Compositions and methods for producing bicarbonate and minerals

Abstract

Described herein are methods and compositions that utilize plant-integrated microorganisms for improved carbon dioxide conversion and sequestration in soil.

Description

[Technical Field]

[0001] cross reference This application claims the interests of U.S. Provisional Patent Application No. 63 / 094,870, filed on 21 October 2020, and U.S. Provisional Patent Application No. 63 / 257,079, filed on 18 October 2021, both of which are incorporated herein by reference.

[0002] Human activities, including the burning of fossil fuels and deforestation, are major contributors to the increase of major greenhouse gases. Burning coal produces the most CO2 compared to other uses of fossil fuels, releasing approximately 2.5 tons of CO2 for every ton of coal burned. To date, the concentration of CO2 in the environment has risen from 280 ppm in the mid-19th century, when the Industrial Revolution began, to over 400 ppm. It is predicted that CO2 concentrations will reach 600 ppm by the middle of this century and likely reach 700 ppm by the end of this century. Global temperatures are projected to rise by more than 2°C by 2050 and by approximately 3-5°C over the next 50-100 years. This global rise in temperature is exacerbating climate change, resulting in global impacts including the Australian bushfires that have devastated wildlife, re-emerged wildfires in California forests, increased frequency and duration of heat waves, prolonged droughts, rising sea levels, and tsunamis. These natural disasters were not common before industrialization, and they are a result of rising CO2 levels.

[0003] Apart from known CO2 emission sources, there are several unexplored sources that not only continuously add CO2 but could also become important sources depending on the severity of climate change. Soil contains far more carbon than vegetation (1500 Pg of C at a depth of 1m, 2500 Pg of C at 2m; 1 Pg = 1 × 10⁻¹⁶). 15Soil contains twice as much carbon as the atmosphere (750pg of C), making it the largest carbon reservoir. It is estimated that one ton of organic carbon in soil releases 3.66 tons of CO2. Organic carbon in soil is added by plants in the following ways: through root death, root exudates, or other root-derived organic substances released through the rhizosphere and root respiration. Plants utilize CO2 during photosynthesis and convert it into sugars, but it is well known that a large amount of unfixed CO2 is released during respiration, mainly from the plant roots. Of the 120pg of carbon taken in by plants, 50% is lost into the atmosphere through plant respiration. This is further exacerbated by soil organisms and microorganisms (rhizosphere) that live near the roots and release CO2 during respiration. CO2 production by rhizobia is 10 times higher than that of plants that do not have a rhizosphere. Microorganisms living in the soil are supplied with nutrients from root exudates and survive by decomposing complex substances present in the soil. The role of soil microorganisms in climate change has been studied for some time, suggesting that global warming may accelerate the activity of heterotrophic microorganisms, consequently increasing the amount of CO2 flux in the soil that is ultimately released into the environment. Because soil temperature can increase soil respiration, global climate change is expected to increase the net transfer of carbon from soil to the atmosphere. Soil is an excellent source of carbon, storing 3.3 times the size of the atmospheric pool (760 gigatons), but global warming could exacerbate the depletion of this carbon pool. Preventing CO2 release into the atmosphere is necessary, but permanently storing CO2 in soil through effective CO2 sequestration is urgently needed. Sequestrating carbon in soil used for agriculture, forestry, and land reclamation is recognized as a potential option for mitigating global warming.

[0004] CO2 sequestration, whether biological or non-biological, began in the early stages of Earth's history when CO2 levels were much higher than they are today. (Approximately 150,000 x 10⁻¹⁰) 12A massive amount of CO2, amounting to metric tons, was fixed in carbonate minerals such as calcite, aragonite, dolomite, and limestone. While CO2 can normally be converted into solids containing carbonate minerals such as calcium carbonate and magnesium carbonate, the hydration of CO2 to produce bicarbonate is a very slow process (~1.3 × 10⁻⁶). -1 s -1 ). [Overview of the project]

[0005] One aspect of the present disclosure is a composition comprising a plant or plant seed and one or more microorganisms associated with the plant or plant seed, wherein the one or more microorganisms are selected to produce or promote the formation of bicarbonates, carbonates, or one or more minerals, or are derived from the one or more microorganisms. In some embodiments, the plant is a commercial plant, fruit plant, nut plant, shrub plant, bulbous plant, grassland plant, turf plant, or any combination thereof. In some embodiments, the plant seed is a commercial plant seed, fruit plant seed, nut plant seed, shrub plant seed, bulbous plant seed, grassland plant seed, turf plant seed, or any combination thereof. In some embodiments, the one or more microorganisms associated with the plant seed are placed in the space between the seed coat and the seed embryo of the plant seed. In some embodiments, the one or more microorganisms associated with the plant seed are placed as a coating of the plant seed. In some embodiments, the one or more microorganisms associated with the plant seed are applied to the plant seed through an irrigation system. In some embodiments, the irrigation system includes in-furrow treatment techniques. In some embodiments, the irrigation system includes a spraying technique. In some embodiments, bicarbonates sequester carbon. In some embodiments, the carbon is gaseous carbon. In some embodiments, the gaseous carbon is carbon dioxide. In some embodiments, carbonates sequester carbon. In some embodiments, the carbon is gaseous carbon. In some embodiments, the gaseous carbon is carbon dioxide. In some embodiments, one or more minerals sequester carbon. In some embodiments, the carbon is gaseous carbon. In some embodiments, the gaseous carbon is carbon dioxide. In some embodiments, one or more microorganisms associated with plant seeds are located in the space between the seed pericarp and the seed aleurone cell layer of the plant seed. In some embodiments, one or more microorganisms include one or more carbonic anhydrases. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases from the alpha class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases from the beta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases from the gamma class.In some embodiments, one or more carbonic anhydrases include carbonic anhydrases from the delta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases from the zeta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases from the eta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases from the iota class.

