antifungal composition, in particular as mentioned above, which is effective against at least one pathogen chosen from: Altemaria solani, Plasmopara viticola, Septoria tritici, Fusarium graminearum, Erysiphe necator, Pythium ultimum and Rhizoctonia solani.

[0010] Another technical problem that the present invention seeks to solve is to provide an antifungal composition, in particular as mentioned above, which is effective for the preventive and / or curative treatment on a plant chosen in particular from plants of the following families: Brassicaceae including rapeseed, Fabaceae including soybean, Vitaceae including vine, Poaceae including wheat, corn, Rosaceae including apple and pear trees, Solanaceae including potato, tomato, eggplant, Alliaceae including onion, garlic, shallot and leek, Apiaceae including carrots, Asteraceae including lettuce, artichoke and sunflower, and Chenopodiaceae including beetroot.

[0011] Another technical problem which the present invention seeks to solve is to provide an antifungal composition which is effective against at least one of the soil pathogenic fungi and whose contamination occurs via the soil and preferably effective against several soil fungal pathogens, chosen from: Altemaria solani, Septoria tritici, Plasmopara viticola, Fusarium graminearum and Erysiphe necator, Pythium ultimum and Rhizoctonia solani.

[0012] Another technical problem which the present invention seeks to solve is to provide an antifungal composition which is harmless to the environment and easily degradable.

[0013] Another technical problem that the invention seeks to solve is to provide an antifungal composition usable in agriculture which is versatile, i.e. effective against at least two soil and / or foliar fungal pathogens.

[0014] Another problem which the present invention seeks to solve is to provide a composition which can be diluted before use and which, depending on its dilution, can be particularly effective against at least one given pathogen. Summary of the invention

[0015] The present invention relates to an antifungal composition usable in agriculture, which contains as active ingredients at least one compound chosen from resveratrol, smilagenin and hecogenin and at least one trace element chosen from iron, potassium, magnesium, copper, manganese and molybdenum ions and in particular a mixture of these 6 ions.

[0016] Advantageously, whatever the embodiment, the composition contains resveratrol, more particularly trans resveratrol.

[0017] Preferably, the composition contains resveratrol, smilagenin and hecogenin.

[0018] The composition may also contain resveratrol and hecogenin or resveratrol and smilagenin.

[0019] It is indeed the merit of the inventors to have demonstrated that these three compounds are effective, together or not, as antifungal agents and that their antifungal action could be increased by the use of at least one of the aforementioned ions. Yucca extracts which were known as antifungal agents contain many compounds including saponins which were considered to be active. However, many saponins can be harmful to human health. Smilagenin and hecogenin which are steroidal saponins are present only in small quantities in the aqueous extract of yucca. It was therefore not obvious that these particular saponins could have an antifungal effect at very low doses.

[0020] The publication by EG Wulff entitled “Yucca shidigera extract: a potential bio-fungicide against seedborne pathogens of sorghum” and published in the journal Plant Pathology on August 3, 2011 clearly shows that a synthetic mixture of saponins containing 8 to 25% by mass of saponin proves to be less effective than an aqueous solution containing 10% by mass of yucca extract on sorghum seed pathogens. Not all saponins are therefore effective as antifungals.

[0021] Furthermore, resveratrol, smilagenin and hecogenin are known to be harmless to human health. Resveratrol is known for its anti-inflammatory and antioxidant effects. Smilagenin is being studied for its use in the treatment of orphan diseases, particularly neurological diseases. Hecogenin is used to alleviate colitis or to treat atopic dermatitis and bronchial hyperreactivity.

[0022] The composition of the invention therefore proves to be effective as an antifungal and harmless to mammals, including humans. Detailed description

[0023] Smilagenin, hecogenin and resveratrol can be extracted and purified from the juice extracted from plants. Thus, resveratrol is present in fruit juices, especially grape juices. Saponins are present in tea, chickpeas, climbing ivy leaves, soapwort, quinoa, fenugreek leaves, licorice root, ginseng roots, horse chestnut cotyledons, white broth leaves and Dioscorea tubers, for example. It is therefore possible to use juices, infusions, extracts or decoctions of these plants, possibly in a mixture, to obtain the composition of the invention. Smilagenin, hecogenin and Resveratrol can also be synthesized, making it easy to produce a standardized composition.

[0024] The above-mentioned ions can be introduced in the form of salts, in particular sulfate, chloride, iodide, ammoniate, hydrochloride, oxychloride, hydroxide, sulfonate, carbonate, formate, acetate, gluconate, lactate, citrate and sodium and ammonium salts.

[0025] The antifungal composition of the invention may also comprise one or more chelating agents chosen from EDTA, HEEDTA, DTPA, EDDHA, EDDHMA, EDDCHA, EDDHSA, IDHA, HBED, and / or at least one complexing agent chosen from lignosulfonic acid and ethanolamine. These agents increase the solubility of the metal ions and thus stabilize the composition of the invention when it contains a solvent.

[0026] Thus, manganese, iron, magnesium, copper can be introduced in the form of sulfates. Molybdenum can be introduced in the form of sodium molybdate. Potassium is preferably introduced in the form of potassium chloride.

[0027] The composition of the invention may be liquid or viscous or in powder form. In this case, it may consist of the aforementioned ingredients and in particular consist of a mixture of resveratrol, smilagenin, hecogenin and salts of the six aforementioned ions. It may also be in the form of a suspension.

