WO2015130911A1

WO2015130911A1 - Dried fungal spore germinative compound mixtures

Info

Publication number: WO2015130911A1
Application number: PCT/US2015/017703
Authority: WO
Inventors: Tommie Eugene HASHMAN
Original assignee: Envera, Llc
Priority date: 2014-02-27
Filing date: 2015-02-26
Publication date: 2015-09-03

Abstract

In one aspect, the present invention is directed to a dried intimate mixture comprising a fungus spore and a germinative compound, and methods for preparing the intimate mixture. In another aspect, this invention is directed to a composition comprising such an intimate mixture. The invention also relates to methods for increasing the germination, growth, metabolism, and/or enzyme activity of a fungus spore comprising preparing an intimate mixture of a fungus spore and a germinative compound. Plant propagative materials comprising a fungus spore and a germinative compound are also provided.

Description

DRIED FUNGAL SPORE GERMINATIVE COMPOUND MIXTURES

RELATED APPLICATIONS

This application claims priority to U.S. Provisional Patent Application No. 61/945,693 filed on February 27, 2014, the contents of which are incorporated herein in their entirety.

FIELD OF THE INVENTION

In one aspect, the present invention is directed to a mixture comprising a fungal spore and a germinative compound, and methods of preparing the mixture. In another aspect, this invention is directed to a composition comprising such a mixture. The invention also relates generally to methods for increasing the germination, growth, metabolism and/or enzyme activity of fungal spores, comprising preparing a mixture comprising a fungal spore and a germinative compound. BACKGROUND OF THE INVENTION

Fungi are omnipresent in the environment and have many beneficial uses. Fungi and the compounds they produce provide plants with nutrients, biological control agents and enzymes. The beneficial association of fungi with plants is evident in the symbiotic relationship demonstrated by mycorrhizas and plant roots. Fungi are also used in applications such as culturing of food, medicines, environmental remediation and production of industrial acids and useful enzymes. These and other applications are more readily conducted by increasing the speed, sustained growth and metabolism of fungi by influencing their germination and growth from the initial spore stage. Thus a need exists to develop methods of increasing fungal germination and growth for a wide range of commercial applications.

SUMMARY OF THE INVENTION A purpose of the invention is to control pathogens that cause rot or disease of the seed, root, leaves, stalks or fruiting structures of a plant. Another purpose is to increase the yield of plants and plant seed or fruit production. Yet another purpose is to increase fungal productivity in uses for food, medicinal, environmental and industrial applications.

It has been surprisingly found that fungal spore germination and growth are increased by treating the fungal spore with germinative compounds.

In certain embodiments, the present invention relates to a mixture comprising a fungal spore and a germinative compound. In a preferred embodiment, the mixture is a dried intimate mixture comprising a fungal spore and a germinative compound, wherein the fungal spore and the germinative compound are maintained in proximate position until they reach an environment conducive to germination. In certain embodiments of the aforementioned mixture, the germinative compound is adsorbed to or absorbed by the fungal spore.

In certain embodiments of the aforementioned mixture, the fungal spore is selected from the group consisting of Aspergillus flavus, Aureobasidium pullulans, Beauveria bassiana, Coniothyrium minitans, Candida oleophila, Gliocladium roseum, Gliocladium catenulatum, Metarhizium anisopliae, Paecilomyces fumosoroseus, Paecilomyces lilacinus, Pantoea agglomerans, Penicillium bilaiae, Penicillium gaestrivorus, Pseudozyma flocculosa, Pythium oligandrum, Trichoderma asperellum, Trichoderma virens, Trichoderma gamsii, Trichoderma hamatum, Trichoderma harzianum, Trichoderma harizatum, Trichoderma koningii, Trichoderma fertile, Ulocladium oudemansii, Ulocladium atrum, Alternaria destruens, Chondrostereum purpureum, Colletotrichum gloeosporioides, Phoma macrostoma, Typhula phacorrhiza, Penicillium chrysogenum, Penicillium griseofulvum, Penicillium pinophilum, Penicillium canescens, Penicillium glabrum, Penicillium glaucum, Penicillium halgiovense, Penicillium roqueforti, Penicillium simplicissium, Penicillium digitatum, Penicillium janthinellum,

Penicillium italicum, Penicillium spinulosum, Penicillium cyclopium, Penicillium

purpurogenum, Penicillium janczewskii, Penicillium montanense, Penicillium restrictum, Penicillium harzianum, Penicillium terrestre, Penicillium ochrochloron, Penicillium

aurantiogriseum, Penicillium crustosum, Penicillium decumbens, Penicillium griseofulvum, Penicillium janthinellum, Penicillium rugulosum, Penicillium velutinum, Penicillium

frequentans, Penicillium verrucosum, Penicillium implicatum, Penicillium citrinum, Glomus aggregatum, Glomus albidum, Glomus ambisporum, Glomus brazillanum, Glomus caledonium Glomus coremioides, Glomus claroideum, Glomus clarum, Glomus clavisporum, Glomus constrictum, Glomus coronatum, Glomus deserticola, Glomus diaphanum, Glomus eburneum, Glomus etunicatum, Glomus fasciculatum, Glomus flstulosum, Glomus fragilistratum, Glomus geosporum, Glomus globiferum, Glomus heterosporum, Glomus hoi, Glomus intraradices, Glomus lacteum, Glomus lamellosum, Glomus luteum, Glomus manihotis, Glomus

microaggregatum, Glomus monosporum, Glomus mosseae, Glomus multicaule, Glomus pansihalos, Glomus pustulatum, Glomus sinuosum, Glomus spurcum, Glomus tortuosum, Glomus trimurales, Glomus verruculosum, Glomus versiforme, Glomus viscosum, Gigaspora albida, Gigaspora decipiens, Gigaspora gigantean, Gigaspora margarita, and Gigaspora rosea.

In certain embodiments, the germinative compound is selected from the group consisting of an amino acid, a purine nucleoside, a salt, and a sugar. In certain embodiments, the amino acid is selected from the group consisting of glycine, L-alanine, L-arginine, L-aspartate, L- glutamate, L-histidine, L-isoleucine, L-lysine, L-methionine, L-serine, L-tryptophan, L-tyrosine, L-valine, L-proline, L-leucine, L-cysteine, L-threonine, L-glutamine, L-asparagine, L- phenylalanine and analogues thereof. In certain embodiments, the purine nucleoside is selected from the group consisting of inosine and adenosine. In certain embodiments the salt is selected from the group consisting of Calcium Chloride, Potassium Chloride, Potassium Phosphate, Potassium Carbonate, Potassium Propionate and Magnesium Sulphate. In certain embodiments, the sugar is selected from the group consisting of glucose, fructose, mannose and trehalose.

In some aspects, the invention relates to a composition comprising the aforementioned mixtures.

The invention also relates to a method for preparing the aforementioned mixture, comprising mixing a fungal spore and a germinative compound.

In a preferred embodiment, the invention relates to a method for preparing a dried intimate mixture comprising a fungal spore and a germinative compound, the method comprising:

a) preparing a solution comprising a fungal spore and a germinative compound; and

b) drying the solution to obtain a dried intimate mixture comprising a fungal spore and a germinative compound,

wherein the fungal spore and the germinative compound are maintained in proximate position until they reach an environment conducive to germination. In certain embodiments of the aforementioned method, the drying is selected from the group consisting of spray-drying, freeze-drying, vacuum-drying, air drying or drum drying.

In another aspect, the invention also provides a composition comprising:

(a) a fungal spore selected from the group consisting of Aspergillus flavus, Aureobasidium pullulans, Beauveria bassiana, Coniothyrium minitans, Candida oleophila, Gliocladium roseum, Gliocladium catenulatum, Metarhizium anisopliae, Paecilomyces fumosoroseus, Paecilomyces lilacinus, Pantoea agglomerans, Penicillium bilaiae, Penicillium gaestrivorus, Pseudozyma flocculosa, Pythium oligandrum, Trichoderma asperellum, Trichoderma virens, Trichoderma gamsii, Trichoderma hamatum, Trichoderma harzianum, Trichoderma harizatum, Trichoderma koningii, Trichoderma fertile, Ulocladium oudemansii, Ulocladium atrum, Alternaria destruens, Chondrostereum purpureum, Colletotrichum gloeosporioides, Phoma macrostoma, Typhula phacorrhiza, Penicillium chrysogenum, Penicillium griseofulvum, Penicillium pinophilum, Penicillium canescens, Penicillium glabrum, Penicillium glaucum, Penicillium halgiovense, Penicillium roqueforti, Penicillium simplicissium, Penicillium digitatum, Penicillium

janthinellum, Penicillium italicum, Penicillium spinulosum, Penicillium cyclopium, Penicillium purpurogenum, Penicillium janczewskii, Penicillium montanense, Penicillium restrictum, Penicillium harzianum, Penicillium terrestre, Penicillium ochrochloron, Penicillium

microaggregatum, Glomus monosporum, Glomus mosseae, Glomus multicaule, Glomus pansihalos, Glomus pustulatum, Glomus sinuosum, Glomus spurcum, Glomus tortuosum, Glomus trimurales, Glomus verruculosum, Glomus versiforme, Glomus viscosum, Gigaspora albida, Gigaspora decipiens, Gigaspora gigantean, Gigaspora mar gar ita, and Gigaspora rosea; and (b) a germinative compound selected from the group consisting of an amino acid, a purine nucleoside, a salt, and a sugar.

In certain embodiments of the aforementioned compositions, the amino acid is selected from the group consisting of glycine, L-alanine, L-arginine, L-aspartate, L-glutamate, L- histidine, L-isoleucine, L-lysine, L-methionine, L-serine, L-tryptophan, L-tyrosine, L-valine, L- proline, L-leucine, L-cysteine, L-threonine, L-glutamine, L-asparagine, L-phenylalanine and analogues thereof.

