Novel seed treatment methods and compositions for improving plant traits and yield

Abstract

Described herein are methods and compositions for improving plant growth and other characteristics. The methods and compositions utilize bacteria and bacterial exudates incorporated into plant seeds to improve plant growth and other characteristics.

Description

[Technical Field]

[0001] cross reference This application claims the benefit of U.S. Provisional Patent Application No. 62 / 835,281, filed April 17, 2019, which is incorporated herein by reference in its entirety.

[0002] Sequence Listing This application contains a Sequence Listing that has been submitted electronically in ASCII format and is incorporated herein by reference in its entirety. The ASCII copy created on April 17, 2019, is named "54449701101_SL.txt" and is 8,555,480 bytes in size.

[0003] The present invention relates to a seed treatment method that involves introducing plant-beneficial microorganisms or synthetic combinations of two or more microorganisms and / or their exudates and / or their personalized biomolecules into seeds. The method involves controlled and rapid hydration of seeds in an aqueous solution of a chemically inert but systemically active compound supplemented with a specific amount of beneficial microorganisms or synthetic combinations of two or more microorganisms and / or their exudates and / or their personalized biomolecules. Seed hydration at the early post-dormancy stage of the plant embryo and incorporation of plant-beneficial microorganisms can promote rapid and uniform germination, improve seed vigor, promote plant growth, and improve plant traits, even several months after seed treatment. [Background technology]

[0004] By 2050, the world's population is expected to reach 9.8 billion (https: / / www.un.org / development / desa / en / news / population / world-population-prospects-2017.html), while more than 500 million hectares of wildland are expected to be converted to farmland (IRP, 2017). Under the current circumstances, agricultural production must face severe challenges from climate change, which brings extreme weather events, and emerging pathogens, with farmers worldwide having to deal primarily with declining yields and lower operating margins due to the latter (GAP 2017; Sessitsch et al.). Considering both the expected global population growth and environmental degradation, it is clear that significantly increasing agriculture and food production in a sustainable and environmentally friendly manner over the next decade will be a major challenge. Summary of the Invention

[0005] One aspect of the invention described herein is a method of incorporating bacteria into plant seeds, the method comprising: contacting the plant seeds with a solution containing the bacteria, wherein the solution comprises about 0.1% to about 2% salt (w / v); and incubating the plant seeds with the solution, thereby incorporating at least 1 colony forming unit (CFU) of the bacteria into the plant seeds. In some embodiments, (b) comprises incubating the plant seeds with the solution, thereby incorporating at least 500 CFU of the bacteria into the plant seeds. In some embodiments, the bacteria comprises endospore-forming bacteria or endospores thereof. In some embodiments, the solution comprises a microbial exudate. In some embodiments, the microbial exudate is derived from the bacteria. In some embodiments, the microbial exudate is not derived from the bacteria. In some embodiments, the bacteria comprises bacteria from the phyla Firmicutes, Proteobacteria, Actinobacteria, or a combination thereof. In some embodiments, the bacteria are Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribacillus sp., Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus sp.In some embodiments, the solution comprises bacteria from Thermoflavimicrobium sp., Thermovenabulum sp., Tuberibacillus sp., Virgibacillus sp., Vulcanobacillus sp., or a combination thereof. In some embodiments, the bacteria comprises bacteria from Bacillus sp. In some embodiments, the bacteria is incorporated between the seed coat and the embryo of the plant seed. In some embodiments, the method further comprises, prior to (a), disinfecting the plant seed. In some embodiments, the solution is at a concentration of about 0.In some embodiments, the solution further comprises Luria-Bertani (LB) broth. In some embodiments, the solution further comprises 85% of the salt. In some embodiments, the salt comprises NaCl. In some embodiments, the plant seeds comprise corn seeds, wheat seeds, rice seeds, sorghum seeds, barley seeds, rye seeds, sugarcane seeds, millet seeds, oat seeds, soybean seeds, cotton seeds, alfalfa seeds, bean seeds, quinoa seeds, lentil seeds, peanut seeds, lettuce seeds, tomato seeds, pea seeds, or cabbage seeds. In some embodiments, the solution further comprises Luria-Bertani (LB) broth. In some embodiments, the solution further comprises dimethyl sulfoxide (DMSO), 1-dodecylazacycloheptan-2-one, laurocapram, 1-methyl-2-pyrrolidone (NMP), oleic acid, ethanol, methanol, polyethylene glycol (Brij 35, 58, 98), polyethylene glycol monolaurate (Tween 20), Tween 40 (polyoxyethylene acid sorbitol ester), Tween 60, Tween 80 (non-ionic), cetylmethylammonium bromide (CTAB), urea, lecithin (soybean-derived solidifying fatty acid), chitosan, poloxamer 188, poloxamer 237, poloxamer 338, poloxamer 407, or a combination thereof. In some embodiments, the solution further comprises calcium, magnesium, manganese, potassium, iron, or a combination thereof. In some embodiments, the solution is maintained at a temperature between about 4°C and about 40°C, about 20°C and about 40°C, or about 10°C and about 20°C. In some embodiments, the solution is maintained at about 23°C or about 30°C. In some embodiments, the plant seeds are incubated with the solution for about 1 minute to about 960 minutes, about 20 minutes to about 240 minutes, or about 1 minute to about 20 minutes. In some embodiments, the plant seeds are incubated with the solution for about 1 minute, about 5 minutes, about 10 minutes, about 20 minutes, about 240 minutes, or about 960 minutes. In some embodiments, the method further comprises inducing endosporulation of the endospore-forming bacterium.

[0006] Another aspect of the disclosure described herein is a modified plant seed comprising at least 1 CFU of a bacterium incorporated between the seed coat and the embryo of the modified plant seed. In some embodiments, the modified plant seed comprises at least 500 CFU or at least 1000 CFU of the bacterium. In some embodiments, the bacterium comprises an endospore-forming bacterium or an endospore thereof. In some embodiments, the modified plant seed comprises a microbial exudate. In some embodiments, the microbial exudate is derived from the bacterium. In some embodiments, the microbial exudate is not derived from the bacterium. In some embodiments, the bacterium comprises a bacterium from the phyla Firmicutes, Proteobacteria, Actinobacteria, or a combination thereof. In some embodiments, the bacteria are Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribacillus sp., Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus sp., Thermoflavimicrobium sp., Thermovenabulum sp., Tuberibacillus sp., Virgibacillus sp., Vulcanobacillus sp., or a combination thereof. In some embodiments, the bacterium comprises a bacterium from a Bacillus sp. In some embodiments, the modified plant seed is corn seed, wheat seed, rice seed, sorghum seed, barley seed, rye seed, sugarcane seed, millet seed, oat seed, soybean seed, cotton seed, alfalfa seed, bean seed, quinoa seed, lentil seed, peanut seed, lettuce seed, tomato seed, pea seed, or cabbage seed. In some embodiments, the plant seed comprises at least 1000 CFU of the microorganism.

[0007] Another aspect of the disclosure described herein is a method for producing a medicament comprising administering to a subject a subject a medicament comprising administering to ... 3The present invention includes a formulation comprising one or more bacteria at CFU / mL, wherein the formulation comprises about 0.1% to about 2% salt. In some embodiments, the formulation comprises 0.85% of the salt. In some embodiments, the salt comprises NaCl. In some embodiments, the bacteria comprises endospore-forming bacteria or endospores thereof. In some embodiments, the formulation comprises a microbial exudate. In some embodiments, the microbial exudate is derived from the bacteria. In some embodiments, the microbial exudate is not derived from the bacteria. In some embodiments, the bacteria comprises bacteria from the phyla Firmicutes, Proteobacteria, or Actinobacteria. In some embodiments, the bacteria are Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribacillus sp., Thalassobacillus sp.、Thermoacetogenium sp.、Thermoactinomyces sp.、Thermoalkalibacillus sp.、Thermoanaerobacter sp.、Thermoanaeromonas sp.、Thermobacillus sp.、Thermoflavimicrobium sp.、Thermovenabulum sp.、Tuberibacillus sp.、Virgibacillus sp.、Vulcanobacillus sp., or a combination thereof. In some embodiments, the bacteria comprises a bacterium from a Bacillus sp. In some embodiments, the formulation further comprises LB broth. In some embodiments, the formulation further comprises dimethyl sulfoxide (DMSO), 1-dodecylazacycloheptan-2-one, laurocapram, 1-methyl-2-pyrrolidone (NMP), oleic acid, ethanol, methanol, polyethylene glycol (Brij 35, 58, 98), polyethylene glycol monolaurate (Tween 20), Tween 40 (polyoxyethylene acid sorbitol esters), Tween 60, Tween 80 (non-ionic), cetylmethylammonium bromide (CTAB), urea, lecithin (solidifying fatty acid derived from soy), chitosan, poloxamer 188, poloxamer 237, poloxamer 338, poloxamer 407, or a combination thereof. In some embodiments, the formulation further comprises calcium, magnesium, manganese, potassium, iron, or a combination thereof. In some embodiments, the formulation is maintained at a temperature between about 4°C and about 40°C, about 20°C and about 40°C, or about 10°C and about 20°C. In some embodiments, the formulation is maintained at about 23°C or about 30°C. In some embodiments, the formulation contains at least 5 x 10. 5 CFU / mL of the bacteria.

[0008] Another aspect of the disclosure described herein is a method of promoting a plant growth effect in a plant seed, the method comprising: contacting the plant seed with a solution comprising a bacterium, wherein the solution comprises about 0.1% to about 2% salt (w / v); and incubating the plant seed with the solution, thereby incorporating at least 500 colony forming units (CFU) of the bacterium into the plant seed. In some embodiments, the method further comprises sterilizing the plant seed prior to (a). In some embodiments, the bacterium comprises an endospore-forming bacterium or an endospore thereof. In some embodiments, the solution comprises a microbial exudate. In some embodiments, the microbial exudate is derived from the bacterium. In some embodiments, the microbial exudate is not derived from the bacterium. In some embodiments, the bacterium is incorporated between the seed coat and the embryo of the modified plant seed. In some embodiments, the solution comprises about 0.85% of the salt. In some embodiments, the salt comprises NaCl. In some embodiments, the bacteria comprise bacteria from the phyla Firmicutes, Proteobacteria, Actinobacteria, or a combination thereof. In some embodiments, the bacteria are Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgulasp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribacillus sp., Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., ThermoanaerobacterIn some embodiments, the plant seed comprises a bacterium from the genus Thermoanaeromonas, Thermobacillus, Thermoflavimicrobium, Thermovenabulum, Tuberibacillus, Virgibacillus, Vulcanobacillus, or a combination thereof. In some embodiments, the bacterium comprises a bacterium from the genus Bacillus. In some embodiments, the plant seed comprises a corn seed, a wheat seed, a rice seed, a sorghum seed, a barley seed, a rye seed, a sugarcane seed, a millet seed, an oat seed, a soybean seed, a cotton seed, an alfalfa seed, a bean seed, a quinoa seed, a lentil seed, a peanut seed, a lettuce seed, a tomato seed, a pea seed, or a cabbage seed. In some embodiments, the solution further comprises LB broth. In some embodiments, the solution may contain dimethyl sulfoxide (DMSO), 1-dodecylazacycloheptan-2-one, laurocapram, 1-methyl-2-pyrrolidone (NMP), oleic acid, ethanol, methanol, polyethylene glycol (Brij 35, 58, 98), polyethylene glycol monolaurate (Tween 20), Tween 40 (polyoxyethylene acid sorbitol ester), Tween 60, TweenIn some embodiments, the solution further comprises ethanol, ethanolamine, ethanolamine 80 (non-ionic), cetylmethylammonium bromide (CTAB), urea, lecithin (a solidifying fatty acid derived from soy), chitosan, poloxamer 188, poloxamer 237, poloxamer 338, poloxamer 407, or a combination thereof. In some embodiments, the solution further comprises calcium, magnesium, manganese, potassium, iron, or a combination thereof. In some embodiments, the solution is maintained at a temperature between about 4°C and about 40°C, about 20°C and about 40°C, or about 10°C and about 20°C. In some embodiments, the solution is maintained at about 23°C or about 30°C. In some embodiments, the plant seeds are incubated with the solution for about 1 minute to about 960 minutes, about 20 minutes to about 240 minutes, or about 1 minute to about 20 minutes. In some embodiments, the plant seeds are incubated with the solution for about 1 minute, about 5 minutes, about 10 minutes, about 20 minutes, about 240 minutes, or about 960 minutes. In some embodiments, the method further comprises inducing endospore formation of the endospore-forming bacterium. In some embodiments, the plant growth effect comprises increased yield, cellular osmoregulation, ion homeostasis, antioxidant defense, heat stress tolerance, maintenance of photosynthetic capacity, nitrogen fixation, or a combination thereof. In some embodiments, the bacterium is selected for the plant growth effect.

[0009] Another aspect of the disclosure described herein is a method of promoting a plant growth effect in a plant seed, the method comprising: contacting the plant seed with a solution comprising a microbial percolate, wherein the solution comprises about 0.1% to about 2% salt (w / v); and incubating the plant seed with the solution, thereby incorporating the microbial percolate into the plant seed. In some embodiments, the method further comprises disinfecting the plant seed prior to (a). In some embodiments, the microbial percolate is derived from endospore-forming bacteria or endospores thereof. In some embodiments, the microbial percolate is derived from non-endospore-forming bacteria. In some embodiments, the microbial percolate is incorporated between the seed coat and embryo of the modified plant seed. In some embodiments, the solution comprises about 0.85% of the salt. In some embodiments, the salt comprises NaCl. In some embodiments, the microbial exudates are derived from bacteria from the phyla Firmicutes, Proteobacteria, Actinobacteria, or combinations thereof. In some embodiments, the microbial exudate is selected from the group consisting of Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., and Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribacillus sp., Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus sp.、Thermoflavimicrobium sp.、Thermovenabulum sp.、Tuberibacillus sp.、Virgibacillus sp.、Vulcanobacillus sp.In some embodiments, the microbial infusion is derived from bacteria from Bacillus sp., In some embodiments, the microbial infusion comprises bacteria from Bacillus sp. In some embodiments, the plant seed comprises corn seed, wheat seed, rice seed, sorghum seed, barley seed, rye seed, sugarcane seed, millet seed, oat seed, soybean seed, cotton seed, alfalfa seed, bean seed, quinoa seed, lentil seed, peanut seed, lettuce seed, tomato seed, pea seed, or cabbage seed. In some embodiments, the solution further comprises dimethyl sulfoxide (DMSO), 1-dodecylazacycloheptan-2-one, laurocapram, 1-methyl-2-pyrrolidone (NMP), oleic acid, ethanol, methanol, polyethylene glycol (Brij 35, 58, 98), polyethylene glycol monolaurate (Tween 20), Tween 40 (polyoxyethylene acid sorbitol ester), Tween 60, Tween 80 (non-ionic), cetylmethylammonium bromide (CTAB), urea, lecithin (soybean-derived solidifying fatty acid), chitosan, poloxamer 188, poloxamer 237, poloxamer 338, poloxamer 407, or a combination thereof. In some embodiments, the solution is maintained at a temperature between about 4°C and about 40°C, about 20°C and about 40°C, or about 10°C and about 20°C. In some embodiments, the solution is maintained at about 23°C or about 30°C. In some embodiments, the plant seeds are incubated with the solution for about 1 minute to about 960 minutes, about 20 minutes to about 240 minutes, or about 1 minute to about 20 minutes. In some embodiments, the plant seeds are incubated with the solution for about 1 minute, about 5 minutes, about 10 minutes, about 20 minutes, about 240 minutes, or about 960 minutes. In some embodiments, the plant growth effect includes increased yield, cellular osmoregulation, ion homeostasis, antioxidant defense, heat stress tolerance, maintenance of photosynthetic capacity, nitrogen fixation, or a combination thereof. In some embodiments, the microbial exudate is selected with respect to the plant growth effect.

[0010] In one aspect, described herein is a modified seed comprising (i) a seed pericarp and a seed aleurone cell layer having a gap therebetween, and (ii) one or more microorganisms disposed in the gap. In another aspect, described herein is a modified seed comprising (i) a seed pericarp and a seed aleurone cell layer, and (ii) one or more microorganisms disposed between the seed pericarp and the seed aleurone cell layer. In some embodiments, the one or more microorganisms are selected to provide a plant growth-promoting effect. In some embodiments, the seed is a monocotyledonous seed. In some embodiments, the seed is selected from corn, rice, and sorghum seeds. In some embodiments, the seed is a corn seed. In some embodiments, the seed is a corn (Zea maize) seed. In some embodiments, the seed is a dicotyledonous seed. In some embodiments, the seed is selected from soybean, wheat, cotton, alfalfa, lettuce, tomato, and cabbage seeds. In some embodiments, the seed is a lettuce seed. In some embodiments, the seeds are lettuce (Lactuca sativa) seeds. In some embodiments, the seeds are tomato (tomato) seeds. In some embodiments, the seeds are tomato (Solanum lycopersicum) seeds. In some embodiments, the seeds are GMO seeds. In some embodiments, the seeds are non-GMO seeds. In some embodiments, the one or more microorganisms comprise a mixture of Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. In some embodiments, the one or more microorganisms comprise a mixture of Acetobacter cereviseae, Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. In some embodiments, the one or more microorganisms comprise a mixture of Ensifer adhaerens and Bacillus nakamurai. In some embodiments, the one or more microorganisms comprise a mixture of Ensifer adhaerens and Bacillus subtilis. In some embodiments, the one or more microorganisms compriseThe microorganisms include a mixture of Ensifer adhaerens and Bacillus cucumis. In some embodiments, the one or more microorganisms include Microbacterium yannicii. In some embodiments, the one or more microorganisms include Microbacterium chocolatum. In some embodiments, the one or more microorganisms include Serratioa ureilytica. In some embodiments, the one or more microorganisms include Serratioa marcescens. In some embodiments, the one or more microorganisms include Glutamicibacter arilaitensis. In some embodiments, the one or more microorganisms include Glutamicibacter halophytocola. In some embodiments, the one or more microorganisms include Ensifer adhaerens. In some embodiments, the one or more microorganisms include Pantoea allii. In some embodiments, the one or more microorganisms include Bacillus subtilis. In some embodiments, the one or more microorganisms include Bacillus cucumis. In some embodiments, the one or more microorganisms include endospore-forming bacteria. In some embodiments, the one or more microorganisms include Bacillus sp. In some embodiments, the one or more microorganisms are selected from the phyla Firmicutes, Proteobacteria, and Actinobacteria. In some embodiments, the one or more microorganisms are selected from the phyla Firmicutes. In some embodiments, the one or more microorganisms are selected from the phyla Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp.,Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribaccillus sp.,In some embodiments, the one or more microorganisms are selected from Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus sp., Thermoflavimicrobium sp., Thermovenabulum sp., Tuberibacillus sp., Virgibacillus sp., and Vulcanobacillus sp. In some embodiments, the one or more microorganisms are selected from the phylum Proteobacteria. In some embodiments, the one or more microorganisms include Actinomyces sp. In some embodiments, the one or more microorganisms are selected from the phylum Actinobacteria. In some embodiments, the one or more microorganisms include Coxiella sp. In some embodiments, the one or more microorganisms form endospores after being placed within the seed. In some embodiments, the one or more microorganisms include Baccillus sp. In some embodiments, the one or more microorganisms include endospores. In some embodiments, one or more microorganisms comprise a 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, one or more microorganisms comprise a 16S nucleic acid sequence that is at least 99% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, one or more microorganisms comprise a 16S nucleic acid sequence that is at least 98% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, one or more microorganisms comprise a 16S nucleic acid sequence that is at least 95% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, one or more microorganisms comprise a 16S nucleic acid sequence that is at least 90% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, one or more microorganisms comprise a gene encoding one or more compounds that cause systemic induced resistance (IST). In some embodiments, one or more microorganisms comprise a gene encoding one or more compounds that cause systemic induced resistance (ISR). In some embodiments, one or more microorganisms compriseThe engineered seeds further comprise a microbial exudate. In some embodiments, the one or more microorganisms comprise a gene associated with nitrogen fixation. In some embodiments, the one or more microorganisms comprise a gene associated with phosphate solubilization. In some embodiments, the one or more microorganisms comprise a gene associated with plant hormone synthesis. In some embodiments, the engineered seeds further comprise a microbial exudate. In some embodiments, the microbial exudate comprises one or more compounds that cause systemic induced resistance (IST). In some embodiments, the microbial exudate comprises one or more compounds that cause systemic induced resistance (ISR). In some embodiments, the microbial exudate comprises one or more compounds that cause plant development. In some embodiments, the microbial exudate is from an endospore-forming bacterium. In some embodiments, the microbial exudate is from a non-endospore-forming bacterium. In some embodiments, the microbial exudate is from a microorganism comprising a 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the microbial exudate is from a microorganism comprising a 16S nucleic acid sequence that is at least 99% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the microbial infusion is from a microorganism comprising a 16S nucleic acid sequence at least 98% identical to the 16S nucleic acid sequence of any one of SEQ ID NOs: 1-10221. In some embodiments, the microbial infusion is from a microorganism comprising a 16S nucleic acid sequence at least 95% identical to the 16S nucleic acid sequence of any one of SEQ ID NOs: 1-10221. In some embodiments, the microbial infusion is from a microorganism comprising a 16S nucleic acid sequence at least 90% identical to the 16S nucleic acid sequence of any one of SEQ ID NOs: 1-10221.

[0011] In one aspect, a method for treating one or more plant seeds is described herein, the method comprising soaking one or more seeds in a medium comprising salt and one or more microorganisms selected to have a plant growth-promoting effect, and incubating the one or more seeds in the medium for a period of time sufficient to incorporate the one or more microorganisms into the seeds. In another aspect, a method for treating one or more plant seeds is described herein, the method comprising soaking one or more seeds in a medium comprising salt and one or more microorganisms selected to have a plant growth-promoting effect, and incubating the one or more seeds in the medium to incorporate bacteria between the pericarp and the aleurone cell layer. In one embodiment, the one or more plant seeds remain in a dormant stage after treatment. In one embodiment, the one or more plant seeds remain in a dormant stage after treatment. In one embodiment, the one or more microorganisms are incorporated within the pericarp of the seed. In one embodiment, the one or more microorganisms are incorporated between the pericarp and the aleurone cell layer. In one embodiment, the method further comprises removing the one or more seeds from the medium. In some embodiments, the method further comprises drying the one or more seeds. In some embodiments, the one or more seeds are dried to about 10% total seed moisture. In some embodiments, the method further comprises drying the one or more seeds to prevent germination. In some embodiments, the method further comprises sterilizing the surface of the one or more seeds before immersing the one or more seeds in the medium. In some embodiments, the method further comprises sterilizing the surface of the one or more seeds after immersing the one or more seeds in the medium. In some embodiments, the method further comprises adding a fungicide to the surface of the seeds. In some embodiments, the one or more seeds comprise seeds of a monocotyledonous plant. In some embodiments, the seeds are selected from seeds of maize, rice, and sorghum. In some embodiments, the seeds are maize seeds. In some embodiments, the seeds are maize (Zea mays).In some embodiments, the seed is a seed of a dicotyledonous plant. In some embodiments, the seed is selected from soybean, wheat, cotton, alfalfa, lettuce, tomato, and cabbage seeds. In some embodiments, the seed is a lettuce seed. In some embodiments, the seed is a lettuce (Lactuca sativa) seed. In some embodiments, the seed is a tomato (tomato) seed. In some embodiments, the seed is a tomato (Solanum lycopersicum) seed. In some embodiments, the seed is a GMO seed. In some embodiments, the seed is a non-GMO seed. In some embodiments, the medium is an aqueous medium. In some embodiments, the medium further comprises poloxamer 188. In some embodiments, the medium further comprises poloxamer 188 at a concentration of 0.1%. In some embodiments, the medium further comprises Tween 20. In certain embodiments, the medium may contain dimethyl sulfoxide (DMSO), 1-dodecylazacycloheptan-2-one, laurocapram, 1-methyl-2-pyrrolidone (NMP), oleic acid, ethanol, methanol, polyethylene glycol (Brij 35, 58, 98), polyethylene glycol monolaurate (Tween 20), Tween 40 (polyoxyethylene acid sorbitol ester), Tween 60, Tween80 (nonionic), cetylmethylammonium bromide (CTAB), urea, lecithin (solidifying fatty acid derived from soy), chitosan, poloxamer 188, poloxamer 237, poloxamer 338, and poloxamer 407. In some embodiments, the medium further comprises one or more components that promote endosporulation of one or more bacteria. In some embodiments, the medium comprises potassium, ferrous sulfate, calcium, magnesium, manganese, or a combination thereof. In some embodiments, the medium further comprises manganese. In some embodiments, the medium comprises calcium, magnesium, and manganese. In some embodiments, the medium further comprises nutrients for one or more microorganisms. In some embodiments, the medium is at room temperature. In some embodiments, the medium is at a temperature of about 4°C. In some embodiments, the medium is at a temperature of about 10°C. In some embodiments, the medium is at a temperature of about 15°C. In some embodiments, the medium is at a temperature of about 4°C to about 20°C. In some embodiments, the medium is at a temperature of about 30°C to about 40°C. In some embodiments, the medium is at a temperature of about 20°C. In some embodiments, the medium is at a temperature of about 30°C. In some embodiments, the medium is at a temperature of about 20°C to about 24°C. In some embodiments, the medium is at a temperature of about 40°C. In some embodiments, the salt comprises sodium chloride. In some embodiments, the salt is at a concentration of 0.1-0.2%, 0.2-0.3%, 0.3-0.4%, 0.4-0.5%, 0.5-0.6%, 0.6-0.7%, 0.7-0.8%, 0.8-0.9%, 0.9-1.0%, 1.0-1.1%, 1.1-1.2%, 1.2-1.3%, 1.3-1.4%, or 1.4-1.5%. In some embodiments, the salt is at a concentration of about 0.85%. In some embodiments, the salt is at a concentration of about 1.25% or less. In some embodiments, the salt is at a concentration of about 1.25%. In some embodiments, the one or more microorganisms are selected from Acetobacter cereviseae, Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. In some embodiments, the one or more microorganisms are selected from AcetobacterIn some embodiments, the one or more microorganisms include Chryseobacterium lactis, Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. In some embodiments, the one or more microorganisms include Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. In some embodiments, the one or more microorganisms include Ensifer adhaerens and Bacillus nakamurai. In some embodiments, the one or more microorganisms include Ensifer adhaerens and Bacillus subtilis. In some embodiments, the one or more microorganisms include Ensifer adhaerens and Bacillus cucumis. In some embodiments, the one or more microorganisms include Microbacterium yannicii. In some embodiments, the one or more microorganisms include Microbacterium chocolatum. In some embodiments, the one or more microorganisms include Serratioa ureilytica. In some embodiments, the one or more microorganisms include Serratioa marcescens. In some embodiments, the one or more microorganisms include Glutamicibacter arilaitensis. In some embodiments, the one or more microorganisms include Glutamicibacter halophytocola. In some embodiments, the one or more microorganisms include Ensifer adhaerens. In some embodiments, the one or more microorganisms include Pantoea allii. In some embodiments, the one or more microorganisms include Bacillus subtilis. In some embodiments, the one or more microorganisms include Bacillus cucumis. In some embodiments, the one or more microorganisms include endospore-forming bacteria. In some embodiments, the one or more microorganisms include Bacillus sp. In some embodiments, the one or more microorganisms are selected from the phyla Firmicutes, Proteobacteria, and Actinobacteria. In some embodiments, the one or more microorganisms are selected from the phyla Firmicutes. In some embodiments, the one or more microorganisms are selected from the phyla Acetonema.sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillussp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribacillus sp., Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus sp., Thermoflavimicrobium sp., Thermovenabulum sp., Tuberibacillus sp., Virgibacillus sp., and Vulcanobacillus sp. In some embodiments, the one or more microorganisms are selected from the phyla Proteobacteria. In some embodiments, the one or more microorganisms include Actinomyces sp. In some embodiments, The one or more microorganisms are selected from the phyla Actinobacteria. In some embodiments, the one or more microorganisms comprise a Coxiella sp. In some embodiments, the one or more microorganisms form endospores after incorporation into a seed. In some embodiments, the one or more microorganisms comprise endospores. In some embodiments, the one or more microorganisms comprise Bacillus endospores. In some embodiments, the one or more microorganisms comprise a 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the one or more microorganisms comprise a 16S nucleic acid sequence at least 99% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the one or more microorganisms comprise a 16S nucleic acid sequence at least 98% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the one or more microorganisms comprise a 16S nucleic acid sequence at least 95% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the one or more microorganisms comprise a 16S nucleic acid sequence that is at least 90% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the medium further comprises a microbial exudate. In some embodiments, the microbial exudate comprises one or more compounds that cause systemic induced resistance (IST). In some embodiments, the microbial exudate comprises one or more compounds that cause systemic induced resistance (ISR). In some embodiments, the microbial exudate comprises one or more compounds that cause plant development. In some embodiments, the microbial exudate is from endospore-forming bacteria. In some embodiments, the microbial exudate is from non-endospore-forming bacteria. In some embodiments, the microbial exudate is from a microorganism that comprises a 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the microbial exudate is from a microorganism that comprises a 16S nucleic acid sequence that is at least 99% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the microbial exudate is from a microorganism that comprises a 16S nucleic acid sequence that is at least 98% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221.In some embodiments, the microbial infusion is from a microorganism comprising a 16S nucleic acid sequence that is at least 95% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the microbial infusion is from a microorganism comprising a 16S nucleic acid sequence that is at least 90% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the concentration of the one or more microorganisms in the medium is about 1 x 10. 6 ~1×10 17 In one embodiment, the concentration of one or more microorganisms in the medium is in the range of 1 x 10 CFU / mL. 6 ~1×10 7 CFU / mL, 1 × 10 7 ~1×10 8 CFU / mL, 1 × 10 8 ~1×10 9 CFU / mL, 1 × 10 9 ~1×10 10 CFU / mL, 1 × 10 10 ~1×10 11 CFU / mL, 1 × 10 11 ~1×10 12 CFU / mL, 1 × 10 12 ~1×10 13 CFU / mL, 1 × 10 13 ~1×10 14 CFU / mL, 1 × 10 14 ~1×10 15 CFU / mL, 1 × 10 15 ~1×10 16 CFU / mL, or 1 x 10 16 ~1×10 17 In one embodiment, the amount of one or more microorganisms present in the culture medium is 10 CFU / mL. 10 In one embodiment, the amount of one or more microorganisms present in the medium is less than about 10 CFU / gram of seed. 5 ~10 9In some embodiments, the one or more microorganisms are selected to provide a plant growth-promoting effect. In some embodiments, the plant growth-promoting effect of the one or more microorganisms is selected from one or more of the following groups: cellular osmoregulation, ion homeostasis, antioxidant defense, heat stress tolerance, and / or maintenance of photosynthetic capacity. In some embodiments, the one or more microorganisms are selected for compatibility. In some embodiments, the one or more microorganisms are selected to prevent predation or antagonism. In some embodiments, the one or more microorganisms are also selected for stability during storage. In some embodiments, the one or more microorganisms are also selected for rapid plant colonization and survival within associated tissues. In some embodiments, the one or more microorganisms are also selected to stimulate large-scale, long-term physiological responses in the plant. In some embodiments, the one or more microorganisms are selected for optimal incorporation into one or more seeds. In some embodiments, at least one of the microorganisms persists throughout the plant life cycle. In some embodiments, the incubation time is less than 1 minute. In some embodiments, the incubation time is about 1 minute. In some embodiments, the incubation time is less than 20 minutes. In some embodiments, the incubation time is less than 4 hours. In some embodiments, the incubation time is less than 16 hours. In some embodiments, the incubation time is less than several days. In some embodiments, the incubation time is less than 12 hours. In some embodiments, the incubation time is less than 1 x 10 6 In one embodiment, greater than 1 x 10 bacterial cells are incorporated into each of one or more seeds. 5 ~1×10 8 In some embodiments, the one or more microorganisms are stably incorporated into the one or more seeds. In some embodiments, the one or more incorporated microorganisms are stable for more than 30 days. In some embodiments, the one or more incorporated microorganisms are stable for more than 6 months. In some embodiments, the one or more incorporated microorganisms are stable for at least 1 year. In some embodiments, the one or more incorporated microorganisms are stable for at least 2 years.