[0006] In some embodiments, one or more microorganisms include bacteria, archaea, fungi, or viruses. In some embodiments, one or more microorganisms include bacteria. In some embodiments, bacteria include endospore-forming bacteria. In some embodiments, the bacteria belong to the genera Acetonema, Actinomyces, Alkalibacillus, Ammoniphilus, Amphibacillus, Anaerobacter, Anerospora, Aneurinibacillus, Anoxybacillus, Bacillus, Brevibacillus, Caldanerobacter, Caloramator, Caminicella, and Cerasibacillus. 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., Gerria Geobacillus sp., Geosporobactersp.), Gracilibacillus sp., Halobacillus sp., Halonatronum sp., Heliobacterium sp., Heliophilum sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorela sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus 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 Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter sp., Sporobacterium sp., Sporohalobacter sp., Sporolactobacillus sp., SporomusaSporosarcina sp., Sporotalea sp., Sporotomaculum sp., Syntrophomonas sp., Syntrophospora sp., Tenuibacillus sp., Tepidibacter sp., Terribacillus sp., Thalassobacillus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter The bacteria include those from the genera Thermoanaeromonas sp., Thermobacillus sp., Thermoflavimicrobium sp., Thermovenabylum sp., Tuberibacillus sp., Virgibacillus sp., Vulcanobacillus sp., or combinations thereof. In some embodiments, the bacteria include those belonging to the phylum Firmicutes. In some embodiments, the bacteria include rhizosphere bacteria. In some embodiments, the rhizosphere bacteria include those from the genera Bacillus sp., PaenibacillusBacteria include B. amyloliquefaciens, B. laterosporus, B. licheniformis, B. macerans, B. cereus, B. circulans, B. firmus, B. subtilis, B. megaterium, B. coagulans, B. brevis This includes B. sphaericus, B. thuringiensis, B. mycoides, B. cucumis, B. endophyticus, B. pumilus, B. velezensis, B. mucilaginosus, B. tequilensis, B. methylotrophicus, or any combination thereof. In some embodiments, the bacteria include Bacillus subtilis S3C23, Bacillus subtilis MP2, Bacillus subtilis RO2C15, Bacillus subtilis RO2C22, Bacillus megaterium 6, Bacillus megaterium S3C21, Bacillus megaterium RO2C12, Bacillus cucumis S3C14, Bacillus endophyticus 5, or any combination thereof. In some embodiments, the bacteria include Bacillus subtilis S3C23. In some embodiments, the bacteria include Bacillus subtilisThe subtilis includes MP2. In some embodiments, the bacteria include Paenibacillus polymyxa, Paenibacillus taohuashanense, Paenibacillus pocheonensis, Paenibacillus aceris, Paenibacillus catalpa, Paenibacillus rigui, Paenibacillus pabuli, Paenibacillus brasiliensis, or any combination thereof. In some embodiments, the bacteria include Paenibacillus polymyxa RO3C16, Paenibacillus taohuashanense TY4D5, Paenibacillus pocheonensis S2C3, Paenibacillus aceris VF2D2, Paenibacillus catalpa TY2B5, Paenibacillus rigui TY2D5, Paenibacillus pabuli PG2A8, or any combination thereof. In some embodiments, the bacteria include non-endospore-forming bacteria. In some embodiments, the bacteria include bacteria belonging to the Proteobacteria phylum. In some embodiments, the bacteria belong to the genera Klebsiella, Rhizobium, Bradyrhizobium, Ochrobactrum, Sinorhizobium, Xanthobacter, Methylobacterium, and Actinomyces.This includes the genera Kosakonia sp., Azotobacter sp., Acetobacter sp., Herbaspirillum sp., Pseudomonas sp., Paraburkholderia sp., Ralstonia sp., Geobacter sp., Serratia sp., Pantoea sp., Ensifer sp., Enterobacter sp., or any combination thereof. In some embodiments, the bacteria include Ensifer adhaerens S3C10. In some embodiments, the bacteria include bacteria belonging to the phylum Actinomycetes. In some embodiments, the bacteria include the genera Streptomyces sp., Coxiella sp., and Frankia sp. In some embodiments, the bacteria include bacteria belonging to the phylum Cyanobacteria. In some embodiments, the bacteria include the genera Cyanobacteria sp. In some embodiments, the bacteria include bacteria belonging to the phylum Chloroflexi phylum. In some embodiments, one or more microorganisms include one or more fungi associated with plant seeds. In some embodiments, one or more fungi associated with plant seeds are placed in the space between the seed coat and the seed embryo of the plant seed. In some embodiments, one or more fungi associated with plant seeds are placed as a coating on the plant seed. In some embodiments, one or more fungi associated with plant seeds are applied to the plant seed by an in-furrow treatment technique. In some embodiments, one or more fungi associated with plant seeds are applied to the plant seed by a spraying technique. In some embodiments, one or more fungi associated with plant seeds are applied to the plant seeds via irrigation. In some embodiments, one or more fungi associated with plant seeds are located in the space between the seed pericarp and the seed alleuron cell layer of the plant seed. In some embodiments, one or more fungi include arbuscular mycorrhizal fungi. In some embodiments, one or more fungi include ectomycorrhizal fungi. In some embodiments, one or more fungi include fungi from the genus Trichoderma. In some embodiments, one or more fungi include fungi from the genus Penicillium. In some embodiments, one or more minerals include calcite, aragonite, dolomite, limestone, or any combination thereof. In some embodiments, one or more minerals include CaCO3, MgCO3, CaMg(CO3)2, or any combination thereof. In some embodiments, the promotion of the production of one or more minerals includes the production of ammonia and an increase in the pH of the medium in which the plant derived from the plant seed is grown. In some embodiments, one or more microorganisms are not naturally present in the space between the seed pericarp and the seed alleuron cell layer of the plant seed.In some embodiments, the plant seeds are monocotyledonous or dicotyledonous plant seeds. In some embodiments, the commercial plant seeds are corn seeds, wheat seeds, rice seeds, sorghum seeds, barley seeds, rye seeds, sugarcane seeds, millet seeds, oat seeds, soybean seeds, cotton seeds, alfalfa seeds, legume seeds, quinoa seeds, lentil seeds, peanut seeds, sunflower seeds, canola seeds, cassava seeds, palm oil seeds, potato seeds, sugar beet seeds, cocoa seeds, coffee seeds, lettuce seeds, tomato seeds, pea seeds, or cabbage seeds.