[0028] Advantageously, it contains a vehicle. The vehicle is not limited according to the invention. It is preferably water. The composition of the invention is preferably sprayable and / or capable of coating a solid. The composition may be a solution, a two-phase mixture, an emulsion or a micellar solution. It is advantageously an aqueous solution.

[0029] Advantageously, the composition contains a mass percentage of trans resveratrol greater than or equal to 0.01 and / or a mass percentage of hecogenin greater than or equal to 0.00032 and / or a mass percentage of smilagenin greater than or equal to 0.004.

[0030] The composition may contain resveratrol and hecogenin or resveratrol and smilagenin. It may also contain all three compounds.

[0031] According to an embodiment that can be combined with each of the other embodiments, the composition contains the aforementioned ions with the exception of the copper ion and / or the manganese ion. According to a particular embodiment that can be combined with each of the other embodiments, the composition contains molybdenum, magnesium and potassium as trace elements.

[0032] According to a particular variant, some of the active compounds are contained in yucca trunk juice and the composition therefore contains yucca trunk juice.

[0033] According to a variant combinable with each of the embodiments or variants aforementioned, in its diluted form, the composition of the invention contains a mass percentage of yucca trunk juice greater than or equal to 0.0012.

[0034] The composition of the invention can be presented in two forms, a diluted form usable as is or a concentrated form to be diluted or dissolved before use.

[0035] In its concentrated form, according to particular embodiments, combinable with each of the aforementioned embodiments, the composition of the invention may contain a mass percentage of yucca juice greater than or equal to: 1; 2; 2.5; 3; 3.5; 4; 4.5; 5; 5.5; 6; 6.5 or 10.

[0036] Thus, the mass percentage of yucca juice can, whatever the embodiment of the invention, be equal to: 0.025; 0.03; 0.05; 0.06; 0.075; 0.09; 0.1; 0.15; 0.18; 0.2; 0.25; 0.3; 0.6; 1 (diluted form); 1.5; 2; 2.5; 3; 3.5; 4; 4.5; 5; 5.5; 6; 6.5; or 10 (concentrated form).

[0037] Whatever the embodiment, the mass percentage of the yucca juice may be less than 30 or 20.

[0038] Whatever the embodiment, the composition of the invention may contain a mass percentage of iron greater than or equal to 0.1 and less than or equal to 4 or 5 and / or a mass percentage of copper greater than or equal to 0.1 and less than or equal to 4 or 5 and / or a mass percentage of potassium greater than or equal to 0.05 and less than or equal to 0.5 and / or a mass percentage of molybdenum greater than or equal to 0.01 and less than or equal to 0.05 and / or a mass percentage of manganese greater than or equal to 0.1 and less than or equal to 5 and / or a mass percentage of magnesium greater than or equal to 0.01 and less than or equal to 1.

[0039] Whatever the embodiment, the composition of the invention may contain more particularly a mass percentage of iron greater than or equal to 0.6 and less than or equal to 0.9 and in particular equal to 0.77 and / or a mass percentage of copper greater than or equal to 0.3 and less than or equal to 0.45 and in particular equal to 0.375 and / or a mass percentage of magnesium greater than or equal to 0.12 and less than or equal to 0.19 and in particular equal to 0.15, and / or a mass percentage of potassium greater than or equal to 0.09 and less than or equal to 0.13 and in particular equal to 0.109 and / or a mass percentage of molybdenum greater than or equal to 0.0035 and less than or equal to 0.005 and in particular equal to 0.004 and / or a mass percentage of manganese greater than or equal to 0.9 and less than or equal to 2.7 and notably equal to 1.18 or 2.24.

[0040] Preferably, the composition of the invention in its concentrated or diluted form contains the 6 ions in combination, each present in particular in the aforementioned ranges.

[0041] Preferably, the composition contains iron, magnesium, potassium and molybdenum.

[0042] Preferably, according to an embodiment combinable with each of the embodiments aforementioned embodiments, the composition further contains, as active ingredient, humic acids and / or boron and / or zinc.

[0043] Boron may be in the form of boron ethanolamine, for example.

[0044] According to another embodiment, combinable with each of the embodiments, it contains as trace element zinc and / or iron and / or manganese and / or copper.

[0045] Advantageously, the mass percentage of boron is equal to or greater than 0.01 and less than or equal to 1 and more particularly greater than or equal to 0.04 and equal to or less than 0.06 and in particular equal to 0.05. The mass percentage of humic acids is equal to or greater than 0.01 and less than or equal to 10 and more particularly greater than or equal to 0.17 and equal to or less than 0.26 and in particular equal to 0.21.

[0046] Zinc can be in the form of zinc sulfate.

[0047] According to an embodiment which can be combined with each of the aforementioned embodiments, it contains a mass percentage of zinc greater than or equal to 0.1 and less than or equal to 10 and more particularly greater than or equal to 1.54 and less than or equal to 1.84 and in particular equal to 1.54.

[0048] The present invention also relates to a sprayable antifungal composition, usable in agriculture, which contains the composition of the invention diluted in a solvent and in particular water; more particularly the sprayable composition may contain from 0.05% to 10% by mass of the composition of the invention and in particular 0.05%, 0.2%, 0.4%, 0.5%, 1%, 2%, 3%, 6% or 10% by mass of the aforementioned composition, diluted in a solvent and in particular water.

