Antifungal formulations for seed dressing, treated seeds and methods

Abstract

The present invention relates to formulations for seed dressing, treated seeds (seed dressing), and dressing methods for protecting seeds and emerging root systems against fungal growth. In particular, the seed dressing embodiments described herein are capable of substantially reducing and / or eliminating fungal contaminants from seeds, root systems, seedlings, and the surrounding environment, where such fungal contaminants include but are not limited to Fusarium, Septoria, Pythium and Rhizoctonia species.

Description

[0001] ANTIFUNGAL FORMULATIONS FOR SEED DRESSING, TREATED SEEDS AND METHODS

[0002] ***** ***** *****

[0003] FIELD OF THE INVENTION

[0004] The present invention refers to the field of products for agricultural crops and, in particular, to the field of formulations for seed dressing.

[0005] STATE OF THE ART

[0006] The growing demand for cereals needed to satisfy human nutrition and the increased demand for vegetables, including horticultural plants for the health and well-being of consumers, has caused a significant increase in cereal and vegetable crops grown throughout the world, particularly in Europe and the United States. Unfortunately, persistent pathogenic infections in both cereals and green leafy vegetables at early stages of development can cause yield losses of up to 70% [see for example Larsson et al., “Disease progression and yield losses from root diseases caused by soilborne pathogens of spinach” Phytopathology 82:403-406 (1992)], resulting in substantial economic losses for growers and increased food prices for consumers. In particular, green leafy vegetables (horticultural plants) are very susceptible to infections due to both poor germination and root rot diseases, which can cause difficult rooting of the roots in infected fields during the first week of development. In infected soils, for example, during the early stages of plant development the yield may decrease due to rapid weakening or death of seedlings, before or after emergence, as well as poor overall growth and yellowing of the leaves. Susceptibility to poor germination and root rot can be exacerbated by poor field conditions, including over-watering, inadequate drainage and / or restricted spacing between planting plants. Various pathogenic species found in soil can cause poor germination or root rot and the most common pathogens include fungal and fungal-like species (oomycetes) such as, for example, Fusarium, Pythium, Septoria and Rhizoctonia. The poor growth of seedlings due to poor germination and / or root rot can also affect other cereal crops, such as wheat, barley, rice, and horticultural crops such as pea, soybean, tomato, alfalfa and other cereals.

[0007] In general, control of lower germination or / and root rot has been attempted with various methods. One method, for example, is to sow the seeds in a sterilized growing medium; such a method, however, may prove ineffective as fungal spores may be introduced into the growing medium on the seeds themselves or after sowing (e.g. in water or by wind). Additionally, methods that require sterile growing environments may be impractical and / or increase costs, particularly for growers managing significant acreage.

[0008] Another method widely used in the agricultural sector to control fungal infections is the use of fungicides. Fungicides such as, for example, Metalaxyl (N- (methoxyacetyl)-N-(2,6-xylyl)-DL-alaninate), Thiram

[0009] (dimethylcarbamothioylsulfanyl-N,N-dimethylcarbamodithioate), Ipconazole (2- [4(chlorophenyl)methyl]-5-(1 -methylethyl)-1 -(1 H-1 ,2,4- triazolylmethyl)cyclopentanol), Fludioxonil (4-(2,2-difluoro-2 / - / -1 ,3-benzodioxol-4- yl)-1 / - / -pyrrole-3-carbonitrile) and Fluxapyroxad (1 H-Pyrazole-4-carboxamide, 3- (difluoromethyl)-l -methyl-N-(3',4',5'-trifluoro[1 , 1 '-biphenyl]-2-yl)), have been shown to effectively control fungal infections in emerging root systems, but the application of conventional chemicals such as those reported makes these materials unsuitable for commercial organic crop production.

[0010] Copper salts such as copper(ll) hydroxide and tribasic copper(ll) sulfate have found wide application in the agricultural industry as fungicides and bactericides. Formulations of copper(ll) hydroxide and tribasic copper(ll) sulfate (Champ®, Kocide® and Cuproxat®) are commercially available in granular, liquid and water- dispersible forms and are traditionally used in crops by foliar application, spraying or otherwise coating the leaf surface; in this way the application takes place directly on the plant in post-emergency. Thus, such contact fungicides in their foliar application are ineffective to treat or prevent fungal conditions present in the soil, infections that invade the emerging root system in the early stages of development (e.g. in the early stages of planting).