[0007] Another aspect of the present disclosure is a composition comprising a plant or a part thereof and one or more microorganisms associated with the plant or a part thereof, wherein the one or more microorganisms are selected to produce or promote the formation of bicarbonates, carbonates, or one or more minerals, or are derived from the one or more microorganisms. In some embodiments, the plant or a part thereof is a plant root, a plant stem, a plant leaf, a plant seed, a plant fruit, a plant tuber, or a plant root nodule. In some embodiments, the plant or a part thereof includes a commercial plant or a part thereof. In some embodiments, the commercial plant or a part thereof is maize, wheat, rice, sorghum, barley, rye, sugarcane, millet, oats, soybeans, cotton, alfalfa, beans, quinoa, lentils, peanuts, sunflowers, canola, cassava, palm oil, potatoes, sugar beets, cocoa, coffee, lettuce, tomatoes, peas, cabbage, fruit trees, nut trees, forest trees, grasslands, or turfgrass. In some embodiments, part of which is a plant seed, one or more microorganisms associated with the plant seed are located in the space between the seed coat and the seed embryo of the plant seed. In some embodiments, one or more microorganisms associated with the plant or part of which are located as a coating on the plant or part of which. In some embodiments, one or more microorganisms associated with the plant or part of which are applied to the plant seed through an irrigation system. In some embodiments, the irrigation system includes in-furrow treatment techniques. In some embodiments, the irrigation system includes spraying techniques. In some embodiments, bicarbonates sequester carbon. In some embodiments, the carbon is gaseous carbon. In some embodiments, the gaseous carbon is carbon dioxide. In some embodiments, carbonates sequester carbon. In some embodiments, the carbon is gaseous carbon. In some embodiments, the gaseous carbon is carbon dioxide. In some embodiments, one or more minerals sequester carbon. In some embodiments, the carbon is gaseous carbon. In some embodiments, the gaseous carbon is carbon dioxide. In some embodiments, part of which is a plant seed, one or more microorganisms associated with the plant seed are located in the space between the seed pericarp and the seed allureon cell layer of the plant seed. In some embodiments,One or more microorganisms contain one or more carbonic anhydrases. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the alpha class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the beta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the gamma class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the delta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the zeta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the eta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the iota class. In some embodiments, one or more microorganisms include bacteria, archaea, fungi, or viruses. In some embodiments, one or more microorganisms include bacteria. In some embodiments, bacteria include endospore-forming bacteria. In some embodiments, the bacteria belong to the genera Acetonema, Actinomyces, Alkalibacillus, Ammoniphilus, Amphibacillus, Anaerobacter, Anerospora, Aneurinibacillus, Anoxybacillus, Bacillus, Brevibacillus, Caldanerobacter, Caloramator, Caminicella, and Cerasibacillus. sp.), Clostridium sp., Clostridiisalibacter sp., Cohnella sp., Coxiella sp.,Genus 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., Moorela 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.,The genera 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., Thalassobacilus sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus sp.), Thermoflavimicrobium sp., Thermovenabylum sp., Tuberibacillus sp., Virgibacillus sp., Vulcanobacillus sp.,The bacteria include those belonging to the phylum Firmicutes, or combinations thereof. In some embodiments, the bacteria include rhizosphere bacteria. In some embodiments, the rhizosphere bacteria include those belonging to the genera Bacillus, Paenibacillus, or both. In some embodiments, the bacteria include B. amyloliquefaciens, B. laterosporus, B. licheniformis, B. macerans, B. cereus, B. circulans, B. firmus, B. subtilis, B. megaterium, B. coagulans, B. brevis This includes B. sphaericus, B. thuringiensis, B. mycoides, B. cucumis, B. endophyticus, B. pumilus, B. velezensis, B. mucilaginosus, B. tequilensis, B. methylotrophicus, or any combination thereof. In some embodiments, the bacteria are Bacillus subtilis S3C23, Bacillus subtilis MP2, Bacillus subtilis RO2C15, Bacillus subtilis RO2C22, Bacillus megaterium 6, Bacillus megaterium S3C21, Bacillus megaterium RO2C12, Bacillus cucumis S3C14,The bacterium includes Bacillus endophyticus 5, or any combination thereof. In some embodiments, the bacterium includes Bacillus subtilis S3C23. In some embodiments, the bacterium includes Bacillus subtilis MP2. In some embodiments, the bacterium includes Paenibacillus polymic, This includes Paenibacillus polymyxa, Paenibacillus taohuashanense, Paenibacillus pocheonensis, Paenibacillus aceris, Paenibacillus catalpa, Paenibacillus rigui, Paenibacillus pabuli, Paenibacillus brasiliensis, or any combination thereof. In some embodiments, the bacteria include Paenibacillus polymyxa RO3C16, Paenibacillus taohuashanense TY4D5, Paenibacillus pocheonensis S2C3, Paenibacillus aceris VF2D2, Paenibacillus catalpa TY2B5, Paenibacillus rigui TY2D5, Paenibacillus pabuli PG2A8, or any combination thereof. In some embodiments, the bacteria include non-endospore-forming bacteria. In some embodiments, the bacteria include bacteria belonging to the Proteobacteria phylum.In some embodiments, the bacteria belong to the genera Klebsiella, Rhizobium, Bradyrhizobium, Ochrobactrum, Sinorhizobium, Xanthobacter, Methylobacterium, Actinomyces, Kosakonia, Azotobacter, Acetobacter, Herbaspirillum, Pseudomonas, Paraburkholderia, and Ralstonia. The bacteria include species of the following genera: Geobacter sp., Serratia sp., Pantoea sp., Ensifer sp., Enterobacter sp., or any combination thereof. In some embodiments, the bacteria include Ensifer adhaerens S3C10. In some embodiments, the bacteria include bacteria belonging to the phylum Actinomycetes. In some embodiments, the bacteria include species of the genera Streptomyces sp., Coxiella sp., and Frankia sp. In some embodiments, the bacteria include bacteria belonging to the phylum Cyanobacteria. In some embodiments, the bacteria include species of Cyanobacteria. In some embodiments, the bacteria include bacteria belonging to the phylum Chloroflexi. In some embodiments, one or more microorganisms include one or more fungi associated with plants or parts thereof. In some embodiments, one or more fungi associated with a plant or part thereof are located in the space between the seed coat and the seed embryo of the plant or part thereof.In some embodiments, one or more fungi associated with a plant or part thereof are placed as a coating on the plant or part thereof. In some embodiments, one or more fungi associated with a plant or part thereof are applied to the plant or part thereof by an in-furrow treatment technique. In some embodiments, one or more fungi associated with a plant or part thereof are applied to the plant or part thereof by a spraying technique. In some embodiments, one or more fungi associated with a plant or part thereof are applied to the plant or part thereof through an irrigation system. In some embodiments, one or more fungi associated with a plant or part thereof are placed in the space between the seed pericarp and the seed allureon cell layer of the plant or part thereof. In some embodiments, one or more fungi include arbuscular mycorrhizal fungi. In some embodiments, one or more fungi include ectomycorrhizal fungi. In some embodiments, one or more fungi include fungi from the genus Trichoderma. In some embodiments, one or more fungi include fungi from the genus Penicillium. In some embodiments, one or more minerals include calcite, aragonite, dolomite, limestone, or any combination thereof. In some embodiments, one or more minerals include CaCO3, MgCO3, CaMg(CO3)2, or any combination thereof. In some embodiments, the promotion of the production of one or more minerals includes the production of ammonia and an increase in the pH of the culture medium in which a plant derived from the plant or a part thereof is grown. In some embodiments, the part thereof is a plant seed, and one or more microorganisms are not naturally present in the space between the seed pericarp and the seed allureon cell layer of the plant or a part thereof. In some embodiments, the plant or a part thereof is a monocotyledonous or dicotyledonous plant.