[0049] Advantageously, the sprayable composition has an acidic pH greater than or equal to 3.2. At these pH values, the sprayable composition proves, at the effective doses, to be non-phytotoxic.

[0050] The present invention also relates to the use of a composition or a sprayable composition according to the invention as an antifungal against at least one pathogen chosen from Alternaria solani, Septoria tritici, Plasmopara viticola, Fusarium graminearum and Erysiphe necator, Pythium ultimum and Rhizoctonia solani.

[0051] The present invention also relates to a method for treating a plant, in particular chosen from plants of the following families: Brassicaceae including rapeseed, Fabaceae including soybean, Vitaceae including vine, Poaceae including wheat, corn, Rosaceae including apple trees, pear trees, Solanaceae including potato, tomato, eggplant, Alliaceae including onion, garlic, shallot or leek, Apiaceae including carrots, Asleraceae including lettuce, artichoke or sunflower and Chenopodiaceae including beetroot.

[0052] According to the invention, according to which the sprayable composition is applied to the leaves of said plant or the composition is applied to the seeds of said plant or said sprayable composition according to the invention.

[0053] The present invention also relates to the use of resveratrol and / or smilagenin and / or hecogenin and in particular of a mixture of trans resveratrol and smilagenin and / or hecogenin as an antifungal and in particular as an antifungal against a foliar fungal pathogen or a soil fungal pathogen chosen in particular from Fusarium graminearum, Pythium ultimum and Rhizoctonia solani.

[0054] The present invention also relates to the use of a yucca juice for the treatment of a plant chosen in particular from the vine, the solanaceae including the potato, the tomato and the eggplant, the cereals including wheat and corn against at least one fungal pathogen living in the soil and chosen in particular from Altemaria solani, Septoria tritici, plasmopara viticola, Fusarium graminearum and Erysiphe necator, Pythium ultimum and Rhizoctonia solani.

[0055] The present invention also relates to an antifungal composition which contains as active ingredients at least one compound chosen from resveratrol, smilagenin and hecogenin and preferably which contains these three compounds and humic acids and / or boron. This composition can be declined according to the same embodiments as the aforementioned composition, with regard to the concentrations of active agents, the vehicles or solvents and the pH, in particular. It can also be diluted in water according to the same dilutions as for the aforementioned sprayable composition. It can also be sprayed on the same plants as those described with reference to the composition described in the present application and containing at least one ion. The boron can be in the form of boron ethanolamine.

[0056] The present invention also relates to an antifungal composition which contains as active ingredients at least one compound chosen from resveratrol, smilagenin and hecogenin and preferably which contains these three compounds and zinc, in particular zinc sulfate, for example. This composition can be declined according to the same embodiments as the aforementioned composition with regard to the concentrations of active agents, the vehicles or solvent and the pH, in particular. It can also be diluted in water according to the same dilutions as for the aforementioned sprayable composition. It can also be sprayed on the same plants as those described with reference to the composition described in the application and containing at least one ion. DEFINITIONS

[0057] The term "trans-resveratrol" refers to 5-[(E)-2-(4-hydroxyphenyl)-ethenyl] benzene-1,3-diol.

[0058] The term “smilagenin” refers to (25R)-5beta-Spirostan-3beta-ol.

[0059] The term "hecogenin" refers to (3[3,5a,25R)-3-Hydroxyspirostan-12-one

[0060] The terms “mass percentage of A” designate the mass fraction of A in a mixture of at least two constituents.

[0061] The terms “volume percentage of A” designate the volume fraction of A in a mixture of at least two constituents.

[0062] The term "yucca trunk juice" refers to an aqueous composition containing, in particular, resveratrol, saponins including smilagenin, trans-3,3',5,5'-tetrahydroxy-4'-methoxystilbene, larixinol and phenol derivatives designated by the terms yuccaols A, B, C, D and E of yuccaone A and hecogenin.... Steroidal saponins represent only 6.8% by mass of the saponins. Yucca trunk juice may also contain yucca leaf juice but preferably it contains only the juice from the trunk.

[0063] The term "yucca" refers to all plants of the genus yucca and preferably yucca schidigera or yucca gloriosa. The juice may be a mixture of juices from yucca of different genera.

[0064] The yucca trunk juice is preferably heated to a temperature between 80°C and 120°C for a time equal to or greater than 1 min and equal to or less than 10 min and preferably heated to 100°C for 3 min.

[0065] The term "boron ethanolamine" refers to the complex formed by ethanolamine and boron.

[0066] The term "humic acids" refers to high molecular weight, negatively charged, black to dark brown polymers resulting from an oxidative condensation process of phenolic compounds and bound to amino acids, peptides and polysaccharides. They are present in humus.

[0067] Throughout the application, when mass percentages are given with reference to several ingredients, it should be understood that these mass percentages of these ingredients are independent of each other. Figures

[0068] [Fig.l] represents the percentage of sporulating leaf surface as a function of the treatments, the reference GA342 designates the preferred mixture which makes it possible to obtain the composition of the invention by dilution, the volume percentage is indicated alongside, the control is pure water. EXPERIMENTAL EXAMPLES Yucca juice

[0069] The yucca juice used is obtained by crushing and pressing Yucca schidigera trunks. The liquid obtained was heated to 100°C for 3 min in order to sterilize it. This juice is used in all the experiments documented in the application. Direct tests against Alternaria Solani Identification of active compounds

[0070] Yucca juice is a complex mixture. It contains a large number of saponins, including steroidal saponins. It also contains resveratrol, particularly in its trans form.