[0011] Currently, there are formulations for the treatment of cereal seeds, commercially available (e.g. commercial formulations based on fluxapyroxad (e.g. Systiva® and Rubin® Plus), fludioxonil (e.g. Celest®, Maxim® 4 FS and Spirato® 480 FS , Sedaxane®, Vibrance Gold®), suitable for the prevention of seed, root and seedling diseases, but having the drawback to be not suitable for use in organic cultivation. For organic crops, in particular, there are formulations for the treatment of seeds comprising microbial species, known as biopesticides (e.g. Micosat F® and Cerall®) and which are intended to mitigate the effects of fungal diseases. However, biopesticides can be difficult to control under environmental conditions such as changes in temperature of soil and air, changes in ambient and soil moisture, and changes in soil nutrient content. The slow growth of these microbial species is equally problematic, as the microbes may not develop appropriately to counteract soil pathogens as regards to root colonization. This slow microbial growth is particularly disadvantageous with respect to Pythium and other fungal species, which establish themselves in the emerging root system within just 7-10 days of planting. Furthermore, some biopesticides have problems in maintaining their activity and effectiveness as they require to be kept at low temperatures (for example, products based on Pseudomonas Chlororaphis).

[0012] WO2015075645A1 and WO2015075644A1 describe formulations for the treatment of plants comprising copper(ll) as a bioactive agent, a carrier consisting of carbonated hydroxyapatite ( / .e. Microsap®) and, optionally, a plant extract or an active component thereof with antifungal / antibacterial activity.

[0013] There are several literature references for seed dressing suitable for organic cultivation and carried out in the presence of copper salts, but in the absence of organic fungicides, such as for example CN113735650, CN10824389, GB503915, US4859466, FR2140269, US20190169080, but the proposed solutions are not completely satisfactory. Furthermore, the formulations currently available for seed dressing have disadvantages; therefore, the need for new seed dressing formulations for use in the production of organic crops is particularly felt. SUMMARY OF THE INVENTION

[0014] The Applicant has surprisingly and unexpectedly found that tribasic copper(ll) sulfate, a carrier endowed also with micronutrient characteristics, such as carbonated hydroxyapatite in aqueous suspension, and optionally but not necessarily plant extracts and / or active components thereof, when combined in a formulation for seed dressing, and in particular for seed coating, act synergistically to fight fungal pathogens and strengthen the emerging root system.

[0015] The object of the present invention is, therefore, the use of a formulation comprising: a) a fungicidal agent consisting of copper from tribasic copper(ll) sulfate; b) a carrier consisting of carbonated hydroxyapatite; and c) optionally, a plant extract or an active component thereof with antifungal / antibacterial activity; as a seed dressing agent to protect seeds and emerging root system from fungal infection.

[0016] Such a formulation has proven surprisingly effective in efficiently and cost-effectively protecting seeds and emerging root systems from fungal infections.

[0017] Aspects of the present invention, therefore, also concern a seed coated with the aforementioned formulation, and a seed dressing method.

[0018] A further object of the present invention is also a seed dressed with the aforementioned antifungal formulation, said seed comprising a coating at least partially surrounding the seed, said coating comprising a) a fungicidal agent comprising copper from tribasic copper(ll) sulfate; b) a carrier, also acting as a micronutrient, consisting of carbonated hydroxyapatite; and c) optionally, a plant extract or an active component with antifungal / antibacterial activity.

[0019] A further object of the present invention is also a seed dressing method comprising dispensing an antifungal formulation as described above onto the surface of a seed or set of seeds.

[0020] A further object of the present invention is also a method of protecting a seed or its emerging root system from a fungal infection, comprising: i. providing at least one seed; ii. coating the at least one seed with the antifungal formulation as described above; iii. drying, then, the formulation to form said at least one coated seed, iv. planting said at least one coated seed under conditions suitable for germination.

[0021] DETAILED DESCRIPTION OF THE INVENTION

[0022] For the purposes of the present invention, carbonated hydroxyapatite means a carbonate substituted hydroxyapatite having formula (I):

[0023] Ca(10-y / 2)(PO4)(6-y)(CO3)y(OH)2 (I) wherein y is between 0.01 and 2.