[0008] Another aspect of the present disclosure is a composition comprising one or more microorganisms, wherein one or more microorganisms are located in the space between the membrane and cell layer of a plant or part thereof, or are selected to produce or promote the formation of one or more bicarbonates, carbonates, or minerals, or are derived from the one or more microorganisms. In some embodiments, the plant or part thereof is a plant root, plant stem, plant leaf, plant seed, plant fruit, plant tuber, or plant root nodule. In some embodiments, the plant or part thereof comprises a commercial plant or part thereof. In some embodiments, the commercial plant or part thereof is maize, wheat, rice, sorghum, barley, rye, sugarcane, millet, oats, soybeans, cotton, alfalfa, beans, quinoa, lentils, peanuts, sunflowers, canola, cassava, palm oil, potatoes, sugar beets, cocoa, coffee, lettuce, tomatoes, peas, cabbage, fruit trees, nut trees, forest trees, grasslands, or turfgrass. In some embodiments, part of which is a plant seed, one or more microorganisms associated with the plant seed are placed in the space between the seed coat and the seed embryo of the plant seed. In some embodiments, one or more microorganisms associated with the plant or part of it are placed as a coating on the plant or part of it. In some embodiments, one or more microorganisms associated with the plant or part of it are applied to the plant seed through an irrigation system. In some embodiments, the irrigation system includes in-furrow treatment techniques. In some embodiments, the irrigation system includes spraying techniques. In some embodiments, bicarbonates sequester carbon. In some embodiments, the carbon is gaseous carbon. In some embodiments, the gaseous carbon is carbon dioxide. In some embodiments, carbonates sequester carbon. In some embodiments, the carbon is gaseous carbon. In some embodiments, the gaseous carbon is carbon dioxide. In some embodiments, one or more minerals sequester carbon. In some embodiments, the carbon is gaseous carbon. In some embodiments, the gaseous carbon is carbon dioxide. In some embodiments, part of which is a plant seed, one or more microorganisms associated with the plant seed areIt is located in the space between the seed pericarp and the seed allureon cell layer of a plant seed. In some embodiments, one or more microorganisms include one or more carbonic anhydrases. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the alpha class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the beta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the gamma class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the delta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the zeta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the eta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the iota class. In some embodiments, one or more microorganisms include bacteria, archaea, fungi, or viruses. In some embodiments, one or more microorganisms include bacteria. In some embodiments, bacteria include endospore-forming bacteria. In some embodiments, the bacteria belong to the genera Acetonema, Actinomyces, Alkalibacillus, Ammoniphilus, Amphibacillus, Anaerobacter, Anerospora, Aneurinibacillus, Anoxybacillus, Bacillus, Brevibacillus, Caldanerobacter, Caloramator, Caminicella, and 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 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 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., Thermovenablum sp., Tuberibacillus sp., Virgibacillus sp.,This includes bacteria from the genus Vulcanobacillus sp., or combinations thereof. In some embodiments, the bacteria include bacteria belonging to the phylum Firmicutes. In some embodiments, the bacteria include rhizosphere bacteria. In some embodiments, the rhizosphere bacteria include Bacillus sp., Paenibacillus sp., or both. In some embodiments, the bacteria are B. amyloliquefaciens, B. laterosporus, B. licheniformis, B. macerans, B. cereus, B. circulans, B. firmus, B. subtilis, B. megaterium, B. coagulans, and B. brevis. This includes B. sphaericus, B. thuringiensis, B. mycoides, B. cucumis, B. endophyticus, B. pumilus, B. velezensis, B. mucilaginosus, B. tequilensis, B. methylotrophicus, or any combination thereof. In some embodiments, the bacteria are Bacillus subtilis S3C23, Bacillus subtilis MP2, Bacillus subtilis RO2C15, Bacillus subtilis RO2C22, Bacillus megaterium 6, Bacillus megaterium S3C21, Bacillus megaterium RO2C12,The bacteria include Bacillus cucumis S3C14, Bacillus endophyticus 5, or any combination thereof. In some embodiments, the bacteria include Bacillus subtilis S3C23. In some embodiments, the bacteria include Bacillus subtilis MP2. In some embodiments, the bacteria include Paeniva, This includes Paenibacillus polymyxa, Paenibacillus taohuashanense, Paenibacillus pocheonensis, Paenibacillus aceris, Paenibacillus catalpa, Paenibacillus rigui, Paenibacillus pabuli, Paenibacillus brasiliensis, or any combination thereof. In some embodiments, the bacteria include Paenibacillus polymyxa RO3C16, Paenibacillus taohuashanense TY4D5, Paenibacillus pocheonensis S2C3, Paenibacillus aceris VF2D2, Paenibacillus catalpa TY2B5, Paenibacillus rigui TY2D5, Paenibacillus pabuli PG2A8, or any combination thereof. In some embodiments, the bacteria include non-endospore-forming bacteria. In some embodiments, the bacteria include bacteria belonging to the Proteobacteria phylum.In some embodiments, the bacteria belong to the genera Klebsiella, Rhizobium, Bradyrhizobium, Ochrobactrum, Sinorhizobium, Xanthobacter, Methylobacterium, Actinomyces, Kosakonia, Azotobacter, Acetobacter, Herbaspirillum, Pseudomonas, Paraburkholderia, and Ralstonia. The bacteria include species of the following genera: Geobacter sp., Serratia sp., Pantoea sp., Ensifer sp., Enterobacter sp., or any combination thereof. In some embodiments, the bacteria include Ensifer adhaerens S3C10. In some embodiments, the bacteria include bacteria belonging to the phylum Actinomycetes. In some embodiments, the bacteria include species of the genera Streptomyces sp., Coxiella sp., and Frankia sp. In some embodiments, the bacteria include bacteria belonging to the phylum Cyanobacteria. In some embodiments, the bacteria include species of Cyanobacteria. In some embodiments, the bacteria include bacteria belonging to the phylum Chloroflexi. In some embodiments, one or more microorganisms include one or more fungi associated with plants or parts thereof. In some embodiments, one or more fungi associated with a plant or part thereof are located in the space between the seed coat and the seed embryo of the plant or part thereof.In some embodiments, one or more fungi associated with a plant or part thereof are placed as a coating on the plant or part thereof. In some embodiments, one or more fungi associated with a plant or part thereof are applied to the plant or part thereof by an in-furrow treatment technique. In some embodiments, one or more fungi associated with a plant or part thereof are applied to the plant or part thereof by a spraying technique. In some embodiments, one or more fungi associated with a plant or part thereof are applied to the plant or part thereof through an irrigation system. In some embodiments, one or more fungi associated with a plant or part thereof are placed in the space between the seed pericarp and the seed allureon cell layer of the plant or part thereof. In some embodiments, one or more fungi include arbuscular mycorrhizal fungi. In some embodiments, one or more fungi include ectomycorrhizal fungi. In some embodiments, one or more fungi include fungi from the genus Trichoderma. In some embodiments, one or more fungi include fungi from the genus Penicillium. In some embodiments, one or more minerals include calcite, aragonite, dolomite, limestone, or any combination thereof. In some embodiments, one or more minerals include CaCO3, MgCO3, CaMg(CO3)2, or any combination thereof. In some embodiments, the promotion of the production of one or more minerals includes the production of ammonia and an increase in the pH of the culture medium in which a plant derived from the plant or a part thereof is grown. In some embodiments, the part thereof is a plant seed, and one or more microorganisms are not naturally present in the space between the seed pericarp and the seed allureon cell layer of the plant or a part thereof. In some embodiments, the plant or a part thereof is a monocotyledonous or dicotyledonous plant.