[0071] The inventors have identified compounds in yucca juice that are effective as antifungal agents: these are resveratrol, smilagenin and hecogenin.

[0072] Aqueous solutions containing a given concentration of resveratrol or smilagenin were tested on a calibrated suspension of A. maria solani spores.

[0073] In vitro evaluations were carried out with a 96-well microtiter assay. A range of concentrations of each compound was tested against a calibrated suspension of A. maria solani spores. Three replicates (wells) were carried out. The EC50 value of each concentration towards A. solani was determined from its impact on the reduction of fungal growth measured using optical density. The results are presented in the following Table 1 which shows the % decrease in the growth of A. solani upon contact with a solution containing a given concentration of resveratrol or smilagenin.

[0074] Throughout the application, the % indicated in the tables with reference to the compositions tested are % by mass; the inhibition produced by the compositions tested is indicated in % in the tables.

[0075] [Tables 1] Concentration (pg / ml or ppm) resveratrol inhibition (%) smilagenin inhibition (%) hecogenin inhibition (%) yucca juice inhibition (%) 0.0064 6.20 7.9 2.90 2.9 0.032 13.60 10.1 5.00 5.00 0.16 13.70 19.6 2.70 2.70 0.8 16.10 22.6 1.00 1.00 4 11.20 28.0 6.60 6.60 20 28.70 29.3 2.50 2.50 40 30.00 6.80 6.80 100 54.70%

[0076] Furthermore, the inhibition on the aforementioned pathogen of various compositions of yucca juice diluted in water were tested. The results are grouped in Table 2 below.

[0077] [Tables2] Concentrations % yucca juice inhibition (2.5%) yucca juice inhibition (10%) 0.05% 2.9 14.4 0.5% 5.00 28 1% 2.70 40.4 2% 1.00 42.1 3% 6.60 45.9 6% 2.50 51.2 10% 6.80 54.9

[0078] It appears from the results in Table 1 that trans-3,4',5-trihydroxystilbene (resveratrol trans form) from the stilbene family at a concentration of 100 ppm in water is capable of reducing the growth of A. solani by 54.7%. It also appears that smilagenin, which is a steroidal saponin, can reduce the growth of A. solani by 30% at a concentration of 40 ppm in water. Hecogenin, on the other hand, has no antifungal action on the pathogen studied. It is likely that the antifungal action of yucca juice comes mainly from the combined presence of resveratrol and smilagenin in yucca juice. The lower antifungal action of yucca juice is due to the lower concentration of resveratrol and smilagenin in yucca juice. In fact, in the table above resveratrol and smilagenin are used diluted in water.Since the concentration of smilagenin and resveratrol in yucca juice is lower than in the tested compositions of smilagenin and resveratrol, the antifungal activity of yucca juice is lower at a dose of 2.5%. By increasing the concentration of yucca, the amounts of resveratrol and smilagenin increase in the same proportions and the effectiveness of Yucca juice becomes notable at 10%.

[0079] Synergy of active compounds with trace elements

[0080] All the compositions described below have a density equal to or greater than 1.15 and equal to or less than 1.2. The pH of each of the solutions is acidic. It is for example adjusted by adding citric acid and / or acetic acid.

[0081] The following compositions were prepared: Composition V1

[0082] Composition VI contains the ingredients indicated in the following Table 3.

[0083] [Tables3] Ingredients Mass (g) Water 75.231 Humic acids 0.486 Potassium salts 0.243 Magnesium salts 0.486 Molybdenum salts 0.010 Copper salts 1.500 Iron salts 4.049 Zinc salts 7.000 Boron salts 0.486 Manganese salts 3.706 Yucca juice 2.500 100.000 Composition V6

[0084] Composition V6 contains the ingredients indicated in the following Table 4.

[0085] [Tables4] Products % by mass Water 84.198 Yucca juice 2.5 Manganese salts 7.000 Potassium salts 0.243 Iron salts 4.049 Magnesium salts 0.500 Copper salts 1.500 Molybdenum salts 0.010

[0086] Dilutions of some of the above compositions were made as shown in Table 5 below.

[0087] The solutions obtained were tested directly on the pathogen A. solani as previously indicated. The results are grouped in Table 5. The pro- percentages in reference to the tested compositions are mass percentages.

[0088] [Tables 5] % by mass in tested composition in water tested composition 0.05% 0.50% 1.00% 2.00% 3.00% 6.00% 10.00% Composition V1 0.00 41.26 67.19 86.25 100.0 100.0 100.0 Manganese salts 1.5% + water + yucca juice 2.5% 0.00 10.0 20.7 53.38 100 100 100 Copper salts 1.5% + water + yucca juice 2.5% 0.0 0.0 13.2 36.0 74.9 100.0 100.0 Iron salts 4.05% + water + yucca juice 2.5% 0.0 0.0 1.2 12.3 15.0 40.1 84.6 Potassium salts 0.24% + water + yucca juice 2.5% 3.7 13.5 10.3 16.0 18.1 18.5 16.8 Magnesium salts 0.5% + water + yucca juice 2.5% -2.1 5.5 8.1 6.5 4.8 3.1 -2.6 Molybdenum salts 0.01% + water + yucca juice 2.5% 9.1 29.0 28.4 25.8 27.0 17.5 19.2 Water + yucca juice 2.5% 0.0 0.0 0.0 0.0 7.1 29.9 65.5 Composition V6 0.00 77.10 109.80 102.90 103.30 97.8 102.2

[0089] Table 6 below groups together the EC50 and EC95 values ​​of the different mixtures tested on A. solani.

