Nothing Special   »   [go: up one dir, main page]

WO2024054959A1 - Formulation granulaire d'inoculum de carbone du sol et ses procédés d'utilisation - Google Patents

Formulation granulaire d'inoculum de carbone du sol et ses procédés d'utilisation Download PDF

Info

Publication number
WO2024054959A1
WO2024054959A1 PCT/US2023/073709 US2023073709W WO2024054959A1 WO 2024054959 A1 WO2024054959 A1 WO 2024054959A1 US 2023073709 W US2023073709 W US 2023073709W WO 2024054959 A1 WO2024054959 A1 WO 2024054959A1
Authority
WO
WIPO (PCT)
Prior art keywords
carbon
dmtr
ctr
pta
accession
Prior art date
Application number
PCT/US2023/073709
Other languages
English (en)
Inventor
Neeraj PURUSHOTHAM
Guy WEBB
Frank Oly
Ahsanul HAQUE
Cassandra O'BRIEN
Dillon HOLTON
Anders CLAASSENS
Pratishtha Sharma
Original Assignee
Loam Bio Pty Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Loam Bio Pty Ltd filed Critical Loam Bio Pty Ltd
Publication of WO2024054959A1 publication Critical patent/WO2024054959A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F11/00—Other organic fertilisers
    • C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
    • A01N63/38—Trichoderma
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P21/00—Plant growth regulators
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F11/00—Other organic fertilisers
    • C05F11/02—Other organic fertilisers from peat, brown coal, and similar vegetable deposits
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F5/00—Fertilisers from distillery wastes, molasses, vinasses, sugar plant or similar wastes or residues, e.g. from waste originating from industrial processing of raw material of agricultural origin or derived products thereof
    • C05F5/002—Solid waste from mechanical processing of material, e.g. seed coats, olive pits, almond shells, fruit residue, rice hulls
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
    • C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
    • C05G3/80—Soil conditioners
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
    • C05G5/00—Fertilisers characterised by their form
    • C05G5/10—Solid or semi-solid fertilisers, e.g. powders
    • C05G5/12—Granules or flakes
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K17/00—Soil-conditioning materials or soil-stabilising materials
    • C09K17/40—Soil-conditioning materials or soil-stabilising materials containing mixtures of inorganic and organic compounds
    • C09K17/42—Inorganic compounds mixed with organic active ingredients, e.g. accelerators
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
    • C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H17/00—Symbiotic or parasitic combinations including one or more new plants, e.g. mycorrhiza
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi

Definitions

  • the present invention generally relates to a soil carbon inoculum granular formulation and uses thereof to increase organic carbon in soil, sequester atmospheric carbon for storage as organic carbon in a soil, and enhance at least one plant characteristic in an inoculated plant.
  • methane is much more shortlived than carbon dioxide, the effect of carbon dioxide is often considered more important than that of methane to the greenhouse effect.
  • the life cycle of carbon includes the removal of carbon dioxide from the atmosphere by plants through photosynthesis. During the process of photosynthesis, the carbon dioxide gets absorbed through the stroma of leaves, and the carbon dioxide is further converted into sugars. Such sugars become nutrients for plants and microbes present in the soil. Carbon enters back into the atmosphere in the form of carbon dioxide by respiration and combustion. Hence, a balanced amount of release and absorption of carbon dioxide is an essential step for balancing the ecosystem.
  • the present invention provides a soil carbon inoculum granular formulation which in some aspects at least achieve the stated objective.
  • the formulations and methods disclosed herein provide cost-efficient solutions that can address current challenges with application of fungal strains as seed coating and facilitate their deployment in the field resulting in increased soil organic carbon and improved agronomic traits in target crops.
  • the disclosed formulations and methods increase carbon sequestration in soil and are compatible with commercial farming equipment.
  • the present disclosure provides a soil carbon inoculum granular formulation comprising: a) a carbon sequestering fungal strain; b) a solid carrier selected from the group consisting of cellulose, a non-viable grain, a non-viable seed, a vegetable fiber, a wood fiber, zeolite, pumice, sand, attapulgite, perlite, vermiculite, peat, corn cob, carbon, activated carbon, and combinations and derivatives thereof; and c) a coating composition comprising a wax agent, chitin, gelatin, alginate, pectin, carrageenan, collagen, xyloglucan, polyvinyl acetate, polyacrylate, methylacrylate, hypromellose (HPMC), polyvinylpyrrolidone, polyvinyl alcohol, polyacrylic acid, polyethylene glycols, vinyl acetate copolymer, vinyl acetate homopolymer, vinyl acetate-acrylic copolymer
  • the carbon sequestering fungal strain is a dark septate endophyte (DSE).
  • DSE belongs to the order of Pleosporales, Microascales, Xylariales, Pezizales, Dothideales, Leotiales, Chaetothyriales, Elaphomycetales, Eurotiales, Onygenales, Saccharomycetales, Neolectales, Taphrinales, Mitosporic, or Hypocreales.
  • the carbon sequestering fungal strain belongs to a genus selected from the group consisting of Clohesyomyces, Darksidea, Phialocephala, Acrocalymma, Clonostachys, Leptodontidium, Periconia, Phaeosphaeria, Thozetella, Trichoderma, and Beauveria.
  • the carbon sequestering fungal strain belongs to a species selected from the group consisting of Clohesyomyces aquaticus, Phialocephala fortinii sd - Acephala applanata species complex (PAC), Darksidea zeta, Acrocalymma vagum, Clonostachys rosea, Leptodontidium orchidicola, Periconia macrospinosa, Periconia circinata, Phaeosphaeria luctuosa, Phaeosphaeria vagans, Thozetella nivea, Trichoderma hamatum, Trichoderma longipile, Trichoderma spirale, and Beauveria bassiana
  • PAC Phialocephala fortinii sd - Acephala applanata species complex
  • the carbon sequestering fungal strain is Clohesyomyces aquaticus DMTR-CTR-7800 (NMI Accession No. V21/002328), Phialocephala fortinii sd - Acephala applanata species complex (PAC) DMTR-CTR-7788 (NMI Accession No. V21/002327), Darksidea zeta DMTR-CTR-6853 (NMI Accession No. V21/002326), Darksidea zeta DMTR- CTR-4796 (NMI Accession No. V21/003117), Darksidea sp. DMTR-CTR-360 (NMI Accession No.
  • V22/006358 Periconia macrospinosa AU-7083 (NMI Accession No. V22/019796), Phaeosphaeria luctuosa / vagans DMTR-CTR-3044 (NMI Accession No. V22/006355), Thozetella nivea DMTR-CTR-2359 (NMI Accession No. V22/003496), Trichoderma hamatum DMTR-CTR-US-73 (ATCC Accession No. PTA-127301), Trichoderma hamatum US-724 (ATCC Accession No. PTA-127448), Trichoderma longipile / spirale DMTR-CTR-1291 (NMI Accession No.
  • Trichoderma longipile / spirale US-77 (ATCC Accession No. PTA-127440), Beauveria bassiana AU-16727 (NMI Accession No. V23/003855), B. bassiana US-52 (ATCC Accession No. PTA-127541), B. bassiana US- 675 (ATCC Accession No. PTA-127540), B. bassiana US-699 (ATCC Accession No. PTA- 127538), B. bassiana US-707 (ATCC Accession No. PTA-127542), and B. bassiana US-803 (ATCC Accession No. PTA-127539), or a mutant thereof having all identifying characteristics of the respective strain.
  • Clohesyomyces aquaticus DMTR-CTR-7800 (NMI Accession No. V21/002328) is also known as Clohesyomyces aquaticus AU-7800 (NMI Accession No. V21/002328).
  • Phialocephala fortinii s.l - Acephala applanata species complex (PAC) DMTR-CTR- 7788 (NMI Accession No. V21/002327) is also known as Phialocephala fortinii s.l - Acephala applanata species complex (PAC) AU-7788 (NMI Accession No. V21/002327).
  • Darksidea zeta DMTR-CTR-6853 (NMI Accession No. V21/002326) is also known as Darksidea zeta AU-6853 (NMI Accession No. V21/002326).
  • Darksidea zeta DMTR-CTR-4796 (NMI Accession No. V21/003117) is also known as Darksidea zeta AU-4796 (NMI Accession No. V21/003117).
  • Darksidea sp. DMTR-CTR-360 (NMI Accession No. V21/003116) is also known as Darksidea sp. AU-360 (NMI Accession No. V21/003116).
  • Acrocalymma vagum DMTR-CTR-11556 (NMI Accession No. V22/006357) is also known as Acrocalymma vagum AU-11556 (NMI Accession No. V22/006357).
  • Clonostachys rosea DMTR-CTR-US-173 (ATCC Accession No. PTA-127299) is also known as Clonostachys rosea US-173 (ATCC Accession No. PTA-127299).
  • Clonostachys rosea DMTR-CTR-1081 (NMI Accession No. V22/003495) is also known as Clonostachys rosea AU-1081 (NMI Accession No. V22/003495).
  • Leptodontidium orchidicola DMTR-CTR-4873 (NMI Accession No. V22/003497) is also known as Leptodontidium orchidicola AU-4873 (NMI Accession No. V22/003497.
  • Periconia circinata DMTR-CTR-6649 (NMI Accession No. V22/006356) is also known as Periconia circinata AU-6649 (NMI Accession No. V22/006356).
  • Periconia macrospinosa DMTR-CTR-US-125 (ATCC Accession No. PTA-127300) is also known as Periconia macrospinosa US-125 (ATCC Accession No. PTA-127300).
  • Periconia macrospinosa DMTR-CTR-1852 (NMI Accession No. V22/006358) is also known as Periconia macrospinosa AU-1852 (NMI Accession No. V22/006358).
  • Phaeosphaeria luctuosa / vagans DMTR-CTR-3044 (NMI Accession No. V22/006355) is also known as Phaeosphaeria luctuosa / vagans AU-3044 (NMI Accession No. V22/006355).
  • Thozetella nivea DMTR-CTR-2359 (NMI Accession No. V22/003496) is also known as Thozetella nivea AU-2359 (NMI Accession No. V22/003496).
  • Trichoderma hamatum DMTR-CTR-US-73 (ATCC Accession No. PTA-127301) is also known as Trichoderma hamatum US-73 (ATCC Accession No. PTA-127301).
  • Trichoderma longipile / spirale DMTR-CTR-1291 (NMI Accession No. V22/006354) is also known as Trichoderma longipile / spirale AU-1291 (NMI Accession No. V22/006354).
  • the carbon sequestering fungal strain comprises fungal spores, fungal hyphae, or a combination thereof. In other aspects, the carbon sequestering fungal strain comprises fungal spores, fungal conidia, fungal hyphae, or a combination thereof.
  • the carbon sequestering fungal strain comprises any one of hyphae, conidia, chlamydospore, zygospores, (micro) sclerotia, ascospores, basidiospores, chlamydospores, oospores, ascospores, uredospores, teleutospores, ustospores, blastospores or hyphal fragments.
  • the solid carrier comprises a cellulose agglomerate.
  • the solid carrier is a non-viable grain or non-viable seed selected from the group consisting of rice, wheat, millet, com, barley, oats, buckwheat, quinoa, sorghum, spelt, triticale, rye, clover, medic, lucerne, teff or flaxseed.
  • the carbon sequestering fungal strain is cultured on the cellulose agglomerate, non-viable grain, or non-viable seed as a substrate prior to applying the coating composition.
  • the solid carrier is zeolite.
  • the soil carbon inoculum granular formulation further comprises a filler selected from the group consisting of com starch powder, talc, mica, oyster shell calcium, diatomaceous earth, calcium carbonate, sodium bicarbonate, silica, silicates, barium sulfate, titanium dioxide, silicon dioxide, calcium sulfate, kaolin, bentonite, montmorillonite, red clay, biochar fines, and combinations thereof.
  • a filler selected from the group consisting of com starch powder, talc, mica, oyster shell calcium, diatomaceous earth, calcium carbonate, sodium bicarbonate, silica, silicates, barium sulfate, titanium dioxide, silicon dioxide, calcium sulfate, kaolin, bentonite, montmorillonite, red clay, biochar fines, and combinations thereof.
  • the soil carbon inoculum granular formulation further comprises a binder selected from the group consisting of a styrene/butadiene copolymer, a butadiene/acrylonitrile copolymer, a styrene/butadiene/acrylonitrile copolymer, a styrene/methacrylic ester copolymer, a vinyl acetate/acrylic copolymer, a vinyl acetate/methacrylic ester copolymer, a vinyl acetate/ethylene copolymer, a vinyl acetate homopolymer, a methacrylic ester/acrylic ester copolymer, and a combination thereof.