[0009] Another aspect of the present disclosure is a method for promoting mineralization, the method comprising the step of culturing a plant or a part thereof and one or more microorganisms associated with the plant or a part thereof, wherein the one or more microorganisms are selected to produce or promote the formation of bicarbonates, carbonates, or one or more minerals, or are derived from such microorganisms. In some embodiments, the plant or a part thereof is a commercial plant, plant roots, plant stems, plant leaves, plant seeds, plant fruits, plant tubers, or plant root nodules. In some embodiments, the one or more microorganisms associated with the plant or a part thereof are placed in the plant roots or rhizosphere of the plant or a part thereof. In some embodiments, the one or more microorganisms associated with the plant or a part thereof are placed in the plant roots or rhizosphere of the plant or a part thereof by an irrigation system. In some embodiments, the irrigation system includes in-furrow treatment techniques. In some embodiments, the irrigation system includes spraying techniques. In some embodiments, the plant or a part thereof is derived from a seedling that is integrated with microorganisms via an irrigation system to stimulate the production of one or more minerals by the plant or a part thereof. In some embodiments, the bicarbonates sequester carbon. In some embodiments, the carbon is gaseous carbon. In some embodiments, gaseous carbon is carbon dioxide. In some embodiments, carbonates sequester carbon. In some embodiments, carbon is gaseous carbon. In some embodiments, gaseous carbon is carbon dioxide. In some embodiments, one or more minerals sequester carbon. In some embodiments, carbon is gaseous carbon. In some embodiments, gaseous carbon is carbon dioxide. In some embodiments, one or more microorganisms include bacteria, archaea, fungi, or viruses. In some embodiments, one or more microorganisms include bacteria. In some embodiments, bacteria include endospore-forming bacteria. In some embodiments, bacteria include rhizosphere bacteria. In some embodiments, rhizosphere bacteria include Bacillus sp., Paenibacillus sp., or both.In some embodiments, the bacteria are B. amyloliquefaciens, B. laterosporus, B. licheniformis, B. macerans, B. cereus, B. circulans, B. firmus, B. subtilis, B. sphaericus, B. megaterium, B. coagulans. This includes B. agulans, B. brevis, B. thuringiensis, B. mycoides, B. cucumis, B. endophyticus, B. pumilus, B. velezensis, B. mucilaginosus, B. tequilensis, B. methylotrophicus, or any combination thereof. In some embodiments, the bacteria include Bacillus subtilis S3C23, Bacillus subtilis MP2, Bacillus subtilis RO2C15, Bacillus subtilis RO2C22, Bacillus megaterium 6, Bacillus megaterium S3C21, Bacillus megaterium RO2C12, Bacillus cucumis S3C14, Bacillus endophyticus 5, or any combination thereof. In some embodiments, the bacteria include Bacillus subtilis S3C23. In some embodiments, the bacteria include Bacillus subtilis MP2.In some embodiments, one or more microorganisms include one or more fungi associated with a plant or a part thereof. In some embodiments, one or more fungi associated with a plant or a part thereof are placed in the plant roots or rhizosphere of the plant or a part thereof by an irrigation system. In some embodiments, one or more fungi include arbuscular mycorrhizal fungi. In some embodiments, one or more fungi include ectomycorrhizal fungi. In some embodiments, one or more fungi include fungi from the genus Trichoderma. In some embodiments, one or more fungi include fungi from the genus Penicillium. In some embodiments, one or more microorganisms result in the formation of one or more carbonic anhydrases. In some embodiments, one or more microorganisms result in the formation of one or more carbonic anhydrases belonging to the alpha class. In some embodiments, one or more microorganisms result in the formation of one or more carbonic anhydrases belonging to the beta class. In some embodiments, one or more microorganisms result in the formation of one or more carbonic anhydrases belonging to the gamma class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the delta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the zeta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the eta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the iota class. In some embodiments, one or more minerals include calcite, aragonite, dolomite, limestone, or any combination thereof. In some embodiments, the promotion of the production of one or more minerals includes the production of ammonia and an increase in the pH of the medium in which the plant or a part thereof is grown. In some embodiments, one or more microorganisms are not naturally present in one or more roots. In some embodiments, the plant or a part thereof is a monocotyledonous or dicotyledonous plant. In some embodiments, the plant or a part thereof includes a commercial plant or a part thereof.In some embodiments, commercial plants or a portion thereof consist of a group essentially comprising corn, wheat, rice, sorghum, barley, rye, sugarcane, millet, oats, soybeans, cotton, alfalfa, beans, quinoa, lentils, peanuts, sunflowers, canola, cassava, palm oil, potatoes, sugar beets, cocoa, coffee, lettuce, tomatoes, peas, cabbage, fruit trees, nut trees, forest trees, grasslands, or turfgrasses.