In certain embodiments of the aforementioned compositions, the purine nucleoside is selected from the group consisting of inosine and adenosine. In certain embodiments the salt is selected from the group consisting of Calcium Chloride, Potassium Chloride, Potassium

Phosphate, Potassium Carbonate, Potassium Propionate and Magnesium Sulphate. In certain embodiments the sugar is selected from the group consisting of glucose, fructose, mannose and trehalose.

In certain embodiments, the composition further comprises an agriculturally acceptable or pharmaceutically acceptable carrier. In certain embodiments, the composition further comprises a carrier that is acceptable for use in bioremediation or food production.

In a further aspect, the invention provides a method for increasing the germination of fungus spores, comprising:

a) preparing a mixture comprising a fungus spore and a germinative compound; and b) exposing the mixture to an environment conducive to germination of the fungus spores, wherein the germination of the fungus spores in the mixture is increased relative to a corresponding fungus spore formulation that lacks a germinative compound.

In a preferred embodiment, the invention relates to a method for increasing the germination of fungus spores, comprising:

a) preparing a solution comprising a fungus spore and a germinative compound; and b) drying the solution to obtain a dried intimate mixture comprising a fungus spore and a germinative compound, wherein the fungus spore and the germinative compound are maintained in proximate position until they reach an environment conducive to germination of the fungal spores; and c) exposing the intimate mixture to an environment conducive to germination of the fungus spores, wherein the germination of the fungus spores in the intimate mixture is increased relative to a corresponding fungus spore formulation that lacks a germinative compound.

The invention also provides a plant propagative material coated with a composition comprising a fungal spore and a germinative compound. In a preferred embodiment, the plant propagative material is a seed.

In certain embodiments of the aforementioned plant propagative material, the fungal spore is selected from the group consisting of Aspergillus flavus, Aureobasidium pullulans, Beauveria bassiana, Coniothyrium minitans, Candida oleophila, Gliocladium roseum,

Gliocladium catenulatum, Metarhizium anisopliae, Paecilomyces fumosoroseus, Paecilomyces lilacinus, Pantoea agglomerans, Penicillium bilaiae, Penicillium gaestrivorus, Pseudozyma flocculosa, Pythium oligandrum, Trichoderma asperellum, Trichoderma virens, Trichoderma gamsii, Trichoderma hamatum, Trichoderma harzianum, Trichoderma harizatum, Trichoderma koningii, Trichoderma fertile, Ulocladium oudemansii, Ulocladium atrum, Alternaria destruens, Chondrostereum purpureum, Colletotrichum gloeosporioides, Phoma macrostoma, Typhula phacorrhiza, Penicillium chrysogenum, Penicillium griseofulvum, Penicillium pinophilum, Penicillium canescens, Penicillium glabrum, Penicillium glaucum, Penicillium halgiovense, Penicillium roqueforti, Penicillium simplicissium, Penicillium digitatum, Penicillium

frequentans, Penicillium verrucosum, Penicillium implicatum, Penicillium citrinum, Glomus aggregatum, Glomus albidum, Glomus ambisporum, Glomus brazillanum, Glomus caledonium Glomus coremioides, Glomus claroideum, Glomus clarum, Glomus clavisporum, Glomus constrictum, Glomus coronatum, Glomus deserticola, Glomus diaphanum, Glomus eburneum, Glomus etunicatum, Glomus fasciculatum, Glomus fistulosum, Glomus fragilistratum, Glomus geosporum, Glomus globiferum, Glomus heterosporum, Glomus hoi, Glomus intraradices, Glomus lacteum, Glomus lamellosum, Glomus luteum, Glomus manihotis, Glomus

In certain embodiments of the aforementioned plant propagative material, the

germinative compound is selected from the group consisting of an amino acid, a purine nucleoside, a salt, and a sugar. In certain embodiments the amino acid is selected from the group consisting of glycine, L-alanine, L-arginine, L-aspartate, L-glutamate, L-histidine, L-isoleucine, L-lysine, L-methionine, L-serine, L-tryptophan, L-tyrosine, L-valine, L-proline, L-leucine, L- cysteine, L-threonine, L-glutamine, L-asparagine, L-phenylalanine and analogues thereof. In certain embodiments the purine nucleoside is selected from the group consisting of inosine and adenosine. In certain embodiments the salt is selected from the group consisting of Calcium Chloride, Potassium Chloride, Potassium Phosphate, Potassium Carbonate, Potassium Propionate and Magnesium Sulphate. In certain embodiments the sugar is selected from the group consisting of glucose, fructose, mannose and trehalose.

In certain aspects, the invention also relates to a method for enhancing the growth of a plant propagative material comprising treating a plant propagative material with any of the aforementioned compositions, wherein growth of the plant propagative material is enhanced relative to a corresponding control plant propagative material that is not treated with the composition.

In certain embodiments of the aforementioned method, the composition is applied to the growth medium. In certain embodiments, the growth medium is soil.

In certain aspects, the invention also relates to a food product or pharmaceutical product comprising any of the aforementioned compositions.

DETAILED DESCRIPTION OF THE INVENTION

The mixtures of this invention are comprised of a fungal spore and a germinative compound. The fungal species employed may be any species which forms spores. In a preferred embodiment, the fungal species include any spore-forming fungus that enhances the growth of plants. Such enhancement may be in the form of making additional resources available to the growing plant, for example by enhancing nitrogen availability, e.g., as provided by root colonizing fungi (such as a mycorrhizae), or providing plant growth promoting materials such as amino acids, organic acids, enzymes or hormones . Alternatively, the fungi may provide plant propagative material with protection from the harmful effects of plant pathogenic fungi or bacteria and/or soil born animals such as those belonging to the phylum Nematoda or

Aschelminthes. Protection against plant parasitic nematodes and parasitic microorganisms can occur through chitinolytic, proteolytic, collagenolytic, or other activities detrimental to these soil borne animals and/or detrimental to microbial populations.

Preferred fungal species that are used for insecticidal, fungicidal, nematicidal control and nutrient cycling include, but are not limited to, Aspergillus flavus, Aureobasidium pullulans, Beauveria bassiana, Coniothyrium minitans, Candida oleophila, Gliocladium roseum,

Gliocladium catenulatum, Metarhizium anisopliae, Paecilomyces fumosoroseus, Paecilomyces lilacinus, Pantoea agglomerans, Penicillium bilaiae, Penicillium gaestrivorus, Pseudozyma flocculosa, Pythium oligandrum, Trichoderma asperellum, Trichoderma virens, Trichoderma gamsii, Trichoderma hamatum, Trichoderma harzianum, Trichoderma harizatum, Trichoderma koningii, Trichoderma fertile, Ulocladium oudemansii, Ulocladium atrum, Glomus aggregatum, Glomus albidum, Glomus ambisporum, Glomus brazillanum, Glomus caledonium Glomus coremioides, Glomus claroideum, Glomus clarum, Glomus clavisporum, Glomus constrictum, Glomus coronatum, Glomus deserticola, Glomus diaphanum, Glomus eburneum, Glomus etunicatum, Glomus fasciculatum, Glomus fistulosum, Glomus fragilistratum, Glomus geosporum, Glomus globiferum, Glomus heterosporum, Glomus hoi, Glomus intraradices, Glomus lacteum, Glomus lamellosum, Glomus luteum, Glomus manihotis, Glomus

Preferred fungal species which are used for herbicidal control include, but are not limited to, Alternaria destruens, Chondrostereum purpureum, Colletotrichum gloeosporioides, Phoma macrostoma, and Typhula phacorrhiza. Preferred fungal species that are used for production of medicine or used in health applications include, but are not limited to, Penicillium chrysogenum, Penicillium griseofulvum, Penicillium pinophilum, Penicillium canescens, and Penicillium glabrum.

Preferred fungal species that are used for production of food include, but are not limited to, Penicillium camemberti, Penicillium glaucum, Penicillium halgiovense, and Penicillium roqueforti.

Preferred fungal species that are used for bioremediation include, but are not limited to, Penicillium sp., most preferably, P. simplicissium, P. chyrysogenum, P. digitatum, P.

janthinellum, P. italicum, P. canescens, P. spinulosum, P. cyclopium, P. purpurogenum, P.

janczewskii, P. montanense, P. restrictum, P. harzianum, P. terrestre, P. ochrochloron, P.

aurantiogriseum, P. crustosum, P. decumbens, P. griseofulvum, P. janthinellum, P. rugulosum, P. velutinum, P. frequentans, P. verrucosum, P. implicatum, P. citrinum, and P. cyclopium. Preferred fungal species that are used for bioremediation also include, but are not limited to, Glomus aggregatum, Glomus albidum, Glomus ambisporum, Glomus brazillanum, Glomus caledonium Glomus coremioides, Glomus claroideum, Glomus clarum, Glomus clavisporum, Glomus constrictum, Glomus coronatum, Glomus deserticola, Glomus diaphanum, Glomus eburneum, Glomus etunicatum, Glomus fasciculatum, Glomus fistulosum, Glomus fragilistratum, Glomus geosporum, Glomus globiferum, Glomus heterosporum, Glomus hoi, Glomus

intraradices, Glomus lacteum, Glomus lamellosum, Glomus luteum, Glomus manihotis, Glomus micro aggregatum, Glomus monosporum, Glomus mosseae, Glomus multicaule, Glomus pansihalos, Glomus pustulatum, Glomus sinuosum, Glomus spurcum, Glomus tortuosum, Glomus trimurales, Glomus verruculosum, Glomus versiforme, Glomus viscosum, Gigaspora albida, Gigaspora decipiens, Gigaspora gigantean, Gigaspora margarita, and Gigaspora rosea.