  • a binder selected from the group consisting of a styrene/butadiene copolymer, a butadiene/acrylonitrile copolymer, a styrene/butadiene/acrylonitrile copolymer, a
  • the soil carbon inoculum granular formulation further comprises an arbuscular mycorrhiza fungus (AMF) selected from the group consisting of: Acaulospora mellea, Claroideoglomus etunicatum, Dominikia aurea, Funneliformis mosseae, Glomus aggregatum, Glomus heterosporum, Rhizophagus clarus, Rhizophagus custos, Rhizophagus diaphanous, Rhizophagus irregularis, Paraglomus brasilianum, and combinations thereof.
  • AMF arbuscular mycorrhiza fungus
  • the soil carbon inoculum granular formulation further comprises plant growth promoting-rhizobacteria (PGPR) belonging to a genus selected from the group consisting of Actinobacter, Alcaligenes, Bacillus, Burkholderia, Buttiauxella, Enterobacter, Klebsiella, Kluyvera, Pantoea, Pseudomonas, Rahnella, Ralstonia, Rhizobium, Serratia, Stenotrophomonas, Paenibacillus, and Lysinibacillus.
  • PGPR plant growth promoting-rhizobacteria
  • the soil carbon inoculum granular formulation further comprises a signaling compound selected from the group consisting of gibberellin hormone, dexamethasone, abscisic acid (ABA), P-aminobutyric acid (BABA), ethanol, auxin, cytokinin (CK), apocarotenoid, flavonoid, jasmonate, strigolactone, salicylic acid, protocatechuic acid (PCA), vanillic acid (VA), phloretin, 1 -naphthaleneacetic acid (NAA), and a combination thereof.
  • a signaling compound selected from the group consisting of gibberellin hormone, dexamethasone, abscisic acid (ABA), P-aminobutyric acid (BABA), ethanol, auxin, cytokinin (CK), apocarotenoid, flavonoid, jasmonate, strigolactone, salicylic acid, protocatechuic acid (PCA), vanillic acid (VA), p
  • the soil carbon inoculum granular formulation further comprises photoproteins, vitamins, chelation agents such as fulvic and humic acids, amino acids, or a combination thereof.
  • the soil carbon inoculum granular formulation further comprises a microbial fermentation supernatant containing bioactive plant and microbial growth stimulating compounds including but not limited to flavonoids.
  • the soil carbon inoculum granular formulation is a granule or pellet with an average diameter of between about 0.1 mm and about 10.0 mm, between about 0.1 mm and about 8.0 mm, between about 0.1 mm and about 6.0 mm, between about 0.1 mm and about 4.0 mm, between about 0.1 mm and about 3.0 mm, between about 0.1 mm and about 2.0 mm, or between about 0.1 mm and about 1.0 mm.
  • the soil carbon inoculum granular formulation further comprises a fungicide that is compatible and does not significantly inhibit the growth of the carbon sequestering fungal strain.
  • This fungicide can function to reduce the soil-bome fungal competition including fungal plant pathogens.
  • the fungicide may be an azole fungicide (e.g., difenoconazole, epoxiconazole, fluquinconazole, flutriafol, imazalil, metconazole, prochloraz, propiconazole, prothioconazole, tebuconazole, triadimenol, triticonazole); or a strobilurin fungicide (e.g., azoxystrobin, dimoxystrobin, fluoxastrobin, kresoxim-methyl, methaminostrobin, orysastrobin, picoxystrobin, pyraclostrobin and trifloxystrobin).
  • azole fungicide e.g., difenoconazole, epoxiconazole, fluquinconazole, flutriafol, imazalil, metconazole, prochloraz, propiconazole, prothioconazole,
  • the fungicide is an inhibitor of the respiratory chain at complex I or II selected from the group consisting of benzovindiflupyr, bixafen, boscalid, fluopyram, fluxapyroxad, isofetamid, isopyrazam (anti-epimeric enantiomer 1R,4S,9S), isopyrazam (anti- epimeric enantiomer 1S,4R,9R), isopyrazam (anti-epimeric racemate 1RS,4SR,9SR), isopyrazam (mixture of syn-epimeric racemate 1RS,4SR,9RS and anti-epimeric racemate 1RS,4SR,9SR), isopyrazam (syn-epimeric enantiomer 1R,4S,9R), isopyrazam (syn-epimeric enantiomer 1S,4R,9S), isopyrazam (syn-epimeric enantiomer 1S,4
  • the fungicide is an inhibitor of nucleic acid synthesis selected from the group consisting of benalaxyl, benalaxyl-M (kiralaxyl), metalaxyl, and metalaxyl-M (mefenoxam).
  • the present disclosure provides a soil carbon inoculum granular formulation
  • a soil carbon inoculum granular formulation comprising: a) a carbon sequestering fungal strain; b) a solid carrier selected from the group consisting of a non-viable grain, a non-viable seed, a vegetable fiber, a wood fiber, peat, corn cob, carbon, activated carbon, and combinations and derivatives thereof; wherein the solid carrier is colonized by the fungal strain and subsequently fragmented and fractionated to enrich the formulation with granules having a higher propagule: substrate ratio compared to the formulation prior to fragmentation and fractionation.
  • the solid carrier is a non-viable grain or non-viable seed selected from the group consisting of rice, wheat, millet, com, barley, oats, buckwheat, quinoa, sorghum, spelt, triticale, rye, clover, medic, lucerne, teff, or flaxseed.
  • the solid carrier is colonized by the fungal strain and subsequently fragmented and fractionated to enrich for granules having a size between 50 pm and 250 pm, between 75 pm and 250 pm, between 100 pm and 250 pm, between 50 pm and 200 pm, between 75 pm and 200 pm, between 100 pm and 200 pm, between 50 pm and 150 pm, between 75 pm and 150 pm, or between 100 pm and 150 pm.
  • the disclosure provides a method of increasing organic carbon in a soil, comprising applying to a plant, seedling, plant propagation material, or the locus surrounding the plant material an effective amount of a soil carbon inoculum granular formulation disclosed herein, wherein the soil carbon inoculum granular formulation is in an amount effective to increase organic carbon in the soil compared to a non-inoculated control soil.
  • the disclosure provides a method for sequestering atmospheric carbon for storage as organic carbon in a soil, comprising applying to a plant, seedling, plant propagation material, or the locus surrounding the plant material an effective amount of a soil carbon inoculum granular formulation disclosed herein, wherein the soil carbon inoculum granular formulation is in an amount effective to increase sequestered atmospheric carbon in the soil compared to a non-inoculated control soil.
  • the disclosure provides a method of plant enhancement comprising applying to a plant, seedling, plant propagation material, or the locus surrounding the plant material an effective amount of a soil carbon inoculum granular formulation disclosed herein to enhance at least one plant characteristic; wherein the plant characteristic is selected from the group consisting of accelerated seed germination, accelerated seedling emergence, improved seedling emergence, improved leaf formation, accelerated leaf formation, improved plant maturation, accelerated plant maturation, increased plant yield, increased plant growth, increased plant quality, increased plant health, increased fruit yield, increased fruit growth, increased fruit quality, improved root health, increased root length, increased root mass, increased root branching, increased root hair density, increased root nodule formation, plant health, plant resistance to salt stress, plant resistance to heat stress, plant resistance to heavy metal stress, plant resistance to drought, and combinations thereof; and wherein the soil carbon inoculum granular formulation is in an amount effective to enhance the at least one plant characteristic compared to a non-inoculated plant.
  • the soil carbon inoculum granular formulation is applied pre-planting, at the time of sowing, and/or post-planting.
  • the plant is a legume, wheat, rice, com (maize), rye, oats, barley, sorghum, millet, flax, hemp, jute, sugarcane, or cotton.
  • the legume is alfalfa, clover, peas, cowpeas, beans, mung beans, lentils, lupins, mesquite, carob, soybeans, peanuts, tamarind, wisteria, siratro, plants from the Lespedeza genus, Genistoid legumes, or serradella.
  • the soil carbon inoculum granular formulation is applied at a rate of 0.01 g to 10 kg per 100 kg of plant propagation or at a rate of 0.01-10 kg per hectare.
  • the soil carbon inoculum granular formulation can be applied at a rate based on the number of seeds sown. For example, application rates include 5 granules per 1 seed, 4 granules per 1 seed, 3 granules per 1 seed, 2 granules per 1 seed, 1 granule per 1 seed, 1 granule per 2 seed, 1 granule per 3 seed, 1 granule per 4 seed, and 1 granule per 5 seed.
  • the soil carbon inoculum granular formulation is applied in a ratio of 25 granules per 1 seed to 1 seed per 25 granules, a ratio of 20 granules per 1 seed to 1 seed per 20 granules, a ratio of 15 granules per 1 seed to 1 seed per 15 granules, a ratio of 10 granules per 1 seed to 1 seed per 10 granules, or a ratio of 5 granules per 1 seed to 1 seed per 5 granules.
  • the soil carbon inoculum granular formulation may be applied at a specific distance from seed (e.g., at a distance of between 0.1 and 20 mm, between 0.1 and 15 mm, between 0.1 and 10 mm, or between 0.1 and 5 mm from the seed).