[0090] [Tableauxô] Tested composition ec50 (% by mass) ec95 (% by mass) Composition V1 0.65% 2.53 Composition V6 0.31 1.30 iron salts 4.05% + yucca juice 2.5g + water 5.85 >10% KC1 0.24% + water + yucca juice 2.5% >10% >10% magnesium salts 0.5% + water + yucca juice 2.5% >10 >10 Copper salts 1.5% + water + 2.5% yucca juice 2.11 2.80 Molybdenum salts 0.01% + water + yucca juice 2.5% >10 >10 yucca juice 3% + water 7.96 >10

[0091] In view of the results in Tables 5 and 6, it is noted that compositions VI and V6 diluted in water are effective against A. solani, which is not the case for the composition containing only 2.5% yucca juice or only yucca juice and one of the trace elements.

[0092] The presence of the mixture of trace elements and in particular copper reduces the EC50 value. In vitro effect on R. solani

[0093] Dilutions of some of the above compositions were made as shown in Table 7 below.

[0094] The solutions obtained were tested directly on the pathogen R. solani as previously indicated. The results are grouped in Table 7. The percentages are mass percentages.

[0095] [Tables?] R. solani % by mass VI V6 Yucca juice 1 Yucca juice 2 0.05% 101.3 0 0.3 8.6 0.50% 100.5 0 -5.3 95.2 1.00% 101.7 72.1 3.6 98.0 2.00% 96.8 101.5 32.5 101.3 3.00% 97.9 93.5 40.4 101.1 6.00% 90.9 96.0 56.7 98.7 10.00% 91.5 92.7 62.0 1003 EC50 (%) <0.05 0.93 0.15 0.93 ec95 (%) <0.05 1.65 0.49 1.65

[0096] It can be seen from the results of the above-mentioned table that composition V1 is effective as soon as it is at a mass percentage of 0.05% while composition V6 is effective against R. solani from 1% or 2%. It can also be seen that yucca juices 1 and 2, which are yucca schidigera juices from different producers, do not have the same effectiveness on R. Solani. At a dose of 0.5%, “Yucca juice 1” has an effectiveness of 0 while “Yucca juice 2” records a performance of 95.2%. This difference in effectiveness clearly shows that it is not possible to produce a standard effective composition in a reproducible manner using products derived from plants directly.

[0097] In vitro tests - direct tests against Plasmopara viticola - Septoria tritici - Fusarium graminearum - Erysiphe necator

[0098] The evaluations were made with a 96-well microtiter test to evaluate the in vitro fungicidal activity of each solution.

[0099] A range of concentrations (up to 8) of each solution was tested against a calibrated suspension of Fusarium graminearum spores and against a calibrated suspension of Zymoseptoria tritici.

[0100] Three repetitions (wells) were carried out. The concentration value which allows 50% inhibition of the growth of each pathogen (EC50) was determined from the measurement of the optical density.

[0101] a) Fusarium graminearum

[0102] Fusarium graminearum belongs to the species Gibberella zeae, ascomycete fungi of the Nectriaceae family. This fungus is the pathogen of fusarium blight of cereals and maize causing heavy yield losses. It causes contamination of harvested grains by mycotoxins (fusariotoxins) dangerous to human and animal health.

[0103] This disease develops both on seeds in the soil and at the foliar level during the season.

[0104] The experimental results obtained according to the same protocol as described above are shown in Table 8 below. The percentages are mass percentages.

[0105] [Tables8] F. graminearum % mass of the mixture tested in water V6 VI Mixture water + 2.5% mass of yucca juice 0.05% 0 19.2 3.4 0.5% 0 57.3 9.7 1% 49.8 76.1 24.2 2% 107.1 80.8 30.3 3% 107.3 87.5 36.6 6% 109.3 100.0 38.9 10% 109.3 100.0 40.7 ec50 (%) 1.05 0.34 >10 ec95 (%) 1.65 4.40 >10

[0106] It can be seen from the results in Table 8 that the addition of trace elements makes it possible to reduce by almost 3 times the quantity of yucca juice to be added to water to obtain 100% efficiency.

[0107] The effect of the active compounds was tested on Fusarium graminearum.

[0108] The results are shown in Table 9 below. Concentrations (pg / ml or ppm) Resveratrol Hecogenin Smilagenin 0.0064 0.30 2.40 5.00 0.032 -2.90 -4.50 -2.60 0.16 -3.90 -1.40 -0.80 0.8 -1.60 -1.50 2.40 4 -3.80 0.00 1.50 20 18.20 -2.70 -8.70 100 53.30 -6.40 2.00 EC50 (pg / ml or ppm) 29.49 >100 >100 EC95 (pg / ml or ppm) >100 >100 >100

[0110] Resveratrol (trans) at 100 ppm can reduce Fusarium graminearum infestation by more than 50%. b) Septoria tritici

[0111] Septoria tritici is an ascomycete fungus responsible for septoria blight of wheat. Its average damage is estimated at 17 q / ha (up to 50 q / ha in the most exposed situations). Due to its frequency and the extent of the potential damage, S. tritici is the most important disease of soft wheat.