Typically, such germinative compounds are amino acids such as glycine and L-amino acids including L-alanine, L-arginine, L-aspartate, L-glutamate, L-histidine, L-isoleucine, L- lysine, L-methionine, L-serine, L-tryptophan, L-tyrosine, L-valine, L-proline, L-leucine, L- cysteine, L-threonine, L-glutamine, L-asparagine, L-phenylalanine and analogues thereof. Such analogues can be created by one of ordinary skill in the art by making substitutions on or within groups of a base chemistry. Additional germinative compounds which may be employed include purine nucleosides (e.g. inosine and adenosine), salts (e.g. Calcium Chloride, Potassium

Chloride, Potassium Phosphate, Potassium Carbonate, Potassium Propionate and Magnesium Sulphate), sugars (e.g. glucose, fructose, mannose & trehalose). Nutrients, including dextrose, starches, and micronutrients which will aid in the growth of fungi once the spores have germinated may also be included.

Such amino acids can be employed as individual compounds, or in the form of polypeptides. In one embodiment the polypeptides are protein hydrolysates, for example casein hydrolysate. Useful polypeptides will typically comprise at least fifty percent amino acids which will function as germinants, such as L-alanine, L-valine, L-proline, L-leucine, L-cysteine, L- threonine, L-glutamine, L-asparagine, and L-phenylalanine, such percentages being based upon the number of amino acids in the polypeptide.

In a preferred embodiment, the germinative compound is selected from the group consisting of an amino acid, a purine nucleoside, a salt, and a sugar.

In another preferred embodiment, the amino acid is selected from the group consisting of glycine, L-alanine, L-arginine, L-aspartate, L-glutamate, L-histidine, L-isoleucine, L-lysine, L- methionine, L-serine, L-tryptophan, L-tyrosine, L-valine, L-proline, L-leucine, L-cysteine, L- threonine, L-glutamine, L-asparagine, L-phenylalanine and analogues thereof.

In a further preferred embodiment, the purine nucleoside is selected from the group consisting of inosine and adenosine.

In a still further preferred embodiment, the salt is selected from the group consisting of Calcium Chloride, Potassium Chloride, Potassium Phosphate, Potassium Carbonate, Potassium Propionate and Magnesium Sulphate.

In yet another preferred embodiment, the sugar is selected from the group consisting of glucose, fructose, mannose and trehalose.

The germinative compound is present in an amount sufficient to cause the fungal spore employed to germinate. This amount can be readily determined for any particular fungal spore/germinative compound mixture by routine experimentation. In embodiments in which the fungal spore and germinative compound are mixed in solution and then dried, the germinative compounds are typically formulated, prior to drying, at concentrations of from 0.0001 mg/mL to 170 mg/mL. In some embodiments, the germinative compounds are formulated, prior to drying, at concentrations of 0.0003 mg/mL to 170 mg/mL, 0.0003 mg/mL to 30 mg/mL, 0.001 mg/mL to 100 mg/mL, or 0.001 mg/mL to 10 mg/mL. In a preferred embodiment, the germinative compound is formulated, prior to drying, at concentrations of from 0.001 mg/mL to 1 mg/mL. In other embodiments in which the fungal spore and germinative compound are mixed in dry form, such germinative compounds are typically mixed dry with the solid fungal spores at concentrations of from 0.0001 mg/mg to 170 mg/mg. In some embodiments, the germinative compounds are mixed dry with the solid fungal spores at concentrations of 0.0003 mg/mg to 170 mg/mg, 0.0003 mg/mg to 30 mg/mg, 0.001 mg/mg to 100 mg/mg, or 0.001 mg/mg to 10 mg/mg. In a preferred embodiment, the germinative compound is mixed dry with the solid fungal spores at concentrations of from 0.001 mg/mg to 1 mg/mg.

In some embodiments, the germinative compound (e.g. an amino acid, a purine nucleoside, a salt, or a sugar) is present in a mixture or composition of the invention at a concentration of about 0.01, 0.05, 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 12.5, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, or 1000 mM. Any of these values may be used to define a range for the concentration of the germinative compound in the mixture or composition. For example, in some embodiments, the concentration of the germinative compound in the mixture or composition is from about 0.01 mM to about 1000 mM, from about 0.01 mM to about 100 mM, or from about 0.1 mM to about 20 mM.

As is employed herein, the term "intimate mixture" refers to a mixture in which the spores and germinative compounds are maintained in proximate position until they reach an environment conducive to germination.

Such an intimate mixture can be achieved employing processes such as spray-drying, freeze-drying, vacuum-drying, air drying or drum drying. In a preferred embodiment, the intimate mixture is produced by spray-drying or freeze-drying. When forming such intimate mixtures, it is important that the spore and germinative compound are not mixed together under conditions which would allow the germinative compound to cause the spore to germinate, as this could cause premature germination with an adverse effect upon the storage life of the mixture. This can be avoided by employing separate streams in a spray-dryer, either by using two nozzles or a single nozzle which permits the simultaneous spraying of two separate streams; or by freeze- drying under conditions (for example temperatures) which are not conducive to germination. Premature germination may also be avoided by introducing the spore mass to a solution containing the germinative compound immediately prior to drying. In a preferred embodiment, the germinative compound is adsorbed to or absorbed by the fungus spore in the intimate mixture. In a further embodiment, the fungus spore and germinative compound are finely dispersed throughout the intimate mixture. In a still further embodiment, the fungus spore and germinative compound are microscopically dispersed throughout the intimate mixture, such that individual particles consisting essentially of fungus spores and individual particles consisting essentially of germinative compounds are not visible to the naked eye.

In one embodiment the intimate mixture is prepared by combining the fungus spore and germinative compound in a solution prior to drying. Preferably, the fungus spore and germinative compound are combined in a solution immediately prior to drying.

Although not wishing to be held to any theory, it is believed that the formation of an intimate mixture places the germinative compound in a proximate position where it can more preferably bind to the germination initiator sites of the spore when the mixture reaches an appropriate environment for germination. Such proximate position permits the germinative compound to outcompete any germination interfering compounds which may be present, with the result that a higher percentage of spores will be germinated. Due to the logarithmic growth of fungus once they enter the vegetative stage; such an increased percentage can quickly result in a several log increase in culture formation.

The present invention also relates to compositions comprising a mixture of a fungal spore and a germinative compound. In a preferred embodiment, the composition comprises an intimate mixture comprising (a) a fungal spore and (b) a germinative compound, wherein the spore and germinative compound are maintained in an inactive form such that the germinative compound will not induce germination of the spore until such composition is subjected to an activation environment.

As is employed herein, the term "activation environment" refers to an environment which permits the germinative compound and the spore to interact, such that the spore is induced to enter into a vegetative state. Such an activation environment may involve the addition of water (through rain, moisture in the soil, or water added to the coated plant propagation material prior to planting), or it may require a combination of factors such as temperature, moisture, pH or salinity. The compositions of this invention may further comprise an acceptable carrier. As is employed herein, the term "acceptable carrier" refers to a carrier which is typically employed in the field involved (for example a carrier typically employed in industrial, agricultural, aquacultural, environmental and/or probiotic applications) which does not adversely affect the bacterial strain involved. Such carriers are well known to those of ordinary skill in the art.

In a preferred embodiment, the carrier is an agriculturally acceptable carrier. In another preferred embodiment, the carrier is an aquaculturally acceptable carrier. In a further preferred embodiment, the carrier is suitable for environmental administration or bioremediation. For many industrial and/or agricultural applications, carriers include porous solid carriers such as talc, bentonite, clay, kaolin, diatomaceous earth, white carbon, vermiculite, slaked lime, siliceous sand, ammonium sulfate, and the like; as well as liquid carriers such as water, isopropyl alcohol, xylene, cyclohexanone, methylnaphthalene, and alkyl glycol and the like. For probiotic use, pharmaceutically acceptable carriers such as sugars and starches which are typically employed may be utilized.

The invention also provides a food product or pharmaceutical product comprising the aforementioned compositions.

The formation of a mixture of a germinative compound and a fungus spore (e.g. an intimate mixture) may enhance the utility of the fungus spore by increasing one or more traits of interest of the fungus spore, including but not limited to germination, growth, metabolism, enzyme activity and tolerance to environmental stresses such as low pH, high salt concentration, and exposure to toxic metals. In some embodiments, the mixture increases the trait of interest by at least 5, 10, 20, 30, 40 , 50, 60, 70, 80, 90, 100, 200, 300, 400 or 500% relative to a

corresponding fungus spore formulation that lacks a germinative compound.

In order to further enhance germination, it is preferred that the spore be shocked prior to the formation of the mixture. Spores may be shocked in a variety of standard methods, e.g., osmotic shock, heat shock, cold shock, pressure shock, deprivation of nutrients, and/or exposure to certain acids. Heat shock involves heating the spores for a sufficient period of time at a temperature sufficient to induce the production of heat shock proteins.

When employed in agricultural or industrial uses, such compositions may further comprise standard formulation aids such as surfactants, emulsifiers, other active ingredients, etc. so long as such other components do not interfere with germination or adversely affect the viability of the germinated spores.

The invention also provides a method for preparing a mixture of preparing a fungus spore and a germinative compound. In a preferred embodiment, the invention relates to a method for preparing a dried intimate mixture comprising a fungus spore and a germinative compound, the method comprising:

a) preparing a solution comprising a fungus spore and a germinative compound; and

b) drying the solution to obtain a dried intimate mixture comprising a fungus spore and a germinative compound, wherein the fungus spore and the germinative compound are maintained in proximate position until they reach an environment conducive to germination.