[0012] In another aspect, described herein is a plant seed treatment medium comprising a salt and one or more microorganisms. In some embodiments, the one or more microorganisms are selected to provide a plant growth-promoting effect. In some embodiments, the medium is an aqueous medium. In some embodiments, the medium further comprises poloxamer 188. In some embodiments, the medium further comprises poloxamer 188 at a concentration of 0.1%. In some embodiments, the medium further comprises Tween 20. In some embodiments, the medium further comprises one or more agents from the group consisting of dimethyl sulfoxide (DMSO), 1-dodecylazacycloheptan-2-one, laurocapram, 1-methyl-2-pyrrolidone (NMP), oleic acid, ethanol, methanol, polyethylene glycol (Brij 35, 58, 98), polyethylene glycol monolaurate (Tween 20), Tween 40 (polyoxyethylene acid sorbitol ester), Tween 60, Tween 80 (non-ionic), cetylmethylammonium bromide (CTAB), urea, lecithin (solidifying fatty acid derived from soybean), chitosan, poloxamer 188, poloxamer 237, poloxamer 338, and poloxamer 407. In some embodiments, the medium further comprises one or more components that promote endosporulation of one or more bacteria. In some embodiments, the medium further comprises potassium, ferrous sulfate, calcium, magnesium, manganese, or a combination thereof. In some embodiments, the medium further comprises manganese. In some embodiments, the medium further comprises calcium, magnesium, and manganese. In some embodiments, the medium further comprises nutrients for the one or more selected microorganisms. In some embodiments, the salt comprises sodium chloride. In some embodiments, the salt is at a concentration of 0.1-0.2%, 0.2-0.3%, 0.3-0.4%, 0.4-0.5%, 0.5-0.6%, 0.6-0.7%, 0.7-0.8%, 0.8-0.9%, 0.9-1.0%, 1.0-1.1%, 1.1-1.2%, 1.2-1.3%, 1.3-1.4%, or 1.4-1.5%. In some embodiments, the salt is at a concentration of about 0.85%. In some embodiments, the salt is at a concentration of about 1.25% or less. In some embodiments, the salt is at a concentration of about 1.25%. In some embodiments, the one or more microorganisms areThe microorganisms are selected from Acetobacter cereviseae, Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. In some embodiments, the one or more microorganisms include Acetobacter cereviseae, Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. In some embodiments, the one or more microorganisms include Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. In some embodiments, the one or more microorganisms include Ensifer adhaerens and Bacillus nakamurai. In some embodiments, the one or more microorganisms include Ensifer adhaerens and Bacillus subtilis. In some embodiments, the one or more microorganisms include Ensifer adhaerens and Bacillus cucumis. In some embodiments, the one or more microorganisms include Microbacterium yannicii. In some embodiments, the one or more microorganisms include Microbacterium chocolatum. In some embodiments, the one or more microorganisms comprise Serratioa ureilytica. In some embodiments, the one or more microorganisms comprise Serratioa marcescens. In some embodiments, the one or more microorganisms comprise Glutamicibacter arilaitensis. In some embodiments, the one or more microorganisms comprise Glutamicibacter halophytocola. In some embodiments, the one or more microorganisms comprise Ensifer adhaerens. In some embodiments, the one or more microorganisms comprise Pantoea allii. In some embodiments, the one or more microorganisms comprise Bacillus subtilis. In some embodiments, the one or more microorganisms comprise Bacillus cucumis. In some embodiments, the one or more microorganisms comprise endospore-forming bacteria. In some embodiments, the one or more microorganisms comprise Bacillus sp. In some embodiments,The one or more microorganisms are selected from the phyla Firmicutes, Proteobacteria, and Actinobacteria. In certain embodiments, the one or more microorganisms are selected from the phyla Firmicutes. In certain embodiments, the one or more microorganisms are Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp.,Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribacillus sp., Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus sp., Thermoflavimicrobium sp., Thermovenabulum sp., Tuberibacillus sp., Virgibacillus sp., and Vulcanobacillus sp. In some embodiments, the one or more microorganisms are selected from the phylum Proteobacteria. In some embodiments, the one or more microorganisms include Actinomyces sp. In some embodiments, the one or more microorganisms are selected from the phylum Actinobacteria. In some embodiments, the one or more microorganisms include Coxiella sp. In some embodiments, the one or more microorganisms form endospores after incorporation into the seed. In some embodiments, the one or more microorganisms include endospores. In some embodiments, the one or more microorganisms include Bacillus endospores. In some embodiments,The one or more microorganisms comprise a 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the one or more microorganisms comprise a 16S nucleic acid sequence at least 99% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the one or more microorganisms comprise a 16S nucleic acid sequence at least 98% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the one or more microorganisms comprise a 16S nucleic acid sequence at least 95% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the one or more microorganisms comprise a 16S nucleic acid sequence at least 90% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the medium further comprises a microbial permeate. In some embodiments, the microbial permeate comprises one or more compounds that cause systemic induced resistance (IST). In some embodiments, the microbial permeate comprises one or more compounds that cause systemic induced resistance (ISR). In some embodiments, the microbial permeate comprises one or more compounds that cause plant development. In some embodiments, the microbial exudate is from endospore-forming bacteria. In some embodiments, the microbial exudate is from non-endospore-forming bacteria. In some embodiments, the microbial exudate is from a microorganism comprising a 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the microbial exudate is from a microorganism comprising a 16S nucleic acid sequence at least 99% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the microbial exudate is from a microorganism comprising a 16S nucleic acid sequence at least 98% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the microbial exudate is from a microorganism comprising a 16S nucleic acid sequence at least 95% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the microbial exudate is from a microorganism comprising a 16S nucleic acid sequence at least 90% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the concentration of one or more microorganisms in the medium is about 1 x 10, 6 ~1×10 17In one embodiment, the one or more microorganisms in the culture medium are in the range of about 1 x 10 CFU / mL. 6 ~1×10 17 In one embodiment, the concentration of one or more microorganisms in the medium is in the range of 1 x 10 CFU / mL. 6 ~1×10 7 CFU / mL, 1 × 10 7 ~1×10 8 CFU / mL, 1 × 10 8 ~1×10 9 CFU / mL, 1 × 10 9 ~1×10 10 CFU / mL, 1 × 10 10 ~1×10 11 CFU / mL, 1 × 10 11 ~1×10 12 CFU / mL, 1 × 10 12 ~1×10 13 CFU / mL, 1 × 10 13 ~1×10 14 CFU / mL, 1 × 10 14 ~1×10 15 CFU / mL, 1 × 10 15 ~1×10 16 CFU / mL, or 1 x 10 16 ~1×10 17 CFU / mL.

[0013] In another aspect, described herein is a method for treating one or more plant seeds, the method comprising soaking one or more seeds in a medium comprising salt and one or more microbial permeates selected to provide a plant growth-promoting effect, and incubating the one or more seeds in the medium for a period of time sufficient to allow the one or more microbial permeates to become incorporated into the seeds. In some embodiments, the microbial permeate comprises one or more compounds that cause systemic induced resistance (IST). In some embodiments, the microbial permeate comprises one or more compounds that cause systemic induced resistance (ISR). In some embodiments, the microbial permeate comprises one or more compounds that cause plant development. In some embodiments, the microbial permeate is from an endospore-forming bacterium. In some embodiments, the microbial permeate is from a non-endospore-forming bacterium. In some embodiments, the microbial permeate is from a microorganism comprising a 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the microbial permeate is from a microorganism comprising a 16S nucleic acid sequence at least 99% identical to the 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. In some embodiments, the microbial infusion is from a microorganism comprising a 16S nucleic acid sequence that is at least 98% identical to the 16S nucleic acid sequence of any one of SEQ ID NOs: 1-10221. In some embodiments, the microbial infusion is from a microorganism comprising a 16S nucleic acid sequence that is at least 95% identical to the 16S nucleic acid sequence of any one of SEQ ID NOs: 1-10221. In some embodiments, the microbial infusion is from a microorganism comprising a 16S nucleic acid sequence that is at least 90% identical to the 16S nucleic acid sequence of any one of SEQ ID NOs: 1-10221. In some embodiments, the salt comprises sodium chloride. In some embodiments, the salt is present at a concentration of 0.1-0.2%, 0.2-0.3%, 0.3-0.4%, 0.4-0.5%, 0.5-0.6%, 0.6-0.7%, 0.7-0.8%, 0.8-0.9%, 0.9-1.0%, 1.0-1.1%, 1.1-1.2%, 1.2-1.3%, 1.3-1.4%, or 1.4-1.5%. In some embodiments, the salt is present at a concentration of about 0.85%. In some embodiments, the salt is present at a concentration of about 1.25% or less. In some embodiments, the salt is present at a concentration of about 1.In some embodiments, the microbial infusion is from Microbacterium yannicii. In some embodiments, the microbial infusion is from Microbacterium chocolatum. In some embodiments, the microbial infusion is from Serratioa ureilytica. In some embodiments, the microbial infusion is from Serratioa marcescens. In some embodiments, the microbial infusion is from Glutamicibacter arilaitensis. In some embodiments, the microbial infusion is from Glutamicibacter halophytocola. In some embodiments, the microbial infusion is from Ensifer adhaerens. In some embodiments, the microbial infusion is from Acetobacter cerevisiae. In some embodiments, the microbial infusion is from Pantoea allii. In some embodiments, the microbial infusion is from Bacillus subtilis. In some embodiments, the microbial infusion is from Bacillus cucumis. In some embodiments, the microbial exudates are derived from microorganisms selected from the phyla Firmicutes, Proteobacteria, and Actinobacteria. In some embodiments, the microbial exudates are derived from microorganisms selected from the phyla Firmicutes, Proteobacteria, and Actinobacteria. In some embodiments, the microbial exudates are derived from Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribaccillus sp., Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus sp., Thermoflavimicrobium sp., Thermovenabulum sp., Tuberibacillus sp., Virgibacillus sp., and Vulcanobacillus sp. In some embodiments, the microbial exudate is derived from a microorganism selected from the phylum Proteobacteria. In some embodiments, the microbial exudate is derived from an Actinomyces sp. In some embodiments, the microbial exudate is derived from a microorganism selected from the phylum Actinobacteria. In some embodiments, the microbial exudate is derived from a Coxiella sp. In some embodiments, the microbial exudate is derived from a Bacillus sp. Incorporated by reference.

[0014] All publications, patents, and patent applications mentioned herein are incorporated by reference herein to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. To the extent that a patent publication, patent, or patent application incorporated by reference conflicts with the disclosure contained herein, the present specification is intended to supersede and / or supersede such conflicting material. [Brief explanation of the drawings]

[0015] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0016] The novel features of the methods and compositions described herein are set forth with particularity in the appended claims. The features and advantages of the methods and compositions described herein will be better understood by reference to the following detailed description that sets forth specific embodiments, in which the principles of the methods and compositions described herein are utilized, and the accompanying drawings. [Figure 1] 1 depicts a proposed two-stage plant fortification strategy according to the practice of the present invention. (101) illustrates a monocotyledonous seed. The seed treatment process described herein incorporates bacteria (or endospores thereof) (106) between the aleurone layer (103) and the pericarp (102). The aleurone layer (103) separates the endosperm (104) from the outer layers. The enhanced plant growth effects described herein result from genetic modification of the plant embryo (105) with the bacteria (106) and / or bacterial exudate. [Figure 2] FIG. 1 is a process diagram showing a seed treatment process (Microprime™ seed treatment process). [Figure 3] 1 illustrates a method of the present invention for obtaining a stable microbial seed treatment (Microprime™). [Figure 4A] Figure 1 shows the spaces inside corn seeds (Zea mays) where bacteria are located after Microprime™ seed treatment. [Figure 4B] 1 shows a magnified image of the space inside a corn seed (Zea mays) where bacteria are located after Microprime™ seed treatment. [Figure 5] Figure 1 shows the loading kinetics of a synthetic bacterial consortium (Lascar) into lettuce seeds (Lactuca sativa) using Microprime™ seed treatment. [Figure 6] 1 shows molecular detection in the roots of corn plants (Zea mays) of specific strains preloaded into seeds by Microprime™ seed treatment. [Figure 7] FIG. 1 shows the dual-tube growth chamber used to study and quantify bacteria from Microprime™ seeds. [Figure 8] Figure 1 Quantification of plant root colonization by bacteria loaded onto seeds by Microprime™ seed treatment. [Figure 9] Figure 1 shows the stability of bacterial consortia harbored inside lettuce seeds (Lactuca sativa) over time. [Figure 10] 1 shows the stability of Bacillus endospores housed inside corn seeds (Zea mays) over time. [Figure 11] FIG. 1 shows the stability of corn seeds (Zea mays) over time after Microprime™ seed treatment. [Figure 12] Shown is the effect of Microprime™ seed treatment on lettuce seeds (Lactuca sativa) using a proprietary composition of bacteria internally called Lascar. [Figure 13] Figure 1 shows the temporal effect of Microprime™ seed treatment on lettuce seeds (Lactuca sativa) using a proprietary composition of bacteria, domestically referred to as JIT. [Figure 14] Figure 1 shows the effect of Microprime™ seed treatments on lettuce seeds (Lactuca sativa) grown under salt stress. [Figure 15] Figure 1 shows the effect of Microprime™ seed treatments on the growth and development of tomato (Solanum lycopersicum). [Figure 16A] 1 shows the effect of Microprime™ seed treatment with a single bacteria (S3C1) on the growth and development of corn (Zea mays) as measured by shoot weight. [Figure 16B] Figure 1 shows the effect of Microprime™ seed treatment with a single bacteria (S3C1) on the growth and development of corn (Zea mays) as measured by root weight. [Figure 17A]1 shows the effect of Microprime™ seed treatment with a synthetic consortium containing two bacteria (strains S3C10 and S3C23) on the growth and development of corn (Zea mays) as measured by shoot weight. [Figure 17B] 1 shows the effect of Microprime™ seed treatment with a synthetic consortium containing two bacteria (strains S3C10 and S3C23) on the growth and development of corn (Zea mays) as measured by root weight. [Figure 17C] 1 shows the effect of Microprime™ seed treatment with a synthetic consortium containing two bacteria (strains S3C10 and S3C23) on the growth and development of corn (Zea mays) as measured by shoot length. [Figure 18] The performance of nitrogen-fixing bacteria (strain S3C14) in an acetylene reduction assay is shown. [Figure 19] 1 shows the performance of corn plants (Zea mays) from seeds treated with Microprime™ with endospores of nitrogen-fixing bacteria (strain S3C14) under nitrogen deficiency. [Figure 20] 1 shows the expression patterns of maize (Zea mays) genes in response to seed treatment with Microprime™. [Figure 21] 1 shows the performance of Microprime™ corn plants (Zea mays) under water stress conditions. [Figure 22A] 1 shows the effect of Microprime™ on the vegetative development of corn plants (Zea mays) under field conditions, as measured by shoot weight. [Figure 22B] 1 shows the effect of Microprime™ on the vegetative development of corn plants (Zea mays) under field conditions, as measured by total plant weight. [Figure 23A]Figure 1 shows the effect of Microprime™ seed treatments on the life cycle of corn (Zea mays) under field conditions as a graph showing the percentage of ears at various reproductive stages. [Figure 23B] FIG. 1 shows the effect of Microprime™ seed treatments on the life cycle of corn (Zea mays) under field conditions as shown in the corn images. [Figure 24] Figure 1 shows the effect of Microprime™ seed treatment on yield of corn (Zea mays) under field conditions. [Figure 25] 1 shows the grain protein content of field-grown Microprime™ corn plants (Zea mays). [Figure 26] The performance of nitrogen-fixing bacteria (strain S3C23) in an acetylene reduction assay in the presence of nitrogen is shown. [Figure 27] Figure 1 shows the effect of Microprime™ seed treatment on yield of corn (Zea mays) under nitrogen deficiency in the field. [Figure 28] 1 shows the success rate of a summary of field trial data for corn (Zea mays) using Microprime™ seed treatment technology. DETAILED DESCRIPTION OF THE INVENTION

[0017] While embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Many variations, changes, and substitutions may occur to those skilled in the art without departing from the invention. It will be understood that various alternatives to the embodiments of the invention described herein may be utilized.

[0018] Agriculture is one of the human activities that contributes most to environmental degradation, with potential risks to human health. Agricultural activities cause soil erosion, nutrient depletion, acidification, salinization, compaction, and chemical contamination, resulting in moderate to severe soil degradation. Conventional farming, in particular, relies heavily on the use of synthetic fertilizers and pesticides to improve crop yields. These products are primarily used in agriculture, but are also used in forestry, home gardens, and recreation. These chemical pollutants can pose environmental risks in both their production and use. Excessive use can disrupt the balance of essential nutrients in the soil, adversely affecting soil microflora and fauna, and ultimately render the land unsuitable for agriculture.

[0019] Our society is currently seeking more sustainable production systems, and some countries' regulatory frameworks do not accept the use of genetic modification to improve crop traits. Furthermore, many chemicals are scheduled to be removed from the market in the coming years (Sessitsch et al., 2018; Timmusk et al., 2017). This consumer pressure favors the withdrawal of many synthetic compounds and the lowering of maximum residue limits imposed by regulatory environments. For example, there has been an increase in regulations worldwide to restrict and reduce the use of nitrogen-based fertilizers and pesticides due to their proven negative impacts on human and environmental health (e.g., greenhouse gas emissions, global warming, water pollution, reduced biological nitrogen fixation in soil, and loss of soil biodiversity and genetic resources). Furthermore, the rising costs of developing and registering synthetic pesticides have significantly reduced the development and launch of new chemicals.

[0020] One possible way to address this issue is to use microbial technologies for agriculture to more effectively improve the productivity and yield of important cultivars (Timmusk et al., 2017). Plant-associated microbiota have been extensively investigated in recent decades (Sessitsch et al., 2018), and colonizing microbiota have been shown to play a fundamental role in determining the rate and extent of plant growth, in some cases providing nutrients and conditions necessary for survival and / or directly stimulating plant development and responses to environmental challenges. Microbial colonization of the phytosphere has been shown to begin with germination and continue throughout the plant life cycle, spreading across the plant surface and concentrating in the rhizosphere, where high nutrient and water supplies from root exudates create a favorable environment for microbial growth (Bais et al., 2006). Furthermore, root exudates containing diverse aromatic compounds have been demonstrated to inhibit the growth of certain microorganisms while stimulating the proliferation of others, creating a selective environment for the rhizosphere (Badri et al., 2013, Ledger et al., 2012, Sasse et al., 2018). Furthermore, associated members of the microbiota respond to changes in root exudate composition throughout plant development and exert catabolic functions important for plant growth (Chaparro et al., 2013). Conversely, microbial colonization can alter the flow and pattern of root exudates, suggesting ongoing communication with the host (Bais et al., 2006). A more intimate relationship is established between plants and microorganisms that colonize their internal tissues without causing any signs of harm or infection (Sturz et al., 2000). These endophytic microorganisms have been shown to include a large number of diverse bacteria ( Ryan et al., 2008 ) found on plant roots, stems, leaves, seeds, fruits, tubers, and root nodules ( Rosenblueth and Martinez-Romero, 2006 ).Beneficial or mutualistic bacteria, commonly known as plant growth-promoting bacteria (PGPB), frequently colonize not only the rhizosphere and internal tissues of plants ( Lugtenberg and Kamilova, 2009 ), but also the surface of leaves and stems, commonly known as the phytosphere, where the microbial populations are referred to as epiphytic microorganisms.

[0021] The ability of PGPB to increase plant growth is well established and can proceed through different molecular mechanisms: for example, improving nutrient supply (nitrogen fixation, phosphate solubilization, etc.), regulating plant hormone balance (through the production of auxins, cytokinins, nitric oxide (NO), etc., or the deamination of ACC), enhancing plant defenses triggered by fungi, bacteria, viruses, herbivores, and nematodes (through the induction of systemic resistance pathways and / or the production of antimicrobial secondary metabolites, extracellular lytic enzymes, surfactants, or volatile organic compounds, etc.), and improving tolerance to abiotic stresses (salt, drought, high and low temperatures, heavy metals, etc.) (Dimkpa et al., 2009; Kloepper et al., 2004; Lugtenberg and Kamilova, 2009; Ledger et al., 2016; Timmermann et al., 2017; Vacheron et al., 2013; Van Loon, 2007; Yang et al., 2016). al., 2009). In this context, PGPB has gained popularity as a microbial inoculant, and many new products have been formulated recently (PCT / US2016 / 017204; US2016 / 0338360A1; US2016 / 0330976A1; US2017 / 0223967A1; US2018 / 0020677A1; Sessitsch et al., 2018).

[0022] Improvement of agricultural plant traits through the effects of plant-associated bacteria Over the past two decades, PGPB inoculation has been widely adopted in conventional agricultural practices. However, due to a lack of scientific knowledge on the ecology, physiology, and biochemistry of associated plant-bacterial interactions, this biotechnology is still lacking for industrial application. In this sense, to enhance PGPB inoculation techniques for non-legume crops, a broader understanding of the physiological state of the inoculated microorganisms and / or their high survival rate under adverse conditions in soil and during storage is necessary. Similarly, when bacteria collaborate with agricultural crops, the expression of genes involved in plant growth promotion may be fundamental for achieving beneficial effects. These genes may be switched on or off depending on environmental conditions, affecting their expression in the field. This could explain why some bacteria improve plant growth under controlled laboratory conditions but frequently fail or show variable results in field conditions, as shown in other cases (Baez-Rogelio et al., 2017).

[0023] Despite the importance of plant-PGPB interactions, only a few capabilities of these microorganisms are clearly and directly linked to plant growth promotion and protection, such as solubilizing growth-limiting inorganic nutrients, fixing atmospheric nitrogen, stimulating nutrient supply and uptake by plant roots, and modulating plant regulatory mechanisms through the production of auxin and / or hormones such as ethylene, gibberellins, cytokinins, volatile organic compounds (VOCs), and other metabolites.

[0024] Agricultural plants are typically exposed to multiple stresses simultaneously. As stressors have detrimental effects on the functionality and productivity of agricultural systems, the role of rhizosphere microorganisms is crucial in helping plants thrive in unfavorable conditions (Barea, 2015). As a strategy for survival or reproduction, stressed plants can induce changes in plant morphology, physiology, transporter activity, and root exudation profiles, and recruit microorganisms with stress-alleviating capabilities. However, the mechanisms involved in plant-microbe interactions under stressful conditions are poorly understood (Barea, 2015). This understanding is relevant for designing biotechnological strategies to optimize plant adaptation mechanisms and for improving the capacity of soil microorganisms for crop stress alleviation (Pozo et al., 2010).

[0025] Various types of stress, including salinity, drought, nutrient deficiency, high and low temperatures, diseases, and pests, can alter plant-microbe interactions in the rhizosphere and severely impact agricultural productivity. For example, much of the world's land mass is gradually becoming more arid due to drought, salinity, and high temperatures. Drought, in particular, is one of the abiotic stresses that most threatens global food production and is expected to cause serious plant growth problems for crops on over 50% of the world's cultivated land by 2050 (Ngumbi and Kloepper, 2016). Meanwhile, salinity is another major constraint in agriculture, affecting approximately 20% of the world's irrigated land in over 100 countries. This proportion is increasing due to natural phenomena, conventional farming practices, and global climate change. Salt-affected soils can be classified as saline, saline-sodic, or sodic depending on the amount of salt, type of salt, amount of sodium present, and soil alkalinity. Different types of saline soils have different characteristics, which dictate different management approaches. In 1995, it was estimated that salinization of irrigated lands caused annual revenue losses of approximately US$12 billion worldwide. Furthermore, the decline in water availability, mainly caused by changing climatic conditions, misuse and overuse of available freshwater sources, and the use of saline water sources (both marine-derived or inland high-conductivity sources, and mainly induced by limited freshwater availability), makes abiotic salt stress a highly relevant problem for current agricultural procedures.

[0026] To cope with osmotic stressors (salt and drought), plants must develop many adaptive mechanisms, primarily through fine-tuning their water uptake capacity and transpiration rate, as well as activating antioxidant mechanisms to overcome stress-induced reactive oxygen species (ROS) overproduction. Maintaining water and ROS balance can be improved by inoculation with PGPB, which can act through a variety of specific mechanisms: i) cellular osmoregulation (related to the accumulation of compatible solutes such as proline, glycine, betaine, soluble sugars, pinitol, and mannitol), ii) ion homeostasis (based on maintaining the delicate balance and ratio of potassium, sodium, and calcium), iii) antioxidant defense (compensating for the production of harmful reactive oxygen species (ROS)), and iv) maintaining photosynthetic capacity. Alterations in root system architecture are also important adaptive traits that plants possess to tolerate drought. A positive correlation between PGPB inoculation and drought tolerance has been reported in several crops, such as soybean, chickpea, and wheat (Ngumbi and Kloepper, 2016). Therefore, there is renewed interest in finding solutions to water-related problems. In particular, there is a need to find solutions that enhance plant tolerance to drought and salt stress and contribute to enhanced growth of crops that can meet food demands under limited water resources. Improving plant salt stress tolerance using PGPB has emerged as a promising strategy to help overcome this limitation, but only a few reports have focused on plant-PGPB interactions under salt stress (Ledger et al., 2016; Pinedo et al., 2015). As a result, plants inoculated with microorganisms can better control their water status and achieve high transpiration and photosynthetic rates even under water-deficit conditions. A Bacillus megaterium strain inoculated into maize roots improved root water uptake under saline conditions (Marulanda et al., 2010). Similar behavior was observed when maize roots were inoculated with Pantoea agglomerans (Gond et al., 2015).Furthermore, under salt stress, inoculation with Azospirillum sp. improves the quality and shelf life of lettuce ( Fasciglione et al., 2015 ).

[0027] Drought imposes abiotic stress on plants through high temperatures. Plant responses to high temperatures are complex, involving changes at the physiological, molecular, and biochemical levels, a complex series of signaling pathways, and altered gene expression that limit plant growth, productivity, and grain quality and yield (Vejan et al., 2016). Specifically, heat stress affects protein denaturation and aggregation, membrane lipid fluidity, chloroplast and mitochondrial enzyme inactivation, protein synthesis inhibition, and loss of membrane integrity (Howarth, 2005). These damages ultimately lead to starvation, growth inhibition, reduced ion flux, toxic compounds, and the production of reactive oxygen species (ROS). To overcome high-temperature-induced productivity and yield losses, improving heat tolerance through PGPB inoculation strategies is a cost-effective biotechnology tool that could potentially be adopted by farmers worldwide. For example, inoculation with the plant growth-promoting Pseudomonas sp. strain AKM-P6 and the thermotolerant P. putida strain c improved the tolerance of sorghum and wheat seedlings to high temperature stress by increasing the synthesis of high-molecular-weight proteins and the levels of intracellular metabolites, respectively (Ali et al., 2009; Zulfikar Ali et al., 2011). However, the effectiveness of microbial inoculants under field conditions remains a challenge due to host specificity, short efficacy period, low predictability, and / or poor survival rate of inoculants under environmental conditions, which severely limit their application to alleviate heat and other abiotic stresses in crops (de Freitas and Germida, 1992; Kaur et al.). Some conflicting results have been reported regarding the effect of PGPB inoculation on crop yield enhancement in field trials under various temperature and climatic regions, cropping systems, and agronomic management conditions (Leggett et al., 2015, 2017). Thus, crop yield statistics need to be approached conservatively due to both the logistical constraints and costs associated with sampling, as well as the underlying complexity of the environmental factors that determine achievable yields (e.g., in relation to theoretical / potential yields).

[0028] A synthetic consortium of plant-associated bacteria A promising strategy for improving the field performance of plant-stimulating microbial inoculants is the design of synthetic microbial consortia, which may overcome the limited efficacy of isolated microorganisms. Several studies have reported that inoculating seeds or plants with a combination of multiple beneficial bacteria has greater potential than inoculating a single bacterial species in terms of resulting plant growth promotion and biological control (Kumar, 2016; Sundaramoorthy et al., 2012) (Oliveira et al., 2009). Using such consortia as inoculants may offer advantages, as different plant growth-promoting bacteria have been shown to interact synergistically with the plant host to provide nutrients, remove inhibitory products, or stimulate growth (Barea et al., 2002; Zoppellari et al., 2014). Furthermore, they have been shown to stimulate each other's survival through metabolic complementarity, predator suppression, biofilm protection, and / or quorum sensing.

[0029] A concrete example of the superior performance of synthetic consortia in terms of nutrient acquisition is the co-inoculation of chickpea with Serratia marcescens (SF3), Serratia spp. (ST9), and Mesorhizobium ciceri, which increased the number of nodules per plant, nodule dry weight, number of pods per plant, grain yield, protein content, and total chlorophyll content under irrigated and rainfed conditions compared to inoculation with a single bacterial strain (Shahzad et al., 2014). On the other hand, inoculation of sugarcane with a consortium of five nitrogen-fixing bacteria (Gluconacetobacter diazotrophicus, Herbaspirillum seropedicae, Herbaspirillum rubrisubalbicans, Azospirillum amazonense, and Paraburkholderia tropica) also showed higher stem production in two soils with low to moderate levels of chemical fertilizer compared to mono-inoculated plants (Oliveira et al. 2009).