  • the disclosure provides a method of producing a soil carbon inoculum granular formulation, the method comprising: providing a solid carrier selected from the group consisting of cellulose, a non-viable grain, a non-viable seed, a vegetable fiber, a wood fiber, zeolite, pumice, sand, attapulgite, perlite, vermiculite, peat, com cob, carbon, activated carbon, and combinations and derivatives thereof; optionally, supplementing the solid carrier with a nutrient supporting fungal growth; culturing a carbon sequestering fungal strain on a medium comprising the solid carrier; and applying a coating composition to the fungal strain and solid carrier, wherein the coating composition comprises a wax agent, chitin, gelatin, alginate, pectin, carrageenan, collagen, xyloglucan, polyvinyl acetate, polyacrylate, methylacrylate, hypromellose (HPMC), polyvinylpyrrolidone,
  • the solid carrier is cellulose, a non-viable grain, a non-viable seed, or a combination or derivative thereof.
  • the non-viable grain or non-viable seed is selected from the group consisting of rice, wheat, millet, corn, barley, oats, buckwheat, quinoa, sorghum, spelt, triticale, rye, or flaxseed.
  • the nutrient comprises casamino acids, ammonium sulfate, glucose, peptone, yeast extract, potato extract, casein, or a combination thereof.
  • FIG. 1A depicts an embodiment of a soil carbon inoculum granular formulation comprising a solid carrier, a fungal inoculum, and a polymer.
  • FIG. IB depicts a further embodiment of a soil carbon inoculum granular formulation similar to that in FIG. IB but with additional components including a soil carbon inoculum package (SCIP) powder inoculum.
  • SCIP soil carbon inoculum package
  • FIG. 2 depicts another embodiment of a soil carbon inoculum granular formulation comprising arbuscular mycorrhiza fungus (AMF) and plant growth promoting (PGP) chemical compounds.
  • FIG. 3 depicts another embodiment of a soil carbon inoculum granular formulation wherein one fungal inoculum is applied as a hyphal powder and another fungal inoculum is applied as a spore powder.
  • FIG. 4 depicts images of BIODAC® (cellulose) granules colonized with carbon sequestering fungal strains in Erlenmeyer flasks.
  • FIG. 5 depicts images of fungal outgrowth on agar plates with BIODAC® (cellulose) granules colonized with carbon sequestering fungal strains after storage at 4°C for about 3 months.
  • BIODAC® cellulose
  • FIG. 6 depicts images of a soil carbon inoculum granular formulation on a zeolite core with a ruler indicating relative diameters of the granules.
  • FIGs. 7A and 7B depict scanning electron microscope (SEM) images of the surface of untreated control BIODAC® (cellulose) granules.
  • FIGs. 8A and 8B depict SEM images of the surface of BIODAC® (cellulose) granules colonized with Periconia macrospinosa DMTR-CTR-1852 (NMI Accession No. V22/006358).
  • FIG. 9 depicts depict SEM images of the surface of BIODAC® (cellulose) granules colonized with Acrocalymma vagum DMTR-CTR-11556 (NMI Accession No. V22/006357).
  • FIG. 10 depicts the experimental setup for a greenhouse experiment measuring total carbon in soil with wheat plants treated with BIODAC® (cellulose) granules colonized with different fungal strains
  • FIG. 11 depicts increases in soil carbon in a greenhouse experiment with wheat plants treated with BIODAC® (cellulose) granules colonized with Phialocephala fortinii s.l - Acephala applanata species complex (PAC) DMTR-CTR-7788 (NMI Accession No. V21/002327), Acrocalymma vagum DMTR-CTR-11556 (NMI Accession No. V22/006357), Clonostachys rosea DMTR-CTR-1081 (NMI Accession No. V22/003495), Periconia sp. DMTR-CTR-6649 (NMI Accession No.
  • FIG. 12A and 12B depicts increases in soil carbon resulting from application of Periconia macrospinosa DMTR-CTR-US-125 (ATCC Accession No. PTA-127300) (also known as “US-125”) or Leptodontidium orchidicola US-210 (ATCC Accession No. PTA- 127441) (also known as “US-210”) to winter wheat as a seed treatment produced with solid state fermentation (SSF) or as colonized BIODAC® (cellulose) granules in field trials conducted in Bradford, Texas, USA, and Hutchinson, Kansas, USA.
  • SSF solid state fermentation
  • BIODAC® BIODAC®
  • FIG. 13 depicts winter wheat yield with plants treated with US- 125 or US-210 as a seed treatment produced with SSF or as colonized BIODAC® (cellulose) granules in field trials conducted in Bradford, Texas, USA, and Hutchinson, Kansas, USA.
  • BIODAC® cellulose
  • FIG. 14A, 14B, and 14C depict the shelf-life at 4 degrees C and 25 degrees C of BIODAC® (cellulose) granules colonized with Thozetella nivea DMTR-CTR-2359 (NMI Accession No. V22/003496), Leptodontidium orchidicola DMTR-CTR-4873 (NMI Accession No. V22/003497), and Acrocalymma vagum AU-11556 (NMI Accession No. V22/006357), respectively.
  • BIODAC® cellulose
  • FIG. 15 depicts the results of an analysis of fragmented millet grains colonized with Thozetella nivea DMTR-CTR-2359 (NMI Accession No. V22/003496) to identify those granules with the highest active propagule density.
  • FIG. 16 depicts more efficient use of available real estate on a seed surface occupied by high potency, biologically active fungal granules produced with fractionation.
  • carrier refers to a substance that physically or chemically binds or combines with a target or active substance (e.g., a carbon sequestering fungal strain) to facilitate the use, storage or application of the target or active substance.
  • Carriers are often inert materials, but they can also include non-inert materials when compatible with the target or active substance.
  • a “granule” as used in this application is an agglomeration of at least a solid carrier or a fragment thereof and a fungal strain.
  • the granule further comprises a coating composition, a filler, a signaling compound, an arbuscular mycorrhiza fungus (AMF), and/or plant growth promoting-rhizobacteria (PGPR).
  • AMF arbuscular mycorrhiza fungus
  • PGPR plant growth promoting-rhizobacteria
  • progule refers to a fungal cell with the ability to reproduce and propagate as daughter cells.
  • progenitor: substrate ratio refers to the ratio of the number of propagules to unit mass of solid carrier (i.e., substrate) colonized by a fungal strain.
  • coating is meant to refer to applying material to a surface of a carrier, for instance as a layer of a material around a carrier. Coating includes film coating, pelleting, and encrusting or a combination of these techniques as known in the art. It will be understood that the term “film coating” refers to a concentrated composition which can be diluted and formed into a slurry with other components added, such as agrochemical actives, in order to make a “carrier coating” which is then applied to the carrier.
  • the culturing process differs from the culturing process that microbial cells experiences in nature.
  • the microbial cell culture produced as a whole and used in the described inventive compositions differs from the culture that results from a microbial cell culturing process that occurs in nature.
  • the soil carbon inoculum granular formulation disclosed herein comprises a carbon sequestering fungal strain, a solid carrier, and a coating composition.
  • the carbon sequestering fungal strain is a dark septate endophyte (DSE).
  • DSEs are a group of endophytic fungi identified by their morphology of melanized, septate, hyphae.
  • the carbon sequestering fungi are DSEs belonging to the order of Pleosporales, Microascales, Xylariales, Pezizales, Dothideales, Leotiales, Chaetothyriales, Elaphomycetales, Eurotiales, Onygenales, Saccharomycetales, Neolectales, Taphrinales, or Mitosporic.
  • the carbon sequestering fungal strain belongs to a genus selected from the group consisting of Clohesyomyces, Darksidea, Phialocephala, Acrocalymma, Clonostachys, Leptodontidium, Periconia, Phaeosphaeria, Thozetella, and Trichoderma.
  • the carbon sequestering fungal strain belongs to a species selected from the group consisting of Clohesyomyces aquaticus, Phialocephala fortinii s.l - Acephala applanata species complex (PAC), Darksidea zeta, Acrocalymma vagum, Clonostachys rosea, Leptodontidium orchidicola, Periconia macrospinosa, Periconia circinata, Phaeosphaeria luctuosa, Phaeosphaeria vagans, Thozetella nivea, Trichoderma hamatum, Trichoderma longipile, and Trichoderma spirale
  • PAC Phialocephala fortinii s.l - Acephala applanata species complex
  • the solid carrier is cellulose.
  • a preferred carrier is agglomerated cellulosic granules sold by Kadant GranTek Inc. in Green Bay, Wisconsin, under its trademark BIODAC®. Methods of agglomeration are disclosed, for example, in U.S. Pat. No. 4,560,527. These and other agglomerated granules preferably contain at least 30% by weight of cellulosic fibers.
  • BIODAC® 8/30, 12/20, 16/30, 20/40, and 30/50 in which the numbers represent the U.S. mesh size of the two screens used may be used as the solid carrier.
  • the size of the BIODAC® (cellulose) granules used depends on the size of the seed with which the soil carbon inoculum granular formulation is sown and the desired application rate for the carbon sequestering fungal strain. For example, a smaller granule (e.g., a granule of between 0.1 mm and 1.0 mm) may be used for a smaller seed (e.g., canola seed) to facilitate application at the time of sowing.
  • the carbon sequestering fungal strain is cultured on the cellulose agglomerates as a substrate prior to applying the coating composition.
  • the BIODAC® (cellulose) granules may be supplemented with nutrients to support fungal growth (e.g., casamino acids, ammonium sulfate, glucose, peptone, yeast extract, potato extract, casein, etc.)
  • BIODAC® cellulose