In a preferred embodiment the drying in the aforementioned method is spray-drying, freeze-drying, air drying or drum drying. In another preferred embodiment, the spore in the aforementioned method is selected from the group consisting of fungal species that are used for insecticidal, fungicidal, nematicidal control and nutrient cycling including, but not limited to, Aspergillus flavus, Aureobasidium pullulans, Beauveria bassiana, Coniothyrium minitans, Candida oleophila, Gliocladium roseum, Gliocladium catenulatum, Metarhizium anisopliae, Paecilomyces fumosoroseus, Paecilomyces lilacinus, Pantoea agglomerans, Penicillium bilaiae, Penicillium gaestrivorus, Pseudozyma flocculosa, Pythium oligandrum, Trichoderma

asperellum, Trichoderma virens, Trichoderma gamsii, Trichoderma hamatum, Trichoderma harzianum, Trichoderma harizatum, Trichoderma koningii, Trichoderma fertile, Ulocladium oudemansii, Ulocladium atrum, Glomus aggregatum, Glomus albidum, Glomus ambisporum, Glomus brazillanum, Glomus caledonium Glomus coremioides, Glomus claroideum, Glomus clarum, Glomus clavisporum, Glomus constrictum, Glomus coronatum, Glomus deserticola, Glomus diaphanum, Glomus eburneum, Glomus etunicatum, Glomus fasciculatum, Glomus fistulosum, Glomus fragilistratum, Glomus geosporum, Glomus globiferum, Glomus

heterosporum, Glomus hoi, Glomus intraradices, Glomus lacteum, Glomus lamellosum, Glomus luteum, Glomus manihotis, Glomus microaggregatum, Glomus monosporum, Glomus mosseae, Glomus multicaule, Glomus pansihalos, Glomus pustulatum, Glomus sinuosum, Glomus spurcum, Glomus tortuosum, Glomus trimurales, Glomus verruculosum, Glomus versiforme, Glomus viscosum, Gigaspora albida, Gigaspora decipiens, Gigaspora gigantean, Gigaspora margarita, and Gigaspora rosea. Additional preferred fungal species for use in the aforementioned method include fungal species which are used for herbicidal control including Alternaria destruens, Chondrostereum purpureum, Colletotrichum gloeosporioides, Phoma macrostoma, and Typhula phacorrhiza, fungal species that are used for production of medicine or used in health applications including Penicillium chrysogenum, Penicillium griseofulvum, Penicillium pinophilum, Penicillium canescens, and Penicillium glabrum, fungal species that are used for production of food including Penicillium camemberti, Penicillium glaucum, Penicillium halgiovense, and

Penicillium roqueforti, and fungal species that are used for bioremediation including Glomus aggregatum, Glomus albidum, Glomus ambisporum, Glomus brazillanum, Glomus caledonium Glomus coremioides, Glomus claroideum, Glomus clarum, Glomus clavisporum, Glomus constrictum, Glomus coronatum, Glomus deserticola, Glomus diaphanum, Glomus eburneum, Glomus etunicatum, Glomus fasciculatum, Glomus fistulosum, Glomus fragilistratum, Glomus geosporum, Glomus globiferum, Glomus heterosporum, Glomus hoi, Glomus intraradices, Glomus lacteum, Glomus lamellosum, Glomus luteum, Glomus manihotis, Glomus

microaggregatum, Glomus monosporum, Glomus mosseae, Glomus multicaule, Glomus pansihalos, Glomus pustulatum, Glomus sinuosum, Glomus spurcum, Glomus tortuosum, Glomus trimurales, Glomus verruculosum, Glomus versiforme, Glomus viscosum, Gigaspora albida, Gigaspora decipiens, Gigaspora gigantean, Gigaspora margarita, and Gigaspora rosea, and Penicillium sp., most preferably P. simplicissium, P. chyrysogenum, P. digitatum, P.

janthinellum, P. italicum, P. canescens, P. spinulosum, P. cyclopium, P. purpurogenum, P. janczewskii, P. montanense, P. restrictum, P. harzianum, P. terrestre, P. ochrochloron, P.

aurantiogriseum, P. crustosum, P. decumbens, P. griseofulvum, P. janthinellum, P. rugulosum, P. velutinum, P. frequentans, P. verrucosum, P. implicatum, P. citrinum, and P. cyclopium.

Typically, the germinative compounds used in the aforementioned method are amino acids such as glycine and L-amino acids including L-alanine, L-arginine, L-aspartate, L- glutamate, L-histidine, L-isoleucine, L-lysine, L-methionine, L-serine, L-tryptophan, L-tyrosine, L-valine, L-proline, L-leucine, L-cysteine, L-threonine, L-glutamine, L-asparagine, L- phenylalanine and analogues thereof. Such analogues can be created by one of ordinary skill in the art by making substitutions on or within groups of a base chemistry. Additional germinative compounds which may be employed include purine nucleosides (e.g. inosine and adenosine), salts (e.g. Calcium Chloride, Potassium Chloride, Potassium Phosphate, Potassium Carbonate, Potassium Propionate and Magnesium Sulphate), and sugars (e.g. glucose, fructose, mannose and trehalose). Nutrients, including dextrose, starches, and micronutrients which will aid in the growth of fungi once the spores have germinated may also be included.

In embodiments of the aforementioned method in which the fungal spore and germinative compound are mixed in solution and then dried, the germinative compounds are typically formulated, prior to drying, at concentrations of from 0.0001 mg/mL to 170 mg/n L. In some embodiments, the germinative compounds are formulated, prior to drying, at concentrations of 0.0003 mg/mL to 170 mg/mL, 0.0003 mg/mL to 30 mg/mL, 0.001 mg/mL to 100 mg/mL, or 0.001 mg/mL to 10 mg/mL. In a preferred embodiment, the germinative compound is formulated, prior to drying, at concentrations of from 0.001 mg/mL to 1 mg/mL.

In some embodiments of the aforementioned method in which the fungal spore and germinative compound are mixed in solution and then dried, the germinative compound (e.g. an amino acid, a purine nucleoside, a salt, or a sugar) is present in the mixture at a concentration of about 0.01, 0.05, 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 12.5, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, or 1000 mM. Any of these values may be used to define a range for the concentration of the germinative compound in the mixture. For example, in some embodiments, the concentration of the germinative compound in the mixture is from about 0.01 mM to about 1000 mM, from about 0.01 mM to about 100 mM, or from about 0.1 mM to about 20 mM.

In other embodiments of the aforementioned method in which the fungal spore and germinative compound are mixed in dry form, such germinative compounds are typically mixed dry with the solid fungal spores at concentrations of from 0.0001 mg/mg to 170 mg/mg. In some embodiments, the germinative compounds are mixed dry with the solid fungal spores at concentrations of 0.0003 mg/mg to 170 mg/mg, 0.0003 mg/mg to 30 mg/mg, 0.001 mg/mg to 100 mg/mg, or 0.001 mg/mg to 10 mg/mg. In a preferred embodiment, the germinative compound is mixed dry with the solid fungal spores at concentrations of from 0.001 mg/mg to 1 mg/mg. In some embodiments, the germinative compound (e.g. an amino acid, a purine nucleoside, a salt, or a sugar) is present in the mixture at a concentration of about 0.01, 0.05, 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 12.5, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, or 1000 mM. Any of these values may be used to define a range for the concentration of the germinative compound in the mixture. For example, in some embodiments, the concentration of the germinative compound in the mixture is from about 0.01 mM to about 1000 mM, from about 0.01 mM to about 100 mM, or from about 0.1 mM to about 20 mM.

In certain embodiments, the germinative compound in the aforementioned method is a polypeptide. In a preferred embodiment of the aforementioned method, the spore has been shocked.

The invention also provides a mixture, for example a dried intimate mixture, produced by the aforementioned methods.

In another aspect, the invention provides a method for increasing the germination, growth, metabolism and/or enzyme activity of fungal spores comprising:

a) preparing a mixture comprising a fungus spore and a germinative compound; and

b) exposing the intimate mixture to an environment conducive to germination of the fungus spores, wherein the germination, growth, metabolism and/or enzyme activity of the fungus spores in the mixture is increased relative to a corresponding fungus spore formulation that lacks a germinative compound.

In a preferred embodiment, the invention provides a method for increasing the germination, growth, metabolism and/or enzyme activity of fungus spores, comprising:

b) drying the solution to obtain a dried intimate mixture comprising a fungus spore and a germinative compound, wherein the fungus spore and the germinative compound are maintained in proximate position until they reach an environment conducive to germination of the fungal spores; and

c) exposing the intimate mixture to an environment conducive to germination of the fungus spores, wherein the germination, growth, metabolism and/or enzyme activity of the fungus spores in the intimate mixture is increased relative to a corresponding fungus spore formulation that lacks a germinative compound.

In some embodiments of the aforementioned methods, the intimate mixture increases the trait of interest by at least 5, 10, 20, 30, 40 , 50, 60, 70, 80, 90, 100, 200, 300, 400 or 500% relative to a corresponding fungus spore formulation that lacks a germinative compound.

In a preferred embodiment of the aforementioned method, the percent germination, growth, metabolism and/or enzyme activity of fungus spores in the intimate mixture is increased by at least 10% relative to the corresponding fungus spore formulation that lacks a germinative compound. In a further preferred embodiment of the aforementioned method, the drying is spray-drying, freeze-drying, vacuum-drying, air drying or drum drying.