[0010] Another aspect of the present disclosure is a method for sequestrating carbon, the method comprising the step of culturing a plant or a part thereof and one or more microorganisms associated with the plant or a part thereof, wherein the one or more microorganisms are selected to produce or promote the formation of one or more carbonaceous minerals, thereby sequestrating carbon, or are derived from such microorganisms. In some embodiments, the plant or a part thereof is a plant root, a plant stem, a plant leaf, a plant seed, a plant fruit, a plant tuber, or a plant root nodule. In some embodiments, the plant or a part thereof includes a commercial plant or a part thereof. In some embodiments, the commercial plant or a part thereof consists of a group essentially comprising maize, wheat, rice, sorghum, barley, rye, sugarcane, millet, oats, soybeans, cotton, alfalfa, beans, quinoa, lentils, peanuts, sunflowers, canola, cassava, palm oil, potatoes, sugar beets, cocoa, coffee, lettuce, tomatoes, peas, cabbage, fruit trees, nut trees, forest trees, grasslands, or turfgrass. In some embodiments, one or more carbonaceous minerals include one or more gaseous carbonic acid species. In some embodiments, one or more gaseous carbonic acid species include carbon monoxide, methane, or carbon dioxide. In some embodiments, one or more gaseous carbonic acid species is carbon dioxide. In some embodiments, one or more plant-related microorganisms are placed in the plant roots or rhizosphere of the plant or a part thereof. In some embodiments, one or more plant-related microorganisms are placed in the plant roots or rhizosphere of the plant or a part thereof by an irrigation system. In some embodiments, the irrigation system includes in-furrow treatment techniques. In some embodiments, the irrigation system includes spraying techniques. In some embodiments, the plant or a part thereof is derived from a seedling that is integrated with microorganisms via an irrigation system to stimulate the production of one or more minerals by the plant or a part thereof. In some embodiments, one or more microorganisms include bacteria, archaea, fungi, or viruses. In some embodiments, one or more microorganisms include bacteria. In some embodiments, bacteria include endospore-forming bacteria. In some embodiments, bacteria include rhizosphere bacteria.In some embodiments, the rhizosphere bacteria include the genera Bacillus sp, Paenibacillus sp, or both. In some embodiments, the bacteria include B. amyloliquefaciens, B. laterosporus, B. licheniformis, B. macerans, B. cereus, B. circulans, B. firmus, B. subtilis, B. megaterium, B. coagulans, and B. brevis. This includes B. sphaericus, B. thuringiensis, B. mycoides, B. cucumis, B. endophyticus, B. pumilus, B. velezensis, B. mucilaginosus, B. tequilensis, B. methylotrophicus, or any combination thereof. In some embodiments, the bacteria include Bacillus subtilis S3C23, Bacillus subtilis MP2, Bacillus subtilis RO2C15, Bacillus subtilis RO2C22, Bacillus megaterium 6, Bacillus megaterium S3C21, Bacillus megaterium RO2C12, Bacillus cucumis S3C14, Bacillus endophyticus 5, or any combination thereof.In some embodiments, the bacteria include Bacillus subtilis S3C23. In some embodiments, the bacteria include Bacillus subtilis MP2. In some embodiments, one or more microorganisms include one or more fungi associated with a plant or part thereof. In some embodiments, one or more fungi associated with a plant or part thereof are placed in the plant roots or rhizosphere of the plant or part thereof by an irrigation system. In some embodiments, one or more fungi include arbuscular mycorrhizal fungi. In some embodiments, one or more fungi include ectomycorrhizal fungi. In some embodiments, one or more fungi include fungi from the genus Trichoderma. In some embodiments, one or more fungi include fungi from the genus Penicillium. In some embodiments, one or more microorganisms are not naturally present in one or more roots. In some embodiments, the plant or part thereof is a monocotyledonous or dicotyledonous plant. In some embodiments, one or more microorganisms result in the formation of one or more carbonic anhydrases. In some embodiments, one or more microorganisms result in the formation of one or more carbonic anhydrases belonging to the alpha class. In some embodiments, one or more microorganisms result in the formation of one or more carbonic anhydrases belonging to the beta class. In some embodiments, one or more microorganisms result in the formation of one or more carbonic anhydrases belonging to the gamma class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the delta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the zeta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the eta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the iota class.