  • the BIODAC® (cellulose) granules are coated with a polymer disclosed herein that adds weight and improved ballistics to the resulting pellet.
  • the polymer provides better heat and chemical shock resistance and also affords improved shelf life and inoculum performance.
  • the solid carrier is a non-viable grain, a non-viable seed, or a derivative thereof.
  • the non-viable grain, non-viable seed, or derivative thereof may be barley, brown rice, buckwheat, bulgur (cracked wheat), flaxseed, grano, millet, oats, oat bread, oat cereal, oatmeal, popcorn, whole wheat cereal flakes, muesli, rolled oats, quinoa, rye, sorghum, spelt, triticale, whole grain barley, wheat berries, whole grain cornmeal, whole rye, whole wheat bread, whole wheat couscous or wild rice.
  • the solid carrier is a non-viable grain or non-viable seed selected from the group consisting of rice, wheat, millet, com, barley, oats, buckwheat, quinoa, sorghum, spelt, triticale, rye, or flaxseed.
  • the non-viable grain or a non-viable seed is autoclaved to ensure nonviability.
  • the solid carrier is a zeolite.
  • Zeolites are microporous, aluminosilicate minerals commonly used as commercial adsorbents and catalysts. They are tetrahedral, three dimensional, crystalline minerals of aluminosilicate earth metals and belong to the acidic catalysts. Zeolites have a porous structure that can accommodate a wide variety of cations, such as Na + , K + , Ca 2+ , Mg 2+ and others. These positive ions are rather loosely held and can readily be exchanged for others in a contact solution. Some of the more common mineral zeolites are analcime, chabazite, clinoptilolite, heulandite, natrolite, phillipsite, and stilbite.
  • Zeolite powder has the desirable chemo-physical characteristics of water imbibition and toxin absorption. Zeolite powder can be sieved to produce granules or pellets of a specific size (e.g., 0.1 mm - 1.0 mm; 0.5 mm - 1.0 mm; 0.8 mm - 1.5 mm; 1.0 mm - 2.0 mm, 1.0 mm - 3.0 mm; 2.0 mm - 5.0 mm; 4.0 mm - 8.0 mm; etc.)
  • Zeolite has many useful benefits for microbial inoculums in that it adsorbs compounds and maintains their molecular integrity, has a large cation exchange capacity (CEC), and possesses a honeycomb structure.
  • the zeolite is coated with a polymer (e.g., methylcellulose) prior to application of a carbon sequestering fungal strain.
  • the carbon sequestering fungal strain is cultured or grown on bed of zeolite chips, granules, or pellets.
  • the zeolite is pre-loaded with flavonoids and other fungal and plant root enhancement compounds prior to application of the carbon sequestering fungal strain.
  • the solid carrier is a fibrous material.
  • the fibrous material may comprise any suitable organic or inorganic fibers or fiber particles.
  • Suitable fibers include vegetable fibers and wood fibers. Vegetable fibers are usually of cellulose, often in combination with lignin. Suitable examples include cotton, bamboo, hemp, jute, flax, ramie, sisal, bagasse, and banana. Wood fiber is distinguished from vegetable fiber, as being from tree sources. Forms include groundwood, lacebark, thermomechanical pulp (TMP), and bleached or unbleached kraft or sulfite pulps. Lignin is removed in the Kraft and sulfite type of pulping process.
  • the solid carrier may comprise about 60 wt.% or less, such as about 55 wt.% or less, such as about 50 wt.% or less, such as about 45 wt.% or less, such as about 40 wt.% or less, such as about 35 wt.% or less, such as about 30 wt.% or less, such as about 25 wt.% or less, such as about 20 wt.% or less, such as about 15 wt.% or less, such as about 13 wt.% or less, such as about 12 wt.% or less, such as about 1 1 wt.% or less, such as about 10 wt.% or less, such as about 7.5 wt.% or less, such as about 6 wt.% or less, such as about 5.5 wt.% or less, such as about 5 wt.% or less, such as about 4 wt.% or less, such as about 3 wt.% or less of the composition.
  • the solid carrier may comprise about 0.5 wt.% or more, such as about 1 wt.% or more, such as about 1 .5 wt.% or more, such as about 2 wt.% or more, such as about 2.5 wt.% or more, such as about 3 wt.% or more, such as about 4 wt.% or more, such as about 4.5 wt.% or more, such as about 5 wt.% or more, such as about 7.5 wt.% or more, such as about 9 wt.% or more, such as about 10 wt.% or more, such as about 12.5 wt.% or more, such as about 15 wt.% or more, such as about 17.5 wt.% or more, such as about 20 wt.% or more, such as about 25 wt.% or more, such as about 30 wt.% or more, such as about 35 wt.% or more, such as about 40 wt.% or more,
  • a coating composition is applied to the solid carrier before application of a carbon sequestering fungal strain, after application of a carbon sequestering fungal strain, and/or between layers of a carbon sequestering fungal strain and another agricultural agent (e.g., plant or fungal signaling compounds, plant growth-promoting rhizobacteria (PGPR), arbuscular mycorrhiza fungi (AMF), etc.).
  • another agricultural agent e.g., plant or fungal signaling compounds, plant growth-promoting rhizobacteria (PGPR), arbuscular mycorrhiza fungi (AMF), etc.
  • the coating composition comprises a water-soluble polymer.
  • Suitable water soluble polymers that may be mentioned herein include, but are not limited to, carrageenan lambda, hyaluronic acid, pullulan, polyvinylpyrrolidone, polyacrylic acid, gum acaia, gelatin, alginate, pectin, carrageenan, collagen, xyloglucan, carboxymethylcellulose, microcrystalline cellulose, hydroxypropylmethylcellulose, ethylcellulose, methylcellulose, xanthan gum, maltodextrin, chitosan, carrageenan iota, carrageenan kappa, starch, pectins, salts of alginic acid, lignins, tragacanth, guar gum, polyacrylamide, and poly(2-ethyl-2-oxazoline), salts thereof, copolymers thereof and blends thereof.
  • More particular polymers that may be mentioned in embodiments of the invention include one or more of those selected from the group consisting of carrageenan lambda, hyaluronic acid, pullulan, polyvinylpyrrolidone, polyacrylic acid, gum acaia, gelatin, alginate, pectin, carrageenan, collagen, xyloglucan, carboxymethylcellulose, microcrystalline cellulose, hydroxypropylmethylcellulose, ethylcellulose, methylcellulose, xanthan gum, maltodextrin, chitosan, carrageenan iota, carrageenan kappa, starch, pectins, salts of alginic acid, lignins, tragacanth, guar gum, polyacrylamide, and poly(2-ethyl-2-oxazoline), salts thereof and copolymers thereof (e.g.,
  • polyvinylpyrrolidone polyvinyl alcohol
  • polyacrylic acid polyethylene glycols
  • carboxymethylcellulose microcrystalline cellulose
  • hydroxypropylmethylcellulose ethylcellulose
  • methylcellulose ethylcellulose
  • the coating composition comprises a polyvinyl alcohol.
  • the polyvinyl alcohol suitably has a molecular weight (weight average) in the range from 2,000 to 100,000, preferably 25,000 to 60,000, more preferably 35,000 to 45,000, particularly 38,000 to 41,000, and especially 39,000 to 40,000.
  • the coating composition comprises a vinyl acetate copolymer.
  • the vinyl acetate copolymer preferably vinyl acetate- Veova copolymer, suitably has a molecular weight (weight average) in the range from 2,000 to 100,000, preferably 20,000 to 70,000.
  • the coating composition comprises a wax agent.
  • the wax agent may be a natural wax such as a petroleum wax (e.g., paraffin wax), a mineral wax (e.g., a montan wax), a vegetable wax (e.g., a carnauba wax), a synthetic wax (e.g., a polyethylene wax), or a combination thereof.
  • the wax agent comprises a natural wax, a mineral wax, a vegetable wax, or a combination thereof.
  • the wax agent comprises a petroleum wax.
  • the wax agent comprises a vegetable wax.
  • the wax agent comprises a synthetic wax.
  • the wax agent excludes synthetic waxes.
  • the wax agent may comprise a carnauba wax, a paraffin wax, or a combination thereof. In one particular embodiment, the wax agent may comprise a carnauba wax. In one particular embodiment, the wax agent may comprise a paraffin wax. In one embodiment, the wax agent may be provided as a microcrystalline wax. In one embodiment, the wax agent may be provided as a nanoscale wax, such as a nanoscale wax emulsion.
  • paraffin waxes are natural, petroleum-based waxes which are solid, firm materials that are typically mixtures of saturated straight-chain hydrocarbons obtained from refining waxy distillates derived from paraffinic crude oils.
  • the paraffin waxes may contain an average of 20 carbon atoms or more, such as about 30 carbon atoms or more, such as about 35 carbon atoms or more, such as about 40 carbon atoms or more to about 80 carbon atoms or less, such as about 70 carbon atoms or less, such as about 60 carbon atoms or less, such as about 50 carbon atoms or less, such as about 40 carbon atoms or less per molecule.
  • carnauba waxes are natural, vegetable-based waxes which typically comprise a mixture of esters of fatty acids and high-molecular weight alcohols and unsaponifiable materials.
  • the coating composition may comprise about 40 wt.% or less, such as about 30 wt.% or less, such as about 25 wt.% or less, such as about 20 wt.% or less, such as about 15 wt.% or less, such as about 13 wt.% or less, such as about 12 wt.% or less, such as about 1 1 wt.% or less, such as about 10 wt.% or less, such as about 7.5 wt.% or less, such as about 6 wt.% or less, such as about 5.5 wt.% or less, such as about 5 wt.% or less, such as about 4 wt.% or less, such as about 3 wt.% or less of the composition.