In certain embodiments, the spore in the aforementioned methods is selected from the group consisting of fungal species that are used for insecticidal, fungicidal, nematicidal control and nutrient cycling including, but not limited to, Aspergillus flavus, Aureobasidium pullulans, Beauveria bassiana, Coniothyrium minitans, Candida oleophila, Gliocladium roseum,

Gliocladium catenulatum, Metarhizium anisopliae, Paecilomyces fumosoroseus, Paecilomyces lilacinus, Pantoea agglomerans, Penicillium bilaiae, Penicillium gaestrivorus, Pseudozyma flocculosa, Pythium oligandrum, Trichoderma asperellum, Trichoderma vir ens, Trichoderma gamsii, Trichoderma hamatum, Trichoderma harzianum, Trichoderma harizatum, Trichoderma koningii, Trichoderma fertile, Ulocladium oudemansii, Ulocladium atrum, Glomus aggregatum, Glomus albidum, Glomus ambisporum, Glomus brazillanum, Glomus caledonium Glomus coremioides, Glomus claroideum, Glomus clarum, Glomus clavisporum, Glomus constrictum, Glomus coronatum, Glomus deserticola, Glomus diaphanum, Glomus eburneum, Glomus etunicatum, Glomus fasciculatum, Glomus fistulosum, Glomus fragilistratum, Glomus geosporum, Glomus globiferum, Glomus heterosporum, Glomus hoi, Glomus intraradices, Glomus lacteum, Glomus lamellosum, Glomus luteum, Glomus manihotis, Glomus

microaggregatum, Glomus monosporum, Glomus mosseae, Glomus multicaule, Glomus pansihalos, Glomus pustulatum, Glomus sinuosum, Glomus spurcum, Glomus tortuosum,

Glomus trimurales, Glomus verruculosum, Glomus versiforme, Glomus viscosum, Gigaspora albida, Gigaspora decipiens, Gigaspora gigantean, Gigaspora margarita, and Gigaspora rosea.

Additional preferred fungal species for use in the aforementioned methods include fungal species which are used for herbicidal control including Alternaria destruens, Chondrostereum purpureum, Colletotrichum gloeosporioides, Phoma macrostoma, and Typhula phacorrhiza, fungal species that are used for production of medicine or used in health applications including Penicillium chrysogenum, Penicillium griseofulvum, Penicillium pinophilum, Penicillium canescens, and Penicillium glabrum, fungal species that are used for production of food including Penicillium camemberti, Penicillium glaucum, Penicillium halgiovense, and

Penicillium roqueforti, and fungal species that are used for bioremediation including Glomus aggregatum, Glomus albidum, Glomus ambisporum, Glomus brazillanum, Glomus caledonium Glomus coremioides, Glomus claroideum, Glomus clarum, Glomus clavisporum, Glomus constrictum, Glomus coronatum, Glomus deserticola, Glomus diaphanum, Glomus eburneum, Glomus etunicatum, Glomus fasciculatum, Glomus fistulosum, Glomus fragilistratum, Glomus geosporum, Glomus globiferum, Glomus heterosporum, Glomus hoi, Glomus intraradices, Glomus lacteum, Glomus lamellosum, Glomus luteum, Glomus mamhotis, Glomus

Typically, the germinative compounds used in the aforementioned methods are amino acids such as glycine and L-amino acids including L-alanine, L-arginine, L-aspartate, L- glutamate, L-histidine, L-isoleucine, L-lysine, L-methionine, L-serine, L-tryptophan, L-tyrosine, L-valine, L-proline, L-leucine, L-cysteine, L-threonine, L-glutamine, L-asparagine, L- phenylalanine and analogues thereof. Such analogues can be created by one of ordinary skill in the art by making substitutions on or within groups of a base chemistry. Additional germinative compounds which may be employed include purine nucleosides (e.g. inosine and adenosine), salts (e.g. Calcium Chloride, Potassium Phosphate, Potassium Carbonate and Magnesium Sulphate), and sugars (e.g. glucose and fructose). Nutrients, including dextrose, starches, and micronutrients which will aid in the growth of fungi once the spores have germinated may also be included.

In some embodiments of the aforementioned method, the germinative compound is a polypeptide.

In embodiments of the aforementioned methods in which the fungal spore and germinative compound are mixed in solution and then dried, the germinative compounds are typically formulated, prior to drying, at concentrations of from 0.0001 mg/mL to 170 mg/mL. In some embodiments, the germinative compounds are formulated, prior to drying, at concentrations of 0.0003 mg/mL to 170 mg/n L, 0.0003 mg/mL to 30 mg/mL, 0.001 mg/mL to 100 mg/mL, or 0.001 mg/mL to 10 mg/mL. In a preferred embodiment, the germinative compound is formulated, prior to drying, at concentrations of from 0.001 mg/mL to 1 mg/mL. In some embodiments, the germinative compound (e.g. an amino acid, a purine nucleoside, a salt, or a sugar) is present in the mixture at a concentration of about 0.01, 0.05, 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 12.5, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, or 1000 mM. Any of these values may be used to define a range for the concentration of the germinative compound in the mixture. For example, in some embodiments, the concentration of the germinative compound in the mixture is from about 0.01 mM to about 1000 mM, from about 0.01 mM to about 100 mM, or from about 0.1 mM to about 20 mM.

In other embodiments of the aforementioned method in which the fungal spore and germinative compound are mixed in dry form, such germinative compounds are typically mixed dry with the solid fungal spores at concentrations of from 0.0001 mg/mg to 170 mg/mg. In some embodiments, the germinative compounds are mixed dry with the solid fungal spores at concentrations of 0.0003 mg/mg to 170 mg/mg, 0.0003 mg/mg to 30 mg/mg, 0.001 mg/mg to 100 mg/mg, or 0.001 mg/mg to 10 mg/mg. In a preferred embodiment, the germinative compound is mixed dry with the solid fungal spores at concentrations of from 0.001 mg/mg to 1 mg/mg. In some embodiments, the germinative compound (e.g. an amino acid, a purine nucleoside, a salt, or a sugar) is present in the mixture at a concentration of about 0.01, 0.05, 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 12.5, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 600, 700, 800, 900, or 1000 mM. Any of these values may be used to define a range for the concentration of the germinative compound in the mixture. For example, in some embodiments, the concentration of the germinative compound in the mixture is from about 0.01 mM to about 1000 mM, from about 0.01 mM to about 100 mM, or from about 0.1 mM to about 20 mM.

In another aspect, the present invention relates to a plant propagative material coated with a composition comprising: (a) a fungal spore; and (b) a germinative compound. The fungal spores include species which enhance the growth of a plant, typically either by acting as a pesticide (for examples, a fungicide, nematocide or insecticide); or by enhancing the

environment around the propagative material for growth, (for example, by increasing the amount of nutrients available, such as nitrogen). The germinative material aids in the rapid

transformation of the fungus from its spore to it vegetative state, thereby enabling the fungus to more quickly enhance the growth of the plant.

As is employed herein, the term "plant propagative material" is intended to include all the generative parts of the plant such as seeds and vegetative plant material such as cuttings and tubers (e. g. potatoes), which can be used for the multiplication of the plant. This includes seeds, roots, fruits, tubers, bulbs, rhizomes, shoots, sprouts and other parts of plants, including seedlings and young plants, which are to be transplanted after germination or after emergence from soil. These young plants may also be protected before transplantation by a total or partial treatment by immersion or pouring. Preferably, the term plant propagative material denotes seeds.

Plants that are particularly useful in the present invention include monocotyledonous and dicotyledonous plants including but not limited to fodder or forage legumes, ornamental plants, food crops, trees, or shrubs selected from Acer spp., Allium spp., Amaranthus spp., Ananas comosus, Apium graveolens, Arachis spp, Asparagus officinalis, Beta vulgaris, Brassica spp. (e.g. Brassica napus, Brassica rapa ssp. [canola, oilseed rape, turnip rape]), Camellia sinensis, Canna indica, Cannabis saliva, Capsicum spp., Castanea spp., Cichorium endivia, Citrullus lanatus, Citrus spp., Cocos spp., Coffea spp., Coriandrum sativum, Corylus spp., Crataegus spp., Cucurbita spp., Cucumis spp., Daucus carota, Fagus spp., Ficus carica, Fragaria spp., Ginkgo biloba, Glycine spp. (e.g. Glycine max, Soja hispida or Soja max), Gossypium hirsutum,

Helianthus spp. (e.g. Helianthus annuus), Hibiscus spp., Hordeum spp. (e.g. Hordeum vulgare), Ipomoea batatas, Juglans spp., Lactuca sativa, Linum usitatissimum, Litchi chinensis, Lotus spp., Luffa acutangula, Lupinus spp., Lycopersicon spp. (e.g. Lycopersicon esculenturn,

Lycopersicon lycopersicum, Lycopersicon pyriforme), Malus spp., Medicago sativa, Mentha spp., Miscanthus sinensis, Morus nigra, Musa spp., Nicotiana spp., Olea spp., Oryza spp. (e.g. Oryza sativa, Oryza latifolia), Panicum miliaceum, Panicum virgatum, Passiflora edulis,

Petroselinum crispum, Phaseolus spp., Pinus spp., Pistacia vera, Pisum spp., Poa spp., Populus spp., Prunus spp., Pyrus communis, Quercus spp., Raphanus sativus, Rheum rhabarbarum, Ribes spp., Ricinus communis, Rubus spp., Saccharum spp., Salix sp., Sambucus spp., Secale cereale, Sesamum spp., Sinapis sp., Solanum spp. (e.g. Solanum tuberosum, Solanum integrifolium or Solanum lycopersicum), Sorghum bicolor, Sorghum halepense, Spinacia spp., Tamarindus indica, Theobroma cacao, Trifolium spp., Triticosecale rimpaui, Triticum spp. (e.g. Triticum aestivum, Triticum durum, Triticum turgidum, Triticum hybernum, Triticum macha, Triticum sativum or Triticum vulgare), Vaccinium spp., Vicia spp., Vigna spp., Viola odorata, Vitis spp., and Zea mays. Especially preferred are rice, oilseed rape, canola, soybean, corn (maize), cotton, sugarcane, alfalfa, sorghum, and wheat.