[0030] Biological control and induced plant defenses are also enhanced when co-inoculated consortia are compared with the application of individual strains, as shown by the protective endophytic strains Bacillus subtilis EPCO16 and Bacillus subtilis EPC5 when combined with the compatible rhizobia strain Pseudomonas fluorescens Pf1 in terms of protection against chili wilt disease caused by Fusarium solani and induction of systemic induced resistance (ISR) responses in the plant host. The combination of the three bacteria was shown to be more effective than each individual strain by inducing metabolic pathways involved in the synthesis of plant defense enzymes and phytoalexins (Sundaramoorthy et al., 2012). Furthermore, a comparison was made between the separate and combined inoculations of fluorescent Pseudomonas aeruginosa (PHU094), Trichoderma harzianum (THU0816), and Mesorhizobium sp. (RL091) for plant growth promotion and defense induction in chickpea plants exposed to the pathogen Sclerotium rolfsii. The results demonstrated that the most effective treatment was the combined application of PHU094, THU0816, and RL091, with or without pathogen exposure (Sigh et al., 2014).

[0031] Furthermore, inoculation with a synthetic consortium has proven more effective than individual strains under abiotic stress conditions. For example, four compatible and drought-tolerant PGPB strains, including Pseudomonas putida (KT2440), Sphingomonas sp. (OF178), Azospirillum brasilense (Sp7), and Acinetobacter sp. (EMM02), were tested as growth promoters for maize plants. Plants inoculated with the bacterial consortium generally outperformed those inoculated with individual bacteria. This advantage was also observed when inoculated seeds were subjected to drought stress before germination, demonstrating the strong protective potential of the synthetic consortium in dryland agricultural applications (Molina-Romero et al., 2017). Additionally, Pseudomonas putida (NBRIRA) and Bacillus amyloliquefaciens (NBRISN13) carrying several PGPB traits were evaluated for their synergistic effect to ameliorate drought stress in chickpea, which showed that plant growth parameters were significantly higher in plants inoculated with the consortium compared to the effect of individual PGPBs ( Kumar, 2016 ).

[0032] In general, isolated microorganisms appear to have limited plant growth-promoting activity due to a) a restricted host range relative to their beneficial effects, as shown for Aeromonas, Pseudomonas, Bacillus, and Enterobacter strains isolated from tomato plants (Vaikuntapu et al., 2014) and for Azospirillum strains obtained from different rice varieties (Chamam et al., 2013); b) low resilience to changing environmental conditions (reviewed in (Mahmood et al., 2016)); c) high susceptibility to antagonism or predation by the native microbiota (Savka et al., 2002); and d) poor competitive ability with respect to the native, well-adapted host microbiota, as demonstrated in comparison to a native Pseudomonas strain that improved the growth of Mentha piperita (Santoro et al., 2015). Furthermore, the rhizosphere environment tends to favor the association of different microorganisms harboring a few or single functions that complement each other and promote plant growth, rather than a single bacterium expressing many complementary functions ( Vacheron et al., 2016 ).

[0033] The next generation of microbial inoculants Despite the fact that inoculating plants with beneficial bacteria is a century-old technique, agricultural microbe-based technologies are currently being put forward as the most revolutionary and environmentally friendly biotechnology to increase agricultural production, based on a balance between economic costs and economic benefits for the conservation of agroecosystems ( Berninger et al., 2018 ; Cassan and Diaz-Zorita, 2016 ; Dunham Trimmer, 2017 ).

[0034] Although a range of microbial inoculants have appeared on the commercial market, the application of PGPB to crops still poses substantial technical challenges. Several factors have been described that limit the effectiveness of isolated microbial or synthetic consortia as agricultural products designed to enhance plant growth and / or induce systemic resistance to environmental stresses (such as soil type, climatic conditions, crop variety, bacterial genotype, bacterial isolate efficacy, poor inoculant quality, or limited availability of appropriate inoculation or production techniques). Therefore, bacterial formulations using PGPB typically fail to achieve the desired efficacy in field applications and are often incompatible with standard agricultural practices (Bashan, 1998; Bashan et al., 2014). Therefore, next-generation microbial technologies for traditional and organic agriculture must overcome these significant current limitations. The formulations and methods described herein overcome these limitations.

[0035] Microbial seed inoculation techniques for improved crop performance For sustainable and precision agriculture, a current challenge is to better manage microorganisms in order to develop more robust and effective biological inoculants. Regardless of the purpose for which beneficial microorganisms are applied to crops, they must be applied in a way that optimizes and ensures their functionality. Several reports have demonstrated various techniques for delivering PGPB microorganisms, such as liquid (for spray application, drenching, or root dipping) or dry formulations (Barea, 2015; O'Callaghan, 2016) (and references therein). However, many of these approaches are not economically efficient or feasible in large-scale scenarios due to the amount of microbial inoculant required (especially for large-scale crops) and other environmental and operational factors that reduce microbial survival and functionality (e.g., drought, high temperatures, contamination, field soil microbiome, microbiologically unsuitable management). Well-known limitations in the field application of biological inoculants include:

[0036] On-farm handling quality: A major concern for growers is the ease of handling the inoculant and its application with standard planting equipment, where possible. Additionally, it is uncommon for farm practices to change to accommodate high-quality inoculant technology using specialized machinery (Date, 2001).

[0037] Long-term shelf life: Inoculants must have a sufficient shelf life, often 1–2 years at room temperature, for microbial technologies to be successfully integrated into current agricultural distribution systems (Deaker et al., 2011).

[0038] Inoculant Performance: Microbial formulations must be stable during production, distribution, storage, and transport to farmers, especially when the main components are viable and mutable, compared to pesticides. Formulating microorganisms into affordable products for use by farmers who are not microbiologically trained is challenging primarily for the following reasons: i) Microbial strains may perform optimally under precise laboratory conditions, and similar results are expected under field conditions, but on-farm conditions may not be ideal. ii) Biological inoculants are typically liquid or solid formulations, and if stored incorrectly (e.g., at high or room temperature) or incorrectly mixed or diluted, microbial viability may decrease, reducing beneficial effects in the field. Furthermore, cross-contamination with other microorganisms can negatively affect the original biological inoculant (Bashan et al., 2014; Mahmood et al., 2016), ultimately negatively impacting the crop.

[0039] Inoculation method: Applying biological inoculants directly to seeds contributes to the survival and efficiency of bacteria in the soil and on the plant. In some embodiments, the effectiveness of the beneficial effects of microorganisms on plants is limited by certain biotic and abiotic factors (including soil temperature and moisture, the presence of nutrients, and pH), the product's storage conditions, and its shelf life (Calabi-Floody et al., 2018; Mahmood et al., 2016; Taylor et al., 1998).

[0040] Direct application of beneficial microorganisms onto seeds has been proposed as an efficient mechanism to overcome some of these drawbacks, as it promotes the colonization of microbial inoculants into the soil and / or plant. Thus, direct seed treatment with beneficial microorganisms aids in the colonization of plants by microorganisms during the early stages of development and throughout their life cycle. Generally, these methods have been reported as the best options for applying various beneficial microorganisms to seeds. However, they have primarily been described for research purposes (O'Callaghan, 2016) (and references therein). Most research on microbial seed inoculation has been developed by agrochemical and seed companies, and the techniques and processes used are rarely published and are kept as "in-house knowledge" (US2010 / 0154299A1; US2015 / 0289515A1; US2018 / 0064116A1; US2018 / 098483A1; US2018 / 0064116A1; US2018 / 0132486A1). Current seed treatment using PGPB as a biological inoculant involves the following steps:

[0041] Coating: A precise amount of active ingredient (bacteria, pesticide, fungicide, etc.) is applied to the seed surface using a liquid medium, forming a thin layer on the seed that prevents it from changing shape. There are two main types of coating: "film coating" and "slurry coating." In film coating, the inoculant is applied as an aqueous cell suspension using a polymer or adhesive material (e.g., methylcellulose, vegetable or paraffin oil, polysaccharides, etc.). In contrast, in slurry coating seed treatment, the inoculant is formulated as a dry powder or attached to a specific carrier (commonly peat, charcoal, lignite, farmyard manure, etc.) and applied to the outside of the seed using various sealants. While film coating is primarily used experimentally, slurry coating is widely used in farms. Although these methods have proven to reduce the issue of inconsistent reproducibility in the field, issues such as seed shelf life and cell viability still remain with commercial formulations ( Calabi-Floody et al., 2018 ; Taylor et al., 1998 ; Taylor and Harman, 1990 ).

[0042] Pelletization: This process involves the addition of inert materials with the intention of enlarging seeds and creating standard-sized spherical units. This procedure has gained popularity in precision agriculture because it can alter the shape and size of small, irregular seeds, facilitating mechanical handling for precision sowing. Seed pelleting involves two main components: a bulking coating material and a binder. The bulking material can be a mixture of several different minerals and / or organic substances, or it can be a single component. The second component, the binder, holds the coating material together. Many different compounds have been used as binders, including various starches, sugars, gum arabic, clays, cellulose, and vinyl polymers (O'Callaghan, 2016; Taylor et al., 1998; Taylor and Harman, 1990) (and references therein).

[0043] Priming: This method involves soaking seeds in an aqueous suspension (without the use of liquid polymers or adhesives) for a predetermined period of time, followed by drying to prevent the onset of germination. Given the labor-intensive nature of this process, it is best suited for low- to medium-volume, high-value crops, such as vegetable seeds (O'Callaghan, 2016; Taylor et al., 1998; Taylor and Harman, 1990). Among various priming techniques, hydration with any biological compound is referred to as "biopriming" (Ashraf and Foolad, 2005; Bennett and Whipps, 2008b, 2008a; Wright et al., 2003; Yadav et al., 2018).

[0044] Current limitations in implementing microbial techniques on seeds A comprehensive review of microbial technology, biological inoculant formulation, and practical aspects has been published (Bashan et al., 2014). The authors reported that many overriding challenges for PGPB inoculants must be carefully analyzed and overcome, considering improvements in delivery system implementation, detailed evaluation of carriers, improved microbial survival in inoculants, increased shelf life of inoculant products, the use of multi-strain inoculants, development of lower-cost techniques, and the implementation of seedbed inoculation for transplant crops. Recently, the most common biopriming techniques were reviewed (O'Callaghan, 2016). While not comprehensive, the cited studies agree on identifying major constraints limiting the commercial development of microbial seed inoculants.

[0045] To date, significant technical challenges must be addressed before a commercially viable seed treatment based on microbial inoculants can be achieved, including: 1) the effective viability of the microbial inoculant on the seeds throughout all stages of seed treatment, processing, and storage to ultimately achieve the desired PGPB effect on plants in the field after sowing; 2) the effective viability of seeds after treatment, as it is well known that seeds treated with liquid imbibition-based techniques such as hydropriming and osmopriming rapidly decline in shelf life, measured as germination ability and vigor (Wang et al., 2018; Schwember and Bradford, 2011; Hill and Cunningham, 2007; Tarquis and Bradford, 1992); and 3) the positive effects resulting from the interaction of the microbial inoculant with the host plant must be successfully realized across different soil types and environmental conditions.

[0046] Biopriming and seed treatment methodology: Microprime (商標) A typical seed priming protocol involves soaking seeds in any solution containing the required priming agents (inorganic and organic salts, nanoparticles, plant growth regulators, and / or plant growth-promoting bacteria) and then re-drying the seeds. This initiates the germination process beyond radicle emergence (Heydecker et al., 1973; Mahakham et al., 2017; McDonald, 1999; Song et al., 2017; Wright et al., 2003). Seed priming using osmotic solutions (osmopriming) has been practiced for decades (Heydecker et al., 1973) and is now common commercial practice for selected high-value horticultural seeds. This concept has been extended to hydropriming in cereals and legumes, reviving the "on-farm" priming technique (Harris et al., 2001). In recent years, several metal- and carbon-based nanoparticles (e.g., AgNPs, AuNPs, CuNPs, ZnNPs, fullerene, and carbon nanotubes) have been applied as seed priming agents to promote seed germination, seedling growth, and stress tolerance in several crops (Mahakham et al., 2017). Among different priming techniques (e.g., hydropriming, osmopriming, nanopriming, etc.), the internal space within seeds potentially possesses ideal conditions for bacterial inoculation and colonization when this procedure is performed with microbial cells (McQuilken et al., 1998; Ashraf and Foolad, 2005; Bennett et al., 2009; Tabassum et al., 2018; Wright et al., 2003).

[0047] Since the early 1990s, biopriming has been widely used on a wide range of crops and is undoubtedly recognized as an environmentally friendly agricultural technique (O'Callaghan, 2016; Taylor and Harman, 1990). Biopriming is sometimes mistakenly defined as the application of whole microorganisms, their exudates, or several biologically active compounds to the exterior of seeds (El-Mougy and Abdel-Kader, 2008; Muller and Berg, 2008; Song et al., 2017; Saber et al., 2012). More precisely, biopriming incorporates biological (seed inoculation with beneficial microorganisms) and physiological (seed hydration) components into seeds by promoting seedling emergence and uniformity and improving plant traits. Microbial-treated seeds are fundamentally different from other biological seed treatments in that colonization and proliferation of the added microorganisms must occur internally, since cells may be viable during the microbial seed treatment. However, the literature to date has yet to provide detailed explanations of these differences. Specifically, results and studies have yet to be reported on: 1) survival and / or proliferation of the biological agent (PGPB strain or consortium) in seeds over the relevant time frame (several months); 2) seed shelf life and effective germination several months after treatment; 3) effective microbial inoculum and plant interaction after the relevant time of seed storage; and 4) an economically viable methodology (taking into account relevant factors such as the time, inputs, and energy required for seed treatment) that is scalable and therefore feasible within traditional seed business models. Furthermore, bioosmopriming, when coupled with a bacterial coating procedure, has been demonstrated to significantly improve germination uniformity and plant growth traits (Bennett et al., 2009; Raj et al., 2004; Sharifi, 2011; Sharifi et al., 2011; Shariffi et al., 2012).Several researchers have reported incubation times ranging from 20 minutes to several days (Bennett et al., 2009; Bennett and Whipps, 2008b, 2008a; Murunde and Wainwright, 2018). Similarly, cell suspensions should be diluted to between 10 and 10 per gram of seed. 9 The range of cells and the type of biological agent (i.e., spores, endospores, or vegetative cells) were dependent on the type of biological agent (i.e., spores, endospores, or vegetative cells) (Wright et al., 2003; Saber et al., 2012; Raj et al., 2004; Murunde and Wainwright, 2018). Indeed, biopriming has been practiced and described in several ways by various researchers, but it remains an ambiguous approach that requires exploration and discussion (Bennett et al., 2009; Callan et al., 1990, 1991; Chakraborty et al., 2011; Mirshekari et al., 2012; Moeinzadeh et al., 2010; Raj et al., 2004; Reddy, 2013; Sharifi, 2011; Sharifi et al., 2011; Sharifi et al., 2012).

[0048] According to the latest technology, Song et al. described the use of Bacillus sp. exudates to induce immunity in cucumber plants (Song et al., 2017). This approach has several misleading results, both in terms of method and scope. 1) It does not use bacterial inoculants or plant growth promoters derived from them; instead, seeds are biostimulated with peptide-based compounds; 2) It does not incorporate live microorganisms into seeds to expose the embryos to the microorganisms or their exudates during the early post-dormancy stage of seed germination; 3) It does not confirm whether the biological agent (e.g., cyclodipeptide) entered the seeds and triggered the PGP effect (changes in gene expression) during the early embryonic stage (before the pericarp breaks); and 4) It does not report on the stability of the plant immune trigger elicitor over time. This last issue is particularly important because commercially viable agricultural microbial technologies must be stable over relatively long periods (e.g., >6 months) to fit into current agricultural distribution systems. In addition, the biological priming agents used in this referenced study are particularly unstable over time and susceptible to change due to biological and abiotic environmental factors (e.g., temperature, pH, biodegradation activity by other microorganisms, etc.).

[0049] The Serratia plymuthica strain HRO-C48 has also been reported as a biological agent for seed inoculation procedures (Muller and Berg, 2008). This study attempted to compare three different techniques: pelleting, membrane coating, and bioosmopriming. Despite measuring the cell count per seed immediately after seed treatment and storage, the authors were unable to accurately quantify the shelf life of a commercially viable product in the agricultural field. In fact, the viability of the HRO-C48 strain was only measured during a very short storage period (30 days). Other unclear topics reported by the authors include: 1) a high initial cell density was adjusted for seed soaking; and 2) a long incubation time of seeds in the presence of the biological agent (reportedly 12 hours) was used, which depends on the optimization of the biopriming procedure. Admittedly, all of these aspects are often not feasible parameters for industrial and commercial implementation of this method (Muller and Berg, 2008).

[0050] Several other publications have also been reported in the state of the art that point to incorporating synthetic microbial preparations into seeds. For example, U.S. Patent Application Nos. 2016 / 0338360A1 and 2016 / 0330976A1 refer to seeds containing beneficial bacteria. The methods presented in both of these referenced publications are based on the direct inoculation of different parts of flowers or plants to ultimately obtain seeds containing the desired microorganisms (Mitter et al., 2016a, 2016b).

[0051] The present invention proposes a new, effective, and reliable alternative to conventional biopriming techniques that directly addresses the problems described above. (商標)The proposed seed treatment method, named "Bio- ... The survival of biological agents, the genetic modification of embryos and the extension of the shelf life of treated seeds are controlled by Microprime (商標) This is guaranteed by seed technology. Finally, the new methodology reported here has certain checkpoints throughout the process, ensuring a reproducible and reliable end product. Some of these checkpoints include confirmation of molecular priming of the seeds and measurement of the expression of specific sets of genes related to development, abiotic stress tolerance, and defense responses against pathogens.

[0052] The present invention proposes a new strategy to enable plant traits and enhance their yield. This strategy is based on a specific seed treatment method (Microprime) as described below. (商標) ) is based on two effects on plant seeds:

[0053] 1. Loading of seed functional bacteria: Current seed treatment methods load seeds with endospore-forming bacteria and / or endospores. Microprime (商標) As a result of the seed treatment, endospore-forming bacteria and / or endospores are allocated to the seeds in the gaps located between the seed pericarp and its aleurone cell layer, as shown in Figures 1 and 4. The endospore-forming bacteria and / or endospores incorporated into the seeds correspond to strains that have the ability to effectively colonize the plant rhizosphere and have the ability to fix nitrogen, solubilize phosphate, and / or synthesize plant hormones. The endospore transformation ability of the selected bacteria and their allocation within the seeds allows Microprime (商標) Stability is guaranteed after seed treatment and throughout the entire commercial storage period. This process is confirmed by bacterial cell counts in time (Examples 1-5).

[0054] 2. Regulation of plant gene expression: This is Microprime (商標) This is achieved through the action of selected bacteria and / or bacterial exudates during the seed treatment process. The bacteria and / or bacterial exudates penetrate the seed and reach the embryo, where they can induce systemic induced resistance (IST) and / or systemic induced resistance (ISR) and / or plant development programs. These three processes enable plants to tolerate abiotic stresses (heat, drought, salinity, etc.), resist biotic stresses (pathogen attack), and strengthen root systems to enhance nutrient and water acquisition, directly improving plant performance. This effect is confirmed by transcriptional analysis of key genes involved in the aforementioned processes using qRT-PCR (Example 11).

[0055] For this invention to be valuable and practically applicable on an industrial scale, it is necessary that the method be cost-effective and scalable. A seed treatment process such as this one involves multiple steps that require time, input, and energy. The proposed method and invention prioritizes keeping the processing cost and time of seed treatment as low as possible. Microprime (商標)The method aims to make the seed treatment process effective at room temperature (20-24°C) while achieving seed imbibition in less than 20 minutes to 16 hours. The latter is not an easy feat to achieve because Microprime (商標) In addition to the desired minimum number of bacteria, endospore-forming bacteria, and / or endospores in the seeds after seed treatment, it is also necessary for the bacteria to remain stable and viable over time. (商標) This is because the seeds (as a product) are not affected by the storage, packaging, logistics and sowing processes in the same way as conventional seeds that have not been treated.

[0056] The proposed methodology involves immersing previously disinfected seeds in a seed treatment medium (hereafter referred to as micropriming solution) containing nutrients, surfactants, and salts. Figure 2 shows a process diagram depicting the seed treatment process.

[0057] Stability of bacterial seed treatments The stability of bacteria in seeds over time is not a simple or obvious problem to address. (商標) When bacteria are incorporated into seeds using seed treatments, the location of the bacteria within the seed is shown in Figure 4 for corn seeds.

[0058] In Figure 4A, Microprime (商標)After seed treatment, we can see the space within the corn seed where bacteria marked with red fluorescent protein (RFP) reside (pink fibers). This location is the space between the seed's pericarp and the seed's aleurone cell layer, which separates the endosperm and embryo from the outer layers. This location allows certain microorganisms to live comfortably for a limited period of time, after which the cells inevitably die due to the depletion of available nutrients or, in the case of certain microorganisms, initiate the process of endosporulation (bacteria from the Firmicutes, Proteobacteria, and Actinobacteria phylum). The advantage of accumulating in this location is that the microorganisms are protected from external factors that could affect their immediate integrity, such as other microorganisms or dehydration.

[0059] To address the issue of long-term viability due to nutrient starvation, the methodology varies depending on the type of bacteria being addressed. Under certain conditions (mainly in scenarios where feasibility is at risk), some bacteria have the ability to cease growth and enter a physical state known as endospores. As endospores, bacteria enter a dormant scenario where they do not consume nutrients for an extended period of time. Microprime has been developed for bacteria, primarily from the Firmicutes, Proteobacteria, and Actinobacteria phylum, that have the ability to produce endospores. (商標) The solution is supplemented with specific salts, which, when incorporated into the seeds, induce the bacteria to enter this dormant state. By doing the latter, it is possible to ensure the long-term viability of the bacteria within the seeds. When the endospores are again found in favorable conditions with moisture and nutrients (e.g., at the time of sowing), they revert to an active bacterial state (vegetative cell) and begin normal functioning and vegetative reproduction.

[0060] Another strategy is to use Microprime rather than forcing the bacteria to transform during the seed treatment process. (商標) The latter is the direct addition of endospores to the solution. (商標)showed better yield in terms of endospores per seed found after seed treatment with

[0061] Table 1 below lists some bacterial genera that are of particular interest for this proposed novel seed treatment due to their ability to transform into endospores.

[0062] [Table 1]

[0063] Bacillus species are among the most abundant bacteria capable of endospore formation, and extensive research has been conducted on several Bacillus strains with PGPB traits. Microprime is a suitable medium for sufficient bacterial growth in seeds. (商標) The solution can be supplemented with nutrients specifically compatible with the selected bacteria, or the endospores of the desired bacteria to be incorporated into the seeds can be added to Microprime. (商標) It is necessary to add it directly to the solution. Biological priming of seed embryos

[0064] The method of the present disclosure involves treating plant seeds with a bacterial composition designed as described above (Microprime (商標)The present invention contemplates the use of bacterial treatments (seed treatments). Such treatments can employ osmotic pressure of the seed to enable bacterial incorporation; a treatment first described by Smith et al. to introduce a chemical priming agent into seeds is now called osmopriming. However, the disclosed method is adapted to incorporate bacterial populations into dormant seeds, aiming to provide early conditioning of the emerging plantlet through direct biological priming of the embryo once dormancy ends and appropriate environmental or agronomic conditions trigger the first stage of germination. This novel approach offers unprecedented advantages over conventional bacterial formulations designed to generate biological priming, as the incorporated bacteria are protected within the dormant seed and are favorably positioned to generate permanent priming of the embryo from the earliest possible developmental stage by themselves or by the action of their exudates (Example 11). Furthermore, the treated seeds are subjected to normal transport, storage, coating pelleting, and sowing treatments in accordance with standard agricultural practices, without additional requirements for handling, nutrient additives, preservatives, or irrigation, and without incompatible restrictions on pest or plant disease control agents. Thus, the methods described herein (Microprime (商標) Seed treatment) also offers a distinct advantage over seed biopriming (Mahmood et al., 2016) because the method involves seed pre-germination and a dormancy-terminating reaction that reduces storage viability and limits the feasibility of manipulation and processing (Examples 5 and 6).

[0065] Additionally, the disclosed method differs from previously reported methods of inoculating seeds by using parent plants as reactors for microbial growth or by inoculating plant sexual organs (Mitter et al., 2016a; Mitter et al., 2016b). Such methods imply an inherent bias in the types of bacteria that can ultimately be incorporated into seeds, because a successful inoculant must be able to survive within the target plant organ or tissue, compete with endogenous microorganisms, and access the seed's internal space. The Microprime method presented herein (商標) The strategy is not hindered by endoparasitic ability or tissue survival, because the engineered bacteria do not need to be endosymbionts, face the defense responses of mature plants, or outcompete the endoparasitic microbiota, but only need to survive long enough or have their exudates reach the plant embryo to produce molecular priming for the plant.

[0066] We believe that the above-mentioned inherent advantages and differentiating features make the disclosed method non-obvious to those skilled in the art of bacterial plant growth stimulation. Indeed, for this strategy to be successful, certain critical conditions must be met by the candidate bacteria in the treatment composition, which are not always considered in standard formulations of plant growth-promoting microorganisms. First, the bacterial composition must be designed using the efficacy and compatibility criteria described above to avoid competition and / or antagonism within the seed. The absence of these effects must be experimentally evaluated before the composition is formulated. Seed internalization must be evaluated for each bacterium included in the designed composition, determining saturation curves and bacterial survival during storage time and seed treatment procedures (Figure 5, Examples 1-5). Furthermore, analysis of internal seed tissue must also be performed to assess the presence and viability of the desired bacteria and the relative abundance of each constituent strain relative to the others (Figure 4, Examples 1-5).

[0067] The plant material must also be conditioned before treatment with the specific bacterial composition. (商標) The seeds may be sterilized to eliminate background noise while determining the effectiveness of the seed treatment.

[0068] Microprime (商標) Once the bacteria emerge, transcriptional analysis of marker genes related to pathogen defense, abiotic stress tolerance, and development must be performed to confirm the beneficial bacterial effects on the treated seeds (Example 11). This analysis must be performed after the seed dormancy stage and before the seed pericarp and endosperm rupture and radicle emergence. Evaluation of transcriptional changes in the developing embryo resulting from previous bacterial treatment of dormant seeds is also an important step in validating the methodology because it provides rapid confirmation of the priming effect and the results are not affected by external factors that appear after seed rupture, including the access of other microorganisms to the developing plant tissue from outside the seed and / or the chemical composition of the surrounding soil or growth substrate.

[0069] The methods and compositions proposed in this invention are (商標) These methods can be summarized as a seed treatment method, in which seeds are incorporated into a physiological saline solution containing a seed-compatible bacterial composition, bacterial-compatible nutrients (if non-endospore-forming bacteria are used), and surfactants to increase the cell load loaded onto the seeds at room temperature and during short-term seed soaking, as well as supplemented minerals to increase the conversion rate of endospore-forming bacteria to endospores (Figure 2).

[0070] FIG. 3 illustrates the methodology of the present invention for obtaining a stable microbial seed treatment.

[0071] Modified plant seeds In one aspect, provided herein are modified plant seeds comprising a microorganism or a microbial exudate incorporated into the seed. In some embodiments, the microorganism or exudate confers a beneficial trait to the plant. In some embodiments, the beneficial trait improves plant growth. In some embodiments, the beneficial trait has improved plant growth. In some preferred embodiments, the microorganism is an endospore-forming bacterium or an endospore thereof.

[0072] The microorganisms or exudates incorporated into the modified plant seeds can improve various plant characteristics that promote plant growth. In some embodiments, the microorganisms or exudates impart a plant growth effect. In some embodiments, the plant growth effect includes cellular osmoregulation, ion homeostasis, antioxidant defense, heat stress tolerance, maintenance of photosynthetic capacity, or a combination thereof. In some embodiments, the plant growth effect includes cellular osmoregulation. In some embodiments, the plant growth effect includes ion homeostasis. In some embodiments, the plant growth effect includes antioxidant defense. In some embodiments, the plant growth effect includes heat stress tolerance. In some embodiments, the plant growth effect includes maintenance of photosynthetic capacity. In some embodiments, the plant growth effect is causing systemic induced resistance. In some embodiments, the plant growth effect is causing systemic induced resistance.

[0073] In some embodiments, the microorganisms or exudates incorporated into seeds improve seed germination rate. In some embodiments, seed germination rate is improved by at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, or at least about 50%. In some embodiments, seed germination rate is improved by at least about 1%. In some embodiments, seed germination rate is improved by at least about 2%. In some embodiments, seed germination rate is improved by at least about 3%. In some embodiments, seed germination rate is improved by at least about 4%. In some embodiments, seed germination rate is improved by at least about 5%. In some embodiments, seed germination rate is improved by at least about 10%. In some embodiments, seed germination rate is improved by at least about 20%. In some embodiments, seed germination rate is improved by at least about 30%. In some embodiments, seed germination rate is improved compared to seeds not incorporating the microorganisms or exudates.

[0074] In some embodiments, the microorganisms or exudates incorporated into the seeds improve plant growth under stress conditions. In some embodiments, the microorganisms or exudates incorporated into the seeds improve drought tolerance. In some embodiments, the improved drought tolerance is the ability to grow under drought. In some embodiments, plant growth is improved under drought conditions compared to plants grown from seeds that do not incorporate the microorganisms or exudates. In some embodiments, plant growth is improved by at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, or at least about 50% under drought conditions.