  • the coating composition may comprise about 0.5 wt.% or more, such as about 1 wt.% or more, such as about 1 .5 wt.% or more, such as about 2 wt.% or more, such as about 2.5 wt.% or more, such as about 3 wt.% or more, such as about 4 wt.% or more, such as about 4.5 wt.% or more, such as about 5 wt.% or more, such as about 7.5 wt.% or more, such as about 9 wt.% or more, such as about 10 wt.% or more, such as about 12.5 wt.% or more, such as about 15 wt.% or more, such as about 17.5 wt.% or more, such as about 20 wt.% or more, such as about 25 wt.% or more of the composition.
  • the coating composition is applied as a liquid composition and/or emulsion and/or dispersion and/or latex composition and thereafter solidified (including cured and/or dried) to form a coating on the solid carrier.
  • liquid coating composition as used in this application is meant to include coating compositions in the form of a suspension, emulsion, and/or dispersion, preferably a dispersion.
  • the coating composition is applied to the solid carrier and/or the carbon sequestering fungal strain by a rotary coater, a rotary dry coater, a pan coater, or a continuous treater.
  • the amount of coating composition applied to the solid carrier and/or the carbon sequestering fungal strain can be in the range of 10 to 1,000 g dry wt. per kg, such as 30 to 650 g dry wt. per kg, 100 to 400 g dry wt. per kg, or 150 to 250 g dry wt. per kg.
  • the coating composition can, for instance, be applied by encrusting, fdm coating, spraying, dipping, or brushing of the coating composition.
  • it is applied at a temperature of 2 to 50° C., for instance 5 to 35° C., more often 15 to 30° C., for instance at room temperature, such as 18 to 25° C.
  • An additional film coat layer may optionally be applied over the top of the coating composition, preferably by encrustment, to provide additional benefits, including but not limited to cosmetics, coverage, actives, nutrients, and processing improvements such as faster drying, flow, durability and the like.
  • the soil carbon inoculum granular formulation comprises a filler.
  • the filler may be any suitable organic or inorganic material.
  • the filler component excludes any fibrous material.
  • a suitable organic filler material is com starch powder.
  • Suitable inorganic filler materials include at least one selected from the group consisting of corn starch powder, talc, mica, oyster shell calcium, diatomaceous earth, calcium carbonate, sodium bicarbonate, silica, silicates, barium sulfate, titanium dioxide, silicon dioxide, calcium sulfate, kaolin, bentonite, montmorillonite, red clay, biochar fines, and combinations thereof.
  • the filler comprises, consists essentially of, or consists of talc and/or corn starch powder, more preferably talc.
  • the filler is in particulate form and may, for example, be irregularly shaped, spherical, approximately spherical, disc, platelet or rod shaped.
  • the filler is preferably platy in particle shape.
  • the filler component is non-fibrous.
  • the filler preferably talc, suitably has a median particle size as determined by x-ray sedimentation using a Sedigraph III Plus Particle Size Analyzer, in the range from 0.1 to 50 pm, preferably 3 to 25 pm, more preferably 8 to 18 pm, particularly 11 to 14 pm, and especially 12 to 13 pm.
  • the amount of filler, preferably talc, in the soil carbon inoculum granular formulation is suitably in the range from 10% to 90%, 20% to 80%, 35% to 70%, or 40% to 60% by weight based on the total weight of the composition.
  • the presently claimed soil carbon inoculum granular formulation may further include a binder.
  • the binder may assist in binding particles to a desired surface, such as the surface of a solid carrier. Without intending to be limited by theory, such binding may be the result of a physical or chemical binding.
  • any binder known in the art may be employed.
  • the binder may include a styrene/butadiene copolymer, a butadiene/acrylonitrile copolymer, a styrene/butadiene/acrylonitrile copolymer, a styrene/methacrylic ester copolymer, a vinyl acetate/acrylic copolymer, a vinyl acetate/methacrylic ester copolymer, a vinyl acetate/ethylene copolymer, a vinyl acetate homopolymer, a methacrylic ester/acrylic ester copolymer, etc.
  • the binder comprises a styrene, butadiene, or a combination thereof. In one particular embodiment, the binder comprises a styrene/butadiene copolymer. In particular, the binder comprises a styrene/butadiene copolymer latex. In one particular embodiment, the binder comprises a carboxylated styrene/butadiene copolymer. In particular, the binder comprises a carboxylated styrene/butadiene copolymer latex.
  • binders may include, but are not limited to, copolymers of methyl vinyl ether with maleic anhydride or monoalkyl esters of maleic anhydride; polyvinylpyrrolidone; copolymers of vinyl pyrrolidone with vinyl acetate; copolymers of vinyl pyrrolidone with vinyl alkyls; polyvinyl acetate; ethylene/vinyl acetate copolymers; vinyl acetate acrylic copolymers; A-B block copolymers of ethylene oxide and propylene oxide; A-B-A triblock copolymers of EO-PO-EO; and polyvinyl alcohol.
  • the binder may be a latex binder.
  • the binder may be in the form of a dispersion or aqueous carrier of polymer particles.
  • the binder may comprise about 1 5 wt.% or less, such as about 1 0 wt. or less, such as about 7.5 wt.% or less, such as about 6 wt.% or less, such as about 5 wt.% or less, such as about 4 wt.% or less, such as about 3.5 wt.% or less, such as about 3 wt.% or less, such as about 2.5 wt.% or less, such as about 2 wt.% or less, such as about 1 .5 wt.% or less, such as about 1 .4 wt.% or less, such as about 1 .3 wt.% or less, such as about 1 wt.% or less, such as about 0.9 wt.% or less, such as about 0.75 wt.% or less of the composition.
  • the binder may comprise about 0.1 wt.% or more, such as about 0.2 wt.% or more, , such as about 0.4 wt.% or more, such as about 0.5 wt.% or more, such as about 0.6 wt.% or more, such as about 0.75 wt.% or more, such as about 1 wt.% or more, such as about 1 .1 wt.% or more, such as about 1 .2 wt.% or more, such as about 1 .25 wt.% or more, such as about 1 .5 wt.% or more, such as about 1 .75 wt.% or more, such as about 1 .9 wt.% or more, such as about 2 wt.% or more, such as about 2.5 wt.% or more, such as about 2.75 wt.% or more, such as about 3 wt.% or more, such as about 4 wt.% or more, such as about 5
  • Scalability - Seed coating technology is an industry standard, and the equipment used for seed coating could also be used to produce the disclosed formulations. This equipment is available at an agricultural scale for manufacturing of the soil carbon inoculum granular formulations, and the economics are favourable.
  • the soil carbon inoculum granular formulation helps standardize the product and reduces the inherent variability of a mycelial product.
  • Detoxification powders such as biochar, zeolite, diatomaceous earth, and bentonite help to reduce fungicide toxicity from the surrounding soil.
  • Bentonite and similar fillers imbibe water and have a swelling function that helps the granular formulation “bloom” out into the soil.
  • Additional compounds such as fungal germination stimulants, signalling compounds, (e.g., flavonoids, strigolactones), chitosan, root stimulants, a microbial fermentation supernatant, and nutrients can be integrated into the formulations.
  • Co-inoculants such as AMF, phosphate solubilizing microbes, nitrogen-fixing microbes, and PGPR can be coated in additional layers.
  • a coated pellet applied through an air seeder would occupy the interseed zone where the plant root can find it safely.
  • a coated pellet near a seed coated with fungicides but not in direct contact with the seed preserves the beneficial fungal inoculum from any detrimental impact from the fungicides.
  • a low-rate soil carbon inoculum granular formulation is easy to transport, measure and apply.
  • FIG. 1A Several prototypes were developed for a soil carbon inoculum formulation.
  • the basic prototype presented in FIG. 1A comprises a nucleus or core of a solid carrier that optionally serves as substrate for the growth of a fungal inoculum, which may be in the form of spores, conidia, and/or hyphae.
  • a polymer in a coating composition provides resistance to abiotic stress (e.g., heat, desiccation, toxins, etc.) and/or favorable ballistic properties.
  • FIG. IB presents a prototype with additional features including specific solid carriers (i.e., BIODAC® (cellulose) or zeolite), a blooming agent such as a bentonite filler, and plant root and fungal stimulants.
  • FIGs. 2 and 3 present additional prototypes where the coating composition confers slow-release properties, the fungal inoculum is applied as a spore powder or a hyphal powder, and plant growth promotion chemical compounds are included in the formulation.
  • BIODAC is a source of cellulose and is a registered trademark of Kadant GranTek Inc., Green Bay, Wisconsin, USA.
  • the product sold under the trademark BIODAC is composed of paper fiber and is formulated as about 0.5-2 mm diameter granules that are substantially free of dust.
  • the BIODAC® (cellulose) granules are sorted by size after being sieved through meshes of specific sizes (e.g., BIODAC® 4/10 U.S. MESH, BIODAC® 8/16 U.S. MESH, BIODAC® 10/30 U.S. MESH, BIODAC® 12/20 U.S. MESH, BIODAC® 20/50 U.S. MESH).