[0011] Another aspect of the present disclosure is a composition comprising one or more microorganisms, wherein the one or more microorganisms are selected to produce or promote the formation of bicarbonates, carbonates, or one or more minerals, or are derived from such microorganisms. In some embodiments, the plant or part thereof is a plant root, plant stem, plant leaf, plant seed, plant fruit, plant tuber, or plant root nodule. In some embodiments, the plant or part thereof comprises a commercial plant or part thereof. In some embodiments, the commercial plant or part thereof is maize, wheat, rice, sorghum, barley, rye, sugarcane, millet, oats, soybeans, cotton, alfalfa, beans, quinoa, lentils, peanuts, sunflowers, canola, cassava, palm oil, potatoes, sugar beets, cocoa, coffee, lettuce, tomatoes, peas, cabbage, fruit trees, nut trees, forest trees, grasslands, or turfgrass. In some embodiments, the bicarbonate sequesters carbon. In some embodiments, the carbon is gaseous carbon. In some embodiments, the gaseous carbon is carbon dioxide. In some embodiments, carbonates sequester carbon. In some embodiments, carbon is gaseous carbon. In some embodiments, gaseous carbon is carbon dioxide. In some embodiments, one or more minerals sequester carbon. In some embodiments, carbon is gaseous carbon. In some embodiments, gaseous carbon is carbon dioxide. In some embodiments, one or more microorganisms include bacteria, archaea, fungi, or viruses. In some embodiments, one or more microorganisms include bacteria. In some embodiments, bacteria include endospore-forming bacteria. In some embodiments, bacteria include rhizosphere bacteria. In some embodiments, rhizosphere bacteria include Bacillus sp., Paenibacillus sp., or both.In some embodiments, the bacteria are B. amyloliquefaciens, B. laterosporus, B. licheniformis, B. macerans, B. cereus, B. circulans, B. firmus, B. subtilis, B. sphaericus, B. megaterium, B. coagulans. This includes B. agulans, B. brevis, B. thuringiensis, B. mycoides, B. cucumis, B. endophyticus, B. pumilus, B. velezensis, B. mucilaginosus, B. tequilensis, B. methylotrophicus, or any combination thereof. In some embodiments, the bacteria include Bacillus subtilis S3C23, Bacillus subtilis MP2, Bacillus subtilis RO2C15, Bacillus subtilis RO2C22, Bacillus megaterium 6, Bacillus megaterium S3C21, Bacillus megaterium RO2C12, Bacillus cucumis S3C14, Bacillus endophyticus 5, or any combination thereof. In some embodiments, the bacteria include Bacillus subtilis S3C23. In some embodiments, the bacteria include Bacillus subtilis MP2.In some embodiments, one or more microorganisms include one or more fungi associated with a plant or a part thereof. In some embodiments, one or more fungi associated with a plant or a part thereof are placed in the plant roots or rhizosphere of the plant or a part thereof by an irrigation system. In some embodiments, one or more fungi include arbuscular mycorrhizal fungi. In some embodiments, one or more fungi include ectomycorrhizal fungi. In some embodiments, one or more fungi include fungi from the genus Trichoderma. In some embodiments, one or more fungi include fungi from the genus Penicillium. In some embodiments, one or more minerals include calcite, aragonite, dolomite, limestone, or any combination thereof. In some embodiments, the promotion of the production of one or more minerals includes the production of ammonia and an increase in the pH of the medium in which the plant or a part thereof is grown. In some embodiments, one or more microorganisms include one or more carbonic anhydrases. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the alpha class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the beta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the gamma class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the delta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the zeta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the eta class. In some embodiments, one or more carbonic anhydrases include carbonic anhydrases belonging to the iota class.

[0012] Embedding by citation All publications, patents, and patent applications referenced herein are incorporated by reference to the extent that each individual publication, patent, or patent application is specifically and individually indicated to be incorporated by reference. To the extent that any publications and patents or patent applications incorporated by reference are in conflict with the disclosures contained herein, this Specification is intended to supersede and / or take precedence over such conflicting material. [Brief explanation of the drawing]

[0013] The patent or application file must include at least one drawing made in color. A copy of this patent or patent application publication containing the color drawing will be provided by the relevant office after the required fees have been requested and paid.

[0014] Novel features of the methods and compositions described herein are specifically described in the appended claims. The features and advantages of the methods and compositions described herein will be better understood by referring to the following detailed description of specific embodiments in which the principles of the methods and compositions described herein are applied, and by referring to the appended drawings. [Figure 1] High CO2 concentrations in the plant rhizosphere represent an opportunity for carbon sequestering mediated by microorganisms that produce carbonic anhydrase. [Figure 2] This specification describes a two-stage plant strengthening strategy. (101) illustrates the seed of a monocotyledonous plant. The seed treatment process described herein involves incorporating bacteria (or their endospores) (106) between the aleurone layer (103) and the pericarp (102). The aleurone layer (103) separates the endosperm (104) from the outer layer. [Figure 3] This is a process diagram showing the seed treatment process (Microprime® seed treatment process). [Figure 4] This document describes a method for obtaining stable microbial seed treatment technology (Microprime® trademark). [Figure 5A] This shows the internal spaces of corn seeds (Zea mays) where bacteria are located after Microprime® seed treatment. [Figure 5B] This image shows a magnified view of the internal space of corn seeds (Zea mays) where bacteria are located after Microprime® seed treatment. [Figure 6A]The graphs show the population growth and logarithm of population growth of Bacillus subtilis (S3C23 strain) endospores inside Microprime® corn seeds before germination. Seeds sown on agar plates using Murashige and Skoog 50% medium initially had a colony-forming unit (CFU) of 164,000 at sowing. After 3 days, the population within the seed reached a level of 7,200,000 CFU / seed. Each point represents the mean ± standard error of 3 pools of 5 seeds per pool. [Figure 6B] The graphs show the population growth and logarithm of population growth of Bacillus subtilis (S3C23 strain) endospores inside Microprime® corn seeds before germination. Seeds sown on agar plates using Murashige and Skoog 50% medium initially had a colony-forming unit (CFU) of 164,000 at sowing. After 3 days, the population within the seed reached a level of 7,200,000 CFU / seed. Each point represents the mean ± standard error of 3 pools of 5 seeds per pool. [Figure 7A] The graphs show the population growth and logarithm of population growth of Bacillus subtilis (S3C23 strain) endospores inside Microprime® rice seeds before germination. Seeds sown on agar plates using Murashige and Skoog 50% medium initially had a colony-forming unit (CFU) of 51,167 at sowing. After 3 days, the population within the seed reached a level of 36,666,667 CFU / seed. Each point represents the mean ± standard error of 3 pools of 5 seeds per pool. [Figure 7B]The graphs show the population growth and logarithm of population growth of Bacillus subtilis (S3C23 strain) endospores inside Microprime® rice seeds before germination. Seeds sown on agar plates using Murashige and Skoog 50% medium initially had a colony-forming unit (CFU) of 51,167 at sowing. After 3 days, the population within the seed reached a level of 36,666,667 CFU / seed. Each point represents the mean ± standard error of 3 pools of 5 seeds per pool. [Figure 8A] The graphs show the population growth and logarithm of population growth of Bacillus subtilis (S3C23 strain) endospores inside Microprime™ soybean seeds before germination. Seeds sown on agar plates using Murashige and Skoog 50% medium initially had a colony-forming unit (CFU) of 123,333 at sowing. After 3 days, the population within the seeds reached a level of 674,666,667 CFU / seed. Each point represents the mean ± standard error of 3 pools of 5 seeds per pool. [Figure 8B] The graphs show the population growth and logarithm of population growth of Bacillus subtilis (S3C23 strain) endospores inside Microprime™ soybean seeds before germination. Seeds sown on agar plates using Murashige and Skoog 50% medium initially had a colony-forming unit (CFU) of 123,333 at sowing. After 3 days, the population within the seeds reached a level of 674,666,667 CFU / seed. Each point represents the mean ± standard error of 3 pools of 5 seeds per pool. [Figure 9]This graph shows the survival rate of Bacillus subtilis endospores (S3C23 strain) inside Microprime® corn seeds over a period of 1 to 18 months after treatment with Microprime® corn seeds. Each time point represents the mean ± standard error of three pools of 5 seeds each. The germination rate of S3C23 Microprime® seeds at 18 months was the same as that of untreated seeds, at 98.33% (n=60 seeds / treatment). [Figure 10] This section describes the carbonic anhydrase activity of lysate samples prepared from liquid cultures of Bacillus subtilis (S3C23 strain), measured by enzymatic hydrolysis of 4-nitrophenylacetic acid to 4-nitrophenol and acetic acid, according to the protocol by Zhuang et al, 2018, which is incorporated herein by reference in its entirety. Values ​​are given as carbonic anhydrase (CA) activity / total protein (mg / ml). The activity in the samples shows a sharp increase at pH 8.5 compared to pH 7.5, consistent with the expected pH dependence of carbonic anhydrase. [Figure 11] This diagram illustrates the enzyme-induced precipitation of carbonate and bicarbonate ions as calcium carbonate (CaCO3). Lysates prepared from Bacillus subtilis (S3C23 strain), recombinant bovine carbonic anhydrase, or bovine serum albumin (BSA, negative control) were mixed with CaCl2 (final concentration 100 mM), Tris pH 8 (final concentration 200 mM), and 50% CO2 saturated water (added last to initiate the reaction). The time to calcium carbonate formation was visually evaluated and recorded. The values ​​were normalized to the time to abiotic CaCO3 precipitation. Recombinant bovine CA rapidly induced precipitation in a dose-dependent manner. 500 μl of S3C23 lysate induced precipitation significantly faster than abiotic precipitation. [Modes for carrying out the invention]