Preferred fungal species that are used in the aforementioned plant propagative materials include, but are not limited to, Aspergillus flavus, Aureobasidium pullulans, Beauveria bdssiana, Coniothyrium minitans, Candida oleophila, Gliocladium roseum, Gliocladium catenulatum, Metarhizium anisopliae, Paecilomyces fumosoroseus, Paecilomyces lilacinus, Pantoea agglomerans, Penicillium bilaiae, Penicillium gaest ivorus, Pseudozyma flocculosa, Pythium oligandrum, Trichoderma asperellum, Trichoderma virens, Trichoderma gamsii, Trichoderma hamatum, Trichoderma harzianum, Trichoderma harizatum, Trichoderma koningii, Trichoderma fertile, Ulocladium oudemansii, Ulocladium atrum, Glomus aggregatum, Glomus albidum, Glomus ambisporum, Glomus brazillanum, Glomus caledonium Glomus coremioides, Glomus claroideum, Glomus clarum, Glomus clavisporum, Glomus constrictum, Glomus coronatum, Glomus deserticola, Glomus diaphanum, Glomus eburneum, Glomus etunicatum, Glomus fasciculatum, Glomus flstulosum, Glomus fragilistratum, Glomus geosporum, Glomus globiferum, Glomus heterosporum, Glomus hoi, Glomus intraradices, Glomus lacteum, Glomus lamellosum, Glomus luteum, Glomus manihotis, Glomus microaggregatum, Glomus

monosporum, Glomus mosseae, Glomus multicaule, Glomus pansihalos, Glomus pustulatum, Glomus sinuosum, Glomus spurcum, Glomus tortuosum, Glomus trimurales, Glomus

verruculosum, Glomus versiforme, Glomus viscosum, Gigaspora albida, Gigaspora decipiens, Gigaspora gigantean, Gigaspora margarita, and Gigaspora rosea.

Typically, such germinative compounds that are used in the aforementioned plant propagative materials are amino acids such as glycine and L-amino acids including L-alanine, L- arginine, L-aspartate, L-glutamate, L-histidine, L-isoleucine, L-lysine, L-methionine, L-serine, L-tryptophan, L-tyrosine, L-valine, L-proline, L-leucine, L-cysteine, L-threonine, L-glutamine, L-asparagine, L-phenylalanine and analogues thereof. Additional germinative compounds which may also be employed in the plant propagative materials include purine nucleosides (e.g. inosine and adenosine), salts (e.g. Calcium Chloride, Potassium Phosphate, Potassium Carbonate and Magnesium Sulphate), sugars (e.g. glucose and fructose). Nutrients, including dextrose, starches, and micronutrients which will aid in the growth of fungi once the spores have germinated may also be included.

Such amino acids can be employed in the plant propagative material as individual compounds, or in the form of polypeptides. In one embodiment the polypeptides are protein hydrolysates, for example casein hydrolysate. Useful polypeptides will typically comprise at least fifty percent amino acids which will function as germinants, such as L-alanine, L-valine, L- proline, L-leucine, L-cysteine, L-threonine, L-glutamine, L-asparagine, and L-phenylalanine, such percentages being based upon the number of amino acids in the polypeptide.

Accordingly, such compositions may comprise dry mixtures of the spore and germinative compound; or they may include other materials (for example, low temperature melting waxes) which prohibit the spore and germinative compound from interacting until required

environmental criteria have been reached. Additional materials that may be used in the composition to prevent the spore and germinative compound from interacting include, but are not limited to, surfactants, sequestering agents, plasticizers, colorants and dyes, brighteners, emulsifiers, flow agents, coalescing agents, defoaming agents, thickeners, waxes, fillers, polymers, wetting agents and anti-freezing agents. Further, such compositions may take the form of emulsions wherein the spore and germinative compound are kept from interacting due to their presence in different phases.

Preferably, but not necessarily, the spore and germinative compound are in intimate mixture with one another, which intimate mixture can be produced by spray-drying, freeze - drying, vacuum-drying, air drying or drum drying. In a preferred embodiment, the plant propagative material is coated with an intimate mixture as described above.

Such compositions may further comprise additional components, including co- germinants, nutrients, and formulation aids, provided that such additives do not induce the spore and the germinative compound to prematurely interact.

The plant propagative material may be coated with the fungus and germinative compound by any conventional means typically employed to coat seed or other germinative material, provided that such means does not adversely affect the viability of the spores and that such means does not result in the germinative compound prematurely converting the fungus into its vegetative state. Conventional means which may be employed include spray treatment, drip treatment, drench treatment, painting treatment, film-coat treatment, pellet-coat treatment and the like. Methods of seed coating are known in the art and are described, for example, in U.S. Pat. No. 7,989,391 and U.S. Pat. No. 5,849,320.

When applying such coating, it is important that the spore and germinative compound are not mixed together under conditions which would allow the germinative compound to cause the spore to germinate, as this could cause premature germination with an adverse effect upon the storage life of the mixture. One way in which this can be avoided is by employing separate streams in a spray-dryer, either by using two nozzles or a single nozzle which permits the simultaneous spraying of two separate streams; or by freeze-drying under conditions (for example temperatures) which are not conducive to germination.

In addition to biologically active ingredients, seed coating compositions may include many materials and additives that are either part of the formulations of the active ingredient or contribute to the handling qualities of the seed coating or its functionality and durability on the seed. An example of a coating additive is a coating polymer which binds the active ingredients to the seed. Seed-coating polymers may include, for example, but are not limited to, proteins, polysaccharides, polyesters, polyurethanes, polymers prepared from unsaturated monomers, and combinations thereof.

Other additives contributing to the handling qualities of the seed coating or its functionality and durability on the seed include but are not limited to surfactants, sequestering agents, plasticizers, colorants and dyes, brighteners, emulsifiers, flow agents, coalescing agents, defoaming agents, thickeners, waxes, bactericides, fillers, polymers, wetting agents and anti- freezing agents. The nature and action of such additives are well-known to those skilled in the art of formulation. Additives should not interfere with the action of the fungus.

Binders that are useful in the present invention preferably comprise an adhesive polymer that may be natural or synthetic and is without phytotoxic effect on the seed to be coated. The binder may be selected from polyvinyl acetates; polyvinyl acetate copolymers; ethylene vinyl acetate (EVA) copolymers; polyvinyl alcohols; polyvinyl alcohol copolymers; celluloses, including ethylcelluloses, methylcelluloses, hydroxymethylcelluloses, hydroxypropylcelluloses and carboxymethylcellulose; polyvinylpyrolidones; polysaccharides, including starch, modified starch, dextrins, maltodextrins, alginate and chitosans; fats; oils; proteins, including gelatin and zeins; gum arabics; shellacs; vinylidene chloride and vinylidene chloride copolymers; calcium lignosulfonates; acrylic copolymers; polyvinylacrylates; polyethylene oxide; acrylamide polymers and copolymers; polyhydroxyethyl acrylate, methylacrylamide monomers; and polychloroprene.

The amount of binder in the coating can vary, but will be in the range of about 0.01 to about 25% of the weight of the seed, more preferably from about 0.05 to about 15%, and even more preferably from about 0.1 % to about 10%.

The propagative material coating may optionally include a filler. The filler can be an absorbent or an inert filler, such as are known in the art, and may include woodflours, clays, activated carbon, sugars, diatomaceous earth, cereal flours, fine-grain inorganic solids, calcium carbonate, and the like. Clays and inorganic solids, which may be used, include calcium bentonite, kaolin, china clay, talc, perlite, mica, vermiculite, silicas, quartz powder,

montmorillonite and mixtures thereof. Sugars, which may be useful, include dextrin and maltodextrin. Cereal flours include wheat flour, oat flour and barley flour.

The filler is selected so that it will provide a proper microclimate for the seed, for example the filler is used to increase the loading rate of the active ingredients and to adjust the control-release of the active ingredients. The filler can aid in the production or process of coating the seed. The amount of filler can vary, but generally the weight of the filler components will be in the range of about 0.05 to about 75% of the seed weight, more preferably about 0.1 to about 50%, and even more preferably about 0.5% to 15%.

Optionally, a plasticizer can be used in the coating formulation. Plasticizers are typically used to make the film that is formed by the coating layer more flexible, to improve adhesion and spreadability, and to improve the speed of processing. Improved film flexibility is important to minimize chipping, breakage or flaking during storage, handling or sowing processes. Many plasticizers may be used, however, useful plasticizers include polyethylene glycol, glycerol, butylbenzylphthalate, glycol benzoates and related compounds. The range of plasticizer in the coating layer will be in the range of from about 0.1 to about 20% by weight.

The treated seeds may also be enveloped with a film overcoating to protect the active components coating. Such overcoatings are known in the art and may be applied using conventional fluidized bed and drum film coating techniques.

The invention also provides a method for enhancing the growth of a plant propagative material comprising treating a plant propagative material with the aforementioned compositions, wherein growth of the plant propagative material is enhanced relative to a corresponding control plant propagative material that is not treated with the composition.

In certain embodiments of the aforementioned method, the plant propagative material is treated with the composition by coating the plant propagative material with the composition as described above. In other embodiments of the aforementioned method, the plant propagative material may be treated with the composition by applying the composition to the plant growth medium before or after planting. For example, the composition may be applied to the soil by spraying or through an irrigation system. The composition may also be applied to the plant growth medium in a solid form, for example as a dried powder.