[0112] The same experiments as above were carried out. The results are grouped in Tables 10 and 11 below. The percentages indicated are mass percentages.

[0113] [Tables10] S. tritici Concentrations (%) VI Water + 10% yucca juice water + 2.5% yucca juice 0.05% 19.10 14.3 6.9 0.50% 40.30 67.4 16.2 1.00% 65.50 80.1 33.6 2.00% 91.70 86.4 42.7 3.00% 96.4 85.7 46.5 6.00% 100.0 85.8 50.6 10.00% 100.0 86.3 53.3 EC50 (%) 0.89 0.44 4.42 ec95 (%) 2.58 >10 >10

[0114] [Tables 11] S. tritici Concentrations (%) VI water + 2.5% yucca juice V6 0.05% -7.2 -1.6 -2.6 0.50% 6.0 43.8 89.7 1.00% 44.3 75.6 91.0 2.00% 63.9 90.0 99.2 3.00% 80.0 94.1 97.4 6.00% 100.0 100.8 100.4 10.00% 100.0 107.0 100.4 EC50 (%) 1.25 0.68 0.17 EC95 (%) 4.90 3.05 1.50

[0115] It is found that the solution containing composition V1 shows total effectiveness at a concentration of 6% by mass of composition VI. At this concentration, the mixture of water + yucca juice shows 100% effectiveness. The diluted composition V6 shows effectiveness as soon as it is present at 0.5% or 1% by mass. It proves to be more effective than composition VI. It is therefore found that the addition of trace elements increases the antifungal character of the yucca juice and therefore the antifungal character of at least one of the identified active compounds, namely resveratrol, smilagenin and hecogenin.

[0116] The antifungal character of the active compounds was tested on S. tritici. The results are grouped in Table 12 below. The figures show the % inhibition of the growth of the pathogen.

[0117] [Tablesl2] Concentrations (pg / ml or ppm) Resveratrol Hecogenin Smilagenin 0.0064 0.00 13.50 -3.90 0.032 -0.40 13.70 -2.30 0.16 -0.40 11.50 -1.90 0.8 0.60 12.90 -9.50 4 -2.60 12.30 0.00 20 -2.00 22.80 -2.10 100 104.90 20.50 -0.20 EC5o (pg / ml or ppm) 72.01 >100 >100

[0118] Resveratrol and hecogenin show efficacy against Septoria tritici at very low concentrations, 100 ppm and 20 ppm respectively.

[0119] c) Test on P. ultimum

[0120] The same experiments as above were carried out. The results are grouped in Tables 13 and 14 below. The percentages indicated are mass percentages except for the % inhibition.

[0121] [Tablesl3] P. ultimum % by mass VI Yucca juice V6 0.05 101.9 - - 0.50 104.8 - - 1.00 105.2 - - 2.00 109.7 - - 3.00 109.3 - - 6.00 101.9 - - 10.00 109.4 - - EC50(%) <0.05 - - ec95(%) <0.05 - - Concentrations (pg / ml or ppm)) resveratrol hecogenin smilagenin 0.0064 -8.20 12.80 -6.70 0.032 -0.30 20.70 0.00 0.16 0.00 28.80 -0.60 0.8 -1.70 36.30 0.10 4 23.30 55.10 -3.40 20 72.40 68.40 -0.10 100 101.30 85.20 0.00 EC50 (pg / ml or ppm) 8.89 2.87 >100 EC95 (pg / ml or ppm) 73.85 >100 >100

[0123] Resveratrol and hecogenin are found to be 100% and 85% effective respectively at a dose of 100 ppm. Composition VI is found to provide 100% effectiveness at low concentrations, from 0.05% (Table 13). The combination of the active molecules resveratrol and hecogenin associated with trace elements makes it possible to achieve complete effectiveness at the lowest concentrations, from 0.05%. d) Test on Rhizoctonia solani

[0124] The same experiments as above were carried out. The results are grouped in Table 15 below. The percentages indicated are mass percentages. R. solani % mass VI water + 2.5% yucca juice V6 0.05% 101.3 0.3 -3.6 0.50% 100.5 -5.3 -2.0 1.00% 101.7 3.6 72.1 2.00% 96.8 32.5 101.5 3.00% 97.9 40.4 93.5 6.00% 90.9 56.7 96.0 10.00% 91.5 62.0 92.7 EC50 (%) <0.05 4.60 0.93 ec95 (%) <0.05 >100 1.65

[0126] It is found that composition V1 is more active than composition V6 probably due to the presence of humic acids and / or boron salt. Test on zoospores of Plasmopara viticola

[0127] The objective is to evaluate the direct effect in vitro of the composition of the invention on the mobility of zoospores of grapevine downy mildew (Plasmopara viticola) suspended in water. Downy mildew (Plasmopara viticola) is a strain-population originating from the Burgundy vineyard, maintained by weekly transplanting on plants. The experiment is carried out in vitro: sporangia of Plasmopara viticola are harvested from a sporulating vine leaf (inoculated 7 days previously) and cultured in suspension in ultrapure water at a concentration of at least 106 sporangia / ml. The sporangia release motile zoospores within one hour. This mobility is verified at the end of the experiment on the control suspension.

[0128] Dilutions from sporangium solutions are prepared extemporaneously in ultrapure water to obtain a final zoospore concentration of 0.33% by mass.