  • BIODAC® cellulose
  • cellulose granules assessed for the development of various fungal species applied as a liquid culture can colonize the granules.
  • Acrocalymma, Dictyochaeta, Humicola, Leptodontidium, Periconia, and Mortierella fungal cells successfully colonized BIODAC® (cellulose) granules. This technique allows for the development of a highly concentrated granule, which can be deployed in the furrow, mixed with seed, or applied via an air seeder (see FIG. 4).
  • a shelf-life assessment of colonized BIODAC® (cellulose) granules demonstrated strong results for product viability in cold storage (see Table 1). Viability of the colonized BIODAC® (cellulose) granules was assessed by randomly selecting several granules after storage at 4°C, plating the selected granules on agar plates, and observing fungal outgrowth from granules (see FIG. 5). If outgrowth was observed from every granule, then this indicated 100% viability. Fungal viability of colonized BIODAC® (cellulose) granules was assessed after 1 week, 2 weeks, 4 weeks, and monthly thereafter at 4°C. Viability of the fungal species shown in Table 1 remained at 100% during each of the time points including those presented.
  • BIODAC® cellulose
  • BIODAC® cellulose
  • Table 2 A stability study with BIODAC® (cellulose) granules colonized with each of the strains listed in Table 2 was conducted as outlined in Example 2. Each of the strains remained 100% viable on the BIODAC® (cellulose) granules over the course of the year-long study with the exception of Acrocalymma vagum DMTR-CTR-11556 (NMI Accession No. V22/006357) (see Table 2).
  • Zeolite granules from Castle Mountain Zeolites (Quirindi, Australia) are assessed for their potential as a fungus colonized formulation or as coated granules.
  • Zeolites are natural rock materials and have been used in agricultural formulations.
  • the zeolite granules are evaluated with fungi applied as a liquid culture or as a dry powder. Shelf-life of the fungal cells on the zeolite is assessed after cold storage at 4°C.
  • Typical diameters of the soil carbon inoculum formulations incorporating zeolite range from about 0.5 mm to about 3 mm (see FIG. 6).
  • the size of the granules can be adjusted by the amounts of fillers and other components that are used in the formulations.
  • BIODAC® (cellulose) formulations stored at different temperatures an additional stability study was conducted. Once a week, twenty BIODAC® (cellulose) granules of each particle size colonized with different fungal strains and stored at different temperatures were plated on potato dextrose agar (PDA) plates and incubated at 25°C in a darkened incubator for 1 week. At the end of incubation, the granules were assessed for viability (number of viable vs. non-viable granules) and vitality (growth rate/hyphal diameter).
  • PDA potato dextrose agar
  • BIODAC® cellulose granules inoculated with various fungal strains on soil carbon.
  • BIODAC® (cellulose) granules were inoculated using the procedure outlined in Example 2 with the following fungal strains: Phialocephala fortinii s.l - Acephala applanata species complex (PAC) DMTR-CTR-7788 (NMI Accession No. V21/002327), Acrocalymma vagum DMTR- CTR-11556 (NMI Accession No. V22/006357), Clonostachys rosea DMTR-CTR-1081 (NMI Accession No. V22/003495), Periconia sp.
  • PAC Phialocephala fortinii s.l - Acephala applanata species complex
  • DMTR-CTR-7788 NMI Accession No. V21/002327
  • Acrocalymma vagum DMTR- CTR-11556
  • DMTR-CTR-6649 NMI Accession No. V22/006356
  • Thozetella nivea DMTR-CTR-2359 NMI Accession No. V22/003496
  • Beauveria bassiana AU-16727 NI Accession No. V23/003855
  • Periconia macrospinosa DMTR-CTR-1852 NMI Accession No. V22/006358.
  • Each pot in the greenhouse experiment contained 2 kg of soil into which 10 g of inoculated BIODAC® (cellulose) granules were added to the top layer of soil near the wheat seed (see FIG. 10).
  • Control pots included soil without inoculated BIODAC® (cellulose) granules and were separated into one group in which wheat seed was sown and another group without wheat seed. All treatment groups contained wheat seed. The pots were watered and exposed to native sunlight until the wheat plants had reached maturity. At wheat maturity, total carbon in soil samples from each pot was measured with a LECO® instrument using combustion of carbon. The total carbon measured in the two control groups was not significantly different.
  • TC total carbon
  • Yield was also determined.
  • a total of five replicates were evaluated for each treatment group and the untreated control group.
  • the mean values and related standard errors were calculated for TC and yield values, and these values along with related percentages of untreated control values are presented in Tables 4 and 5.
  • Table 4. Total carbon (TC) measured in soil collected near sorghum plants treated with fungal inoculum applied on BIODAC® (cellulose) granules. The mean TC values, standard error (SE), and percentage of the untreated control are presented.
  • TC in soil samples collected was measured with a LECO® instrument using combustion of carbon. Yield was determined at harvest.
  • the relative increase in soil carbon was calculated as the ratio of TC at harvest to the TC prior to planting (see FIG. 12A).
  • the relative increase in soil carbon compared to control was calculated as the difference between the relative increase in soil carbon of a treatment group and the relative increase of the untreated control (see FIG. 12B). All TC and yield values represent the mean of five replicates.
  • FIGs. 12A and 12B a similar increase in TC in the soil was observed with winter wheat treated with the fungal strains applied on BIODAC® (cellulose) granules or applied as a seed treatment.
  • FIG. 13 indicates that yield was maintained across treatment groups and untreated control without any significant difference.
  • BIODAC® cellulose
  • cellulose granules colonized with each of the fungal strains were stored at 4 degrees C and or 25 degrees C. Samples from each group were then plated on PDA plates and incubated in a darkened incubator for 1 week to allow outgrowth of fungal cells from the granules.
  • the granules were assessed for viability (i.e., number of viable vs. non-viable granules), and the viability rate was recorded as a percentage of viable granules over the course of 22 to 30 weeks.
  • BIODAC® cellulose granules colonized with Thozetella nivea DMTR-CTR-2359 (NMI Accession No. V22/003496) or Leptodontidium orchidicola DMTR-CTR-4873 (NMI Accession No. V22/003497) remained viable at both temperatures through week 22 (see FIGs 14A and 14B). Contamination prevented further measurement after this time point.
  • the granules colonized with Acrocalymma vagum AU-11556 (NMI Accession No. V22/006357) remained viable at both temperatures through week 30 (see FIG. 14C).
  • Millet grains were colonized with Thozetella nivea DMTR-CTR-2359 (NMI Accession No. V22/003496), which were then ground to a fine granular formulation. Upon inspection of this fine granular formulation under the microscope, it was observed that a heterogenous mixture of different-sized particles were present.
  • the fine granular formulation was cleared in potassium hydroxide to reduce substrate pigments and then stained with trypan blue to reveal fungal components. Particles composed of high proportions of fungal hyphae and embedded fungal propagules were clearly visible. The size and cohesion of propagule-dense particles appeared to be determined by the degree of hyphal ramification of the substrate. In other words, particles with a favorable propagule: substrate ratio disproportionately fell within a well- defined size class. Larger fractions lost cohesion and dissipated. Smaller fractions included disproportionately high amounts of particles with a low propagule: substrate ratio. These smaller fractions contained greater amounts of organo-mineral composites (see FIG. 15 capturing observations from microscopy).
  • the size and structural integrity of particles appeared to be determined by the hyphal density of particles which behave as the binding agent.
  • particles in the 106 pm and 125 pm fractions had characteristics of interest, specifically a favorably high active propagule: substrate ratio and reliable particle cohesion, enabling more effective product quality control and standardization.
  • on-seed real estate for biologically active fungal granules is at a premium, and smaller, inert particles may more readily adhere and occupy space on seed, limiting adhesion of particles with high biological activity.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Pest Control & Pesticides (AREA)
  • General Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Environmental Sciences (AREA)
  • Biotechnology (AREA)
  • Mycology (AREA)
  • Soil Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Inorganic Chemistry (AREA)
  • Dentistry (AREA)
  • Genetics & Genomics (AREA)
  • Botany (AREA)
  • Virology (AREA)
  • Biochemistry (AREA)
  • Agronomy & Crop Science (AREA)
  • General Engineering & Computer Science (AREA)
  • Materials Engineering (AREA)
  • Molecular Biology (AREA)
  • General Life Sciences & Earth Sciences (AREA)
  • Mechanical Engineering (AREA)
  • Environmental & Geological Engineering (AREA)
  • Biomedical Technology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Cultivation Of Plants (AREA)