EXAMPLES

The following examples are intended to further illustrate the invention, but are not intended to limit the invention in any manner whatsoever.

Example 1

Metarhizium anisopliae spore germination (Microscope studies)

• Spores were obtained from approximately 3 week PDA plate cultures

• Spores were resuspended in 0.01 M Potassium Phosphate and germinant blend

• Incubated at 30°C

• Observed under microscope at 18h, 20h, 24h and 44h

Tables 1 A and IB show the germinative compounds used in each blend. "+" indicates that the germinative compound was added to the sample, and "-" indicates that the germinative compound was not added.

Table 1A

Table IB

The best germinant blends for M. anisopliae spore germination contain L-methionine or L-methionine and sugars. For example, blends 5, 6, 9 & 10 provided greater than 30% germination at 20 hours versus <2% germination in the buffer control treatment. Example 2

Metarhizium anisopliae spore germination (Microscope studies)

• Spores were obtained from approximately 3 week PDA plate cultures

· Spores were resuspended in 0.01 M Potassium Phosphate and germinant blend

• Incubated at 30°C

• Observed under microscope at 18h, 20h, 24h and 44h

Table 2. Germinative compounds and concentrations used in each sample. "+" indicates that the germinative compound was added to the sample at the concentration listed, and "-" indicates that the germinative compound was not added.

Addition of L-Glutamic acid to the germinant blend improved germination of M.

anisopliae. For example, samples 6 and 10 had an increase in germination of greater than 50% as compared to the buffer (untreated) water control.

Example 3

Testing different germinant blends for P. bilaiae and Penicillium sp. spore germination

• Spores were obtained from approximately 3 week PDA plate cultures

• Spores were resuspended in 0.01 M Potassium Phosphate and germinant blend

• Incubated at 30°C

• Observed under microscope at 18h, 20h, 24h and 44h

Table 3. Concentrations of germinative compounds for each sample. "+" indicates that the germinative compound was added to the sample at the concentration listed, and "-" indicates that the germinative compound was not added.

There were more P. bilaiae germination tubes emerging in samples 4, 6, 8, and 10, than in the other blends, indicating that L-arginine and sugars are important for P. bilaiae germination and growth. The complex blend in treatment #10 performed the best with greater than 70% germination and germ tube growth compared to the untreated control. Example 4

Trichoderma harzianum germination experiments

• Spores of Trichoderma harzianum were obtained from PDA plate cultures using

Potassium Phosphate buffer and 0.05% Tween

• Spores were resuspended in 0.01 M Potassium Phosphate and germinant blend

• Incubated at 30°C

• Observed under microscope at 18h, 20h, 24h and 44h

Table 4. Concentrations of germinative compounds for each sample. "+" indicates that the germinative compound was added to the sample at the concentration listed, and "-" indicates that the germinative compound was not added.

Sample 4 containing a complex mixture of amino acids and sugars resulted in the best germination and growth of Trichoderma harzianum. Germination was greater than 70% compared to the untreated control.

Example 5: Comparison of germination rates between intimate mixtures and conventional mixing of fungal spores and germinative compounds

In order to compare the germination rate of the spores of intimate mixtures with that of spores conventionally mixed with a germinative compound, the following treatments are performed:

A. Formation of an intimate mixture: Spores of Aspergillus flavus, Aureobasidium pullulans,

Beauveria bassiana, Coniothyrium minitans, Candida oleophila, Gliocladium roseum,

microaggregatum, Glomus monosporum, Glomus mosseae, Glomus multicaule, Glomus pansihalos, Glomus pustulatum, Glomus sinuosum, Glomus spurcum, Glomus tortuosum, Glomus trimurales, Glomus verruculosum, Glomus versiforme, Glomus viscosum, Gigaspora albida, Gigaspora decipiens, Gigaspora gigantean, Gigaspora margarita, and Gigaspora rosea are dried with various combinations of the germinative compounds L-alanine, glycine, L-leucine, L-tryptophan, L-methionine, L-glutamic acid, arginine, glucose, fructose, L-asparagine, potassium propionate, inosine, magnesium sulfate, calcium chloride, adenosine, and 4- aminobenzoic acid (PABA) introduced to the spore mass immediately prior to drying as a solution containing 1, 10, 12. 5 or 20 mM of the germinative compound. The intimate mixture produced is germinated by subsequent introduction in a solution consisting of 0.01 M potassium phosphate buffer in distilled water. B. Conventional mixing of spores with a germinative compound: Spores of Aspergillus flavus, Aureobasidium pullulans, Beauveria bassiana, Coniothyrium minitans, Candida oleophila, Gliocladium roseum, Gliocladium catenulatum, Metarhizium anisopliae, Paecilomyces fumosoroseus, Paecilomyces lilacinus, Pantoea agglomerans, Penicillium bilaiae, Penicillium gaestrivorus, Pseudozyma flocculosa, Pythium oligandrum, Trichoderma asperellum,

Trichoderma virens, Trichoderma gamsii, Trichoderma hamatum, Trichoderma harzianum, Trichoderma harizatum, Trichoderma koningii, Trichoderma fertile, Ulocladium oudemansii, Ulocladium atrum, Alternaria destruens, Chondrostereum purpureum, Colletotrichum gloeosporioides, Phoma macrostoma, Typhula phacorrhiza, Penicillium chrysogenum,

Penicillium griseofulvum, Penicillium pinophilum, Penicillium canescens, Penicillium glabrum, Penicillium glaucum, Penicillium halgiovense, Penicillium roqueforti, Penicillium simplicissium, Penicillium digitatum, Penicillium janthinellum, Penicillium italicum, Penicillium spinulosum, Penicillium cyclopium, Penicillium purpurogenum, Penicillium janczewskii, Penicillium montanense, Penicillium restrictum, Penicillium harzianum, Penicillium terrestre, Penicillium ochrochloron, Penicillium aurantiogriseum, Penicillium crustosum, Penicillium decumbens, Penicillium griseofulvum, Penicillium janthinellum, Penicillium rugulosum, Penicillium velutinum, Penicillium frequentans, Penicillium verrucosum, Penicillium implicatum,

Penicillium citrinum, Glomus aggregatum, Glomus albidum, Glomus ambisporum, Glomus brazillanum, Glomus caledonium Glomus coremioides, Glomus claroideum, Glomus clarum, Glomus clavisporum, Glomus constrictum, Glomus coronatum, Glomus deserticola, Glomus diaphanum, Glomus eburneum, Glomus etunicatum, Glomus fasciculatum, Glomus fistulosum, Glomus fragilistratum, Glomus geosporum, Glomus globiferum, Glomus heterosporum, Glomus hoi, Glomus intraradices, Glomus lacteum, Glomus lamellosum, Glomus luteum, Glomus manihotis, Glomus microaggregatum, Glomus monosporum, Glomus mosseae, Glomus multicaule, Glomus pansihalos, Glomus pustulatum, Glomus sinuosum, Glomus spurcum, Glomus tortuosum, Glomus trimurales, Glomus verruculosum, Glomus versiforme, Glomus viscosum, Gigaspora albida, Gigaspora decipiens, Gigaspora gigantean, Gigaspora margarita, and Gigaspora rosea are hydrated and dried. Such spores are germinated by the introduction in a 0.01 M potassium phosphate buffer in distilled water containing 1, 10, 12. 5 or 20 mM of various combinations of the germinative compounds L-alanine, glycine, L-leucine, L-tryptophan, L-methionine, L-glutamic acid, arginine, glucose, fructose, L-asparagine, potassium propionate, inosine, magnesium sulfate, calcium chloride, adenosine, and 4-amino benzoic acid (PABA).

C. Germination of spores alone: Spores of Aspergillus flavus, Aureobasidium pullulans, Beauveria bassiana, Coniothyrium minitans, Candida oleophila, Gliocladium roseum,

microaggregatum, Glomus monosporum, Glomus mosseae, Glomus multicaule, Glomus pansihalos, Glomus pustulatum, Glomus sinuosum, Glomus spurcum, Glomus tortuosum, Glomus trimurales, Glomus verruculosum, Glomus versiforme, Glomus viscosum, Gigaspora albida, Gigaspora decipiens, Gigaspora gigantean, Gigaspora mar garita, and Gigaspora rosea are hydrated and dried. The spores are subsequently introduced into a solution consisting of .01 M potassium phosphate buffer in distilled water.

The germination of spores resulting for each such treatment is measured by counting the number of germinated spores under a microscope. It is found that the intimately mixed compositions germinate more rapidly than their corresponding conventionally mixed equivalent or than spores that are not treated with a germinative compound.

Example 6: Evaluation of germination, plant growth, and yield for maize seeds coated with a composition comprising a fungal spore and a germinative compound

Maize seeds are coated with compositions comprising various combinations of the fungal spores and germinative compounds listed in Example 5. Seed coating is performed by conventional means, such as those described in U.S. Pat. No. 7,989,391 and U.S. Pat. No.

5,849,320. The following treatments are evaluated: A. Seed coated with an intimate mixture of the fungal spores and the germinative compound

B. Seed coated with a conventional mixture of the fungal spores and the germinative compound

C. Seed coated with the fungal spores alone

D. Uncoated seed

Seed germination rates are measured in greenhouse and field trials by measuring seedling emergence each day beginning on the first day that plants emerge and continuing for three weeks after the first date of emergence. Yield is measured by harvesting the mature ears and drying them for three days in an oven at 37°C. The ears are then threshed and the seeds are collected and counted. Total seed weight is also measured by weighing the seed harvested from each plant.