[0024] Without wishing to be bound by any theory, it is believed that the seed dressing formulation object of the present invention is capable of providing protection against fungal species present in the soil in which the seeds are planted. In particular, in one aspect, the formulation may be capable of both acting as a fungicide and promoting root growth to ensure healthy plant growth from the planted seed. It is also believed that the absorption of the antifungal and antibacterial agent represented by tribasic copper(ll) sulfate, and possibly by plant extracts and / or the active components thereof, is significantly improved by the presence of an inorganic carrier such as carbonated hydroxyapatite, which aids the transport of tribasic copper(ll) sulfate and that of plant extracts and / or the active components thereof across the fungal cell membrane, while improving the absorption of metallic micronutrients also present in the soil by the emerging plant. As a result, the formulations provide a means to both attack the fungal pathogen at the cellular level and strengthen the root system in a manner that allows that root system to better defend itself against pathogen invasion.

[0025] Seed dressing according to the present invention can be defined as the application of a material to a seed before planting it in soil to improve the characteristics of the seed, protect it before germination, support its germination and / or support the growth of the resulting plant. In the embodiments of the present invention, the seed dressing or treatment includes agricultural active ingredients bound to (e.g. coated onto) a seed or a plurality of seeds. “Treating” as used herein refers to any effect, for example, attenuation, reduction or modulation, resulting in the improvement of the condition, disease, disorder and the like.

[0026] The seed dressing formulation according to the present invention can be effective in reducing severity or preventing various diseases. In particular, the seed dressing formulation can mitigate the effects of, or prevent fungal diseases of the emerging root system. In some embodiments, the dressing formulation according to the present invention may be effective in mitigating or preventing fungal infection of an emerging root system by one or more of the following species: Fusarium, Microdochium, Tilletia tritici, Ustilago nuda, Ustilago tritici, Ustilago hordei, Septoria tritici, Drechslera sorokiniana, Gaeumannomyces graminis, Lapschocita, Botrytis, Rhizoctonia, Alternaria, Colletotrichum, Macrophomina, Phyllosticta, Phytophthora, Pseudomonas, Pythium, Sclerotium, Thielaviopsis, Ascochyta; preferably Fusarium, Microdochium, Tilletia tritici, Ustilago tritici, Septoria tritici, Rhizoctonia, Alternaria, Colletotrichum, Pythium, Ascochyta; or preferably Fusarium, Microdochium, Tilletia tritici, Ustilago tritici, Septoria, Drechslera sorokiniana, Botrytis, Rhizoctonia. Alternaria, Colletotrichum, Pythium, Ascochyta.

[0027] Commercially available forms of tribasic copper(ll) sulfate that may be employed in the seed dressing formulation of the present invention include, but are not limited to, Cuproxat® SDI and Cuprital® SDI containing 26.95% of tribasic copper sulfate. Various concentrations of tribasic copper(ll) sulfate may be used in the seed dressing formulation in amounts sufficient to mitigate or prevent fungal infection of an emerging root system. Such amounts can be determined in light of this invention and according to methods known to those skilled in the art.

[0028] Preferably, the antifungal formulation for seed dressing to protect seeds and emerging root systems from fungal infections, is comprising: a) 1 -9% w / w of a copper fungicidal agent consisting of copper from tribasic copper(ll) sulfate; b) 1 -7% w / w of a carrier consisting of carbonated hydroxyapatite; and c) 0-3% w / w of a plant extract or an active component thereof with antifungal / antibacterial activity; wherein the % w / w are based on the total weight of the formulation.

[0029] Preferably, the formulation according to the use of the present invention comprises 2-8% w / w of copper from tribasic copper (II) sulfate, more preferably 2-5% w / w.

[0030] The seed dressing formulation according to the present invention comprises a carrier consisting of carbonated hydroxyapatite which also acts as a micronutrient. Carbonated hydroxyapatite is a source of micronutrients, such as calcium and phosphorus, that remain available to the emerging root system, providing additional strength and vitality. Preferably, the formulation of the present invention comprises 1 -5% w / w of carbonated hydroxyapatite, more preferably 1.1 -4.8% w / w.

[0031] According to a preferred embodiment of the present invention, the amount of carbonated hydroxyapatite in the formulation is equal to 1 -14% w / w with respect to tribasic copper(ll) sulfate present in the formulation, more preferably 2-12% w / w, even more preferably 4-8% w / w.

[0032] According to another preferred embodiment of the present invention, the carrier consists of an aqueous suspension of carbonated hydroxyapatite at 10-30% w / w with respect to the weight of the suspension.