[0075] In some embodiments, plant growth is improved by about 1% to about 50%. In some embodiments, plant growth is improved by about 1% to about 2%, about 1% to about 3%, about 1% to about 4%, about 1% to about 5%, about 1% to about 10%, about 1% to about 20%, about 1% to about 30%, about 1% to about 40%, about 1% to about 50%, about 2% to about 3%, about 2% to about 4%, about 2% to about 5%, about 2% to about 10%, about 2% to about 20%, about 2% to about 30%, about 2% to about 40%, about 2% to about 50%, about 3% to about 4%, about 3% to about 5%, about 3% to about 10%, about 3% to about 20%, about 3% to about 30%, or about 3% to about 40%. , about 3% to about 50%, about 4% to about 5%, about 4% to about 10%, about 4% to about 20%, about 4% to about 30%, about 4% to about 40%, about 4% to about 50%, about 5% to about 10%, about 5% to about 20%, about 5% to about 30%, about 5% to about 40%, about 5% to about 50%, about 10% to about 20%, about 10% to about 30%, about 10% to about 40%, about 10% to about 50%, about 20% to about 30%, about 20% to about 40%, about 20% to about 50%, about 30% to about 40%, about 30% to about 50%, or about 40% to about 50% improvement. In some embodiments, plant growth is improved by about 1%, about 2%, about 3%, about 4%, about 5%, about 10%, about 20%, about 30%, about 40%, or about 50%. In some embodiments, plant growth is improved by at least about 1%, about 2%, about 3%, about 4%, about 5%, about 10%, about 20%, about 30%, or about 40%. In some embodiments, plant growth is improved by up to about 2%, about 3%, about 4%, about 5%, about 10%, about 20%, about 30%, about 40%, or about 50%. In some embodiments, plant growth is measured relative to plants grown from seeds that have not been treated with the microorganism or leachate.

[0076] Plant growth may be measured by measuring the size of a part of the plant. The part of the plant may depend on the type of plant. For example, if the plant seed is a lettuce plant seed (e.g., Lactuca sativa), the size of the rosette may be measured. In some embodiments, fruit size is used to measure plant growth. In some embodiments, the weight of a plant's shoot is used to measure plant growth. In some embodiments, the weight of a plant's roots is used to measure plant growth. In some embodiments, the plant or plant part is dried before measurement.

[0077] In some embodiments, plant growth is measured after a desired period of time. In some embodiments, plant growth is measured after about 1 week to about 36 weeks. In some embodiments, plant growth is measured after about 1 week to about 2 weeks, about 1 week to about 4 weeks, about 1 week to about 6 weeks, about 1 week to about 8 weeks, about 1 week to about 12 weeks, about 1 week to about 18 weeks, about 1 week to about 36 weeks, about 2 weeks to about 4 weeks, about 2 weeks to about 6 weeks, about 2 weeks to about 8 weeks, about 2 weeks to about 12 weeks, about 2 weeks to about 18 weeks, about 2 weeks to about 36 weeks, or about 4 weeks to about 6 weeks. , about 4 weeks to about 8 weeks, about 4 weeks to about 12 weeks, about 4 weeks to about 18 weeks, about 4 weeks to about 36 weeks, about 6 weeks to about 8 weeks, about 6 weeks to about 12 weeks, about 6 weeks to about 18 weeks, about 6 weeks to about 36 weeks, about 8 weeks to about 12 weeks, about 8 weeks to about 18 weeks, about 8 weeks to about 36 weeks, about 12 weeks to about 18 weeks, about 12 weeks to about 36 weeks, or about 18 weeks to about 36 weeks. In some embodiments, plant growth is measured after about 1 week, about 2 weeks, about 4 weeks, about 6 weeks, about 8 weeks, about 12 weeks, about 18 weeks, or about 36 weeks. In some embodiments, plant growth is measured after at least about 1 week, about 2 weeks, about 4 weeks, about 6 weeks, about 8 weeks, about 12 weeks, or about 18 weeks. In some embodiments, plant growth is measured after up to about 2 weeks, about 4 weeks, about 6 weeks, about 8 weeks, about 12 weeks, about 18 weeks, or about 36 weeks. In some embodiments, plant growth is measured after about 7 days to about 42 days. In some embodiments, plant growth is measured after about 7 days to about 14 days, about 7 days to about 28 days, about 7 days to about 42 days, about 14 days to about 28 days, about 14 days to about 42 days, or about 28 days to about 42 days. In some embodiments, plant growth is measured after about 7 days, about 14 days, about 28 days, or about 42 days. In some embodiments, plant growth is measured after at least about 7 days, about 14 days, or about 28 days, hi some embodiments, plant growth is measured after up to about 14 days, about 28 days, or about 42 days.

[0078] In some embodiments, the microorganisms or exudates are incorporated inside the seed. In some embodiments, the microorganisms or exudates are incorporated into the seed below the pericarp. In some embodiments, the microorganisms or exudates are incorporated into the seed between the pericarp and the aleurone cell layer. In some embodiments, the microorganisms or exudates contact the seed embryo. In some embodiments, the microorganisms or exudates do not contact the seed embryo. In some embodiments, the microorganisms or exudates contact the seed endosperm. In some embodiments, the microorganisms or exudates do not contact the seed endosperm. In some embodiments, the microorganisms or exudates are incorporated into the seed in the space between the seed coat and the seed embryo. In some embodiments, the microorganisms or exudates are incorporated into the space between the seed pericarp and the aleurone cell layer of the seed.

[0079] The modified plant seed may be any type of plant seed. In some embodiments, the modified seed is a monocotyledonous seed. In some embodiments, the plant seed is a corn, wheat, rice, barley, rye, sugarcane, millet, oat, or sorghum seed. In some embodiments, the plant seed is a corn seed. In some embodiments, the plant seed is a corn (Zea maize) seed. In some embodiments, the modified seed is a dicotyledonous seed. In some embodiments, the seed is a soybean, cotton, alfalfa, bean, quinoa, lentil, peanut, lettuce, tomato, or cabbage seed. In some embodiments, the seed is a lettuce (lettuce) seed. In some embodiments, the seed is a lettuce (Lactuca sativa) seed. In some embodiments, the seed is a tomato (tomato) seed. In some embodiments, the seed is a tomato (Solanum lycopersicum) seed. In some embodiments, the seed is a genetically modified organism (GMO) seed. In some embodiments, the seeds are non-GMO seeds.

[0080] The amount of microorganism or exudate incorporated into the seed should be at a level sufficient to impart a plant growth benefit to the plant. In some embodiments, the amount of microorganism incorporated into the seed is from about 250 colony forming units (CFU) to about 5,000 CFU. In some embodiments, the amount of microorganisms incorporated into seeds is from about 250 CFU to about 500 CFU, from about 250 CFU to about 750 CFU, from about 250 CFU to about 1,000 CFU, from about 250 CFU to about 2,000 CFU, from about 250 CFU to about 3,000 CFU, from about 250 CFU to about 4,000 CFU, from about 250 CFU to about 5,000 CFU, from about 500 CFU to about 750 CFU, from about 500 CFU to about 1,000 CFU, from about 500 CFU to about 2,000 CFU, from about 500 CFU to about 3,000 CFU, from about 500 CFU to about 4,000 CFU, from about 500 CFU to about 5,000 CFU, from about 750 CFU to about 1,000 CFU, 50 CFU to about 2,000 CFU, about 750 CFU to about 3,000 CFU, about 750 CFU to about 4,000 CFU, about 750 CFU to about 5,000 CFU, about 1,000 CFU to about 2,000 CFU, about 1,000 CFU to about 3,000 CFU, about 1,000 CFU to about 4,000 CFU, about 1,000 CFU to about 5,000 CFU, about 2,000 CFU to about 3,000 CFU, about 2,000 CFU to about 4,000 CFU, about 2,000 CFU to about 5,000 CFU, about 3,000 CFU to about 4,000 CFU, about 3,000 CFU to about 5,000 CFU, or about 4,000 CFU to about 5,000 CFU. In some embodiments, the amount of microorganisms incorporated into the seeds is about 250 CFU, about 500 CFU, about 750 CFU, about 1,000 CFU, about 2,000 CFU, about 3,000 CFU, about 4,000 CFU, or about 5,000 CFU. In some embodiments, the amount of microorganisms incorporated into the seeds is at least about 250 CFU, about 500 CFU, about 750 CFU, about 1,000 CFU, about 2,000 CFU, about 3,000 CFU, or about 4,000 CFU. In some embodiments, the amount of microorganisms incorporated into the seeds is at most about 500 CFU, about 750 CFU, about 1,000 CFU, about 2,000 CFU, about 3,000 CFU, about 4,000 CFU, or about 5,000 CFU.In some embodiments, at least about 500 CFU are incorporated into the seeds. In some embodiments, at least about 1000 CFU are incorporated into the seeds.

[0081] In some embodiments, the microorganisms or exudates incorporated into the seeds have shelf stability over an extended period of time. In some embodiments, shelf stability indicates that the plant growth-promoting effects of the incorporated microorganisms or exudates persist over an extended period of time. In some embodiments, the modified seeds have shelf stability for about 3 months to about 36 months. In some embodiments, the modified seeds are from about 3 months to about 6 months, from about 3 months to about 9 months, from about 3 months to about 12 months, from about 3 months to about 15 months, from about 3 months to about 18 months, from about 3 months to about 21 months, from about 3 months to about 24 months, from about 3 months to about 30 months, from about 3 months to about 36 months, from about 6 months to about 9 months, from about 6 months to about 12 months, from about 6 months to about 15 months, from about 6 months to about 18 months, from about 6 months to about 21 months, from about 6 months to about 24 months, from about 6 months to about 30 months, from about 6 months to about 36 months, from about 9 months to about 12 months, from about 9 months to about 15 months, from about 9 months to about 18 months, from about 9 months to about 21 months, from about 9 months to about 24 months, from about 9 months to about 30 months, from about 9 months to about 36 months The storage stability is for about 12 months to about 36 months, about 12 months to about 15 months, about 12 months to about 18 months, about 12 months to about 21 months, about 12 months to about 24 months, about 12 months to about 30 months, about 12 months to about 36 months, about 15 months to about 18 months, about 15 months to about 21 months, about 15 months to about 24 months, about 15 months to about 30 months, about 15 months to about 36 months, about 18 months to about 21 months, about 18 months to about 24 months, about 18 months to about 30 months, about 18 months to about 36 months, about 21 months to about 24 months, about 21 months to about 30 months, about 21 months to about 36 months, about 24 months to about 30 months, about 24 months to about 36 months, or about 30 months to about 36 months. In some embodiments, the modified seeds have storage stability for about 3 months, about 6 months, about 9 months, about 12 months, about 15 months, about 18 months, about 21 months, about 24 months, about 30 months, or about 36 months. In some embodiments, the modified seeds have storage stability for at least about 3 months, about 6 months, about 9 months, about 12 months, about 15 months, about 18 months, about 21 months, about 24 months, or about 30 months. In some embodiments, the modified seeds have storage stability for up to about 6 months, about 9 months, about 12 months, about 15 months, about 18 months, about 21 months, about 24 months, about 30 months, or about 36 months.

[0082] In some embodiments, the microorganism incorporated into the seed is stable after incorporation. In some embodiments, the microorganism is stable for more than 30 days, more than 6 months, more than 1 year, or more than 2 years. In some embodiments, the microorganism is stable for more than 30 days. In some embodiments, the microorganism is stable for more than 6 months. In some embodiments, the microorganism is stable for more than 1 year. In some embodiments, the microorganism is stable for more than 2 years.

[0083] The microorganism or exudate thereof incorporated into the plant seeds may be any of the microorganisms provided herein or other microorganisms. In some embodiments, the microorganism is a microbe. In some embodiments, the microorganism is an endospore-forming microorganism. In some embodiments, the microorganism is an endospore-forming microorganism or an endospore thereof. In some embodiments, the microorganism is an endospore of a microorganism provided herein. In some embodiments, the microorganism is an endospore-forming bacterium or an endospore thereof.

[0084] How to incorporate bacteria In one aspect, provided herein are methods for incorporating one or more microorganisms or exudates thereof into one or more plant seeds. In some embodiments, the methods include disinfecting plant seeds. In some embodiments, the methods include contacting the seeds with a solution containing one or more microorganisms or exudates thereof. In some embodiments, the solution further comprises salt. In some embodiments, the methods include incubating the seeds with the solution for a period of time. In some embodiments, the period of time is sufficient to incorporate a desired amount of the microorganisms or exudates thereof into the plant seeds. In some embodiments, the methods incorporate a desired amount of the microorganisms or exudates thereof into the seeds.

[0085] In some embodiments, the method includes contacting the seeds with a solution containing a salt. Any salt may be used. In some preferred embodiments, the salt is NaCl. In some embodiments, the salt is NaCl, LiCl, KCl, MgCl2, CaCl2, NaBr, LiBr, KBr, MgBr2, CaBr2, NaI, LiI, KI, MgI2, or CaI2. In some embodiments, the salt includes sodium, lithium, or potassium ions. In some embodiments, the salt includes alkali metal ions. In some embodiments, the salt includes alkaline earth metal ions. In some embodiments, the salt includes halide ions. In some embodiments, the salt is an alkali or alkaline earth halide salt. In some embodiments, the salt includes chloride, bromide, or iodide ions. In some embodiments, the salt is a sulfate, phosphate, carbonate, or nitrate salt.

[0086] The salt may be present in the solution at any suitable concentration. In some embodiments, the solution comprises about 0.85% salt (w / v). In some embodiments, the solution comprises about 0.1% to about 1.25% salt (w / v). In some embodiments, the solution comprises about 0.1% to about 2.0% salt (w / v). In some embodiments, the solution comprises about 0.1% to about 0.25%, about 0.1% to about 0.5%, about 0.1% to about 0.6%, about 0.1% to about 0.7%, about 0.1% to about 0.75%, about 0.1% to about 0.8%, about 0.1% to about 0.85%, about 0.1% to about 0.9%, about 0.1% to about 0.95%, about 0.1% to about 1%, about 0.1% to about 1.25%, about 0.25% to about 0.5%, about 0.25% to about 0.6%, about 0.25% to about 0.7%, about 0.25% to about 0.75%, about 0.25% to about 0. 0.8%, about 0.25% to about 0.85%, about 0.25% to about 0.9%, about 0.25% to about 0.95%, about 0.25% to about 1%, about 0.25% to about 1.25%, about 0.5% to about 0.6%, about 0.5% to about 0.7%, about 0.5% to about 0.75%, about 0.5% to about 0.8%, about 0.5% to about 0.85%, about 0.5% to about 0.9%, about 0.5% to about 0.95%, about 0.5% to about 1%, about 0.5% to about 1.25%, about 0.6% to about 0.7%, about 0.6% to about 0.75%, about 0.6% to about 0. 8%, about 0.6% to about 0.85%, about 0.6% to about 0.9%, about 0.6% to about 0.95%, about 0.6% to about 1%, about 0.6% to about 1.25%, about 0.7% to about 0.75%, about 0.7% to about 0.8%, about 0.7% to about 0.85%, about 0.7% to about 0.9%, about 0.7% to about 0.95%, about 0.7% to about 1%, about 0.7% to about 1.25%, about 0.75% to about 0.8%, about 0.75% to about 0.85%, about 0.75% to about 0.9%, about 0.75% to about 0.95%, about 0.75% to about 1 %, about 0.75% to about 1.25%, about 0.8% to about 0.85%, about 0.8% to about 0.9%, about 0.8% to about 0.95%, about 0.8% to about 1%, about 0.8% to about 1.25%, about 0.85% to about 0.9%, about 0.85% to about 0.95%, about 0.85% to about 1%, about 0.85% to about 1.25%, about 0.9% to about 0.95%, about 0.9% to about 1%, about 0.9% to about 1.25%, about 0.95% to about 1%, about 0.95% to about 1.25%, or about 1% to about 1.25% salt (w / v).In some embodiments, the solution comprises about 0.1%, about 0.25%, about 0.5%, about 0.6%, about 0.7%, about 0.75%, about 0.8%, about 0.85%, about 0.9%, about 0.95%, about 1%, or about 1.25% salt (w / v). In some embodiments, the solution comprises at least about 0.1%, about 0.25%, about 0.5%, about 0.6%, about 0.7%, about 0.75%, about 0.8%, about 0.85%, about 0.9%, about 0.95%, or about 1% salt (w / v). In some embodiments, the solution contains up to about 0.25%, about 0.5%, about 0.6%, about 0.7%, about 0.75%, about 0.8%, about 0.85%, about 0.9%, about 0.95%, about 1%, or about 1.25% salt (w / v). In some embodiments, the solution contains about 0.85% salt (w / v). In some embodiments, the solution contains about 0.8% to about 0.9% salt (w / v). In some embodiments, the solution contains about 0.75% to about 0.95% salt (w / v). In some embodiments, the solution contains about 0.7% to about 1% salt (w / v). In some embodiments, the solution contains about 0.5% to about 1.25% salt (w / v). In some embodiments, the solution contains about 0.5% to about 2% salt (w / v). In some embodiments, the solution comprises 0.1-0.2%, 0.2-0.3%, 0.3-0.4%, 0.4-0.5%, 0.5-0.6%, 0.6-0.7%, 0.7-0.8%, 0.8-0.9%, 0.9-1.0%, 1.0-1.1%, 1.1-1.2%, 1.2-1.3%, 1.3-1.4%, or 1.4-1.5% salt (w / v).

[0087] In some embodiments, the solution contains additional additives, such as dimethyl sulfoxide (DMSO), 1-dodecylazacycloheptan-2-one, laurocapram, 1-methyl-2-pyrrolidone (NMP), oleic acid, ethanol, methanol, polyethylene glycol (Brij 35, 58, 98), polyethylene glycol monolaurate (e.g., Tween 20), Tween 40 (polyoxyethylene acid sorbitol ester), Tween 60, Tween 80 (non-ionic), cetylmethylammonium bromide (CTAB), urea, lecithin (soybean-derived solidifying fatty acid), chitosan, poloxamer 188, poloxamer 237, poloxamer 338, poloxamer 407, or a combination thereof. In some embodiments, the solution comprises polyethylene glycol monolaurate (e.g., Tween 20), poloxamer 188, poloxamer 237, poloxamer 338, poloxamer 407, or a combination thereof. In some embodiments, the solution comprises a poloxamer. In some embodiments, the solution comprises polyethylene glycol monolaurate (e.g., Tween 20). The additional additives may be present at any concentration. In some embodiments, the additional additives comprise up to about 0.01%, 0.05%, 0.1%, 0.125%, 0.15%, 0.2%, 0.5%, or 1% (v / v) of the solution. In some embodiments, the additional additives comprise between about 0.01% and about 1% (v / v) of the solution. In some embodiments, the additional additives comprise about 0.1% (v / v) of the solution.

[0088] In some embodiments, the solution includes additional metal ions. In some embodiments, the solution includes magnesium, calcium, manganese, or a combination thereof. In some embodiments, the solution includes magnesium. In some embodiments, the solution includes calcium. In some embodiments, the solution includes manganese. In some embodiments, the solution includes magnesium and calcium. In some embodiments, the solution includes magnesium and manganese. In some embodiments, the solution includes calcium and manganese. In some embodiments, the solution includes magnesium, calcium, and manganese.

[0089] In some embodiments, the solution comprises one or more nutrients for the microorganism. In some embodiments, the solution comprises bacterial growth medium. In some embodiments, the solution comprises lysogeny broth (LB), nutrient broth, or a combination thereof. In some embodiments, the solution comprises lysogeny broth. In some embodiments, the solution comprises nutrient broth.

[0090] In some embodiments, the solution comprises a microorganism. In some embodiments, the solution comprises about 10 3 ~about 10 17 In some embodiments, the solution contains about 10 colony forming units (CFU) / mL of microorganisms. 3 ~about 10 4 , about 10 3 ~about 10 5 , about 10 3 ~about 10 6 , about 10 3 ~about 10 7 , about 10 3 ~about 10 8 , about 10 3 ~about 10 9 , about 10 3 ~about 10 10 , about 10 3 ~about 10 12 , about 10 3 ~about 10 15 , about 10 3 ~about 10 17 , about 10 4 ~about 10 5, about 10 4 ~about 10 6 , about 10 4 ~about 10 7 , about 10 4 ~about 10 8 , about 10 4 ~about 10 9 , about 10 4 ~about 10 10 , about 10 4 ~about 10 12 , about 10 4 ~about 10 15 , about 10 4 ~about 10 17 , about 10 5 ~about 10 6 , about 10 5 ~about 10 7 , about 10 5 ~about 10 8 , about 10 5 ~about 10 9 , about 10 5 ~about 10 10 , about 10 5 ~about 10 12 , about 10 5 ~about 10 15 , about 10 5 ~about 10 17 , about 10 6 ~about 10 7 , about 10 6 ~about 10 8 , about 10 6 ~about 10 9 , about 10 6 ~about 10 10 , about 10 6 ~about 10 12 , about 10 6 ~about 10 15 , about 10 6 ~about 10 17 , about 10 7 ~about 10 8 , about 10 7 ~about 10 9 , about 10 7 ~about 10 10 , about 10 7 ~about 10 12 , about 10 7 ~about 10 15 , about 10 7 ~about 10 17 , about 10 8~about 10 9 , about 10 8 ~about 10 10 , about 10 8 ~about 10 12 , about 10 8 ~about 10 15 , about 10 8 ~about 10 17 , about 10 9 ~about 10 10 , about 10 9 ~about 10 12 , about 10 9 ~about 10 15 , about 10 9 ~about 10 17 , about 10 10 ~about 10 12 , about 10 10 ~about 10 15 , about 10 10 ~about 10 17 , about 10 12 ~about 10 15 , about 10 12 ~about 10 17 , or about 10 15 ~about 10 17 In some embodiments, the solution contains about 10 colony forming units (CFU) / mL of microorganisms. 3 , about 10 4 , about 10 5 , about 10 6 , about 10 7 , about 10 8 , about 10 9 , about 10 10 , about 10 12 , about 10 15 , or about 10 17 In some embodiments, the solution contains at least about 10 CFU / mL of microorganisms. 3 , about 10 4 , about 10 5 , about 10 6 , about 10 7 , about 10 8 , about 10 9 , about 10 10 , about 10 12 , or about 10 15 In some embodiments, the solution contains up to about 10 CFU / mL of microorganisms. 4 , about 10 5, about 10 6 , about 10 7 , about 10 8 , about 10 9 , about 10 10 , about 10 12 , about 10 15 , or about 10 17 In some embodiments, the solution contains at least 10 CFU / mL of microorganisms. 6 ~10 7 In some embodiments, the solution contains 1 x 10 CFU / mL of microorganisms. 3 ~1×10 4 CFU / mL, 1 × 10 4 ~1×10 5 CFU / mL, 1 × 10 5 ~1×10 6 CFU / mL, 1 × 10 6 ~1×10 7 CFU / mL, 1 × 10 7 ~1×10 8 CFU / mL, 1 × 10 8 ~1×10 9 CFU / mL, 1 × 10 9 ~1×10 10 CFU / mL, 1 × 10 10 ~1×10 11 CFU / mL, 1 × 10 11 ~1×10 12 CFU / mL, 1 × 10 12 ~1×10 13 CFU / mL, 1 × 10 13 ~1×10 14 CFU / mL, 1 × 10 14 ~1×10 15 CFU / mL, 1 × 10 15 ~1×10 16 CFU / mL, or 1 x 10 16 ~1×10 17 Contains CFU / mL of microorganisms.

[0091] In some embodiments, the solution contains a desired amount of microorganisms per seed mass. In some embodiments, the solution contains about 10 3 ~about 10 17In some embodiments, the solution contains about 10 colony forming units (CFU) per gram of seeds. 3 ~about 10 4 , about 10 3 ~about 10 5 , about 10 3 ~about 10 6 , about 10 3 ~about 10 7 , about 10 3 ~about 10 8 , about 10 3 ~about 10 9 , about 10 3 ~about 10 10 , about 10 3 ~about 10 12 , about 10 3 ~about 10 15 , about 10 3 ~about 10 17 , about 10 4 ~about 10 5 , about 10 4 ~about 10 6 , about 10 4 ~about 10 7 , about 10 4 ~about 10 8 , about 10 4 ~about 10 9 , about 10 4 ~about 10 10 , about 10 4 ~about 10 12 , about 10 4 ~about 10 15 , about 10 4 ~about 10 17 , about 10 5 ~about 10 6 , about 10 5 ~about 10 7 , about 10 5 ~about 10 8 , about 10 5 ~about 10 9 , about 10 5 ~about 10 10 , about 10 5 ~about 10 12 , about 10 5 ~about 10 15 , about 10 5 ~about 10 17 , about 10 6 ~about 10 7 , about 106 ~about 10 8 , about 10 6 ~about 10 9 , about 10 6 ~about 10 10 , about 10 6 ~about 10 12 , about 10 6 ~about 10 15 , about 10 6 ~about 10 17 , about 10 7 ~about 10 8 , about 10 7 ~about 10 9 , about 10 7 ~about 10 10 , about 10 7 ~about 10 12 , about 10 7 ~about 10 15 , about 10 7 ~about 10 17 , about 10 8 ~about 10 9 , about 10 8 ~about 10 10 , about 10 8 ~about 10 12 , about 10 8 ~about 10 15 , about 10 8 ~about 10 17 , about 10 9 ~about 10 10 , about 10 9 ~about 10 12 , about 10 9 ~about 10 15 , about 10 9 ~about 10 17 , about 10 10 ~about 10 12 , about 10 10 ~about 10 15 , about 10 10 ~about 10 17 , about 10 12 ~about 10 15 , about 10 12 ~about 10 17 , or about 10 15 ~about 10 17 In some embodiments, the solution contains about 10 CFU / gram of seeds. 3 , about 10 4 , about 10 5 , about 106 , about 10 7 , about 10 8 , about 10 9 , about 10 10 , about 10 12 , about 10 15 , or about 10 17 In some embodiments, the solution contains at least about 10 CFU / gram of seeds. 3 , about 10 4 , about 10 5 , about 10 6 , about 10 7 , about 10 8 , about 10 9 , about 10 10 , about 10 12 , or about 10 15 In some embodiments, the solution contains up to about 10 CFU / gram of seeds. 4 , about 10 5 , about 10 6 , about 10 7 , about 10 8 , about 10 9 , about 10 10 , about 10 12 , about 10 15 , or about 10 17 In some embodiments, the solution contains 10 CFU / gram of seeds. 10 In some embodiments, the solution contains less than 10 CFU / gram of seeds. 9 In some embodiments, the solution contains less than 10 CFU / gram of seeds. 8 In some embodiments, the solution contains less than 10 CFU / gram of seeds. 11 In some embodiments, the solution contains less than about 10 CFU / gram of seeds. 5 ~about 10 9 Contains CFU / gram of seeds.

[0092] In some embodiments, the solution is about 10 3 ~about 10 17 In some embodiments, the solution contains about 10 colony cells / gram of seeds. 3 ~about 10 4 , about 10 3 ~about 10 5 , about 103 ~about 10 6 , about 10 3 ~about 10 7 , about 10 3 ~about 10 8 , about 10 3 ~about 10 9 , about 10 3 ~about 10 10 , about 10 3 ~about 10 12 , about 10 3 ~about 10 15 , about 10 3 ~about 10 17 , about 10 4 ~about 10 5 , about 10 4 ~about 10 6 , about 10 4 ~about 10 7 , about 10 4 ~about 10 8 , about 10 4 ~about 10 9 , about 10 4 ~about 10 10 , about 10 4 ~about 10 12 , about 10 4 ~about 10 15 , about 10 4 ~about 10 17 , about 10 5 ~about 10 6 , about 10 5 ~about 10 7 , about 10 5 ~about 10 8 , about 10 5 ~about 10 9 , about 10 5 ~about 10 10 , about 10 5 ~about 10 12 , about 10 5 ~about 10 15 , about 10 5 ~about 10 17 , about 10 6 ~about 10 7 , about 10 6 ~about 10 8 , about 10 6 ~about 10 9 , about 10 6 ~about 10 10 , about 10 6 ~about 1012 , about 10 6 ~about 10 15 , about 10 6 ~about 10 17 , about 10 7 ~about 10 8 , about 10 7 ~about 10 9 , about 10 7 ~about 10 10 , about 10 7 ~about 10 12 , about 10 7 ~about 10 15 , about 10 7 ~about 10 17 , about 10 8 ~about 10 9 , about 10 8 ~about 10 10 , about 10 8 ~about 10 12 , about 10 8 ~about 10 15 , about 10 8 ~about 10 17 , about 10 9 ~about 10 10 , about 10 9 ~about 10 12 , about 10 9 ~about 10 15 , about 10 9 ~about 10 17 , about 10 10 ~about 10 12 , about 10 10 ~about 10 15 , about 10 10 ~about 10 17 , about 10 12 ~about 10 15 , about 10 12 ~about 10 17 , or about 10 15 ~about 10 17 In some embodiments, the solution contains about 10 cells / gram of seeds. 3 , about 10 4 , about 10 5 , about 10 6 , about 10 7 , about 10 8 , about 10 9 , about 10 10 , about 10 12 , about 10 15 , or about 1017 In some embodiments, the solution contains at least about 10 cells / gram of seeds. 3 , about 10 4 , about 10 5 , about 10 6 , about 10 7 , about 10 8 , about 10 9 , about 10 10 , about 10 12 , or about 10 15 In some embodiments, the solution contains up to about 10 cells / gram of seeds. 4 , about 10 5 , about 10 6 , about 10 7 , about 10 8 , about 10 9 , about 10 10 , about 10 12 , about 10 15 , or about 10 17 In some embodiments, the solution contains 10 cells / gram of seeds. 10 In some embodiments, the solution contains less than 10 cells / gram of seeds. 9 In some embodiments, the solution contains less than 10 cells / gram of seeds. 8 In some embodiments, the solution contains less than 10 cells / gram of seeds. 11 In some embodiments, the solution contains less than about 10 cells / gram of seeds. 5 ~about 10 9 cells / gram of seeds.