It is found that maize seeds coated with an intimate mixture of the fungal spores and germinative compound have higher seed germination rates and higher yields than seed coated with a conventional mixture of the fungal spores, seed coated with fungal spores alone, or uncoated seed.

Claims

What is claimed is: 1. A mixture comprising a fungal spore and a germinative compound, wherein the mixture is a dried intimate mixture, wherein the fungal spore and the germinative compound are maintained in proximate position until they reach an environment conducive to germination.

2. The mixture of claim 1, wherein the germinative compound is adsorbed to or absorbed by the fungal spore.

3. The mixture of claim 1 or 2, wherein the fungal spore is selected from the group consisting of Aspergillus flavus, Aureobasidium pullulans, Beauveria bassiana, Coniothyrium minitans, Candida oleophila, Gliocladium roseum, Gliocladium catenulatum, Metarhizium anisopliae, Paecilomyces fumosoroseus, Paecilomyces lilacinus, Pantoea agglomerans, Penicillium bilaiae, Penicillium gaestrivorus, Pseudozyma flocculosa, Pythium oligandrum, Trichoderma

asperellum, Trichoderma virens, Trichoderma gamsii, Trichoderma hamatum, Trichoderma harzianum, Trichoderma harizatum, Trichoderma koningii, Trichoderma fertile, Ulocladium oudemansii, Ulocladium atrum, Alternaria destruens, Chondrostereum purpureum,

Colletotrichum gloeosporioides, Phoma macrostoma, Typhula phacorrhiza, Penicillium chrysogenum, Penicillium griseofulvum, Penicillium pinophilum, Penicillium canescens, Penicillium glabrum, Penicillium glaucum, Penicillium halgiovense, Penicillium roqueforti, Penicillium simplicissium, Penicillium digitatum, Penicillium janthinellum, Penicillium italicum, Penicillium spinulosum, Penicillium cyclopium, Penicillium purpurogenum,

Penicillium janczewskii, Penicillium montanense, Penicillium restrictum, Penicillium harzianum, Penicillium terrestre, Penicillium ochrochloron, Penicillium aurantiogriseum, Penicillium crustosum, Penicillium decumbens, Penicillium griseofulvum, Penicillium janthinellum,

Penicillium rugulosum, Penicillium velutinum, Penicillium frequentans, Penicillium verrucosum, Penicillium implicatum, Penicillium citrinum, Glomus aggregatum, Glomus albidum, Glomus ambisporum, Glomus brazillanum, Glomus caledonium Glomus coremioides, Glomus claroideum, Glomus clarum, Glomus clavisporum, Glomus constrictum, Glomus coronatum, Glomus deserticola, Glomus diaphanum, Glomus eburneum, Glomus etunicatum, Glomus fasciculatum, Glomus flstulosum, Glomus fragilistratum, Glomus geosporum, Glomus globiferum, Glomus heterosporum, Glomus hoi, Glomus intraradices, Glomus lacteum, Glomus lamellosum, Glomus luteum, Glomus manihotis, Glomus microaggregatum, Glomus

4. The mixture of any one of claims 1-3 wherein the germinative compound is selected from the group consisting of an amino acid, a purine nucleoside, a salt, and a sugar.

5. The mixture of claim 4, wherein the amino acid is selected from the group consisting of glycine, L-alanine, L-arginine, L-aspartate, L-glutamate, L-histidine, L-isoleucine, L-lysine, L- methionine, L-serine, L-tryptophan, L-tyrosine, L- valine, L-proline, L-leucine, L-cysteine, L- threonine, L-glutamine, L-asparagine, L-phenylalanine and analogues thereof.

6. The mixture of claim 4, wherein the purine nucleoside is selected from the group consisting of inosine and adenosine.

7. The mixture of claim 4, wherein the salt is selected from the group consisting of Calcium Chloride, Potassium Chloride, Potassium Phosphate, Potassium Carbonate, Potassium Propionate and Magnesium Sulphate .

8. The mixture of claim 4, wherein the sugar is selected from the group consisting of glucose, fructose, mannose and trehalose.

9. A method for preparing the mixture of claim 1, comprising mixing a fungal spore and a germinative compound.

10. A composition comprising the mixture of any one of claims 1-8.

1 1. A method for preparing a dried intimate mixture comprising a fungal spore and a germinative compound, the method comprising:

wherein the fungal spore and the germinative compound are maintained in proximate position until they reach an environment conducive to germination.

12. The method of claim 11, wherein the drying is spray-drying, freeze-drying, vacuum-drying, air drying or drum drying.

13. A composition comprising:

microaggregatum, Glomus monosporum, Glomus mosseae, Glomus multicaule, Glomus pansihalos, Glomus pustulatum, Glomus sinuosum, Glomus spurcum, Glomus tortuosum, Glomus trimurales, Glomus verruculosum, Glomus versiforme, Glomus viscosum, Gigaspora albida, Gigaspora decipiens, Gigaspora gigantean, Gigaspora margarita, and Gigaspora rosea; and

(b) a germinative compound selected from the group consisting of an amino acid, a purine nucleoside, a salt, and a sugar.

14. The composition of claim 13, wherein the amino acid is selected from the group consisting of glycine, L-alanine, L-arginine, L-aspartate, L-glutamate, L-histidine, L-isoleucine, L-lysine,

L-methionine, L-serine, L-tryptophan, L-tyrosine, L-valine, L-proline, L-leucine, L-cysteine, L- threonine, L-glutamine, L-asparagine, L-phenylalanine and analogues thereof.

15. The composition of claim 13, wherein the purine nucleoside is selected from the group consisting of inosine and adenosine.

16. The composition of claim 13, wherein the salt is selected from the group consisting of Calcium Chloride, Potassium Chloride, Potassium Phosphate, Potassium Carbonate, Potassium Propionate and Magnesium Sulphate.

17. The composition of claim 13, wherein the sugar is selected from the group consisting of glucose, fructose, mannose and trehalose.

18. A method for increasing the germination of fungus spores, comprising:

a) preparing a mixture comprising a fungus spore and a germinative compound; and b) exposing the mixture to an environment conducive to germination of the fungus spores, wherein the germination of the fungus spores in the mixture is increased relative to a

corresponding fungus spore formulation that lacks a germinative compound.

19. A method for increasing the germination of fungus spores, comprising:

c) exposing the intimate mixture to an environment conducive to germination of the fungus spores, wherein the germination of the fungus spores in the intimate mixture is increased relative to a corresponding fungus spore formulation that lacks a germinative compound.

20. A plant propagative material coated with a composition comprising:

a) a fungal spore; and

b) a germinative compound,

wherein the fungal spore and the germinative compound are in a dried intimate mixture, and wherein the fungal spore and the germinative compound are maintained in proximate position until they reach an environment conducive to germination.

21. The plant propagative material of claim 20, wherein the plant propagative material is a seed.

22. The plant propagative material of claim 20 or 21, wherein the fungal spore is selected from the group consisting of Aspergillus flavus, Aureobasidium pullulans, Beauveria bassiana, Coniothyrium minitans, Candida oleophila, Gliocladium roseum, Gliocladium catenulatum, Metarhizium anisopliae, Paecilomyces fumosoroseus, Paecilomyces lilacinus, Pantoea agglomerans, Penicillium bilaiae, Penicillium gaestrivorus, Pseudozyma flocculosa, Pythium oligandrum, Trichoderma asperellum, Trichoderma virens, Trichoderma gamsii, Trichoderma hamatum, Trichoderma harzianum, Trichoderma harizatum, Trichoderma koningii, Trichoderma fertile, Ulocladium oudemansii, Ulocladium atrum, Alternaria destruens, Chondrostereum purpureum, Colletotrichum gloeosporioides, Phoma macrostoma, Typhula phacorrhiza, Penicillium chrysogenum, Penicillium griseofulvum, Penicillium pinophilum, Penicillium canescens, Penicillium glabrum, Penicillium glaucum, Penicillium halgiovense, Penicillium roqueforti, Penicillium simplicissium, Penicillium digitatum, Penicillium janthinellum,

23. The plant propagative material of any one of claims 20-22, wherein the germinative compound is selected from the group consisting of an amino acid, a purine nucleoside, a salt, and a sugar.

24. The plant propagative material of claim 23, wherein the amino acid is selected from the group consisting of glycine, L-alanine, L-arginine, L-aspartate, L-glutamate, L-histidine, L- isoleucine, L-lysine, L-methionine, L-serine, L-tryptophan, L-tyrosine, L-valine, L-proline, L- leucine, L-cysteine, L-threonine, L-glutamine, L-asparagine, L-phenylalanine and analogues thereof.

25. The plant propagative material of claim 23, wherein the purine nucleoside is selected from the group consisting of inosine and adenosine.

26. The plant propagative material of claim 23, wherein the salt is selected from the group consisting of Calcium Chloride, Potassium Chloride, Potassium Phosphate, Potassium

Carbonate, Potassium Propionate and Magnesium Sulphate.

27. The plant propagative material of claim 23, wherein the sugar is selected from the group consisting of glucose, fructose, mannose and trehalose.

28. A method for enhancing the growth of a plant propagative material comprising treating a plant propagative material with the composition of claim 10, wherein growth of the plant propagative material is enhanced relative to a corresponding control plant propagative material that is not treated with the composition.

29. The method of claim 28, wherein the composition is applied to the growth medium.

30. The method of claim 29, wherein the growth medium is soil.

31. The composition of claim 10, further comprising an agriculturally acceptable or pharmaceutically acceptable carrier.

32. The composition of claim 10, further comprising a carrier that is acceptable for use in bioremediation or food production.

33. A food product or pharmaceutical product comprising the composition of claim 10.