[0129] The effect on zoospore mobility is observed within 5 minutes, using a Malassez cell, under a microscope. A response gradient is observed within this time and noted according to the following scale:

[0130] High efficiency or 100%: all zoospores are immobile (or even destroyed),

[0131] The results are shown in Table 16 below. The percentages are mass percentages. Compositions tested % by mass Test on vine cuttings against Plasmopara viticola Direct test against Plasmopara viticola VI 0.0033 / high eff-100 VI 0.033 / 100 VI 0.33 98-100% 100 VI 0.67 98-100% / Yucca juice 0.0033 / 0 Yucca juice 0.033 / high eff Yucca juice 0.33 71-89 100

[0133] It is noted from the results in Table 16 that the diluted composition VI blocks the mobility of zoospores of Plasmopara viticola at the reference concentration of 0.033% (v / v). Even diluted to 0.0033%, the mixture blocks or disrupts the mobility of zoospores of the pathogen. The density of the product being equal to or greater than 1.15 and equal to or less than 1.2, the mass percentage of composition V6 equivalent to the dilution of 0.0033 is equal to 0.004% which is equivalent to a mass percentage of yucca juice equal to 0.00012%. Here again, it is noted that the presence of the ions increases the antifungal activity of resveratrol and smilagenin contained in the yucca juice. Yucca juice at a dose of 0.33% has an action against zoospore mobility at a concentration 100 times higher (0.33%). TESTING ON PLANTS UNDER CONTROLLED CONDITIONS

[0134] Tests on plants under controlled conditions make it possible to get closer to the real conditions of use of plant protection products.

[0135] In these tests, the product to be tested is no longer placed directly in vitro in contact with the pathogen but sprayed onto the leaves of the plant. The latter is grown in a greenhouse and therefore in the light. This verifies the effectiveness of the product in almost real conditions.

[0136] Controlled trials - Plasmopara viticola on vines

[0137] The plant material consists of 2-month-old herbaceous cuttings of the Marselan variety (Grenache x Cabernet-Sauvignon) propagated in the INRAE ​​greenhouses, grown in 0.5 L pots (2 / 3 potting soil-1 / 3 perlite). At 2 months, the cuttings have 5-6 spread leaves. The mildew inoculum (Plasmopara viticola') is a strain-population originating from the Burgundy vineyard, maintained by weekly transplanting on plants. At least five plants are used per method.

[0138] The composition of the invention to be tested is sprayed onto the cuttings with a hand sprayer below the runoff point, on both sides of the leaves. The droplets are left to dry for 2 to 3 hours before placing the plants back in the greenhouse.

[0139] The plants are inoculated 48 hours post-treatment, by spraying with a suspension of Plasmopara viticola sporangia (104 sporangia / ml) prepared extemporaneously, on the underside of the leaves. After inoculation, the plants are placed in a humid tent for at least 3 hours to allow the zoospores to encyst, then they are placed back in the greenhouse.

[0140] In the vine, very young leaves (approximately 1 / 3 of their final size) and older leaves are resistant to the disease (ontogenetic resistance). For this reason, the disease is generally observed on 2 sensitive leaves designated rank 1 leaf and rank 2 leaf. Rank 1 corresponds to a leaf that has reached approximately 2 / 3 of its final surface area, rank 2 leaf, located immediately below rank 1 leaf, is an adult-sized leaf, still susceptible to mildew.

[0141] Five days after inoculation, at least 6 discs of 1.3 cm in diameter are removed using a punch per leaf and per row, immediately placed adaxial side down on wet paper, in a Plexiglas box. The latter is left for 2 days in a culture chamber (day 10h 20°C / night 14h 18°C) to allow sporulation. The results correspond to the percentage of disc surface covered by sporangiophores, determined by image analysis. The infection percentage values ​​indicated in the graphs for each treatment correspond to the average of the values ​​obtained for 30 discs of "row 1" or 30 discs of "row 2". The observation of the sporulation rate is carried out by image analysis for a rating on the leaves of ranks 1 and 2. A rate of 5% corresponds to an infection already visually marked, a rate higher than 10% indicates a strong infection (the disc is almost entirely covered with sporangia).

[0142] The first trial was carried out with the initially recommended doses of IL, 2L and 3L / ha diluted in 150 L / ha, i.e. 6.67 ml / L, 13.33 ml / L and 20 ml / L. The following 2 experiments were carried out with lower doses: 1.67 ml / L, 3.33 ml / L and 6.67 ml / L.

[0143] Reference GA342 designates composition V1 in [Fig. 1]. The dilution is shown next to it.

[0144] The means were compared with the Tuckey T-test (comparison of the solution containing composition V1 with the water control).

[0145] The average efficacy of composition VI at the dose of 0.33% over 5 tests is 98.7%. The efficacy of composition VI against vine downy mildew is almost total from the dose of 1.7 mL / L or 0.17% by volume. The inhibition under in vitro conditions is well correlated with that under in vivo conditions. The density of the composition being equal or greater than 1.15 and equal to or less than 1.2.