[0033] According to a preferred embodiment of the present invention, it comprises a carrier consisting of carbonated hydroxyapatite whose Ca / P molar ratio is comprised between 1 .5 and 2.5, preferably between 1 .7 and 2.4, more preferably between 1 .8. and 2.3.

[0034] The seed dressing formulation according to the present invention preferably comprises at least one plant extract and / or active components thereof present in an amount comprised between 0.5% and 8% w / w with respect to the total weight of the formulation, preferably in an amount between 1 % and 5% w / w.

[0035] In accordance with the present invention, the plant extract or active component thereof with antifungal / antibacterial activity is preferably selected from the plant extracts of Abies sibirica, Malpighia glabra, Achillea millefolium, Allium sativum, Medicago sativa, Aloe vera Citrus sinensis, Artemisia annua, Arnica Montana, Ocimum basilicum, Betula pendula, Betula pubescens, Calendula officinalis, Matricaria chamomilla, Chamaemelum nobile, Cinnamomum verum, Gotu kola, Chelidonium majus, Syzygium aromaticum, Allium cepa, Equisetum arvense, Curcuma longa, Echinacea purpurea, Echinacea angustifolia, Eucalyptus globulus, Hypericum perforatum, Fucus vesiculosus, Gentiana lutea, Lavandula angustifolia, Citrus limon, Melilotus officinalis, Melissa officinalis, Punica granatum, Peppermint, Mentha spicata, Vaccinium myrtillus, Orthosiphon stamineus, Urtica dioica, Olea europeae, Tabebuia impetiginosa, Plantago lanceolata, Hieracium pilosella, Pinus sibirica, Polypodium leucotomos, Citrus paradise, Quassia amara, Rheum Thais, Rosa canina, Rosmarinus officinalis, Ruscus aculeatus, Salix alba, Salvia officinalis, Camelia sinensis, Tilia tomentosa, Thymus vulgaris, Arctostaphylos uva-ursi, Valeriana officinalis, Solidago virgaurea, Loranthus europaeus, Zingiber officinalis and mixtures thereof.

[0036] According to one embodiment, the plant extract is selected from extracts of Syzygium aromaticum and Mentha spicata. In one embodiment, the active components of the plant extracts are selected from 1,8-cineole, thymol, eugenol, eucalyptol, alpha pinene, beta pinene, d-Hmonene, menthol, camphorene, carvacrol, carvone, caryophyllene, linalool.

[0037] Preferably, the formulation of the present invention further comprises at least one further source of micronutrients selected in the group consisting of zinc, manganese, iron, boron, chromium, cobalt, copper and molybdenum salts, such as phosphates, oxides, sulfates, carbonates, chlorides, nitrates, borates, molybdates and the like, and mixtures thereof. For example, the following representative materials may be used as sources of micronutrients: zinc sulfate, ethylenediaminetetraacetic acid disodium zinc salt, zinc nitrate, calcium sulfate, calcium nitrate, magnesium sulfate, magnesium nitrate, ferrous sulfate, ferrous nitrate, manganese sulfate, manganese nitrate, copper sulfate, copper nitrate, boric acid, sodium borate, sodium molybdate, ammonium molybdate and the like, alone or in combination.

[0038] In a preferred embodiment, the source of micronutrients is zinc sulfate and ethylenediaminetetraacetic acid disodium zinc salt.

[0039] According to a preferred embodiment of the present invention, the amount of micronutrients is comprised between 0.5% and 5% w / w with respect to the total weight of the formulation, preferably in an amount between 1 % and 3% w / w.

[0040] The seed dressing formulation can be used with all types of seeds.

[0041] In one embodiment, the seed dressing formulation is used with straw cereal seeds such as common wheat, durum wheat, barley, triticale, rye, oats, millet, spelt, and rice; preferably common wheat, durum wheat, barley, rice.

[0042] In one embodiment, the seed dressing formulation is used with vegetable seeds such as pea, chickpea, alfalfa, soy, tomato, melon, watermelon, broad bean, field bean, kidney bean, lentil, aubergine, pepper, cucumber, pumpkin, courgette, carrot, parsley, fennel, celery, onion, shallot, leek, spring onion, mustard, radish, rocket, lettuce, endive, chicory, radicchio, lamb’s lettuce, valerianella, spinach, coriander; it is preferably used for pea, chickpea, alfalfa, soy, tomato, melon, watermelon, coriander, onion, cabbage, lettuce; more preferably it is used for pea, alfalfa, soybean, tomato, melon, coriander, onion.