[0093] In some embodiments, the seeds contain a desired amount of microorganisms per seed. In some embodiments, the solution contains about 10 3 ~about 10 17 In some embodiments, the solution contains about 10 colony forming units (CFU) / seed. 3 ~about 10 4 , about 10 3 ~about 10 5 , about 10 3 ~about 10 6 , about 10 3 ~about 10 7 , about 10 3 ~about 108 , about 10 3 ~about 10 9 , about 10 3 ~about 10 10 , about 10 3 ~about 10 12 , about 10 3 ~about 10 15 , about 10 3 ~about 10 17 , about 10 4 ~about 10 5 , about 10 4 ~about 10 6 , about 10 4 ~about 10 7 , about 10 4 ~about 10 8 , about 10 4 ~about 10 9 , about 10 4 ~about 10 10 , about 10 4 ~about 10 12 , about 10 4 ~about 10 15 , about 10 4 ~about 10 17 , about 10 5 ~about 10 6 , about 10 5 ~about 10 7 , about 10 5 ~about 10 8 , about 10 5 ~about 10 9 , about 10 5 ~about 10 10 , about 10 5 ~about 10 12 , about 10 5 ~about 10 15 , about 10 5 ~about 10 17 , about 10 6 ~about 10 7 , about 10 6 ~about 10 8 , about 10 6 ~about 10 9 , about 10 6 ~about 10 10 , about 10 6 ~about 10 12 , about 10 6 ~about 10 15 , about 10 6 ~about 10 17 , about 107 ~about 10 8 , about 10 7 ~about 10 9 , about 10 7 ~about 10 10 , about 10 7 ~about 10 12 , about 10 7 ~about 10 15 , about 10 7 ~about 10 17 , about 10 8 ~about 10 9 , about 10 8 ~about 10 10 , about 10 8 ~about 10 12 , about 10 8 ~about 10 15 , about 10 8 ~about 10 17 , about 10 9 ~about 10 10 , about 10 9 ~about 10 12 , about 10 9 ~about 10 15 , about 10 9 ~about 10 17 , about 10 10 ~about 10 12 , about 10 10 ~about 10 15 , about 10 10 ~about 10 17 , about 10 12 ~about 10 15 , about 10 12 ~about 10 17 , or about 10 15 ~about 10 17 In some embodiments, the solution contains about 10 CFU / seed. 3 , about 10 4 , about 10 5 , about 10 6 , about 10 7 , about 10 8 , about 10 9 , about 10 10 , about 10 12 , about 10 15 , or about 10 17 In some embodiments, the solution contains at least about 10 CFU / seed. 3 , about 10 4 , about 105 , about 10 6 , about 10 7 , about 10 8 , about 10 9 , about 10 10 , about 10 12 , or about 10 15 In some embodiments, the solution contains up to about 10 CFU / seed. 4 , about 10 5 , about 10 6 , about 10 7 , about 10 8 , about 10 9 , about 10 10 , about 10 12 , about 10 15 , or about 10 17 In some embodiments, the solution contains 10 CFU / seed. 10 In some embodiments, the solution contains less than 10 CFU / seed. 9 In some embodiments, the solution contains less than 10 CFU / seed. 8 In some embodiments, the solution contains less than 10 CFU / seed. 11 In some embodiments, the solution contains less than about 10 CFU / seed. 5 ~about 10 9 Contains less than CFU / seed.

[0094] In some embodiments, the seeds contain a desired amount of microorganisms per seed. In some embodiments, the solution contains about 10 3 ~about 10 17 In some embodiments, the solution contains about 10 3 ~about 10 4 , about 10 3 ~about 10 5 , about 10 3 ~about 10 6 , about 10 3 ~about 10 7 , about 10 3 ~about 10 8 , about 10 3 ~about 10 9 , about 10 3 ~about 10 10 , about 10 3 ~about 10 12 , about 103 ~about 10 15 , about 10 3 ~about 10 17 , about 10 4 ~about 10 5 , about 10 4 ~about 10 6 , about 10 4 ~about 10 7 , about 10 4 ~about 10 8 , about 10 4 ~about 10 9 , about 10 4 ~about 10 10 , about 10 4 ~about 10 12 , about 10 4 ~about 10 15 , about 10 4 ~about 10 17 , about 10 5 ~about 10 6 , about 10 5 ~about 10 7 , about 10 5 ~about 10 8 , about 10 5 ~about 10 9 , about 10 5 ~about 10 10 , about 10 5 ~about 10 12 , about 10 5 ~about 10 15 , about 10 5 ~about 10 17 , about 10 6 ~about 10 7 , about 10 6 ~about 10 8 , about 10 6 ~about 10 9 , about 10 6 ~about 10 10 , about 10 6 ~about 10 12 , about 10 6 ~about 10 15 , about 10 6 ~about 10 17 , about 10 7 ~about 10 8 , about 10 7 ~about 10 9 , about 10 7 ~about 10 10 , about 10 7 ~about 1012 , about 10 7 ~about 10 15 , about 10 7 ~about 10 17 , about 10 8 ~about 10 9 , about 10 8 ~about 10 10 , about 10 8 ~about 10 12 , about 10 8 ~about 10 15 , about 10 8 ~about 10 17 , about 10 9 ~about 10 10 , about 10 9 ~about 10 12 , about 10 9 ~about 10 15 , about 10 9 ~about 10 17 , about 10 10 ~about 10 12 , about 10 10 ~about 10 15 , about 10 10 ~about 10 17 , about 10 12 ~about 10 15 , about 10 12 ~about 10 17 , or about 10 15 ~about 10 17 In some embodiments, the solution contains about 10 3 , about 10 4 , about 10 5 , about 10 6 , about 10 7 , about 10 8 , about 10 9 , about 10 10 , about 10 12 , about 10 15 , or about 10 17 In some embodiments, the solution contains at least about 10 3 , about 10 4 , about 10 5 , about 10 6 , about 10 7 , about 10 8 , about 10 9 , about 10 10 , about 10 12 , or about 1015 In some embodiments, the solution contains up to about 10 4 , about 10 5 , about 10 6 , about 10 7 , about 10 8 , about 10 9 , about 10 10 , about 10 12 , about 10 15 , or about 10 17 In some embodiments, the solution contains 10 10 In some embodiments, the solution contains less than 10 cells / seed. 9 In some embodiments, the solution contains less than 10 cells / seed. 8 In some embodiments, the solution contains less than 10 cells / seed. 11 In some embodiments, the solution contains less than about 10 cells / seed. 5 ~about 10 9 Contains less than 100 cells / seeds.

[0095] In some embodiments, the microorganism is a bacterium. In some embodiments, the bacterium is an endospore-forming bacterium. In some embodiments, the method includes inducing endospore formation in the endospore-forming bacterium. In some embodiments, the bacterium incorporated into the seed is an endospore. In some embodiments, the solution includes one or more ingredients for inducing endospore formation. In some embodiments, the solution includes potassium, ferrous sulfate, calcium, magnesium, manganese, or a combination thereof.

[0096] In some embodiments, the method includes sterilizing the seeds. In some embodiments, the method includes sterilizing the surface of the seeds. Any method that produces seeds with a sterilized surface may be employed. In some embodiments, the seeds are sterilized with a bleach solution. In some embodiments, the seeds are sterilized before soaking the seeds in a solution containing one or more microorganisms. In some embodiments, the seeds are sterilized seeds. In some embodiments, the seeds have a sterilized surface. As used herein, "sterilizing," "sterilized," and related terms (e.g., "disinfecting," etc.) refer to the substantial absence of viable microorganisms on something that is sterilized. In some embodiments, the seeds are sterilized before incubating the seeds in a solution containing microorganisms. In some embodiments, the seeds are sterilized after incubating the seeds in a solution containing microorganisms. In some embodiments, a fungicide is added to the surface of the seeds.

[0097] In some embodiments, sterilized or disinfected seeds are substantially free of living microorganisms on the seed (e.g., on the surface of the seed), hi some embodiments, sterile or disinfected seeds contain less than 1 CFU, less than 5 CFU, less than 10 CFU, less than 20 CFU, less than 30 CFU, less than 40 CFU, or less than 50 CFU of microorganisms on the seed.

[0098] In some embodiments, the plant seeds are incubated with a solution containing the microorganism for a time sufficient to incorporate the microorganism into the seeds, ie, the plant seeds are incubated with a solution containing the endospore-forming bacteria or endospores thereof for about 1 minute to about 960 minutes. In some embodiments, the plant is incubated with the solution containing the endospore-forming bacterium or endospores thereof for about 1 minute to about 5 minutes, about 1 minute to about 10 minutes, about 1 minute to about 20 minutes, about 1 minute to about 60 minutes, about 1 minute to about 240 minutes, about 1 minute to about 960 minutes, about 5 minutes to about 10 minutes, about 5 minutes to about 20 minutes, about 5 minutes to about 60 minutes, about 5 minutes to about 240 minutes, about 5 minutes to about 960 minutes, about 10 minutes to about 20 minutes, about 10 minutes to about 60 minutes, about 10 minutes to about 240 minutes, about 10 minutes to about 960 minutes, about 20 minutes to about 60 minutes, about 20 minutes to about 240 minutes, about 20 minutes to about 960 minutes, about 60 minutes to about 240 minutes, about 60 minutes to about 960 minutes, or about 240 minutes to about 960 minutes. In some embodiments, the plant seeds are incubated with a solution containing endospore-forming bacteria or endospores thereof for about 1 minute, about 5 minutes, about 10 minutes, about 20 minutes, about 60 minutes, about 240 minutes, or about 960 minutes. In some embodiments, the plant seeds are incubated with a solution containing endospore-forming bacteria or endospores thereof for at least about 1 minute, about 5 minutes, about 10 minutes, about 20 minutes, about 60 minutes, or about 240 minutes. In some embodiments, the plant seeds are incubated with a solution containing endospore-forming bacteria or endospores thereof for up to about 5 minutes, about 10 minutes, about 20 minutes, about 60 minutes, about 240 minutes, or about 960 minutes. In some embodiments, the plant seeds are incubated with a solution containing endospore-forming bacteria or endospores thereof for about 1 minute. In some embodiments, the plant seeds are incubated with a solution containing endospore-forming bacteria or endospores thereof for about 5 minutes. In some embodiments, the plant seeds are incubated with the solution containing the endospore-forming bacteria or endospores thereof for about 10 minutes, hi some embodiments, the plant seeds are incubated with the solution containing the endospore-forming bacteria or endospores thereof for about 20 minutes.In some embodiments, the plant seeds are incubated with a solution containing endospore-forming bacteria or endospores thereof for about 60 minutes. In some embodiments, the plant seeds are incubated with a solution containing endospore-forming bacteria or endospores thereof for about 240 minutes. In some embodiments, the plant seeds are incubated with a solution containing endospore-forming bacteria or endospores thereof for about 960 minutes.

[0099] In some embodiments, the plant seeds are incubated with the solution containing the microorganism or exudate thereof for about 1 minute to about 960 minutes. In some embodiments, the plant is incubated with the solution containing the microorganism or an exudate thereof for about 1 minute to about 5 minutes, about 1 minute to about 10 minutes, about 1 minute to about 20 minutes, about 1 minute to about 60 minutes, about 1 minute to about 240 minutes, about 1 minute to about 960 minutes, about 5 minutes to about 10 minutes, about 5 minutes to about 20 minutes, about 5 minutes to about 60 minutes, about 5 minutes to about 240 minutes, about 5 minutes to about 960 minutes, about 10 minutes to about 20 minutes, about 10 minutes to about 60 minutes, about 10 minutes to about 240 minutes, about 10 minutes to about 960 minutes, about 20 minutes to about 60 minutes, about 20 minutes to about 240 minutes, about 20 minutes to about 960 minutes, about 60 minutes to about 240 minutes, about 60 minutes to about 960 minutes, or about 240 minutes to about 960 minutes. In some embodiments, the plant seeds are incubated with the solution comprising the microorganism or its exudate for about 1 minute, about 5 minutes, about 10 minutes, about 20 minutes, about 60 minutes, about 240 minutes, or about 960 minutes. In some embodiments, the plant seeds are incubated with the solution comprising the microorganism or its exudate for at least about 1 minute, about 5 minutes, about 10 minutes, about 20 minutes, about 60 minutes, or about 240 minutes. In some embodiments, the plant seeds are incubated with the solution comprising the microorganism or its exudate for up to about 5 minutes, about 10 minutes, about 20 minutes, about 60 minutes, about 240 minutes, or about 960 minutes. In some embodiments, the plant seeds are incubated with the solution comprising the microorganism or its exudate for about 1 minute. In some embodiments, the plant seeds are incubated with the solution comprising the microorganism or its exudate for about 5 minutes. In some embodiments, the plant seeds are incubated with the solution comprising the microorganism or its exudate for about 10 minutes. In some embodiments, the plant seeds are incubated with the solution containing the microorganism or its exudate for about 20 minutes. In some embodiments, the plant seeds are incubated with the solution containing the microorganism or its exudate for about 60 minutes. In some embodiments, the plant seeds are incubated with the solution containing the microorganism or its exudate for about 240 minutes. In some embodiments, the plant seeds are incubated with the solution containing the microorganism or its exudate for about 960 minutes.

[0100] In some embodiments, the seeds are incubated with the solution at a desired temperature. In some embodiments, the seeds are incubated with the solution at a temperature of about 2 to about 40° C. In some embodiments, the seeds are incubated with the solution at a temperature of about 2 to about 4° C., about 2 to about 8° C., about 2 to about 12° C., about 2 to about 16° C., about 2 to about 25° C., about 2 to about 30° C., about 2 to about 35° C., about 2 to about 40° C., about 4 to about 8° C., about 4 to about 12° C., about 4 to about 16° C., about 4 to about 25° C., about 4 to about 30° C., about 4 to about 35° C., about 4 to about 40° C., about 8 to about 12° C., about 8 to about 16° C., about 8 to about 25° C., about 8 to about 30° C., about 8 ...40° C. The solution is incubated at a temperature of about 35° C., about 8 to about 40° C., about 12 to about 16° C., about 12 to about 25° C., about 12 to about 30° C., about 12 to about 35° C., about 12 to about 40° C., about 16 to about 25° C., about 16 to about 30° C., about 16 to about 35° C., about 16 to about 40° C., about 25 to about 30° C., about 25 to about 35° C., about 25 to about 40° C., about 30 to about 35° C., about 30 to about 40° C., or about 35 to about 40° C. In some embodiments, the seeds are incubated with the solution at a temperature of about 2° C., about 4° C., about 8° C., about 12° C., about 16° C., about 25° C., about 30° C., about 35° C., or about 40° C. In some embodiments, the seeds are incubated with the solution at a temperature of at least about 2° C., about 4° C., about 8° C., about 12° C., about 16° C., about 25° C., about 30° C., or about 35° C. In some embodiments, the seeds are incubated with the solution at a temperature of at most about 4° C., about 8° C., about 12° C., about 16° C., about 25° C., about 30° C., about 35° C., or about 40° C.

[0101] In some embodiments, the method includes drying the seeds. In some embodiments, the seeds are dried to about 10% total seed moisture. In some embodiments, the seeds are dried to about 5% to about 25% total seed moisture. In some embodiments, the seeds are dried to about 5% to about 8%, about 5% to about 10%, about 5% to about 12%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 8% to about 10%, about 8% to about 12%, about 8% to about 15%, about 8% to about 20%, about 8% to about 25%, about 10% to about 12%, about 10% to about 15%, about 10% to about 20%, about 10% to about 25%, about 12% to about 15%, about 12% to about 20%, about 12% to about 25%, about 15% to about 20%, about 15% to about 25%, or about 20% to about 25% of the total seed moisture. In some embodiments, the seeds are dried to about 5%, about 8%, about 10%, about 12%, about 15%, about 20%, or about 25% of total seed moisture. In some embodiments, the seeds are dried to at least about 5%, about 8%, about 10%, about 12%, about 15%, or about 20% of total seed moisture. In some embodiments, the seeds are dried to a maximum of about 8%, about 10%, about 12%, about 15%, about 20%, or about 25% of total seed moisture. In some embodiments, the seeds are dried to prevent seed germination. In some embodiments, the seeds are dried to prevent germination before planting the seeds.

[0102] Formulations for incorporating microorganisms In one aspect, provided herein is a formulation for incorporating microorganisms, endospores, or exudates thereof into seeds. In some embodiments, the formulation comprises one or more microorganisms or endospores thereof and salts. The one or more microorganisms may be any of the microorganisms or endospores thereof provided herein. In some embodiments, the one or more microorganisms comprise one or more endospore-forming bacteria or endospores thereof. In some embodiments, the formulation comprises a microbial exudate. The exudate may be from any of the microorganisms provided herein.

[0103] In some embodiments, the formulation is a solution. In some embodiments, the formulation is an aqueous solution.

[0104] In some embodiments, the formulation includes a salt. The salt may be present in the formulation at any suitable concentration. In some embodiments, the formulation includes about 0.85% salt (w / v). In some embodiments, the formulation includes about 0.1% to about 1.25% salt (w / v). In some embodiments, the formulation includes about 0.1% to about 2.0% salt (w / v).In some embodiments, the formulation comprises about 0.1% to about 0.25%, about 0.1% to about 0.5%, about 0.1% to about 0.6%, about 0.1% to about 0.7%, about 0.1% to about 0.75%, about 0.1% to about 0.8%, about 0.1% to about 0.85%, about 0.1% to about 0.9%, about 0.1% to about 0.95%, about 0.1% to about 1%, about 0.1% to about 1.25%, about 0.25% to about 0.5%, about 0.25% to about 0.6%, about 0.25% to about 0.7%, about 0.25% to about 0.75%, about 0.25% to about 0.8%, about 0.25% to about 0.85%, about 0.25% to about 0.9%, about 0.25% to about 0.95%, about 0.25% to about 1%, about 0.25% to about 1.25%, about 0.5% to about 0.6%, about 0.5% to about 0.7%, about 0.5% to about 0.75%, about 0.5% to about 0.8%, about 0.5% to about 0.85%, about 0.5% to about 0.9%, about 0.5% to about 0.95%, about 0.5% to about 1%, about 0.5% to about 1.25%, about 0.6% to about 0.7%, about 0.6% to about 0.75%, about 0.6% to about 0. 8%, about 0.6% to about 0.85%, about 0.6% to about 0.9%, about 0.6% to about 0.95%, about 0.6% to about 1%, about 0.6% to about 1.25%, about 0.7% to about 0.75%, about 0.7% to about 0.8%, about 0.7% to about 0.85%, about 0.7% to about 0.9%, about 0.7% to about 0.95%, about 0.7% to about 1%, about 0.7% to about 1.25%, about 0.75% to about 0.8%, about 0.75% to about 0.85%, about 0.75% to about 0.9%, about 0.75% to about 0.95%, about 0.75% to about 1 , about 0.75% to about 1.25%, about 0.8% to about 0.85%, about 0.8% to about 0.9%, about 0.8% to about 0.95%, about 0.8% to about 1%, about 0.8% to about 1.25%, about 0.85% to about 0.9%, about 0.85% to about 0.95%, about 0.85% to about 1%, about 0.85% to about 1.25%, about 0.9% to about 0.95%, about 0.9% to about 1%, about 0.9% to about 1.25%, about 0.95% to about 1%, about 0.95% to about 1.25%, or about 1% to about 1.25% salt (w / v). In some embodiments, the formulation comprises about 0.1%, about 0.25%, about 0.5%, about 0.6%, about 0.7%, about 0.75%, about 0.8%, about 0.85%, about 0.9%, about 0.95%, about 1%, or about 1.25% salt (w / v).In some embodiments, the formulation contains at least about 0.1%, about 0.25%, about 0.5%, about 0.6%, about 0.7%, about 0.75%, about 0.8%, about 0.85%, about 0.9%, about 0.95%, or about 1% salt (w / v). In some embodiments, the formulation contains up to about 0.25%, about 0.5%, about 0.6%, about 0.7%, about 0.75%, about 0.8%, about 0.85%, about 0.9%, about 0.95%, about 1%, or about 1.25% salt (w / v). In some embodiments, the formulation contains about 0.85% salt (w / v). In some embodiments, the formulation contains about 0.8% to about 0.9% salt (w / v). In some embodiments, the formulation contains about 0.75% to about 0.95% salt (w / v). In some embodiments, the formulation comprises about 0.7% to about 1% salt (w / v). In some embodiments, the formulation comprises about 0.5% to about 1.25% salt (w / v). In some embodiments, the formulation comprises about 0.5% to about 2% salt (w / v). In some embodiments, the formulation comprises 0.1-0.2%, 0.2-0.3%, 0.3-0.4%, 0.4-0.5%, 0.5-0.6%, 0.6-0.7%, 0.7-0.8%, 0.8-0.9%, 0.9-1.0%, 1.0-1.1%, 1.1-1.2%, 1.2-1.3%, 1.3-1.4%, or 1.4-1.5% salt (w / v).

[0105] Any salt may be used. In some preferred embodiments, the salt is NaCl. In some embodiments, the salt is NaCl, LiCl, KCl, MgCl, CaCl, NaBr, LiBr, KBr, MgBr, CaBr, NaI, LiI, KI, MgI, or CaI. In some embodiments, the salt comprises sodium, lithium, or potassium ions. In some embodiments, the salt comprises alkali metal ions. In some embodiments, the salt comprises alkaline earth metal ions. In some embodiments, the salt comprises halide ions. In some embodiments, the salt is an alkali or alkaline earth halide salt. In some embodiments, the salt comprises chloride, bromide, or iodide ions. In some embodiments, the salt is a sulfate, phosphate, carbonate, or nitrate salt.

[0106] In some embodiments, the formulation comprises additional additives, such as dimethyl sulfoxide (DMSO), 1-dodecylazacycloheptan-2-one, laurocapram, 1-methyl-2-pyrrolidone (NMP), oleic acid, ethanol, methanol, polyethylene glycol (Brij 35, 58, 98), polyethylene glycol monolaurate (e.g., Tween 20), Tween 40 (polyoxyethylene acid sorbitol ester), Tween 60, Tween 80 (non-ionic), cetylmethylammonium bromide (CTAB), urea, lecithin (soybean-derived solidified fatty acid), chitosan, poloxamer 188, poloxamer 237, poloxamer 338, poloxamer 407, or a combination thereof. In some embodiments, the formulation comprises polyethylene glycol monolaurate (e.g., Tween 20), poloxamer 188, poloxamer 237, poloxamer 338, poloxamer 407, or a combination thereof. In some embodiments, the formulation comprises a poloxamer. In some embodiments, the formulation comprises polyethylene glycol monolaurate (e.g., Tween 20). The additional additives may be present in any concentration. In some embodiments, the additional additives comprise up to about 0.01%, 0.05%, 0.1%, 0.125%, 0.15%, 0.2%, 0.5%, or 1% (v / v) of the formulation. In some embodiments, the additional additives comprise from about 0.01% to about 1% (v / v) of the formulation. In some embodiments, the additional additives comprise about 0.1% (v / v) of the formulation.

[0107] In some embodiments, the formulation includes an additional metal ion. In some embodiments, the formulation includes magnesium, calcium, manganese, or a combination thereof. In some embodiments, the formulation includes magnesium. In some embodiments, the formulation includes calcium. In some embodiments, the formulation includes manganese. In some embodiments, the formulation includes magnesium and calcium. In some embodiments, the formulation includes magnesium and manganese. In some embodiments, the formulation includes calcium and manganese. In some embodiments, the formulation includes magnesium, calcium, and manganese.

[0108] In some embodiments, the formulation comprises one or more nutrients for the microorganism. In some embodiments, the formulation comprises a bacterial growth medium. In some embodiments, the formulation comprises lysogeny broth (LB), nutrient broth, or a combination thereof. In some embodiments, the formulation comprises lysogeny broth. In some embodiments, the formulation comprises nutrient broth.

[0109] In some embodiments, the formulation includes additional components to promote endosporulation of one or more microorganisms. In some embodiments, the formulation includes potassium, ferrous sulfate, calcium, magnesium, manganese, or a combination thereof. In some embodiments, the formulation includes potassium. In some embodiments, the formulation includes iron sulfide. In some embodiments, the formulation includes calcium. In some embodiments, the formulation includes magnesium. In some embodiments, the formulation includes manganese.

[0110] In some embodiments, the preparation comprises a microorganism. In some embodiments, the preparation comprises about 10 3 ~about 10 17 In some embodiments, the formulation contains at least 1 x 10 colony forming units (CFU) / mL of microorganisms. 6 In some embodiments, the formulation contains about 10 CFU / mL of microorganisms. 3 ~about 104 , about 10 3 ~about 10 5 , about 10 3 ~about 10 6 , about 10 3 ~about 10 7 , about 10 3 ~about 10 8 , about 10 3 ~about 10 9 , about 10 3 ~about 10 10 , about 10 3 ~about 10 12 , about 10 3 ~about 10 15 , about 10 3 ~about 10 17 , about 10 4 ~about 10 5 , about 10 4 ~about 10 6 , about 10 4 ~about 10 7 , about 10 4 ~about 10 8 , about 10 4 ~about 10 9 , about 10 4 ~about 10 10 , about 10 4 ~about 10 12 , about 10 4 ~about 10 15 , about 10 4 ~about 10 17 , about 10 5 ~about 10 6 , about 10 5 ~about 10 7 , about 10 5 ~about 10 8 , about 10 5 ~about 10 9 , about 10 5 ~about 10 10 , about 10 5 ~about 10 12 , about 10 5 ~about 10 15 , about 10 5 ~about 10 17 , about 10 6 ~about 10 7 , about 10 6 ~about 10 8 , about 10 6 ~about 10 9 , about 106 ~about 10 10 , about 10 6 ~about 10 12 , about 10 6 ~about 10 15 , about 10 6 ~about 10 17 , about 10 7 ~about 10 8 , about 10 7 ~about 10 9 , about 10 7 ~about 10 10 , about 10 7 ~about 10 12 , about 10 7 ~about 10 15 , about 10 7 ~about 10 17 , about 10 8 ~about 10 9 , about 10 8 ~about 10 10 , about 10 8 ~about 10 12 , about 10 8 ~about 10 15 , about 10 8 ~about 10 17 , about 10 9 ~about 10 10 , about 10 9 ~about 10 12 , about 10 9 ~about 10 15 , about 10 9 ~about 10 17 , about 10 10 ~about 10 12 , about 10 10 ~about 10 15 , about 10 10 ~about 10 17 , about 10 12 ~about 10 15 , about 10 12 ~about 10 17 , or about 10 15 ~about 10 17 In some embodiments, the formulation contains about 10 CFU / mL of microorganisms. 3 , about 10 4 , about 10 5 , about 10 6 , about 10 7 , about 10 8 , about 10 9 , about 1010 , about 10 12 , about 10 15 , or about 10 17 In some embodiments, the formulation contains at least about 10 CFU / mL of microorganisms. 3 , about 10 4 , about 10 5 , about 10 6 , about 10 7 , about 10 8 , about 10 9 , about 10 10 , about 10 12 , or about 10 15 In some embodiments, the formulation contains up to about 10 CFU / mL of microorganisms. 4 , about 10 5 , about 10 6 , about 10 7 , about 10 8 , about 10 9 , about 10 10 , about 10 12 , about 10 15 , or about 10 17 In some embodiments, the formulation contains at least 10 CFU / mL of microorganisms. 6 ~10 7 In some embodiments, the formulation contains 1 x 10 CFU / mL of microorganisms. 3 ~1×10 4 CFU / mL: 1 x 10 4 ~1×10 5 CFU / mL: 1 x 10 5 ~1×10 6 CFU / mL: 1 x 10 6 ~1×10 7 CFU / mL: 1 x 10 7 ~1×10 8 CFU / mL: 1 x 10 8 ~1×10 9 CFU / mL: 1 x 10 9 ~1×10 10 CFU / mL: 1 x 10 10 ~1×10 11 CFU / mL: 1 x 10 11 ~1×10 12 CFU / mL: 1 x 10 12 ~1×10 13 CFU / mL: 1 x 1013 ~1×10 14 CFU / mL: 1 x 10 14 ~1×10 15 CFU / mL: 1 x 10 15 ~1×10 16 CFU / mL; or 1 × 10 16 ~1×10 17 CFU / mL of the microorganism. The microorganism may be any of the microorganisms provided herein or an endospore of any of the microorganisms provided herein.

[0111] In some embodiments, the formulation is maintained at a desired temperature. In some embodiments, the formulation is maintained at a temperature of about 2 to about 40° C. In some embodiments, the formulation is maintained at a temperature of about 2 to about 4, about 2 to about 8, about 2 to about 12, about 2 to about 16, about 2 to about 25, about 2 to about 30, about 2 to about 35, about 2 to about 40, about 4 to about 8, about 4 to about 12, about 4 to about 16, about 4 to about 25, about 4 to about 30, about 4 to about 35, about 4 to about 40, about 8 to about 12, about 8 to about 16, about 8 to about 25, about 8 to about 30, The formulation is maintained at a temperature of about 8 to about 35° C., about 8 to about 40° C., about 12 to about 16° C., about 12 to about 25° C., about 12 to about 30° C., about 12 to about 35° C., about 12 to about 40° C., about 16 to about 25° C., about 16 to about 30° C., about 16 to about 35° C., about 16 to about 40° C., about 25 to about 30° C., about 25 to about 35° C., about 25 to about 40° C., about 30 to about 35° C., or about 35 to about 40° C. In some embodiments, the formulation is maintained at a temperature of about 2, about 4, about 8, about 12, about 16, about 25, about 30, about 35, or about 40° C. In some embodiments, the formulation is maintained at a temperature of at least about 2, about 4, about 8, about 12, about 16, about 25, about 30, or about 35° C. In some embodiments, the formulation is maintained at a temperature of up to about 4, about 8, about 12, about 16, about 25, about 30, about 35, or about 40°C.

[0112] Microorganisms and leachables The microorganisms provided herein or their exudates can produce plant growth-promoting effects when incorporated into plant seeds. In some embodiments, the microorganisms are bacteria. In some embodiments, the microorganisms are endospore-forming bacteria. In some embodiments, the microorganisms are bacterial endospores. Whenever a microorganism (e.g., bacteria) referred to herein can form endospores, any endospores of the microorganism are also intended to be encompassed. For example, if a plant seed treatment formulation contains Bacillus sp., the formulation may contain endospores of Bacillus sp.

[0113] In some embodiments, the microorganism is a microorganism from the phyla Firmicutes, Proteobacteria, and Actinobacteria. In some embodiments, the microorganism is a microorganism of the phylum Firmicutes. In some embodiments, the microorganism is a microorganism of the phylum Proteobacteria. In some embodiments, the microorganism is a microorganism of the phylum Actinobacteria. In some embodiments, the microorganism is an endospore of any of the microorganisms.

[0114] In some embodiments, the microorganism is Acetonema sp.、Actinomyces sp.、Alkalibacillus sp.、Ammoniphilus sp.、Amphibacillus sp.、Anaerobacter sp.、Anaerospora sp.、Aneurinibacillus sp.、Anoxybacillus sp.、Bacillus sp.、Brevibacillus sp.、Caldanaerobacter sp.、Caloramator sp.、Caminicella sp.、Cerasibacillus sp.、Clostridium sp.、Clostridiisalibacter sp.、Cohnella sp.、Coxiella sp.、Dendrosporobacter sp.、Desulfotomaculum sp.、Desulfosporomusa sp.、Desulfosporosinus sp.、Desulfovirgula sp.、Desulfunispora sp.、Desulfurispora sp.、Filifactor sp.、Filobacillus sp.、Gelria sp.、Geobacillus sp.、Geosporobacter sp.、Gracilibacillus sp.、Halobacillus sp.、Halonatronum sp.、Heliobacterium sp.、Heliophilum sp.、Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribacillus sp., Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus sp., Thermoflavimicrobium sp., Thermovenabulum sp., Tuberibacillus sp., Virgibacillus sp., and Vulcanobacillus sp. In some embodiments, the microorganism is a microorganism selected from Acetobacter sp., Actinomyces sp., Bacillus sp., Chryseobacterium sp., Coxiella sp., Ensifer sp., Glutamicibacter sp., Microbacterium sp., and Serratia sp. In some embodiments, the microorganism is an Acetobacter sp. In some embodiments, the microorganism is an Actinomyces sp. In some embodiments, the microorganism is a Bacillus sp. In some embodiments, the microorganism is a Chryseobacterium sp. In some embodiments, the microorganism is a Coxiella sp. In some embodiments, the microorganism is an Ensifer sp.In some embodiments, the microorganism is a Glutamicibacter sp. In some embodiments, the microorganism is a Microbacterium sp. In some embodiments, the microorganism is a Pantoea sp. In some embodiments, the microorganism is a Serratia sp. In some embodiments, the microorganism is an endospore of any of the microorganisms.