[0146] Furthermore, other mixtures containing water and yucca juice were tested against plasmopara viticola on vines using the same protocol as above (3 repetitions of 2 or 3 foliar sprays at 1-week intervals). The results are grouped in Table 17 below. Tested composition % by volume of the tested composition Plant: vine Effectiveness on Plasmopara viticola (%) VI 0.67-1.33-2 100 0.17 98-100 0.0033 / 0.033 / 0.33 100 0.67 98-100 Yucca juice 3g + water 83.7g 0.33 71 Copper salts 1.5g + water 94.5g + yucca juice 3g 0.33 99 Zinc salts 7g + water 80g + yucca juice 3g 0.33 96 Humic acids 0.486g + water 96.514g + yucca juice 3g 0.33 97 Boron salts 0.486g + water 95.514g +yucca juice 3g 0.33 96 water 83.2g +yucca juice 3g + Mg salts MgSO4 0.5g 0.33 27 Water 90g +yucca juice 3g +Mn salts 7g 0.33 71 Water 96.76g +yucca juice 3g+ K salts 0.24g 0.33 51 Water 92.95g +yucca juice 3g +Fe salts 4.05g 0.33 83

[0148] It can be seen from the results in Table 17 that the addition of the mixture of trace elements increases the antifungal activity of the composition of the invention.

[0149] In conclusion, it is noted that the composition according to the invention used in the light and on a plant remains effective as an antifungal. Field trials (composition VI)

[0150] In field trials, [Fig.l] shows that composition VI significantly reduces the first mildew infestations on leaves and bunches compared to a control. The dose of 3 1 / ha improves the effectiveness compared to the lower doses 1 and 2 1 / ha. The effectiveness level reaches 60% on leaves and bunches. In [Fig.l], the percentage (%) is a volume percentage. Thus, in this figure, 0.17% indicates that 1.7 mL of composition VI was dissolved in 998.3 mL of water. In mass percentage, the tested solution contains 0.20% of composition VL Effect of pH on efficacy and phytotoxicity

[0151] In the above example, the compositions obtained from composition V1 had different pHs, as indicated in Table 18 below. The percentages (%) indicated are volume percentages.

[0152] [Tables 18] 0.17% 0.33% 0.67% PH 3.48 3.28 3.06

[0153] Slight necroses were observed with the 0.67% concentration, identical to those of the first test at the same concentration; the two lower concentrations are less aggressive.

[0154] It is therefore noted that the dilution of composition V1 makes it possible to increase its pH and therefore to reduce its phytotoxicity without reducing its effectiveness.

Claims

Claims

1. Antifungal composition usable in agriculture, containing molybdenum ions as active ingredients, characterized in that it also contains, as active ingredients: - at least one compound chosen from resveratrol and smilagenin and hecogenin, and - at least one trace element chosen from iron, potassium, magnesium, copper, manganese ions and in particular a mixture of these 5 or more ions.

2. Composition according to claim 1, characterized in that it further contains a vehicle and in particular water as vehicle / solvent.

3. Composition according to any one of the preceding claims, characterized in that said resveratrol is predominantly trans-resveratrol.

4. Composition according to one of the preceding claims, characterized in that it contains a mass percentage of trans resveratrol greater than or equal to 0.01 and / or a mass percentage of hecogenin greater than or equal to 0.00032 and / or a mass percentage of smilagenin greater than or equal to 0.

004.

5. Composition according to any one of the preceding claims, characterized in that it contains a mass percentage of yucca juice greater than or equal to 1; 2; 2.5; 3; 3.5; 4; 4.5; 5; 5.5; 6; 6.5 or 10

6. IV. Composition according to any one of the preceding claims, characterized in that it contains a mass percentage of iron greater than or equal to 0.1 and less than or equal to 4 or 5 and / or a mass percentage of copper greater than or equal to 0.1 and less than or equal to 4 or 5 and / or a mass percentage of potassium greater than or equal to 0.05 and less than or equal to 0.5 and / or a mass percentage of molybdenum greater than or equal to 0.01 and less than or equal to 0.05 and / or a mass percentage of manganese greater than or equal to 0.1 and less than or equal to 5 and / or a mass percentage of magnesium greater than or equal to 0.01 and less than or equal to 1.

7. Composition according to any one of the preceding claims, characterized in that it further contains, as active ingredients, humic acids and / or boron and / or zinc.

8. Sprayable antifungal composition usable in agriculture, ca- characterized in that it contains in particular from 0.05% to 10% of the composition according to any one of claims 1 to 7, diluted in a solvent and in particular water and in particular 0.05, 0.2%, 0.4%, 0.5% 1%, 2%, 3%, 6% or 10% by mass of the composition according to any one of claims 1 to 7, diluted in a solvent and in particular water.

9. Sprayable composition according to claim 8, characterized in that it has an acidic pH greater than or equal to 3.

2.

10. Method for antifungal treatment of a plant chosen in particular from plants of the following families: Brassicaceae including rapeseed, Fabaceae including soybean, Vitaceae including vine, Poaceae including wheat, corn, Rosaceae including apple trees, pear trees, Solanaceae including potato, tomato, eggplant, Alliaceae including onion, garlic, shallot or leek, Apiaceae including carrots, Asteraceae including lettuce, artichoke, sunflower, Chenopodiaceae including beetroot, against in particular at least one pathogen chosen from Altemaria solani, Septoria tritici, Plasmopara viticola, Fusarium graminearum and Erysiphe necator, Pythium ultimum and Rhizoctonia solani, characterized in that the sprayable composition according to claim 9 or 10 is applied to the leaves of said plant or in that the sprayable composition according to claim 9 or 10 is applied to the leaves of said plant. seeds of said plant the composition according to any one of the preceding claims.

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