[0043] Generally, and preferably, the seed dressing formulation may comprise one or more additives performing various functions. For example, the seed dressing formulation may include a binding agent for improved adherence of the seed treatment formulation to the seed, a source of humic acid for improved chelation, a soil amendment for improved availability and absorption of micronutrients, and other additives that can confer a beneficial effect onto the emerging root system.

[0044] In some embodiments, the seed dressing formulation includes at least one soil amendment. Soil amendments can be effective in stimulating antimicrobial activity and improving seed growing conditions. A representative soil amendment that can be used is BorreGro® CA (commercially available from LignoTech AGRO), a purified calcium lignosulfonate product. Typically used as a soil based product, a soil amendment for use with the seed dressing formulation according to the present invention is suitable for application to the seed itself (e.g. by binding or coating it) rather than being deposited in the surrounding land.

[0045] Preferably, the seed dressing formulation according to the present invention comprises a soil amendment in an amount not exceeding about 3%, otherwise it may have a negative effect onto the emerging root system due to toxicity, more preferably in an amount ranging from about 0.5% to about 3%, based on the total weight of the formulation; more preferably in an amount ranging from about 1 % to about 2%, based on the total weight of the formulation, or more preferably in an amount ranging from about 0.5% to about 1.5%, based on the total weight of the formulation.

[0046] In some embodiments, the seed dressing formulation according to the present invention may include a binder component. The binding component may consist of an adhesive polymer which can be natural or synthetic and has no phytotoxic effects on the seed to be treated. The binder may be selected from polyvinyl acetates, polyvinyl acetate copolymers, polyvinyl alcohols, polyvinyl alcohol copolymers, celluloses, including ethylcelluloses and methylcelluloses, hydroxymethylcellulose, hydroxypropylcellulose, hydroxymethylpropylcellulose, polyvinylpyrrolidones, dextrins, maltodextrins, polysaccharides, fats, oils, proteins, fiber gums including gum arabic, shellac, vinylidene chloride, vinylidene chloride copolymers, calcium lignosulfonates, acrylic copolymers, starches, polyvinyl acrylates, zeins, gelatin, carboxymethylcellulose, chitosan, polyethylene oxide, acrylimide polymers and copolymers, polyhydroxyethyl acrylate, methylacrylimide monomers, alginate, ethylcellulose, polychloroprene and syrups or mixtures thereof. Preferred binders include polymers and copolymers of vinyl acetate, methylcellulose, polyvinyl alcohol, vinylidene chloride, acrylic, cellulose, polyvinylpyrrolidone and polysaccharide. Particularly preferred classes of binders include polyvinyl acetates (e.g. for use in conventional cultivation) and fiber gums (e.g. for use in organic applications).

[0047] In a preferred embodiment, the seed dressing formulation according to the present invention may comprise a colorant selected from natural colorants such as carotene, anthocyanins, flavonoids, lycopene, chlorophyll, carminic acid, lutein, safflower, spirulina.

[0048] In a preferred embodiment, the seed dressing formulation according to the present invention may comprise a source of humic acid, which functions as a complexing agent for soil nutrients and maintains soil nutrients in forms that are more readily available to the plant. In an exemplary embodiment, the seed dressing formulation comprises humic acid in an amount ranging from about 0.5% to about 3% by weight based on the total weight of the formulation; more preferably in an amount from about 0.5% to about 1.5% of the total seed dressing formulation, even more preferably in an amount of about 1 % humic acid based on the total weight of the formulation. In some preferred embodiments, the humic acid source is substantially free of fulvic acid or fulvic acid additives, which would produce undesirable precipitates in the formulation.

[0049] The seed dressing formulation according to the present invention is applied to the seed and dried to form a coating. The relative amounts of the ingredients in the formulation will be the same as the relative amounts in the coating except for evaporated water. Any of the ingredients described for the seed treatment will also be ingredients in the coating.

[0050] The seed dressing formulation may be applied to a seed or a set of seeds by methods known to those skilled in the art on seeds with a maximum humidity of 12%. The dressing can be applied to the seed or bundle of seeds by various treatment methods, including drum coating, spin coating, pan coating, fluidized bed coating, and the like. In certain preferred embodiments, at least 90% of the seed surface area is coated with the formulation of the invention, more preferably at least 95% of the seed surface area is coated, even more preferably the seed or seeds are substantially encapsulated by the seed dressing coating, i.e. 100% of the surface area of the seeds is coated with the formulation.