[0115] In some embodiments, the microorganism is Acetobacter cerevisiae, Bacillus cucumis, Bacillus endophyticus, Bacillus megaterium, Bacillus nakamurai, Bacillus subtilis, Chryseobacterium lactis, Ensifer adhaerens, Glutamicibacter arilaitensis, Glutamicibacter halophytocola, Microbacterium chocolatum, Microbacterium yannicii, Pantoea allii, Serratia marcescens, or Serratia ureilytica. In some embodiments, the microorganism comprises Acetobacter cerevisiae, Bacillus cucumis, Bacillus endophyticus, Bacillus megaterium, Bacillus nakamurai, Bacillus subtilis, Chryseobacterium lactis, Ensifer adhaerens, Glutamicibacter halophytocola, Microbacterium chocolatum, Pantoea allii, or Serratia marcescens. In some embodiments, the microorganism comprises Acetobacter cerevisiae. In some embodiments, the microorganism comprises Bacillus cucumis. In some embodiments, the microorganism comprises Bacillus endophyticus. In some embodiments, the microorganism comprises Bacillus megaterium. In some embodiments, the microorganism comprises Bacillus subtilis. In some embodiments, the microorganism comprises Chryseobacterium lactis. In some embodiments, the microorganism comprises Ensifer adhaerens. In some embodiments, the microorganism comprises Glutamicibacter halophytocola. In some embodiments, the microorganism comprises Microbacterium chocolatum.In some embodiments, the microorganism comprises Pantoea allii. In some embodiments, the microorganism comprises Serratia marcescens. In some embodiments, the microorganism is an endospore of any microorganism.

[0116] In some embodiments, the microorganism is an endospore-forming bacterium. In some embodiments, the endospore-forming bacterium is from the genus Bacillus. In some embodiments, the endospore-forming bacterium is from a Bacillus sp. In some embodiments, the endospore-forming bacterium includes Bacillus cucumis, Bacillus endophyticus, Bacillus megaterium, Bacillus nakamurai, and Bacillus subtilis. In some embodiments, the endospore-forming bacterium includes Bacillus cucumis, Bacillus endophyticus, Bacillus megaterium, or Bacillus subtilis. In some embodiments, the endospore-forming bacterium includes Bacillus cucumis. In some embodiments, the endospore-forming bacterium includes Bacillus megaterium. In some embodiments, the endospore-forming bacterium includes Bacillus nakamurai. In some embodiments, the endospore-forming bacterium includes Bacillus subtilis. In some embodiments, the microorganism is an endospore of any of the microorganisms.

[0117] In some embodiments, the microorganism is an endospore. In some embodiments, the endospore is from the genus Bacillus. In some embodiments, the endospore is a Bacillus sp. In some embodiments, the endospore comprises Bacillus cucumis, Bacillus endophyticus, Bacillus megaterium, Bacillus nakamurai, or Bacillus subtilis. In some embodiments, the endospore comprises Bacillus cucumis, Bacillus endophyticus, Bacillus megaterium, or Bacillus subtilis. In some embodiments, the endospore comprises Bacillus cucumis. In some embodiments, the endospore comprises Bacillus megaterium. In some embodiments, the endospore comprises Bacillus nakamurai. In some embodiments, the endospore comprises Bacillus subtilis.

[0118] This is the best way to make a slightly less expensive carpet The active ingredient is Acetonema sp. Actinomyces sp. Alkalibacillus sp. Ammoniphilus sp. Amphibacillus sp. Anaerobacter sp. Anaerospora sp. Aneurinibacillus sp. Anoxybacillus sp. Bacillus sp sp. Caldanaerobacter sp. Caloramator sp. Caminicella sp. Cerasibacillus sp. Clostridium sp. Clostridiisalibacter sp. Cohnella sp. Coxiella sp. Dendrosporobacter sp. Desulfotomaculum sp sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Phylobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp. Halonatronum sp. Heliobacterium sp. Heliophilum sp. Laceyella sp. Lentibacillus sp. Lysinibacillus sp. Mahela sp. Metabacterium sp. Moorella sp sp. Ornithinibacillus sp. Oxalophagus sp. Oxobacter sp. Paenibacillus sp. Paraliobacillus sp. Pelospora sp. Pelotomaculum sp. Piscibacillus sp. Planifilum sp.Pontibacillus sp. Propionispora sp. Salinibacillus sp. Salsuginibacillus sp. Seinonella sp. Shimazuella sp. Sporacetigenium sp sp. Sporohalobacter sp. Sporolactobacillus sp. Sporomusa sp. Sporosarcina sp. Sporotalea sp. Sporotomaculum sp. Syntrophomonas sp. Syntrophospora sp. Tenuibacillus sp sp. Thalassobacillus sp. Thermoacetogenium sp. Thermoactinomyces sp. Thermoalkalibacillus sp. Thermoanaerobacter sp. Thermoanaeromonas sp sp., Tuberibacillus sp., Virgibacillus sp., and Vulcanobacillus sp. The 2-year-old sp The main strain is Acetobacter sp., Actinomyces sp., Bacillus sp., Chryseobacterium sp., Coxiella sp., Ensifer sp., Glutamicibacter sp., Microbacterium sp., and Serratia sp.In some embodiments, the consortium comprises two or more bacteria selected from the group consisting of: (a) a bacterium selected from the group consisting of: (b) a bacterium selected from the group consisting of: (c) a bacterium selected from the group consisting of: (d) a bacterium selected from the group consisting of: (e) a bacterium selected from the group consisting of: (i ...

[0119] In some embodiments, the consortium comprises bacteria from the phylum Bacillus and one or more bacteria. In some embodiments, the consortium comprises bacteria from the phylum Bacillus and a subset of bacteria, including Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribacillus sp., Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus sp., Thermoflavimicrobium sp., Thermovenabulum sp., Tuberibacillus sp., Virgibacillus sp., and Vulcanobacillus sp. In some embodiments, the consortium comprises two or more bacteria selected from Acetobacter sp., Actinomyces sp., Bacillus sp., Chryseobacterium sp., Coxiella sp., Ensifer sp., Glutamicibacter sp., Microbacterium sp., and Serratia sp. In some embodiments, the consortium comprises endospores of any of the microorganisms.

[0120] In some embodiments, the consortium comprises a mixture of two or more bacteria selected from Bacillus endophyticus, Bacillus megaterium, Bacillus nakamurai, Bacillus subtilis, Chryseobacterium lactis, Ensifer adhaerens, Glutamicibacter arilaitensis, Glutamicibacter halophytocola, Microbacterium chocolatum, Microbacterium yannicii, Pantoea allii, Serratia marcescens, and Serratia ureilytica. In some embodiments, the consortium comprises a mixture of two or more bacteria selected from Acetobacter cerevisiae, Bacillus cucumis, Bacillus endophyticus, Bacillus megaterium, Bacillus nakamurai, Bacillus subtilis, Chryseobacterium lactis, Ensifer adhaerens, Glutamicibacter halophytocola, Microbacterium chocolatum, Pantoea allii, and Serratia marcescens. In some embodiments, the consortium comprises a mixture of 2, 3, 4, 5, 6, 7, 8, 9, or 10 bacteria. In some embodiments, the consortium comprises two bacteria. In some embodiments, the consortium comprises three bacteria. In some embodiments, the consortium comprises four bacteria. In some embodiments, the consortium comprises five bacteria. In some embodiments, the consortium comprises six bacteria. In some embodiments, the consortium includes endospores of any of the microorganisms.

[0121] In some embodiments, the consortium comprises two or more bacteria selected from Acetobacter cereviseae, Chryseobacterium lactis, Bacillus cucumis, Bacillus endophyticus, Bacillus megaterium, Bacillus subtilis, and Ensifer adhaerens. In some embodiments, the consortium comprises two bacteria. In some embodiments, the consortium comprises three bacteria. In some embodiments, the consortium comprises four bacteria. In some embodiments, the consortium comprises five bacteria. In some embodiments, the consortium comprises six bacteria. In some embodiments, the consortium comprises seven bacteria. In some embodiments, the consortium comprises endospores of any of the microorganisms.

[0122] In some embodiments, the consortium includes two or more bacteria selected from Acetobacter cereviseae, Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. In some embodiments, the consortium includes two bacteria selected from Acetobacter cereviseae, Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. In some embodiments, the consortium includes two bacteria selected from Acetobacter cereviseae, Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. In some embodiments, the consortium includes three bacteria selected from Acetobacter cereviseae, Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. In some embodiments, the consortium comprises a mixture of Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. In some embodiments, the consortium comprises a mixture of Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. In some embodiments, the consortium comprises endospores of any of the microorganisms.

[0123] In some embodiments, the consortium comprises two or more bacteria selected from Bacillus subtilis, Bacillus cucumis, and Ensifer adhaerens. In some embodiments, the consortium comprises Ensifer adhaerens and Bacillus subtilis or Bacillus cucumis. In some embodiments, the consortium comprises Ensifer adhaerens and Bacillus subtilis. In some embodiments, the consortium comprises Ensifer adhaerens and Bacillus subtilis. In some embodiments, the consortium comprises Ensifer adhaerens and Bacillus cucumis. In some embodiments, the consortium comprises endospores of any of the microorganisms.

[0124] In some embodiments, the exudates of any of the microorganisms provided herein are incorporated into cells. In some embodiments, the exudate is Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp.Pontibacillus sp. Propionispora sp. Salinibacillus sp. Salsuginibacillus sp. Seinonella sp. Shimazuella sp. Sporacetigenium sp sp. Sporohalobacter sp. Sporolactobacillus sp. Sporomusa sp. Sporosarcina sp. Sporotalea sp. Sporotomaculum sp. Syntrophomonas sp. Syntrophospora sp. Tenuibacillus sp sp. Thalassobacillus sp. Thermoacetogenium sp. Thermoactinomyces sp. Thermoalkalibacillus sp. Thermoanaerobacter sp. Thermoanaeromonas sp sp., Tuberibacillus sp., Virgibacillus sp sp., the strains of which are Acetobacter sp., Actinomyces sp., Bacillus sp., Chryseobacterium sp., Coxiella sp., Ensifer sp., Glutamicibact sp., Microbacterium sp., Serratia sp.In some embodiments, the exudate is from Acetobacter cerevisiae, Bacillus cucumis, Bacillus endophyticus, Bacillus megaterium, Bacillus nakamurai, Bacillus subtilis, Chryseobacterium lactis, Ensifer adhaerens, Glutamicibacter arilaitensis, Glutamicibacter halophytocola, Microbacterium chocolatum, Microbacterium yannicii, Pantoea allii, Serratia marcescens, or Serratia ureilytica. In some embodiments, the exudate is from Bacillus cucumis, Bacillus endophyticus, Bacillus megaterium, Bacillus nakamurai, or Bacillus subtilis. In some embodiments, the exudate is from the endospores of any of the microorganisms.

[0125] In some embodiments, the microorganisms provided herein comprise genes encoding one or more compounds that induce plant growth. In some embodiments, the compounds induce systemic induced resistance (IST). In some embodiments, the compounds induce systemic induced resistance (ISR). In some embodiments, the microorganisms comprise genes associated with nitrogen fixation, phosphate solubilization, or plant hormone synthesis, or any combination thereof. In some embodiments, the microorganisms comprise genes associated with nitrogen fixation. In some embodiments, the microorganisms comprise genes associated with phosphate solubilization. In some embodiments, the microorganisms comprise genes associated with plant hormone synthesis.

[0126] In some embodiments, the microorganisms are selected for one or more characteristics related to the microorganisms' ability to interact with plants. In some embodiments, the microorganisms are selected for compatibility. In some embodiments, the microorganisms are selected to ensure that predation or antagonism does not occur. In some embodiments, the microorganisms are selected for stability during storage. In some embodiments, the microorganisms are selected for rapid plant colonization and survival within relevant tissues. In some embodiments, the microorganisms are selected for stimulation of large-scale, long-term physiological responses in the plant. In some embodiments, the microorganisms are selected for promotion of the plant's life cycle. In some embodiments, the microorganisms are selected for optimal incorporation into one or more seeds. In some embodiments, the microorganisms persist throughout the plant's life cycle.

[0127] In some embodiments, the microorganisms incorporated into the seeds are stable after incorporation. In some embodiments, the microorganisms are stable for more than 30 days, more than 6 months, more than 1 year, or more than 2 years. In some embodiments, the microorganisms are stable for more than 30 days. In some embodiments, the microorganisms are stable for more than 6 months. In some embodiments, the microorganisms are stable for more than 1 year. In some embodiments, the microorganisms are stable for more than 2 years.

[0128] Whenever the terms "at least," "greater than," or "equivalent to" precede the first number in a series of two or more numbers, the terms "at least," "greater than," or "equivalent to" apply to each and every number in the series. For example, 1, 2, or 3 or more are equivalent to 1 or more, 2 or more, or 3 or more.

[0129] Whenever the terms "no more than," "less than," or "less than or equal to" precede the first number in a series of two or more numbers, the terms "no more than," "less than," or "less than or equal to" apply to each and every number in the series. For example, 3, 2, or 1 or less is equivalent to 3 or less, 2 or less, or 1 or less.

[0130] Numbered Embodiments 1. An engineered seed, the engineered seed comprising: (i) a seed coat and an embryo, the seed coat having a gap therebetween; and (ii) one or more microorganisms disposed in the gap. 2. An engineered seed, the engineered seed comprising: (i) a seed pericarp and a seed aleurone cell layer, the seed coat having a gap therebetween; and (ii) one or more microorganisms disposed in the gap. 3. An engineered seed, the engineered seed comprising: (i) a seed pericarp and a seed aleurone cell layer; and (ii) one or more microorganisms disposed between the seed pericarp and the seed aleurone cell layer. 4. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms are selected to produce a plant growth-promoting effect. 5. The engineered seed of any one of the preceding embodiments, wherein the seed is a monocotyledonous plant seed. 6. The engineered seed of any one of the preceding embodiments, wherein the seed is selected from maize, rice, wheat, and sorghum seeds. 7. The engineered seed of any one of the preceding embodiments, wherein the seed is a corn seed. 8. The engineered seed of any one of the preceding embodiments, wherein the seed is a corn (Zea maize) seed. 9. The engineered seed of any one of the preceding embodiments, wherein the seed is a dicotyledonous seed. 10. The engineered seed of any one of the preceding embodiments, wherein the seed is selected from soybean, cotton, alfalfa, bean, quinoa, lentil, peanut, lettuce, tomato, and cabbage seeds. 11. The engineered seed of any one of the preceding embodiments, wherein the seed is a lettuce seed or a tomato seed. 12. The engineered seed of any one of the preceding embodiments, wherein the seed is a lettuce seed or a tomato seed. 13. The engineered seed of any one of the preceding embodiments, wherein the seed is a GMO seed. 14. The engineered seed of any one of the preceding embodiments, wherein the seed is a non-GMO seed. 15. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise a mixture of Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium.16. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise a mixture of Acetobacter cereviseae, Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. 17. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise a mixture of Ensifer adhaerens and Bacillus nakamurai, Bacillus subtilis, or Bacillus cucumis. 18. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise Microbacterium yannicii or Microbacterium chocolatum. 19. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise Serratia ureilytica or Serratia marcescens. 20. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise Glutamicibacter arilaitensis or Glutamicibacter halophytocola. 21. The engineered seed of any one of the preceding embodiments, wherein the one or more microbial organisms comprise Ensifer adhaerens, Pantoea allii, Bacillus subtilis, or Bacillus cucumis. 22. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise an endospore-forming microorganism. 23. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise a Bacillus sp. 24. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms are selected from the phyla Firmicutes, Proteobacteria, and Actinobacteria. 25. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms are selected from the phylum Firmicutes. 26. The one or more microorganisms are Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribacillus sp., Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus sp., Thermoflavimicrobium sp., Thermovenabulum sp., Tuberibacillus sp., Virgibacillus sp., and Vulcanobacillus sp. 27. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms are selected from the phylum Proteobacteria. 28. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise Actinomyces sp. 29. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms are selected from the phylum Actinobacteria. 30. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise Coxiella sp. 31. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms form endospores after being disposed within the seed. 32. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise Bacillus sp. 33. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise endospores. 34.34. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise a 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. 35. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise a 16S nucleic acid sequence at least 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, or 90% identical to any of SEQ ID NOs: 1-10221. 36. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise a 16S nucleic acid sequence at least 99% identical to any of SEQ ID NOs: 1-10221. 37. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise a 16S nucleic acid sequence at least 98% identical to any of SEQ ID NOs: 1-10221. 38. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise a 16S nucleic acid sequence at least 95% identical to any of SEQ ID NOs: 1-10221. 39. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise a 16S nucleic acid sequence at least 90% identical to any of SEQ ID NOs: 1-10221. 40. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise a gene encoding one or more compounds that cause systemic induced resistance (IST). 41. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise a gene encoding one or more compounds that cause systemic induced resistance (ISR). 42. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise a gene encoding one or more compounds that induce plant growth. 43. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms comprise a gene associated with nitrogen fixation. 44. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms have a gene associated with phosphate solubilization. 45. The engineered seed of any one of the preceding embodiments, wherein the one or more microorganisms have a gene associated with plant hormone synthesis. 46. The engineered seed of any one of the preceding embodiments, further comprising a microbial exudate. 47.48. The engineered seed of any one of the preceding embodiments, wherein the microbial percolate contains one or more compounds that cause systemic induced resistance (IST). 49. The engineered seed of any one of the preceding embodiments, wherein the microbial percolate contains one or more compounds that cause systemic induced resistance (ISR). 50. The engineered seed of any one of the preceding embodiments, wherein the microbial percolate contains one or more compounds that cause plant growth. 50. The engineered seed of any one of the preceding embodiments, wherein the microbial percolate is from a microorganism comprising a 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. 51. The engineered seed of any one of the preceding embodiments, wherein the microbial percolate is from a microorganism comprising a 16S nucleic acid sequence at least 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, or 90% identical to any of SEQ ID NOs: 1-10221. 52. The engineered seed of any one of the preceding embodiments, wherein the microbial percolate is from a microorganism comprising a 16S nucleic acid sequence at least 99% identical to any of SEQ ID NOs: 1-10221. 53. The engineered seed of any one of the preceding embodiments, wherein the microbial percolate is from a microorganism comprising a 16S nucleic acid sequence at least 98% identical to any of SEQ ID NOs: 1-10221. 54. The engineered seed of any one of the preceding embodiments, wherein the microbial percolate is from a microorganism comprising a 16S nucleic acid sequence that is at least 95% identical to any of SEQ ID NOs: 1-10221. 55. The engineered seed of any one of the preceding embodiments, wherein the microbial percolate is from a microorganism comprising a 16S nucleic acid sequence that is at least 90% identical to any of SEQ ID NOs: 1-10221.

[0131] 56. A method for treating one or more plant seeds, the method comprising: soaking the one or more seeds in a formulation, the formulation comprising salt and one or more microorganisms selected to produce a plant growth-promoting effect; and incubating the one or more seeds in the formulation for a period of time sufficient to incorporate the one or more microorganisms into the seeds.

[0132] 57. A method for treating one or more plant seeds, the method comprising: (i) soaking one or more seeds in a formulation, the formulation comprising salt and one or more microorganisms selected to produce a plant growth-promoting effect; and (ii) incubating the one or more seeds to incorporate bacteria between the seed pericarp and the seed aleurone cell layer. 58. The method of embodiment 57, wherein the one or more plants remain dormant after treatment. 59. The method of embodiment 57 or embodiment 58, wherein the one or more plants remain dormant after treatment. 60. The method of any one of embodiments 57-59, wherein the one or more microorganisms are incorporated inside the seed pericarp. 61. The method of any one of embodiments 57-60, wherein the one or more microorganisms are incorporated between the seed pericarp and the aleurone cell layer. 62. The method of any one of embodiments 57-61, further comprising removing the one or more seeds from the formulation. 63. The method of any one of embodiments 57-62, further comprising thoroughly sterilizing the one or more seeds before soaking the one or more seeds in the formulation. 64. The method of any one of embodiments 57-63, further comprising drying the one or more seeds. 65. The method of any one of embodiments 57-64, wherein the one or more seeds are dried to about 10% of their total seed moisture. 66. The method of any one of embodiments 57-65, further comprising drying the one or more seeds to prevent germination. 67. The method of any one of embodiments 57-66, further comprising sterilizing the surface of the one or more seeds before soaking the one or more seeds in the formulation. 68. The method of any one of embodiments 57-67, further comprising sterilizing the surface of the one or more seeds after soaking the one or more seeds in the formulation. 69. The method of any one of embodiments 57-68, further comprising adding a fungicide to the surface of the seeds. 70. The method of any one of embodiments 57-69, wherein the one or more seeds are seeds of a monocotyledonous plant. 71. The method of any one of embodiments 57-70, wherein the seeds are selected from corn, wheat, rice, barley, rye, sugarcane, millet, oats, and sorghum. 72.The method of any one of embodiments 57-71, wherein the seed is a maize seed. 73. The method of any one of embodiments 57-72, wherein the seed is a maize (Zea mays) seed. 74. The method of any one of embodiments 57-73, wherein the seed is a dicotyledonous seed. 75. The method of any one of embodiments 57-74, wherein the seed is selected from soybean, cotton, alfalfa, bean, quinoa, lentil, peanut, lettuce, tomato, and cabbage seeds. 76. The method of any one of embodiments 57-75, wherein the seed is a lettuce seed or a tomato seed. 77. The method of any one of embodiments 57-76, wherein the seed is a lettuce seed or a tomato seed. 78. The method of any one of embodiments 57-77, wherein the seed is a GMO seed. 79. The method of any one of embodiments 57-78, wherein the seed is a non-GMO seed. 80. The method of any one of embodiments 57-79, wherein the formulation is an aqueous formulation. 81. The method of any one of embodiments 57-80, wherein the formulation further comprises poloxamer 188. 82. The method of any one of embodiments 57-81, wherein the formulation further comprises poloxamer 188 at a concentration of 0.1%. 83. The method of any one of embodiments 57-82, wherein the formulation further comprises Tween 20. 84. The method of any one of embodiments 57-83, wherein the formulation further comprises one or more agents selected from the group consisting of dimethyl sulfoxide (DMSO), 1-dodecylazacycloheptan-2-one, laurocapram, 1-methyl-2-pyrrolidone (NMP), oleic acid, ethanol, methanol, polyethylene glycol (Brij 35, 58, 98), polyethylene glycol monolaurate (Tween 20), Tween 40 (polyoxyethylene acid sorbitol ester), Tween 60, Tween 80 (nonionic), cetylmethylammonium bromide (CTAB), urea, lecithin (solidified fatty acid from soy), chitosan, poloxamer 188, poloxamer 237, poloxamer 338, and poloxamer 407.The method of any one of embodiments 57-84, wherein the formulation further comprises one or more components that promote endospore formation of one or more bacteria. 86. The method of any one of embodiments 57-85, wherein the formulation comprises potassium, ferrous sulfate, calcium, magnesium, manganese, or a combination thereof. 87. The method of any one of embodiments 57-86, wherein the formulation further comprises manganese. 88. The method of any one of embodiments 57-87, wherein the formulation comprises calcium, magnesium, and manganese. 89. The method of any one of embodiments 57-88, wherein the formulation comprises nutrients for one or more microorganisms. 90. The method of any one of embodiments 57-89, wherein the formulation is at room temperature. 91. The method of any one of embodiments 57-90, wherein the formulation is at a temperature of about 4°C, 10°C, 15°C, 20°C, or 30°C. 92. The method of any one of embodiments 57-91, wherein the formulation is at a temperature between about 4° C. and 20° C., or between about 30° C. and 40° C. 93. The method of any one of embodiments 57-92, wherein the formulation temperature is between about 20° C. and 24° C. 94. The method of any one of embodiments 57-93, wherein the formulation is at a temperature of about 40° C. 95. The method of any one of embodiments 57-94, wherein the salt comprises sodium chloride. 96. The method of any one of embodiments 57-95, wherein the salt is at a concentration of 0.1-0.2%, 0.2-0.3%, 0.3-0.4%, 0.4-0.5%, 0.5-0.6%, 0.6-0.7%, 0.7-0.8%, 0.8-0.9%, 0.9-1.0%, 1.0-1.1%, 1.1-1.2%, 1.2-1.3%, 1.3-1.4%, or 1.4-1.5%. 97. The method of any one of embodiments 57-96, wherein the salt is at a concentration of about 0.85%. 98. The method of any one of embodiments 57-97, wherein the salt is at a concentration of about 1.25% or less. 99. The method of any one of embodiments 57-68, wherein the salt is at a concentration of about 1.25%.The method of any one of embodiments 57-99, wherein the one or more microorganisms are selected from Acetobacter cereviseae, Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. 101. The method of any one of embodiments 57-100, wherein the one or more microorganisms comprise Acetobacter cereviseae, Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. 102. The method of any one of embodiments 57-101, wherein the one or more microorganisms comprise Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. 103. The method of any one of embodiments 57-102, wherein the one or more microorganisms comprise Ensifer adhaerens and Bacillus nakamurai or Bacillus subtilis. 104. The method of any one of embodiments 57-103, wherein the one or more microorganisms comprise Microbacterium yannicii or Microbacterium chocolatum. 105. The method of any one of embodiments 57-104, wherein the one or more microorganisms comprise Serratia ureilytica or Serratia marcescens. 106. The method of any one of embodiments 57-105, wherein the one or more microorganisms comprise Glutamicibacter arilaitensis or Glutamicibacter halophytocola. 107. The method of any one of embodiments 57-106, wherein the one or more microorganisms comprise Ensifer adhaerens, Pantoea allii, Bacillus subtilis, or Bacillus cucumis. 108. The method of any one of embodiments 57-107, wherein the one or more microorganisms comprise endospore-forming microorganisms. 109. The method of any one of embodiments 57-108, wherein the one or more microorganisms comprise Bacillus sp. 110.110. The method of any one of embodiments 57-109, wherein the one or more microorganisms are selected from the phylum Firmicutes, Proteobacteria, and Actinobacteria. 111. The method of any one of embodiments 57-110, wherein the one or more microorganisms are selected from the phylum Firmicutes. 112. One or more of the above microorganisms may include Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobac. illus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribacillus sp., Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus sp., Thermoflavimicrobium sp., Thermomovenabulum sp., Tuberibacillus sp., Virgibacillus sp., and Vulcanobacillus sp. 113. The method of any one of embodiments 57-112, wherein the one or more microorganisms are selected from the phylum Proteobacteria. 114. The method of any one of embodiments 57-113, wherein the one or more microorganisms comprise Actinomyces sp. 115. The method of any one of embodiments 57-114, wherein the one or more microorganisms are selected from the phylum Actinobacteria. 116. The method of any one of embodiments 57-115, wherein the one or more microorganisms comprise Coxiella sp. 117. The method of any one of embodiments 57-116, wherein the one or more microorganisms form endospores after incorporation into the seed. 118. The method of any one of embodiments 57-117, wherein the one or more microorganisms comprise endospores. 119. The method of any one of embodiments 57-118, wherein the one or more microorganisms comprise Bacillus endospores. 120. The method of any one of embodiments 57-119, wherein the one or more microorganisms comprise a 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221.121. The method of any one of embodiments 57-120, wherein the one or more microorganisms comprise a 16S nucleic acid sequence at least 99% identical to any of the sequences set forth in SEQ ID NOs: 1-10221. 122. The method of any one of embodiments 57-121, wherein the one or more microorganisms comprise a 16S nucleic acid sequence at least 98% identical to any of the sequences set forth in SEQ ID NOs: 1-10221. 123. The method of any one of embodiments 57-122, wherein the one or more microorganisms comprise a 16S nucleic acid sequence at least 95% identical to any of the sequences set forth in SEQ ID NOs: 1-10221. 124. The method of any one of embodiments 57-123, wherein the one or more microorganisms comprise a 16S nucleic acid sequence at least 90% identical to any of the sequences set forth in SEQ ID NOs: 1-10221. 125. The method of any one of embodiments 57-124, wherein the formulation further comprises a microbial exudate. 126. The method of any one of embodiments 57-125, wherein the microbial percolate contains one or more compounds that cause systemic induced resistance (IST). 127. The method of any one of embodiments 57-126, wherein the microbial percolate contains one or more compounds that cause systemic induced resistance (ISR). 128. The method of any one of embodiments 57-127, wherein the microbial percolate contains one or more compounds that induce plant growth. 129. The method of any one of embodiments 57-128, wherein the microbial percolate is from a microorganism comprising a 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. 130. The method of any one of embodiments 57-129, wherein the microbial percolate is from a microorganism comprising a 16S nucleic acid sequence at least 99% identical to any of SEQ ID NOs: 1-10221. 131. The method of any one of embodiments 57-130, wherein the microbial infusion is from a microorganism comprising a 16S nucleic acid sequence that is at least 98% identical to any of SEQ ID NOs: 1-10221. 132. The method of any one of embodiments 57-131, wherein the microbial infusion is from a microorganism comprising a 16S nucleic acid sequence that is at least 95% identical to any of SEQ ID NOs: 1-10221.133. The method of any one of embodiments 57-132, wherein the microbial exudate is from a microorganism comprising a 16S nucleic acid sequence that is at least 90% identical to any of SEQ ID NOs: 1-10221. 134. The concentration of the one or more microorganisms in the formulation is about 1 x 10. 6 ~1×10 17 The method of any one of embodiments 57-133, wherein the concentration of the one or more microorganisms in the formulation is in the range of: 1 x 10 CFU / mL. 3 ~1×10 4 CFU / mL: 1 x 10 4 ~1×10 5 CFU / mL: 1 x 10 5 ~1×10 6 CFU / mL: 1 x 10 6 ~1×10 7 CFU / mL: 1 x 10 7 ~1×10 8 CFU / mL: 1 x 10 8 ~1×10 9 CFU / mL: 1 x 10 9 ~1×10 10 CFU / mL: 1 x 10 10 ~1×10 11 CFU / mL: 1 x 10 11 ~1×10 12 CFU / mL: 1 x 10 12 ~1×10 13 CFU / mL: 1 x 10 13 ~1×10 14 CFU / mL: 1 x 10 14 ~1×10 15 CFU / mL: 1 x 10 15 ~1×10 16 CFU / mL; or 1 × 10 16 ~1×10 17 The method according to any one of embodiments 57-134, wherein the amount of the one or more microorganisms in the formulation is 10 CFU / mL per gram of seeds. 10 The method of any one of embodiments 57-135, wherein the amount of the one or more microorganisms in the formulation is less than about 10 CFU per gram of seeds. 5 ~10 9The method of any one of embodiments 57-136, wherein the microorganisms are cells of the plant. 138. The method of any one of embodiments 57-137, wherein the one or more microorganisms are selected to produce a plant growth-promoting effect. 139. The method of any one of embodiments 57-138, wherein the plant growth-promoting effect of the one or more microorganisms is selected from the group consisting of osmoregulation, ion homeostasis, antioxidant defense, heat stress resistance, and / or maintenance of photosynthetic capacity of the cells. 140. The method of any one of embodiments 57-139, wherein the one or more microorganisms are selected for compatibility. 141. The method of any one of embodiments 57-140, wherein the one or more microorganisms are selected to ensure that predation or antagonism does not occur. 142. The method of any one of embodiments 57-141, wherein the one or more microorganisms are selected for stability during storage. 143. The method of any one of embodiments 57-142, wherein the one or more microorganisms are selected for rapid plant colonization and survival in associated tissues. 144. The method of any one of embodiments 57-143, wherein the one or more microorganisms are selected for stimulating a large-scale, long-term physiological response in the plant. 145. The method of any one of embodiments 57-144, wherein the one or more microorganisms are selected for promoting the plant's life cycle. 146. The method of any one of embodiments 57-145, wherein the one or more microorganisms are selected for optimal incorporation into one or more seeds. 147. The method of any one of embodiments 57-146, wherein at least one of the microorganisms remains present throughout the plant's life cycle. 148. The method of any one of embodiments 57-147, wherein the incubation time is less than 1 minute. 149. The method of any one of embodiments 57-148, wherein the incubation time is about 1 minute. 150. The method of any one of embodiments 57-149, wherein the incubation time is less than 10 minutes. 151. The method of any one of embodiments 57-150, wherein the incubation time is less than 5 minutes. 152. The method of any one of embodiments 57-151, wherein the incubation time is less than 20 minutes.153. The method of any one of embodiments 57-152, wherein the incubation time is less than 4 hours or less than 16 hours. 154. The method of any one of embodiments 57-153, wherein the incubation time is less than several days. 155. The method of any one of embodiments 57-154, wherein the incubation time is less than 12 hours. 156. 1 x 10. 6 157. The method of any one of embodiments 57-155, wherein bacterial cells greater than 157.1 x 10 are incorporated into each of the one or more seeds. 5 ~1×10 8 158. The method of any one of embodiments 57-157, wherein the one or more microorganisms are stably incorporated into one or more seeds. 159. The method of any one of embodiments 57-158, wherein the incorporated one or more microorganisms are stable for more than 30 days. 160. The method of any one of embodiments 57-159, wherein the incorporated one or more microorganisms are stable for more than 6 months. 161. The method of any one of embodiments 57-160, wherein the incorporated one or more microorganisms are stable for at least 1 year. 162. The method of any one of embodiments 57-161, wherein the incorporated one or more microorganisms are stable for at least 2 years.