Abstract

La présente divulgation concerne une formulation granulaire d'inoculum de carbone du sol comprenant : a) une souche fongique séquestrant le carbone ; b) un support solide choisi dans le groupe constitué par la cellulose, une graine non viable, une semence non viable, une fibre végétale, une fibre de bois, de la zéolite, de la pierre ponce, du sable, de l'attapulgite, de la perlite, de la vermiculite, de la tourbe, de la rafle de maïs, du carbone, du charbon actif, et des combinaisons et des dérivés de ceux-ci ; et c) une composition de revêtement comprenant un agent de type cire, de la chitine, de la gélatine, de l'alginate, de la pectine, du carraghénane, du collagène, du xyloglucane, de l'acétate de polyvinyle, du polyacrylate, du méthylacrylate, de l'hypromellose (HPMC), de la polyvinylpyrrolidone, de l'alcool polyvinylique, de l'acide polyacrylique, des polyéthylène glycols, un copolymère d'acétate de vinyle, un homopolymère d'acétate de vinyle, un copolymère acétate de vinyle-acrylique, du vinyl-acrylique, de l'acrylique, du chlorure d'éthylène-vinyle, de l'anhydride maléique d'éther vinylique, du butadiène styrène, de la carboxyméthylcellulose, de la cellulose microcristalline, de l'hydroxypropylméthylcellulose, de l'éthylcellulose, de la méthylcellulose, du phtalate d'acétate de cellulose (CAP), du chitosane, du chlorhydrate de chitosane, de la gomme de guar, de la gomme xanthane, un copolymère de ceux-ci, ou une combinaison de ceux-ci.
PCT/US2023/073709 2022-09-09 2023-09-11 Formulation granulaire d'inoculum de carbone du sol et ses procédés d'utilisation WO2024054959A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US202263375213P 2022-09-09 2022-09-09
US63/375,213 2022-09-09
US202363498251P 2023-04-25 2023-04-25
US63/498,251 2023-04-25

Publications (1)

Publication Number Publication Date
WO2024054959A1 true WO2024054959A1 (fr) 2024-03-14

Family

ID=90191950

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2023/073709 WO2024054959A1 (fr) 2022-09-09 2023-09-11 Formulation granulaire d'inoculum de carbone du sol et ses procédés d'utilisation

Country Status (1)

Country Link
WO (1) WO2024054959A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN118460380A (zh) * 2024-05-09 2024-08-09 云南省林业和草原科学院 一种植物内生真菌菌株的培养方法和应用

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20120304719A1 (en) * 2011-06-06 2012-12-06 Michael Cheiky Method for enhancing soil growth using bio-char
CA2914547A1 (fr) * 2014-12-12 2016-06-12 Avalon Alliance Inc. Fertilisant mineral biodisponible et applications derivees, notamment les processus de produits
US20180177196A1 (en) * 2016-12-23 2018-06-28 The Texas A&M University System Fungal endophytes for improved crop yields and protection from pests
US20220142173A1 (en) * 2014-06-26 2022-05-12 Ait Austrian Institute Of Technology Gmbh Plant-endophyte combinations and uses therefor
WO2022174313A1 (fr) * 2021-02-22 2022-08-25 Loam Bio Pty Ltd Procédés de capture de carbone et d'augmentation du rendement de plantes cultivées
WO2022256275A1 (fr) * 2021-05-31 2022-12-08 Loam Bio Pty Ltd Procédés de capture de carbone et d'augmentation du rendement de plantes

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20120304719A1 (en) * 2011-06-06 2012-12-06 Michael Cheiky Method for enhancing soil growth using bio-char
US20220142173A1 (en) * 2014-06-26 2022-05-12 Ait Austrian Institute Of Technology Gmbh Plant-endophyte combinations and uses therefor
CA2914547A1 (fr) * 2014-12-12 2016-06-12 Avalon Alliance Inc. Fertilisant mineral biodisponible et applications derivees, notamment les processus de produits
US20180177196A1 (en) * 2016-12-23 2018-06-28 The Texas A&M University System Fungal endophytes for improved crop yields and protection from pests
WO2022174313A1 (fr) * 2021-02-22 2022-08-25 Loam Bio Pty Ltd Procédés de capture de carbone et d'augmentation du rendement de plantes cultivées
WO2022256275A1 (fr) * 2021-05-31 2022-12-08 Loam Bio Pty Ltd Procédés de capture de carbone et d'augmentation du rendement de plantes

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN118460380A (zh) * 2024-05-09 2024-08-09 云南省林业和草原科学院 一种植物内生真菌菌株的培养方法和应用

Similar Documents

Publication Publication Date Title
Pirzada et al. Recent advances in biodegradable matrices for active ingredient release in crop protection: Towards attaining sustainability in agriculture
Szopa et al. Encapsulation efficiency and survival of plant growth-promoting microorganisms in an alginate-based matrix–A systematic review and protocol for a practical approach
WO2024054959A1 (fr) Formulation granulaire d'inoculum de carbone du sol et ses procédés d'utilisation
Cortés-Rojas et al. Seed coating as a delivery system for the endophyte Trichoderma koningiopsis Th003 in rice (Oryza sativa)
CN111109093B (zh) 一种复合型猫砂及其制备方法
CA2839535C (fr) Graines enduites
Kangsopa et al. Lettuce seeds pelleting: A new bilayer matrix for lettuce (Lactuca sativa) seeds
CN103224421B (zh) 苹果专用稻壳炭基肥及其制备方法
CN111778032A (zh) 一种土壤改良剂及制备方法与应用
US20230323285A1 (en) Complex of mutualistic microbes designed to increase plant productivity
Dingley et al. Application of superabsorbent natural polymers in agriculture
Jarrar et al. Seed enhancement technologies for sustainable dryland restoration: Coating and scarification
CA3091167A1 (fr) Plateforme de support a base d'argile pour les produits biologiques en agriculture
Pitaktamrong et al. Development of arbuscular mycorrhizal fungi-organic fertilizer pellets encapsulated with alginate film
EP2958415B1 (fr) Enrobage de semence contenant des particules minérales et des micro-organismes secs
US20240225010A1 (en) Encapsulated microbial compositions and methods of making the same
KR100840747B1 (ko) 토양전염성 병해 방제용 서방형 미생물제제 및 그 제조방법
US10961454B2 (en) Method for reducing water stress in plants
Yogeesha et al. Standardization of protocol for seed pelleting in onion (Allium cepa) to improve seed handling
Batista et al. Quality of grain sorghum seeds Coated with different combinations of materials
Feng et al. Development of biochar-impregnated alginate beads for the delivery of biocontrol agents for peanut aflatoxin
WO2023242462A1 (fr) Système et procédé pour l'enrobage de semences
WO2024108059A2 (fr) Compositions microbiennes et procédés
JPH0423989A (ja) 微生物担持用炭材
JPH10225207A (ja) 造粒コーティング種子およびその製造方法

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 23864027

Country of ref document: EP

Kind code of ref document: A1