[0051] The seed dressing formulation is used to produce an effective seed coating in order to protect emerging root systems from fungal infections. In some embodiments, the coating is effective in protecting the emerging root system from fungal infection for at least 30 days. As a further example, the coating is effective for protecting the emerging root system from fungal infection for at least 40 days. As another example, the coating is effective for protecting the emerging root system for at least 50 days. As another example, the coating is effective for protecting the emerging root system for at least 60 days. As a further example, the coating is effective for protecting the emerging root system for at least 70, 80, 90, 100, 110, 120 days. The coating may be effective for protecting the seed from pre-emergence fungal infection, postemergence fungal infection, or both pre- and post-emergence fungal infection.

[0052] In certain exemplary embodiments, the seed dressing formulation is organic (i.e., organic certified). For a seed dressing formulation to be organic, it should meet specific government standards. These government standards vary from country to country. The term “organic” or “certified organic” as used herein is intended to indicate that the seed dressing formulation met the standards to be designated as organic in the European Union and the United States.

[0053] The present invention can be better understood in light of the following examples. EXPERIMENTAL PART

[0054] The following non-limiting examples are provided to further illustrate the present invention. It will be apparent to those skilled in the art that the techniques described in the following examples may be considered examples of modes of practice. However, in light of the present invention, those skilled in the art should understand that modifications can be made to the specific embodiments that are described and still achieve the same or similar result without departing from the spirit and scope of the invention. EXAMPLE 1 - Preparation of a formulation containing 7.5% Cu from tri basic copper sulfate, 20%> Microsap®, 2% zinc

[0055] The formulation of example 1 was prepared by mixing 49.2 g of Cuproxat S.D.I., containing 7.5 g of copper, 8.6 g of zinc sulfate heptahydrate, 20 grams of Microsap containing 16-18% of carbonated hydroxyapatite, and 23 g of water under stirring up to complete homogenization.

[0056] EXAMPLE 2 - Preparation of a formulation containing 7.5 ) of Cu from tribasic copper sulfate, 20% of Microsap®, 2% of zinc, and 1%> of a mixture of origan and bitter orange oils

[0057] Similarly to example 1 , the formulation of example 2 was prepared from 49.2 g of Cuproxat S.D.I. containing 7.5 g of copper, 8.6 g of zinc sulfate heptahydrate, 20 grams of Microsap containing 16-18% of carbonated hydroxyapatite, 21 g of water, 1 g of a mixture of oils consisting of origan and bitter orange oils in a 1 :1 v:v ratio, and 1 g of propylene glycol.

[0058] EXAMPLE 3 - Preparation of a formulation containing 3.5% of Cu from tribasic copper sulfate, 20% of Microsap®, 2% of zinc and 3% of a mixture of origan and bitter orange oils

[0059] Similarly to what was described in example 1 , the formulation of example 3 was prepared from 23.5 g of Cuproxat S.D.I. containing 3.5 g of copper, 8.6 g of zinc sulfate heptahydrate, 20 grams of Microsap containing 16-18% of carbonated hydroxyapatite, 19 g of water, 3 g of a mixture of oils consisting of origan and bitter orange oils in a 1 :1 v:v ratio, and 1 g of propylene glycol.

[0060] EXAMPLE 4 - Preparation of a formulation containing 3.5% of Cu from tribasic copper sulfate, 2% of zinc and 3% of a mixture of origan and bitter orange oils Similarly to what was described in example 1 , the formulation of example 4 was prepared from 23.5 g of Cuproxat S.D.I. containing 3.5 g of copper, 8.6 g of zinc sulfate heptahydrate, 39 g of water, 3 g of a mixture of oils consisting of origan and bitter orange oils in a 1 :1 v:v ratio, and 1 g of propylene glycol. EXAMPLE 5 - Preparation of a formulation containing 3.5% Cu from tri basic copper sulfate, 2% zinc, 20% Microsap® and 3%> clove oil

[0061] Similarly to what was described in example 1 , the formulation of example 5 was prepared from 23.5 g of Cuproxat S.D.I. containing 3.5 g of copper, 8.6 g of zinc sulfate heptahydrate, 20 grams of Microsap containing 16-18% carbonated hydroxyapatite, 19 g of water, 3 g of clove oil, and 1 g of propylene glycol.