[0133] 163. A plant seed treatment formulation comprising a salt and one or more microorganisms. 164. The formulation of embodiment 163, wherein the one or more microorganisms are selected to provide a plant growth-promoting effect. 165. The formulation of embodiment 163 or 164, wherein the formulation is an aqueous formulation. 166. The formulation of any one of embodiments 163-165, wherein the formulation further comprises poloxamer 188. 167. The formulation of any one of embodiments 163-166, wherein the formulation further comprises poloxamer 188 at a concentration of 0.1%. 168. The formulation of any one of embodiments 163-167, wherein the formulation further comprises Tween 20. 169. The formulation of any one of embodiments 163-168, further comprising one or more agents from the group consisting of dimethyl sulfoxide (DMSO), 1-dodecylazacycloheptan-2-one, laurocapram, 1-methyl-2-pyrrolidone (NMP), oleic acid, ethanol, methanol, polyethylene glycol (Brij 35, 58, 98), polyethylene glycol monolaurate (Tween 20), Tween 40 (polyoxyethylene acid sorbitol esters), Tween 60, Tween 80 (nonionic), cetylmethylammonium bromide (CTAB), urea, lecithin (coagulated fatty acid from soy), chitosan, poloxamer 188, poloxamer 237, poloxamer 338, and poloxamer 407. 170. The formulation of any one of embodiments 163-169, further comprising one or more ingredients that promote endospore formation of one or more bacteria. 171. The formulation of any one of embodiments 163-170, further comprising potassium, ferrous sulfate, calcium, magnesium, manganese, or a combination thereof. 172. The formulation of any one of embodiments 163-171, further comprising manganese. 173. The formulation of any one of embodiments 163-172, further comprising calcium, magnesium, and manganese. 174. The formulation of any one of embodiments 163-173, further comprising nutrients for one or more selected microorganisms. 175. The medium of any one of embodiments 163-174, wherein the salt comprises sodium chloride. 176. The salt is present in an amount of 0.1-0.2%, 0.2-0.3%, 0.3-0.4%, 0.4-0.5%, 0.5-0.6%, 0.6-0.7%, 0.176. The formulation of any one of embodiments 163-177, wherein the salt is at a concentration of about 0.7-0.8%, 0.8-0.9%, 0.9-1.0%, 1.0-1.1%, 1.1-1.2%, 1.2-1.3%, 1.3-1.4%, or 1.4-1.5%. 177. The formulation of any one of embodiments 163-176, wherein the salt is at a concentration of about 0.85%. 178. The formulation of any one of embodiments 163-177, wherein the salt is at a concentration of about 1.25% or less. 179. The formulation of any one of embodiments 163-178, wherein the salt is at a concentration of about 1.25%. 180. The formulation of any one of embodiments 163-179, wherein the one or more microorganisms are selected from Acetobacter cereviseae, Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. 181. The formulation of any one of embodiments 163-180, wherein the one or more microorganisms comprise Acetobacter cereviseae, Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. 182. The formulation of any one of embodiments 163-181, wherein the one or more microorganisms comprise Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. 183. The method of any one of embodiments 163-182, wherein the one or more microorganisms comprise Ensifer adhaerens and Bacillus nakamurai or Bacillus subtilis. 184. The formulation of any one of embodiments 163-183, wherein the one or more microorganisms comprise Microbacterium yannicii or Microbacterium chocolatum. 185. The formulation of any one of embodiments 163-184, wherein the one or more microorganisms comprise Serratioa ureilytica or Serratioa marcescens. 186.The formulation of any one of embodiments 163-185, wherein the one or more microorganisms comprise Glutamicibacter arilaitensis or Glutamicibacter arilaitensis. 187. The formulation of any one of embodiments 163-186, wherein the one or more microorganisms comprise Ensifer adhaerens, Pantoea allii, Bacillus subtilis, or Bacillus subtilis. 188. The formulation of any one of embodiments 163-187, wherein the one or more microorganisms comprise an endospore-forming microorganism. 189. The formulation of any one of embodiments 163-188, wherein the one or more microorganisms comprise a Bacillus sp. 190. The formulation of any one of embodiments 163-189, wherein the one or more microorganisms are selected from the phyla Firmicutes, Proteobacteria, and Actinobacteria. 191. A formulation according to any one of embodiments 163-190, wherein the one or more microorganisms are selected from the phylum Firmicutes. 192. One or more of the above microorganisms may include Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribacillus sp., Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus sp., Thermoflavimicrobium sp., Thermovenabulum sp., Tuberibacillus sp., Virgibacillus sp., and Vulcanobacillus sp.. 193. The formulation of any one of embodiments 163-192, wherein the one or more microorganisms are selected from the phylum Proteobacteria. 194. The formulation of any one of embodiments 163-193, wherein the one or more microorganisms comprise Actinomyces sp. 195. The formulation of any one of embodiments 163-194, wherein the one or more microorganisms are selected from the phylum Actinobacteria. 196. The formulation of any one of embodiments 163-195, wherein the one or more microorganisms comprise Coxiella sp. 197. The formulation of any one of embodiments 163-196, wherein the one or more microorganisms form endospores after being incorporated into the seeds. 198. The formulation of any one of embodiments 163-197, wherein the one or more microorganisms comprise endospores. 199. The formulation of any one of embodiments 163-198, wherein the one or more microorganisms comprise Bacillus endospores. 200. The formulation of any one of embodiments 163-199, wherein the one or more microorganisms comprise a 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. 201. The formulation of any one of embodiments 163-200, wherein the one or more microorganisms comprise a 16S nucleic acid sequence that is at least 99% identical to any of SEQ ID NOs: 1-10221. 202. The formulation of any one of embodiments 163-201, wherein the one or more microorganisms comprise a 16S nucleic acid sequence that is at least 98% identical to any of SEQ ID NOs: 1-10221. 203. The formulation of any one of embodiments 163-202, wherein the one or more microorganisms comprise a 16S nucleic acid sequence that is at least 95% identical to any of the sequences in SEQ ID NOs: 1-10221. 204. The formulation of any one of embodiments 163-203, wherein the one or more microorganisms comprise a 16S nucleic acid sequence that is at least 90% identical to any of the sequences in SEQ ID NOs: 1-10221. 205.The formulation of any one of embodiments 163-204, further comprising a microbial percolate.206. The formulation of any one of embodiments 163-205, wherein the microbial percolate contains one or more compounds that cause systemic induced resistance (IST).207. The formulation of any one of embodiments 163-206, wherein the microbial percolate contains one or more compounds that cause systemic induced resistance (ISR).208. The formulation of any one of embodiments 163-207, wherein the microbial percolate contains one or more compounds that induce plant growth.209. The microbial percolate. 210. The formulation of any one of embodiments 163-209, wherein the microbial infusion is from a microorganism comprising a 16S nucleic acid sequence at least 99% identical to any of the sequences of SEQ ID NOs: 1-10221. 211. The formulation of any one of embodiments 163-210, wherein the microbial infusion is from a microorganism comprising a 16S nucleic acid sequence at least 98% identical to any of the sequences of SEQ ID NOs: 1-10221. 212. The formulation of any one of embodiments 163-211, wherein the microbial infusion is from a microorganism comprising a 16S nucleic acid sequence at least 95% identical to any of the sequences of SEQ ID NOs: 1-10221. 213. The formulation of any one of embodiments 163-212, wherein the microbial exudate is from a microorganism comprising a 16S nucleic acid sequence that is at least 90% identical to any of SEQ ID NOs: 1-10221. 214. The concentration of the one or more microorganisms in the formulation is about 1 x 10 3 ~1×10 17 The formulation of any one of embodiments 163-213, wherein the concentration of the one or more microorganisms in the formulation is in the range of: 1 × 10 to 1 × 10 CFU / mL. 4 CFU / mL: 1x104 to 1x10 5 CFU / mL: 1 x 10 5 ~1×10 6 CFU / mL: 1 x 10 6 ~1×10 7 CFU / mL: 1 x 10 7 ~1×108 CFU / mL: 1 x 10 8 ~1×10 9 CFU / mL: 1 x 10 9 ~1×10 10 CFU / mL: 1 x 10 10 ~1×10 11 CFU / mL: 1 x 10 11 ~1×10 12 CFU / mL: 1 x 10 12 ~1×10 13 CFU / mL: 1 x 10 13 ~1×10 14 CFU / mL: 1 x 10 14 ~1×10 15 CFU / mL: 1 x 10 15 ~1×10 16 CFU / mL; or 1 × 10 16 ~1×10 17 The formulation of any one of embodiments 163-214, wherein the CFU / mL.

[0134] 216. A method for treating one or more plant seeds, the method comprising: (i) soaking the one or more seeds in a formulation, the formulation comprising salt and one or more microbial percolate selected to produce a plant growth-promoting effect; and (ii) incubating the one or more seeds in the formulation for a period of time sufficient to incorporate the one or more microbial percolate into the seeds. 217. The method of embodiment 216, wherein the microbial percolate contains one or more compounds that cause systemic induced resistance (IST). 218. The method of embodiment 216 or 217, wherein the microbial percolate contains one or more compounds that cause systemic induced resistance (ISR). 219. The method of any one of embodiments 216-218, wherein the microbial percolate contains one or more compounds that induce plant growth. 220. The method of any one of embodiments 216-219, wherein the microbial percolate is from a microorganism comprising a 16S nucleic acid sequence of any of SEQ ID NOs: 1-10221. 221. The method of any one of embodiments 216-220, wherein the microbial infusion is from a microorganism comprising a 16S nucleic acid sequence at least 99% identical to any one of SEQ ID NOs: 1-10221. 222. The method of any one of embodiments 216-221, wherein the microbial infusion is from a microorganism comprising a 16S nucleic acid sequence at least 98% identical to any one of SEQ ID NOs: 1-10221. 223. The method of any one of embodiments 216-222, wherein the microbial infusion is from a microorganism comprising a 16S nucleic acid sequence at least 95% identical to any one of SEQ ID NOs: 1-10221. 224. The method of any one of embodiments 216-223, wherein the microbial infusion is from a microorganism comprising a 16S nucleic acid sequence at least 90% identical to any one of SEQ ID NOs: 1-10221. 225. The method of any one of embodiments 216-224, wherein the salt comprises sodium chloride. 226. The above salts may be present in concentrations of 0.1-0.2%, 0.2-0.3%, 0.3-0.4%, 0.4-0.5%, 0.5-0.6%, 0.6-0.7%, 0.7-0.8%, 0.8-0.9%, 0.9-1.0%, 1.0-1.1%, 1.1-1.2%, 1.2-1.3%, 1.3-1.4%, or 1.4-1.5%.226. The method of any one of embodiments 216-226, wherein the salt is at a concentration of about 0.85%. 227. The method of any one of embodiments 216-226, wherein the salt is at a concentration of about 0.85%. 228. The method of any one of embodiments 216-227, wherein the salt is at a concentration of about 1.25% or less. 229. The method of any one of embodiments 216-228, wherein the salt is at a concentration of about 1.25%. 230. The method of any one of embodiments 216-229, wherein the microbial exudate is derived from Microbacterium yannicii or Microbacterium chocolatum. 231. The method of any one of embodiments 216-230, wherein the microbial exudate is derived from Serratioa ureilytica or Serratioa marcescens. 232. The method of any one of embodiments 216-231, wherein the microbial exudate is derived from Glutamicibacter arilaitensis or Glutamicibacter halophytocola. 233. The method of any one of embodiments 216-232, wherein the microbial exudate is derived from Ensifer adhaerens. 234. The method of any one of embodiments 216-233, wherein the microbial exudate is derived from Chryseobacterium lactis. 235. The method of any one of embodiments 216-234, wherein the microbial exudate is derived from Acetobacter cerevisiae, Pantoea allii, Bacillus subtilis, or Bacillus cucumis. 236. The method of any one of embodiments 216-235, wherein the microbial exudate is derived from a microorganism selected from the phyla Firmicutes, Proteobacteria, and Actinobacteria. 237. The above microbial exudates include Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribacillus sp., Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus sp., Thermoflavimicrobium sp., Thermovenabulum sp., Tuberibacillus sp., Virgibacillus sp., and Vulcanobacillus sp. 238. The method of any one of embodiments 216-237, wherein the microbial exudate is derived from a microorganism selected from the phylum Proteobacteria. 239. The method of any one of embodiments 216-238, wherein the microbial exudate is derived from an Actinomyces sp. 240. The method of any one of embodiments 216-239, wherein the microbial exudate is derived from the phylum Actinobacteria. 241. The method of any one of embodiments 216-240, wherein the microbial exudate is derived from a Coxiella sp. 242. The method of any one of embodiments 216-241, wherein the microbial exudate is derived from a Bacillus sp.

[0135] 242. A method for incorporating bacteria into plant seeds, the method comprising: a. contacting the plant seeds with a solution containing the bacteria, wherein the solution comprises about 0.1% to about 2% salt (w / v); and b. incubating the plant seeds with the solution, thereby incorporating at least 1 colony forming unit (CFU) of the bacteria into the plant seeds. 243. The method of embodiment 242, wherein (b) comprises incubating the plant seeds with the solution, thereby incorporating at least 500 CFU of the bacteria into the plant seeds. 244. The method of embodiment 242 or 243, wherein the bacteria comprises an endospore-forming bacterium or an endospore thereof. 245. The method of any one of embodiments 242-244, wherein the solution comprises a microbial exudate. 246. The method of embodiment 245, wherein the microbial exudate is derived from the bacteria. 247. The method of embodiment 245, wherein the microbial exudate does not originate from the bacteria. 248. The method of any one of embodiments 242-247, wherein the bacteria comprises bacteria from the phyla Firmicutes, Proteobacteria, Actinobacteria, or a combination thereof. 249. The bacteria include Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribacillus sp., Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus sp., Thermoflavimicrobium sp., Thermovenabulum sp., Tuberibacillus sp., Virgibacillus sp., Vulcanobacillus sp., or a combination thereof. 250. The method of any one of embodiments 242-249, wherein the bacterium comprises a bacterium from a Bacillus sp. 251. The method of any one of embodiments 242-250, wherein the bacterium is incorporated between the seed coat and the embryo of the plant seed. 252. The method of any one of embodiments 242-251, further comprising disinfecting the plant seed prior to (a). 253. The method of any one of embodiments 242-252, wherein the solution comprises about 0.85% of the salt. 254. The method of any one of embodiments 242-253, wherein the salt comprises NaCl. 255. The method of any one of embodiments 242-254, wherein the plant comprises corn seeds, wheat seeds, rice seeds, sorghum seeds, barley seeds, rye seeds, sugarcane seeds, millet seeds, oat seeds, soybean seeds, cotton seeds, alfalfa seeds, bean seeds, quinoa seeds, lentil seeds, peanut seeds, lettuce seeds, tomato seeds, pea seeds, or cabbage seeds. 256. The method of any one of embodiments 242-255, wherein the solution further comprises Luria-Bertani (LB) broth. 257.257. The method of any one of embodiments 242-256, wherein the solution further comprises dimethyl sulfoxide (DMSO), 1-dodecylazacycloheptan-2-one, laurocapram, 1-methyl-2-pyrrolidone (NMP), oleic acid, ethanol, methanol, polyethylene glycol (Brij 35, 58, 98), polyethylene glycol monolaurate (Tween 20), Tween 40 (polyoxyethylene acid sorbitol ester), Tween, Tween 80 (nonionic) 60, cetylmethylammonium bromide (CTAB), urea, lecithin (coagulated fatty acid from soy), chitosan, poloxamer 188, poloxamer 237, poloxamer 338, poloxamer 407, or a combination thereof. 258. The method of any one of embodiments 242-257, wherein the solution further comprises calcium, magnesium, manganese, potassium, iron, or a combination thereof. 259. The method of any one of embodiments 242-258, wherein the solution is maintained at a temperature between about 4°C and about 40°C; about 20°C and about 40°C; or about 10°C and about 20°C. 260. The method of any one of embodiments 242-259, wherein the solution is maintained at about 23°C or about 30°C. 261. The method of any one of embodiments 242-260, wherein the plant seeds are incubated with the solution for about 1 minute to about 960 minutes, about 20 minutes to about 240 minutes, or about 1 minute to about 20 minutes. 262. The method of any one of embodiments 242-261, wherein the plant seeds are incubated with the solution for about 1 minute, about 5 minutes, about 10 minutes, about 20 minutes, about 240 minutes, or about 960 minutes. 263. The method of any one of embodiments 245-262, further comprising inducing endospore formation of the endospore-forming bacterium.

[0136] 264. A modified plant seed, comprising at least one CFU of a bacterium incorporated between the seed coat and the embryo of the modified plant seed. 265. The modified plant seed of embodiment 264, wherein the modified plant seed comprises at least 500 CFU or at least 1000 CFU of the bacterium. 266. The modified plant seed of embodiment 264 or 265, wherein the bacterium comprises an endospore-forming bacterium or an endospore thereof. 267. The modified plant seed of any one of embodiments 264-266, wherein the modified plant seed comprises a microbial exudate. 268. The modified plant seed of embodiment 267, wherein the microbial exudate is derived from the bacterium. 269. The modified plant seed of embodiment 267, wherein the microbial exudate is not derived from the bacterium. 270. The modified plant seed of any one of embodiments 264-269, wherein the bacteria comprise bacteria from the phyla Firmicutes, Proteobacteria, Actinobacteria, or a combination thereof. 271. The bacteria include Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribacillus sp., Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus sp., Thermoflavimicrobium sp., Thermovenabulum sp., Tuberibacillus sp., Virgibacillus sp., Vulcanobacillus sp., or a combination thereof. 272. The modified plant seed of any one of embodiments 264-271, wherein the bacterium comprises a bacterium from a Bacillus sp. 273. The modified plant seed of any one of embodiments 264-272, wherein the modified seed is corn seed, wheat seed, rice seed, sorghum seed, barley seed, rye seed, sugarcane seed, millet seed, oat seed, soybean seed, cotton seed, alfalfa seed, bean seed, quinoa seed, lentil seed, peanut seed, lettuce seed, tomato seed, pea seed, or cabbage seed. 274. The modified plant seed of any one of embodiments 264-273, wherein the plant seed comprises at least 1000 CFU of the microorganism.

[0137] 275.1×10 3A formulation containing at least one or more CFU / mL bacteria, wherein the formulation comprises about 0.1% to about 2% salt.276. The formulation of embodiment 275, comprising 0.85% salt.277. The formulation of embodiment 275 or 276, wherein the salt comprises NaCl.278. The formulation of any one of embodiments 275-277, wherein the bacteria comprises an endospore-forming bacterium or an endospore thereof.279. The formulation of any one of embodiments 275-278, wherein the formulation comprises a microbial exudate.280. The method of embodiment 279, wherein the microbial exudate is derived from the bacterium.281. The method of embodiment 279, wherein the microbial exudate is not derived from the bacterium.282. The formulation of any one of embodiments 275-281, wherein the bacterium comprises a bacterium from the phyla Firmicutes, Proteobacteria, or Actinobacteria. 283. The bacteria include Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribacillus sp., Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus sp., Thermoflavimicrobium sp., Thermovenabulum sp., Tuberibacillus sp., Virgibacillus sp., Vulcanobacillus sp., or a combination thereof. 284. The formulation of any one of embodiments 275-283, wherein the bacteria comprises bacteria from Bacillus sp. 285. The formulation of any one of embodiments 275-284, wherein the formulation further comprises LB broth. 286. The formulation of any one of embodiments 275-285, wherein the formulation further comprises dimethyl sulfoxide (DMSO), 1-dodecylazacycloheptan-2-one, laurocapram, 1-methyl-2-pyrrolidone (NMP), oleic acid, ethanol, methanol, polyethylene glycol (Brij 35, 58, 98), polyethylene glycol monolaurate (Tween 20), Tween 40 (polyoxyethylene acid sorbitol esters), Tween, Tween 80 (nonionic) 60, cetylmethylammonium bromide (CTAB), urea, lecithin (coagulated fatty acid from soy), chitosan, poloxamer 188, poloxamer 237, poloxamer 338, poloxamer 407, or a combination thereof. 287. The formulation of any one of embodiments 275-286, wherein the formulation further comprises calcium, magnesium, manganese, potassium, iron, or a combination thereof. 288. The formulation of any one of embodiments 275-287, wherein the formulation is maintained at a temperature between about 4°C and about 40°C; about 20°C and about 40°C; or about 10°C and about 20°C. 289. The formulation of any one of embodiments 275-288, wherein the formulation is maintained at 23°C or 30°C. 290. The formulation contains at least 5 x 10. 5 The formulation of any one of embodiments 275-289, containing CFU / mL of the bacteria.

[0138] 291. A method for promoting a plant growth effect in a plant seed, the method comprising: a. contacting the plant seed with a solution containing bacteria, wherein the solution comprises about 0.1% to about 2% salt (w / v); and b. incubating the plant seed with the solution, thereby incorporating at least 500 colony forming units (CFU) of the bacteria into the plant seed. 292. The method of embodiment 291, further comprising disinfecting the plant seed before (a). 293. The method of embodiment 291 or 292, wherein the bacteria comprises an endospore-forming bacterium or an endospore thereof. 294. The method of any one of embodiments 291-293, wherein the solution comprises a microbial exudate. 295. The method of embodiment 294, wherein the microbial exudate is derived from the bacteria. 296. The method of embodiment 294, wherein the microbial exudate is not derived from the bacteria. 297. The method of any one of embodiments 291-296, wherein the bacterium is incorporated between the seed coat and the embryo of the modified plant seed. 298. The method of any one of embodiments 291-297, wherein the solution comprises about 0.85% of the salt. 299. The method of any one of embodiments 291-298, wherein the salt comprises NaCl. 300. The method of any one of embodiments 291-299, wherein the bacterium comprises a bacterium from the phyla Firmicutes, Proteobacteria, Actinobacteria, or a combination thereof. 301. The bacteria include Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribaccillus sp.301. The method of any one of embodiments 291-300, wherein the bacterium comprises a bacterium from the species Bacillus sp., Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus sp., Thermoflavimicrobium sp., Thermomovenabulum sp., Tuberibacillus sp., Virgibacillus sp., Vulcanobacillus sp., or a combination thereof. 302. The method of any one of embodiments 291-301, wherein the bacterium comprises a bacterium from the species Bacillus sp. 303. The method of any one of embodiments 291-302, wherein the plant comprises corn seeds, wheat seeds, rice seeds, sorghum seeds, barley seeds, rye seeds, sugarcane seeds, millet seeds, oat seeds, soybean seeds, cotton seeds, alfalfa seeds, bean seeds, quinoa seeds, lentil seeds, peanut seeds, lettuce seeds, tomato seeds, pea seeds, or cabbage seeds. 304. The method of any one of embodiments 291-303, wherein the solution further comprises LB broth. 305. The method of any one of embodiments 291-304, wherein the solution further comprises dimethyl sulfoxide (DMSO), 1-dodecylazacycloheptan-2-one, laurocapram, 1-methyl-2-pyrrolidone (NMP), oleic acid, ethanol, methanol, polyethylene glycol (Brij 35, 58, 98), polyethylene glycol monolaurate (Tween 20), Tween 40 (polyoxyethylene acid sorbitol ester), Tween, Tween 80 (nonionic) 60, cetylmethylammonium bromide (CTAB), urea, lecithin (coagulated fatty acid from soy), chitosan, poloxamer 188, poloxamer 237, poloxamer 338, poloxamer 407, or a combination thereof. 306. The method of any one of embodiments 291-305, wherein the solution further comprises calcium, magnesium, manganese, potassium, iron, or a combination thereof. 307.The method of any one of embodiments 291-306, wherein the solution is maintained at a temperature between about 4°C and about 40°C; about 20°C and about 40°C; or about 10°C and about 20°C. 308. The method of any one of embodiments 291-307, wherein the solution is maintained at about 23°C or about 30°C. 309. The method of any one of embodiments 291-308, wherein the plant seeds are incubated with the solution for about 1 minute to about 960 minutes, about 20 minutes to about 240 minutes, or about 1 minute to about 20 minutes. 310. The method of any one of embodiments 291-309, wherein the plant seeds are incubated with the solution for about 1 minute, about 5 minutes, about 10 minutes, about 20 minutes, about 240 minutes, or about 960 minutes. 311. The method of any one of embodiments 291-310, further comprising inducing endospore formation of the endospore-forming bacterium. 312. The method of any one of embodiments 291-311, wherein the plant growth effect comprises increased yield, cellular osmoregulation, ion homeostasis, antioxidant defense, heat stress tolerance, maintenance of photosynthetic capacity, nitrogen fixation, or a combination thereof. 313. The method of embodiment 312, wherein the bacterium is selected for the plant growth effect.

[0139] 314. A method for promoting a plant growth effect in a plant seed, the method comprising: a. contacting the plant seed with a solution containing a microbial permeate, the solution comprising about 0.1% to about 2% salt (w / v); and b. incubating the plant seed with the solution, thereby incorporating the microbial permeate into the plant seed. 315. The method of embodiment 314, further comprising disinfecting the plant seed before (a). 316. The method of embodiment 314 or 315, wherein the microbial permeate is derived from an endospore-forming bacterium or its endospore. 317. The method of embodiment 314 or 315, wherein the microbial permeate is derived from a non-endospore-forming bacterium. 318. The method of any one of embodiments 314-317, wherein the microbial permeate is incorporated between the seed coat and the embryo of the modified plant seed. 319. The method of any one of embodiments 314-318, wherein the solution comprises about 0.85% of the salt. 320. The method of any one of embodiments 314-319, wherein the salt comprises NaCl. 321. The method of any one of embodiments 314-320, wherein the microbial exudate is from the phyla Firmicutes, Proteobacteria, Actinobacteria, or a combination thereof. 322. The microbial exudates include Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp., Desulfosporomusa sp., Desulfosporosinus sp., Desulfovirgula sp., Desulfunispora sp., Desulfurispora sp., Filifactor sp., Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., Sporomusa sp., Sporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribacillus sp., Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus sp., Thermoflavimicrobium sp., Thermovenabulum sp., Tuberibacillus sp., Virgibacillus sp., Vulcanobacillus sp., or a combination thereof. 323. The method of any one of embodiments 314-322, wherein the microbial exudate is derived from bacteria from Bacillus sp. 324. The method of any one of embodiments 314-323, wherein the plant comprises a corn seed, a wheat seed, a rice seed, a sorghum seed, a barley seed, a rye seed, a sugarcane seed, a millet seed, an oat seed, a soybean seed, a cotton seed, an alfalfa seed, a bean seed, a quinoa seed, a lentil seed, a peanut seed, a lettuce seed, a tomato seed, a pea seed, or a cabbage seed. 325. The method of any one of embodiments 314-324, wherein the solution further comprises dimethyl sulfoxide (DMSO), 1-dodecylazacycloheptan-2-one, laurocapram, 1-methyl-2-pyrrolidone (NMP), oleic acid, ethanol, methanol, polyethylene glycol (Brij 35, 58, 98), polyethylene glycol monolaurate (Tween 20), Tween 40 (polyoxyethylene acid sorbitol ester), Tween, Tween 80 (nonionic) 60, cetylmethylammonium bromide (CTAB), urea, lecithin (solidified fatty acid from soybean), chitosan, poloxamer 188, poloxamer 237, poloxamer 338, poloxamer 407, or a combination thereof. 326. The method of any one of embodiments 314-325, wherein the solution is maintained at a temperature between about 4° C. and about 40° C.; about 20° C. and about 40° C.; or about 10° C. and about 20° C. 327. The method of any one of embodiments 314-326, wherein the solution is maintained at about 23° C. or at about 30° C. 328.The method of any one of embodiments 314-327, wherein the plant seeds are incubated with the solution for about 1 minute to about 960 minutes, about 20 minutes to about 240 minutes, or about 1 minute to about 20 minutes. 329. The method of any one of embodiments 314-328, wherein the plant seeds are incubated with the solution for about 1 minute, about 5 minutes, about 10 minutes, about 20 minutes, about 240 minutes, or about 960 minutes. 330. The method of any one of embodiments 314-329, wherein the plant growth effect comprises increased plant yield, cellular osmoregulation, ion homeostasis, antioxidant defense, heat stress tolerance, maintenance of photosynthetic capacity, nitrogen fixation, or a combination thereof. 331. The method of any one of embodiments 314-330, wherein the microbial exudate is selected for the plant growth effect. 332. The method of any one of embodiments 314-331, wherein the plant growth effect comprises nitrogen fixation. 333. The method of any one of embodiments 314-331, wherein the plant growth effect comprises an increase in plant yield.