[0062] EXAMPLE 6 - Preparation of a formulation containing 7.5% Cu from tribasic copper sulfate, 2% zinc, and 1% clove oil

[0063] Similarly to example 1 , the formulation of example 7 was prepared from 49.2 g of Cuproxat S.D.I. containing 7.5 g of copper, 8.6 g of zinc sulfate heptahydrate, 21 g of water, 1 g of clove oil and 1 g of propylene glycol.

[0064] EXAMPLE 7 (comparative) - Preparation of Example 1 from US20190169080 Similarly to what was described in example 1 of US Patent Application 20190169080, 16.2 g of anhydrous zinc sulfate, 11 .3 g of copper(ll) hydroxide (Idrox WG) corresponding to 7.5% copper, 1.3 g of lignin sulfonate, and 9.5 g of zeolite (Zeocel) were mixed with 59.0 g of water until a homogeneous suspension was obtained. 1.6 g of xanthan gum was then added and stirred for complete homogenization.

[0065] EXAMPLE 8 - In vitro activity on pathogens for examples 1-7 for cereal and horticultural seed pathogens

[0066] The formulations of examples 1 -7 were suitably diluted with water at a 1 :1 dilution equal to 500 ml of formulation with 500 ml of water for the treatment of 100 kg of cereal seeds, and equal to 1000 ml of formulation with 1000 ml of water for the treatment of 100 kg of vegetable seeds.

[0067] The formulations of examples 1 -7 were tested on the following pathogens

[0068] Table 1 - Application of the formulations of examples 1 -7 onto cereal seeds: in vitro efficacy tests on pathogens

[0069] Table 2 - Application of the formulations of examples 1 -7 onto vegetable seeds: in vitro tests of efficacy on pathogens

[0070] The in vitro inhibition tests on the pathogens indicated above were performed by evaluating the growth of the fungal colony on the agar substrate alone and on Potatoes Dextrose Agar (PDA Dlifco) as a reference test, and the products of examples 1 -7 were added to the same substrate at a concentration of 0.16%.

[0071] At the beginning of the test (time 0) a circle of 0.5 cm in diameter made up of agar colonized by the previously cultivated fungal colony was placed in the center of Petri dishes of 8.5 cm in diameter containing 10 ml of substrate. The plate was then closed with parafilm and placed for incubation at 22 + / - 1 °C for the entire duration of the test. 5 repetitions were set up for each product and for each concentration. Starting from two days of incubation until the end of the test, when the colonies of the test sample had colonized the entire agar surface, the diameter of the colonies was measured. The standard error was calculated for the diameter values. The evaluation of the efficacy of each formulation of examples 1 -7 was calculated by comparing the variation x of the average value of the fungal colony growth diameter on the substrate due to the tested formulation and the average value of the growth diameter of the test sample, based on the following formula

[0072] The activity is expressed as a Gl% factor for which the closer to 100 the better the antifungal activity of the formulation. The efficacy was evaluated on the last day of the tests equal to 10 days of incubation Table 3 reports the % activity of the formulations carried out on cereals

[0073] Table 4 reports the % activity of the formulations carried out on horticultural crops

Claims

CLAIMS1 . A use of an antifungal formulation comprising: a) a fungicidal agent consisting of copper from tribasic copper(ll) sulfate; b) a carrier consisting of carbonated hydroxyapatite; and c) optionally, a plant extract or an active component thereof with antifungal / antibacterial activity; as a seed dressing agent to protect seeds and emerging root system from fungal infection.

2. The use according to claim 1 , wherein said fungal infection is caused by one or more species selected from the group consisting of Fusarium, Microdochium, Tilletia tritici, Ustilago nuda, Ustilago tritici, Ustilago hordei, Septoria tritici, Drechslera sorokiniana, Gaeumannomyces graminis, Lapschocita, Botrytis, Rhizoctonia, Alternaria, Colletotrichum, Macrophomina, Phyllosticta, Phytophthora, Pseudomonas, Pythium, Sclerotium, Thielaviopsis, Ascochyta; preferably Fusarium, Microdochium, Tilletia tritici, Ustilago tritici, Septoria tritici, Rhizoctonia, Alternaria, Colletotrichum, Pythium, Ascochyta; preferably Fusarium, Microdochium, Tilletia tritici, Ustilago tritici, Septoria, Drechslera sorokiniana, Botrytis, Rhizoctonia. Alternaria, Colletotrichum, Pythium, Ascochyta.

3. The use according to any one of claims 1 -2, wherein said formulation is comprising: d) 1-9% w / w of a copper fungicidal agent consisting of copper from tribasic copper(ll) sulfate; e) 1 -7% w / w of a carrier consisting of carbonated hydroxyapatite; and f) 0-3% w / w of a plant extract or an active component thereof with antifungal / antibacterial activity; wherein the % w / w are based on the total weight of the formulation.

4. The use according to any one of claims 1 -3, wherein the plant extract or an active component thereof with antifungal / antibacterial activity is selected from the plant extracts of Abies sibirica, Malpighia glabra, Achillea millefolium. Allium sativum, Medicago sativa, Aloe vera Citrus sinensis, Artemisia annua, ArnicaMontana, Ocimum basilicum, Betula pendula, Betula pubescens, Calendula officinalis, Matricaria chamomilla, Chamaemelum nobile, Cinnamomum verum, Gotu kola, Chelidonium majus, Syzygium aromaticum, Allium cepa, Equisetum arvense, Curcuma longa, Echinacea purpurea, Echinacea angustifolia, Eucalyptus globulus, Hypericum perforatum, Fucus vesiculosus, Gentiana lutea, Lavandula angustifolia, Citrus limon, Melilotus officinalis, Melissa officinalis, Punica granatum, Peppermint, Mentha spicata, Vaccinium myrtillus, Orthosiphon stamineus , Urtica dioica, Olea Europeae, Tabebuia impetiginosa, Plantago lanceolata, Hieracium pilosella, Pinus sibirica, Polypodium leucotomos, Citrus paradise, Quassia amara, Rheum Thais, Rosa canina, Rosmarinus officinalis, Ruscus aculeatus, Salix alba, Salvia officinalis, Camelia sinensis, Tilia tomentosa, Thymus vulgaris, Arctostaphylos uva-ursi, Valeriana officinalis, Solidago virgaurea, Loranthus europaeus, Zingiber officinalis, and mixtures thereof.

5. The use according to any one of claims 1 -4, wherein the formulation further comprises at least one further source of micronutrients selected from the group consisting of zinc, manganese, iron, boron, chromium, cobalt, copper and molybdenum salts such as phosphates, oxides, sulfates, carbonates, chlorides, nitrates, borates, molybdates and the like, and mixtures thereof.

6. The use according to any one of claims 1 -5, wherein the formulation comprises a further source of micronutrients selected from the group consisting of zinc salts, preferably ethylenediaminetetraacetic acid disodium zinc salt.

7. The use according to any one of claims 1 -6, wherein the seeds are selected from the group consisting of common wheat, durum wheat, barley, triticale, rye, oats, millet, spelt and rice, or from the group consisting of pea, chickpea, alfalfa, soy, tomato, melon, watermelon, broad bean, field bean, kidney bean, lentil, aubergine, pepper, cucumber, pumpkin, courgette, carrot, parsley, fennel, celery, onion, shallot, leek, spring onion, mustard, radish, rocket, lettuce, endive, chicory, radicchio, lamb’s lettuce, valerianella, spinach, coriander.

8. The use according to any one of claims 1 -7, wherein the formulation comprises a binding agent for improved adherence of the seed treatmentformulation to the seed, and / or a source of humic acid for improved chelation, and / or a soil amendment to improve the availability and absorption of micronutrients and / or other additives that can confer a beneficial effect onto the emerging root system.

9. A seed comprising a coating at least partially surrounding the seed, said coating comprising: a) a fungicidal agent comprising copper from tribasic copper(ll) sulfate; b) a carrier also acting as a micronutrient, consisting of carbonated hydroxyapatite; and c) optionally, a plant extract or an active component thereof with antifungal / antibacterial activity.

10. A seed dressing method comprising dispensing an antifungal formulation, as defined in any one of claims 1 -8, onto the surface of a seed or set of seeds.

11. A method of protecting a seed or the emerging root system thereof from a fungal infection, said method comprising: i. providing at least one seed; ii. coating the at least one seed with the antifungal formulation as defined in any one of claims 1 -8; iii. drying then the formulation to form said at least one coated seed, iv. planting said at least one coated seed under conditions suitable for germination.

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