[0140] 334. The engineered seed of any one of embodiments 1-55, wherein the one or more microorganisms are selected to improve plant yield.

[0141] 335. The method of any one of embodiments 57-161, wherein the plant growth effect comprises increased yield.

[0142] 336. The method of any one of embodiments 291-312, wherein the plant growth effect comprises an increase in yield. [Example]

[0143] The methods and compositions of the present disclosure are designed to enhance the growth, nutritional status, and resistance to environmental and biotic stresses of seed-derived agricultural plants or herbs by treating the seeds with plant beneficial microorganisms, and / or their exudates, and / or their personalized biomolecules during the dormant stage. (商標)Seed treatment allows for biological priming of the seed embryo. At this stage, bacteria and / or endospores and exudates previously incorporated into the seed are favorably positioned to access the plant embryo, stimulate continued plant development, and / or induce resistance responses in the plant host. At the same time, bacteria, if used, have the advantage of successfully colonizing the root niche before the roots are exposed to the soil microbiota. For example, achieving effective root niche colonization with nitrogen-fixing bacteria allows for efficient delivery of ammonium to the plant.

[0144] Definition of Microbial Formulation: To achieve initial conditioning, the present disclosure uses seed treatment compositions containing synthetic consortia or single isolated bacterial species or endospores in a suspension medium. Typically, plant cultivation compositions and methods include diverse, environmentally adaptable, plant-associated bacteria belonging to a wide variety of bacterial genera distributed within the Proteobacteria phylum (α-, β-, γ-, and δ-proteobacteria classes) and different taxa within the phylum Firmicutes, Bacteroidetes, and Actinobacteria. The inventors have isolated and characterized plant growth-promoting bacteria belonging to various genera, including commonly plant-associated microorganisms, within these large taxa, which can be applied to seeds using the methods of the present disclosure to improve plant growth and health. The compositions contain one, two, three, or several different bacterial genera individually cultured and then labeled Microprime. (商標) mixed for seed treatment.

[0145] Example 1. Seed treatment to increase loading of non-endospore-forming bacteria, endospore-forming bacteria, and / or bacterial endospore-forming bacteria: Microprime (商標) technology. The concept of loading bacteria and bacterial endospores inside seeds is illustrated in Figure 1. A process that can be performed at room temperature in a short period of time is always desirable because it is industrially scalable and more economical since it uses fewer resources and energy. To streamline the process of incorporating microorganisms into seeds at room temperature, several surfactants or agents were explored. Surfactants that may improve seed penetration and the placement of desired elements within seeds, either alone or in combination, include dimethyl sulfoxide (DMSO), 1-dodecylazacycloheptan-2-one, laurocapram, 1-methyl-2-pyrrolidone (NMP), oleic acid, ethanol, methanol, polyethylene glycol (Brij 35, 58, 98), polyethylene glycol monolaurate (Tween 20), Tween 40 (polyoxyethylene acid sorbitol ester), Tween 60, Tween 80 (nonionic), cetylmethylammonium bromide (CTAB), urea, lecithin (a solidifying fatty acid derived from soybeans), chitosan, and various poloxamers (188, 237, 338, 407). The table below summarizes the experiments (Table 2) conducted with 0.85% w / v NaCl salt in medium supplemented with two selected surfactants or drugs (Tween 20 and Poloxamer 188). The treatments were carried out with a strain of Serratia sp. on commercial corn seeds (Dekalb DK630) at 23°C for 5 minutes. Microprime (商標) The concentration of cells in the solution was 6.4 x 10 9 CFU / ml.

[0146] [Table 2]

[0147] Microprime (商標) Adding poloxamer 188 (0.1% w / v) to the solution increased bacterial loading inside the seeds by 52%. (Microprime (商標)This effect of significantly increasing bacterial cells within the seeds (while maintaining the parameters of ambient temperature and short imbibition time during the seed treatment process) is highly desirable for its industrial-scale implementation.

[0148] The results of the same previous study are shown in Table 3 below, but Microprime (商標) The solution contained a higher concentration of NaCl (1.25% w / v), and the cell concentration was increased to 2.9 × 10 12 CFU / ml.

[0149] [Table 3]

[0150] When comparing both results, it is clear that the effects of both Tween 20 and Poloxamer 188 continue to aid in incorporating more bacterial cells into seeds when compared to the same imbibition medium without these surfactants. The use of 1.25% (w / v) NaCl dramatically reduces the efficiency of incorporating bacterial cells into seeds.

[0151] Nutrients were added (in the following examples, Microprime) to reduce the standard deviation and standard error of the number of bacterial cells present in the treated seeds. (商標) The same Luria-Bertani (LB) medium was used to grow the bacteria before incorporating them into the solution. Table 4 shows that adding a nutrient source for the bacteria achieves a decrease in the standard deviation and standard error, and therefore, the number of bacterial cells within the seeds can become more homogenous.

[0152] [Table 4]

[0153] The lower standard deviation and standard error when the absorption medium is supplemented with a nutrient source for the bacteria may be explained because, when the bacteria are placed inside the seed, the seed can continue to grow by accessing these nutrients until they are depleted or until viable growth conditions for the bacteria are ceased (e.g., when the seed dries out).

[0154] Nutritional supplements that can improve the conversion of endospore-forming bacteria from their vegetative growth phase to endospores include calcium, magnesium, potassium, manganese, and ferrous sulfate, either alone or in combination. Table 5 reports the number of endospores per milliliter obtained in various mineral-containing media and for various incubation times of isolated Bacillus sp.

[0155] [Table 5]

[0156] To ensure proper growth of the bacteria loaded onto the seeds, Microprime with nutrients specifically compatible with the selected bacteria. (商標) Supplement the solution or add Microprime endospores of the bacteria desired to be incorporated into the seeds. (商標) It will need to be added directly to the solution.

[0157] Loading desired bacteria, endospores, or bacterial consortia into dicotyledonous or monocotyledonous seeds is a complex and nonlinear process. As shown in Table 6, and in the case of monocotyledonous seeds (maize, Zea mays), successful loading of desired amounts of endospores of Bacillus sp. isolates with in-plant efficacy requires Microprime (商標) There is an inflection point in terms of the initial minimum concentration of endospores in solution, after which seeds can be effectively loaded with cells.

[0158] Figure 5 shows the loading kinetics of a synthetic bacterial consortium, internally called Lascar, consisting of four bacterial isolates, into dicotyledonous seeds (lettuce, Lactuca sativa). (商標) The bacterial loading process by seed treatment does not follow a linear behavior, and there are several conditions that must be met to achieve efficient loading of bacteria into seeds. (商標) It is confirmed that there is an initial minimum concentration of colony-forming units (CFU) per milliliter in solution. Furthermore, these kinetic curves indicate the existence of a saturation point, which is approximately 1.00E+5 CFU per seed. It can be observed, however, that loading kinetics and initial minimum concentrations are not generalizable and depend on the type of bacteria.

[0159] [Table 6]

[0160] Microprime (商標) The duration of the seed treatment process is key to industrial scaling. Co-culture times were evaluated using vegetative cells and endospores of two different bacteria, as shown in Table 7. Seeds were externally sterilized, dried for 24 hours, and ground into powder, after which the average colony-forming units (CFU) within the seeds were quantified. For CFU / seed quantification, three biological replicates consisting of three seed pools each were used. In all cases, at least 5 minutes of exposure is sufficient to load thousands of bacterial cells, with the exception of vegetative cells of strain S3C23. Maximum loading is achieved over 20 minutes for all cell types.

[0161] [Table 7]

[0162] Example 2. Microprime with fluorescent bacteria (商標) Seed treatment visualization. To assess the spatial distribution of the loaded bacteria within the seeds, we used an Escherichia coli fluorescent reporter strain. Corn seeds (Dekalb DK630) were transfected with Microprime using E. coli that constitutively expresses red fluorescent protein. (商標) The colony forming units per seed was 1.99E+5, and the Microprime (商標) The treatment conditions were 37°C for 240 minutes (4 hours). Seeds were fixed and cut into 0.5 cm long sections. Samples were then analyzed under a confocal microscope at a wavelength of 558 nm. Bacterial loading and localization were visually confirmed (pink fibers, Figure 4A). Closer observation showed that the endosperm and embryo were separated from the outer layers by specifically localizing the bacteria in the gap between the seed pericarp and the seed aleurone cell layer (Figure 4B).

[0163] These results led to Microprime (商標) Seed treatments have shown that bacteria can be effectively loaded into seeds, and in the case of monocotyledonous plants, into the space between the seed pericarp and the seed aleurone cell layer.

[0164] Example 3. Microprime (商標) Seed loading efficiency of specific bacteria by seed treatment process Microprime (商標) The seed treatment process involves watering seeds in a well-defined liquid solution containing specific bacteria for a period of time sufficient to allow bacterial loading into the seeds. It is necessary to know the efficiency of this process, i.e., the percentage of seeds that can be loaded with bacteria. To determine this value, specific DNA primers are designed that can detect bacteria at the molecular level by using PCR technology, as shown in Figure 6. Microprime uses the bacterial isolate Ensifer adhaerens, internally referred to as strain S3C10. (商標)Treated corn seeds (Dekalb DK630) were germinated in vitro using a culture tube containing 3 ml of Murashige-Skoog liquid medium. After the plant roots emerged from the seeds (3-5 days after sowing), a rapid DNA extraction procedure was performed from the liquid medium, and PCR reactions were carried out using specific primers as shown in Figure 6. Primed seeds (seeds treated under the same conditions and formulations, but excluding bacteria) were used as a control. The colony-forming units per seed was 6.53E+4, and Microprime (商標) The seed treatment conditions were 20 minutes at 23°C.

[0165] Table 8 shows the percentage of seeds in which the loaded bacteria were detectable by PCR. (商標) The seed loading efficiency of the seed treatment is approximately 98%.

[0166] [Table 8]

[0167] Example 4. Microprime (商標) Colonization of plant roots by bacteria loaded into seeds by seed treatment Microprime (商標) Maize seeds (Dekalb DK630) treated with the bacterial isolate Ensifer adhaerens (S3C10) were grown in a double-tube growth chamber using agar as a substrate, as shown in Figure 7, according to the method described in Niu et al., 2018. This protocol was developed to set up a gnotobiotic system for cultivating maize seedlings colonized by simplified root-associated colonies. Untreated seeds (seeds that were neither treated nor inoculated), primed seeds (seeds that were treated and not inoculated), and Microprime (商標)Seeds (treated and inoculated) were germinated directly in a double-tube chamber under the following conditions: 16 hours of light (day) and 8 hours of darkness (night), 25°C, and 54% relative humidity. Maize seedlings were stored under the above conditions for 15 days. After this time, 1 cm-long primary root fragments, smaller than the kernel size, were collected from the germinated seeds by cutting the primary root with a sterile scalpel blade. The root fragments were then weighed on a balance, rinsed in sterile 1x PBS buffer, and crushed with sterile pistils. The mixed bacterial suspension was serially diluted and plated on LB agar. Quantification of colony-forming units (CFU) per gram of root for each treatment is shown in Figure 8 (n=15). Under these experimental conditions, CFUs cannot be detected in untreated primed seedlings. Instead, Microprime (商標) In seed seedlings, an average of 7.00E+7 CFU / gram of roots was detected, indicating that the bacteria loaded into the seeds were able to effectively colonize the plant root structure after germination.

[0168] Example 5. Microprime (商標) Temporal stability of bacteria loaded onto seeds by seed treatment For bacterial seed treatment technology to fit into traditional seed industry logistics and be valuable to farmers, it needs to be able to guarantee bacterial stability, understood as the survival rate of bacteria within the seeds, over several months (typical seed industry logistics involve storing seeds for months or even years before sowing / planting by farmers). Microprime (商標) To evaluate bacterial stability after seed treatment, a bacterial consortium, internally named Lascar, and endospores of the bacterial isolate Bacillus subtilis, internally designated strain S3C23, were used in Microprime. (商標) Lettuce and corn seeds were treated under the seed treatment process. The specific bacterial colony forming units (CFU) were counted using Microprime (商標)The storage time of seeds treated with Microprime was evaluated. In both cases, triplicate pools of seeds were ground at different times after treatment and CFU per seed was recorded. The colony forming units per seed in MP-S3C23 seeds was 7.10E+3, and the number of colony forming units per seed in Microprime was 7.10E+3. (商標) The seed treatment conditions were 20 minutes at 23°C. (商標) 1 shows short-term (<50 days) and long-term (>100 days) survival of bacterial consortia on lettuce seeds.

[0169] In addition, Figure 10 shows the Microprime (商標) Figure 1 shows the survival of endospores of strain S3C23 in maize seeds over a period of 1 to 12 months after seed treatment.

[0170] Both results indicate a shelf life for the inoculum that is compatible with the requirements of the conventional seed industry and agricultural practices.

[0171] Example 6. Microprime (商標) Temporal stability of plant embryos in treated seeds Osmotic or water-based seed treatments significantly affect embryo viability and twinning over time, reducing seed vigor over time, which determines the plant's potential for rapid and uniform emergence. (商標) Tomato, lettuce, and corn seeds were analyzed at various times to demonstrate that the seed treatments did not adversely affect germination rates and seed vigor. Table 9 shows the results of Microprime treatment on tomato (Solanum lycopersicum) embryos (Tomato Ferry-Morse cv. Roma VF). (商標)The effects of seed treatments and conventional seed priming treatments (also known as osmopriming) on ​​vigor are shown. The percentage emergence of 36 plants per treatment was measured 7 days after sowing in peat:perlite (used as soil substrate). Growth conditions were 25°C, 54% humidity, and 16 h / 8 h (light / dark). Microprime was used to culture the bacterial isolate (Pantoea allii, internally referred to as strain P9C1) and two different imbibition medium solutions (sol. 1 and sol. 2). (商標) Seed treatments were performed. Sol. 1 contained only bacteria, and Sol. 2 contained bacteria and their exudates. The colony forming units per seed in the seeds of MP-P9C1_sol.1 were 2.00E+5, and the colony forming units per seed in the seeds of MP-P9C1_sol.2 were 5.00E+5. Microprime (商標) The seed treatment conditions were 30°C for 240 minutes (4 hours). Control seeds were untreated. Primed seeds were treated with the same two imbibition media, sol. 1 and sol. 2, but without bacteria (essentially osmoprimed seed treatment).

[0172] As is well known, after the priming process, in which seeds are immersed in a liquid solution (osmotically or non-osmotically), the vigor of plant embryos is adversely affected over time. (商標) When seed treatments are used, plant embryo vigor is maintained similar to that of untreated seeds, as shown in the table below (Table 9).

[0173] [Table 9]

[0174] Figure 11 shows the Microprime PCR assay using two bacterial isolates, Serratia marcescens and Glutamicibacter halophytocola, internally designated S8C6 and S8C7. (商標) Seed treatment (Microprime (商標)The germination rates of 21 maize seeds (Dekalb's GMO DK630d) were compared using Microprime. (商標) The seeds were measured over several storage times, including 0, 33, 51, and 98 days after seed treatment. Control seeds were untreated. Primed seeds were treated with Microprime (商標) The seeds were treated with the same imbibition medium but without the bacteria (essentially an osmoprimed seed treatment). (商標) It can be seen that seeds treated with showed the same germination rate as the control / untreated seeds, both of which were significantly higher than the primed seeds (Figure 11).

[0175] Additionally, Microprime, which includes a synthetic consortium of bacteria called JIT, (商標) Seed treatments were used to assess the germination rate of lettuce (Lactuca sativa) seeds. The JIT consortium consisted of bacteria 4 (Chryseobacterium lactis), bacteria 5 (Bacillus endophyticus), and bacteria 6 (Bacillus megaterium).

[0176] The germination rate of lettuce seeds 240 days after seed treatment is shown in the table below (Table 10).

[0177] [Table 10]

[0178] These results led to Microprime (商標) The seed treatment has been shown to have no effect on germination rate or seed vigor while allowing tomato, lettuce, and corn to grow normally, and is compatible with commercially available products. (商標) To verify whether the treatments affect seed vigor over long storage times, an accelerated aging (AA) test was performed using Microprime (商標)The AA test was carried out on (MP) corn seeds (Dekalb DK630). AA tests provide valuable information about the storage and seedling emergence potential. Twenty corn seeds from each lot or treatment were exposed to 43°C for 72 hours and approximately 95% relative humidity. After AA treatment, the seeds were sown in peat:perlite soil and incubated in a greenhouse at a temperature of 25°C, 54% humidity, and a 16-hour light / 8-hour dark photoperiod. Seven days after sowing, the emergence of hypocotyls and the number of normal, abnormal, and dead seedlings were determined. Control seeds were untreated. Primed seeds were then incubated in Microprime (商標) The MP-S3C10+S3C23 seeds were treated with the solution but without bacteria (essentially an osmopriming treatment), and then incubated in Microprime, a synthetic consortium containing two bacterial isolates, Ensifer adhaerens (S3C10) and Bacillus subtilis (S3C23). (商標) solution, and MP-S8C7 was cultured in Microprime containing the bacterial isolate Glutamicibacter halophytocola, internally referred to as strain S8C7. (商標) The colony forming units (CFU) per seed of strain S3C10 was 2.20E+3, the colony forming units (CFU) per seed of strain S3C23 was 5.00E+1, and the colony forming units (CFU) per seed of strain S8C7 was 1.98E+4. (商標) The seed treatment conditions were 30°C for 10 minutes.

[0179] The results of the AA test are shown in the table below (Table 11).

[0180] [Table 11]

[0181] The results illustrated in Figure 11, along with Tables 9, 10, and 11, are presented in Microprime (商標)The seed treatment did not affect germination rate, embryo viability, and seed vigor over time, which is comparable to the commercial product (Microprime (商標) This indicates that it is highly compatible with the seeds.

[0182] Example 7. Microprime in lettuce growth and development (商標) Effect of seed treatments.

[0183] One of the main goals of incorporating microorganisms into seeds is to provide an efficient method of bacterial delivery that improves desired agronomic traits, such as enhanced growth and field performance. To evaluate the effects in plants, Microprime (商標) The growth and development of lettuce plants grown from seeds was examined. Figure 12 shows the effect of a proprietary bacterial composition, internally referred to as Lascar, on lettuce seeds (Lactuca sativa). The Lascar consortium consists of Bacterium 1 (Acetobacter cereviseae), Bacterium 4 (Chryseobacterium lactis), Bacterium 5 (Bacillus endophyticus), and Bacterium 6 (Bacillus megaterium). At 67 days after sowing, Microprime (商標)Fresh rosette weights of lettuce (Seminis brand cv. Mohawk) were measured in plants grown from seeds (MP-Lascar), inoculated with Lascar in liquid form (Lascar), treated with a bacterial-free imbibed solution (primed), and untreated (untreated). The Lascar treatment is a positive control corresponding to a synthetic consortium, which is applied externally to seeds before sowing, as is typically done in microbial technology applications. The primed treatment is a negative control consisting of an imbibed solution without a bacterial consortium. Bars represent the mean ± 1 standard error of the mean for at least 15 plants per treatment. Asterisks indicate statistical significance (one-way ANOVA, p<0.05; Dunnett's multiple comparison test, p<0.05). Assays were performed in a greenhouse with a peat:perlite soil substrate and a 67-day growing period. The growth temperature was 19°C, humidity was 54%, and the photoperiod was 16 hours light and 8 hours dark.

[0184] These results led to the launch of Microprime with the Lascar consortium. (商標) Seed treatment was shown to increase vegetative growth of lettuce as much as conventional inoculation in liquid form (Figure 12). However, this novel delivery system (Microprime (商標) Seeds) have several advantages ranging from commercial issues (scalability, long-term storage) to operational issues (product that is easy for farmers to use, reduced chance of contamination), thus lowering the technical risks of using beneficial microorganisms in the field.

[0185] Microprime (商標)The preservation effect of lettuce seeds treated with a proprietary bacterial composition, internally referred to as JIT, was evaluated. The JIT consortium consists of bacteria 4 (Chryseobacterium lactis), 5 (Bacillus endophyticus), and 6 (Bacillus megaterium). Figure 13 shows the fresh weight of Harris Moran lettuce (cv. Desert Storm) seeds 28 days after sowing and storage for 60 days using the following treatments: MP-JIT (treated and inoculated), primed (treated and uninoculated), and untreated. The treatment labeled "primed" refers to negative control seeds that were immersed in a solution without the bacterial consortium. Bars represent the mean ± 1 standard error of the mean for at least 15 plants per treatment. Asterisks indicate statistical significance (Kruskal-Wallis, p < 0.05; Dunnett's multiple comparison test, p < 0.05). Assays were conducted in a greenhouse with a 28-day growth period, peat:perlite soil as substrate, growth temperature of 19°C, humidity of 54%, and a photoperiod of 16 hours light and 8 hours dark.

[0186] Treatment of plants with beneficial bacteria can induce abiotic stress tolerance. Microprime using the JIT consortium (商標) To evaluate whether seed treatments confer tolerance to abiotic stress, lettuce plants (Harris Moran cv. Desert Storm) were grown under salt stress conditions for 14 days, after which whole-plant fresh weights were measured (Figure 14). In vitro assays were performed using square plates containing Murashige-Skoog medium supplemented with 120 / 12 mM NaCl / CaCl2 (salt stress) and control plates without salt (0 / 0 mM NaCl / CaCl2). Plants were grown at 19°C and 54% humidity with a 16-h / 8-h (light / dark) photoperiod. Bars represent the mean ± 1 standard error of the mean for at least 10 plants per treatment. Asterisks indicate statistical significance (one-way ANOVA, p<0.05; Dunnett's multiple comparison test, p<0.05).

[0187] These results suggest that Microprime, a JIT consortium (商標) The treatment was shown to induce salt tolerance in lettuce plants.

[0188] Example 8. Microprime on the growth of tomato plants (商標) Effects of seed treatment Microprime for growth and development of tomato plants (商標) To evaluate the efficacy of seed treatments, tomato (Ferry-Morse cv. Roma VF) seeds were treated with Microprime containing the bacterial isolate Bacillus subtilis, internally referred to as strain S3C23. (商標) The seed treatments were performed using Microprime. As shown in Figure 15, the total plant fresh weights of control (untreated), primed, and MP-S3C23 tomato seedlings were measured 42 days after sowing. The treatment designated "primed" is a negative control consisting of soaking the seeds in a solution containing no bacteria. Bars represent the mean ± 1 standard error of the mean for at least 15 plants per treatment. Asterisks represent statistically significant differences (one-way ANOVA, p-value < 0.05; Dunnett's multiple comparison test, p-value < 0.05). The colony-forming units (CFU) per ml of strain S3C23 in the imbibition medium were 1.20E+9 CFU / ml. After the seeds were dried, the CFU / seed was 4.00E+3. Microprime (商標) Seed treatment conditions were 240 minutes (4 hours) at 30°C. Assays were conducted in a greenhouse with a peat:perlite soil substrate and a 42-day growth period. Growth temperature was 25°C, humidity was 54%, and the photoperiod was 16 hours light and 8 hours dark.

[0189] These results suggest that Microprime using strain S3C23 (商標) It is shown that the treatment significantly increases the fresh weight of the tomato plants.

[0190] Example 9. Microprime in corn growth and development (商標) effect. Microprime in maize plant growth and development (商標) To evaluate the efficacy of seed treatments, maize seeds (Dekalb brand, DK630 non-GMO seeds) were treated with Microprime containing the bacterial isolate Microbacterium chocolatum, internally designated strain S3C1. (商標) Figure 16 shows the fresh weight of the shoots and roots of the plants measured 14 days after sowing in semi-solid substrate. Treatments included liquid inoculated seeds (S3C1) and Microprime (商標) The samples included treated seeds (MP-S3C1), primed seeds (primed), and untreated and uninoculated seeds (control). The treatment labeled S3C1 is a positive control corresponding to external inoculation of the seeds with strain S3C1 (liquid preparation, outside the seeds), as performed in conventional microbiological technology applications. The treatment labeled priming is a negative control consisting of soaking the seeds in a solution without a bacterial consortium. The colony-forming units (CFU) per seed for MP-S3C1 seeds was 1.90E+5, a value comparable to Microprime. (商標) Seed treatment conditions were 30°C for 960 minutes (16 hours). Bars represent the mean ± 1 standard error of at least 15 plants per treatment. Asterisks represent statistically significant differences (Kruskal Wallis, p-value < 0.05; Dunnett's multiple comparison test, p-value < 0.05). Seed treatment was carried out at 30°C for 5 minutes, after which the seeds were dried to achieve a moisture content of 11.2%. Microprime (商標) The colony-forming units (CFU) per ml of the seed treatment was 7.79E+15. After the seeds were dried, the CFU per seed was 1.9E+5. Assays were performed in a greenhouse and on semi-solid substrates. Data were obtained 14 days after sowing (14 DAS). The growth temperature was 25°C, humidity was 54%, and the photoperiod was 16 hours light and 8 hours dark. The seed treatment MP-S3C1 increased the shoot fresh weight of corn plants by 37.2% (Figure 16A), and the seed treatment MP-S3C1 increased the root fresh weight of corn plants by 45.9% (Figure 16B).

[0191] To evaluate the effectiveness of multiple bacteria, a synthetic consortium containing two bacterial isolates, Ensifer adhaerens (S3C10) and Bacillus subtilis (S3C23), was tested (Figure 17). Dekalb non-GMO seeds (DK630) were used. Microprime (商標) The colony-forming units (CFU) per ml of seed treatment solution were 4.52E+10 for strain S3C10 and 3.76E+08 for strain S3C23. Seed treatment was performed at 30°C for 10 minutes, after which the seeds were dried to achieve a moisture content of 11.4%. After simultaneous loading of these two strains, th...

Claims

1. A modified plant seed, comprising at least 10,000 CFU of bacteria incorporated between the seed coat and the aleurone cell layer of said modified plant seed.

2. The modified plant seed of claim 1 , wherein the bacterium comprises an endospore-forming bacterium or an endospore thereof.

3. The modified plant seed of claim 1 , wherein the modified plant seed comprises a microbial exudate.

4. The modified plant seed of claim 3 , wherein the microbial exudate is derived from the bacteria.

5. The modified plant seed of claim 3 , wherein the microbial exudate is not derived from the bacteria.

6. 2. The modified plant seed of claim 1, wherein the bacteria comprises bacteria from Phyla Firmicutes, Proteobacteria, Actinobacteria, or a combination thereof.

7. The above bacteria are Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanaerobacter sp., Caloramator sp., Caminicella sp., Cerasibacillus sp., Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp., Dendrosporobacter sp., Desulfotomaculum sp.、Desulfsporomusa sp.、Desulfsporosinus sp.、Desulfvirgula sp.、Desulffunispora sp.、Desulfspora sp.、Filifactor sp.、Filobacillus sp., Gelria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorella sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp., Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planiphylum sp., Pontibacillus sp., Propionispora sp., Salinibacillussp. , Salsuginibacillus sp. , Seinonella sp. , Shimazuella sp. , Sporacetigenium sp. , Sporoanaerobacter sp. , Sporobacter sp. , Sporobacterium sp. , Sporohalobacter sp. , Sporolactobacillus sp. , Sporomusa sp. , Sporosarcina sp. , Sporotalea sp. , Sporotomaculum sp. , Syntrophonas sp. , Syntrophospora sp. , Tenuibacillus sp. , Tepidibacter sp. , Terribacillus sp. , Thalassobacillus sp. , Thermoacetogenium sp. , Thermoactinomyces sp. , Thermoalkalibacillus sp. , Thermoanaerobacter sp. , Thermoanaeromonas sp. , Thermobacillus sp. , Thermoflavimicrobium sp.

7. The modified plant seed of claim 6, comprising bacteria from Thermococcus sp., Thermovenabulum sp., Tuberibacillus sp., Virgibacillus sp., Vulcanobacillus sp., or combinations thereof.

8. 8. The modified plant seed of claim 7, wherein the bacteria comprises a bacterium from the genus Bacillus sp.

9. 8. The modified plant seed of claim 7, wherein the bacteria comprises bacteria from Paenibacillus sp.

10. 7. The modified plant seed of claim 6, wherein the bacteria comprises bacteria from Actinomyces sp., Acetobacter sp., Herbaspirillum sp., Pseudomonas sp., Paraburkholderia sp., Serratia sp., Pantoea sp., Ensifer sp., Enterobacter sp., or any combination thereof.

11. 11. The modified plant seed of claim 10, wherein the bacteria comprises bacteria from Ensifer sp.

12. 8. The modified plant seed of claim 7, wherein the bacteria comprises bacteria from Coxiella sp.

13. 2. The modified plant seed of claim 1, wherein the modified plant seed is a corn seed, a wheat seed, a rice seed, a sorghum seed, a barley seed, a rye seed, a sugarcane seed, a millet seed, an oat seed, a soybean seed, a cotton seed, an alfalfa seed, a bean seed, a quinoa seed, a lentil seed, a peanut seed, a lettuce seed, a tomato seed, a pea seed, or a cabbage seed.

14. The modified plant seed of claim 1 , wherein the seed coat is the pericarp of the seed.

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