[0015] While various embodiments of the present invention are shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided only as examples. Many modifications, changes, and substitutions can be understood by those skilled in the art without departing from the present invention. It should be understood that various alternatives to the embodiments of the present invention described herein may be used.

[0016] Land used for plant cultivation provides an ideal location for CO2 sequestration because, as a result of the respiration of plant roots and the soil microbiome, the CO2 levels are significantly higher compared to atmospheric CO2 levels.

[0017] As described herein, plant seeds and plants derived from plant seeds can be modified with one or more microorganisms described herein to enhance bicarbonate production and mineral formation in a given plot of soil in which the plants are grown. The slow, rate-limited process of bicarbonate production can be accelerated by using an enzyme called carbonic anhydrase (CA). CA, a zinc-containing metalloenzyme, catalyzes the reversible hydration of CO2 to bicarbonate, and protons are about 10 times faster than in non-enzymatic processes. 7 High. CA is the fastest enzyme known to catalyze the reversible hydration of CO2, and the typical rate of different types of CA is 10⁻¹⁰ 6 s -1 It can reach levels of cations. CA is ubiquitous in animals, plants, archaea, and bacteria. This includes many fungi and bacteria commonly found in soil that produce intracellular and extracellular CA. Due to CA's extremely high selectivity for CO2, it is a perfect candidate for CO2 sequestration in gases mixed with other polluting gases. There is plenty of compelling evidence (20, 21) that CA plays a role in sequestrating atmospheric CO2 into bicarbonates (HCO3-) which can precipitate and form minerals by reacting with various cations.

[0018] According to the present disclosure, one or more microorganisms may be selected based on a sufficiently high CA activity or CA activity to produce a desired amount of HCO3- in a predetermined section of soil. These microorganisms may be associated with various plant seeds, plants, and / or the rhizosphere of roots, as described herein. In some embodiments, microorganisms with low CA expression levels can be genetically modified to enhance CA expression. In some embodiments, microorganisms that do not express CA can be genetically modified to express CA.

[0019] Plants derived from the compositions described herein can be cultivated in a predetermined section of soil to enhance bicarbonate and mineral formation in the soil.

[0020] The mineralization process induced by carbonic anhydrase (CA) begins with the reversible hydration of CO2 to bicarbonate ions (shown in Equation 1). CO2 + H2O ⇔ HCO3 - + H + (1)

[0021] The production of carbonate ions from bicarbonate by hydrolysis is the second step in mineralization (shown in Equation 2). These carbonate ions and bicarbonate ions increase the pH of the medium. The pH of the medium can be further increased up to 9.2 by the production of ammonia by microorganisms. HCO3 - + OH - ⇔ CO3 2- + H2O (2)

[0022] The final stage of mineralization involves the precipitation of carbonate and bicarbonate ions by cations (Equation 3). Ca 2+ + CO3 2- → CaCO3(3)

[0023] Because soils naturally contain different types of cations, CA-expressing soil microorganisms found in roots and / or rhizospheres are well-suited to the sustained conversion of CO2 into minerals. CA can also play a secondary role in increasing the supersaturation of several minerals in liquid culture media by releasing large amounts of carbonates and bicarbonates. The biomineralization process can be further accelerated by a group of genes known as lcfA (consisting of five genes: lcfA, ysiA, ysiB, etfB, and etfA), which play a role in the biomineralization of carbonates from Bacillus subtilis.

[0024] To date, no technology is available that can reliably deliver microorganisms to CO2-releasing sites (roots) for effective CO2 sequestration. Integrating the compositions described herein with Microprime® is highly suitable for agricultural practice not only to maintain soil productivity but also to capture CO2 that could otherwise be released into the atmosphere. Furthermore, the methods and compositions described herein are cost-effective, costing only $0.20 per acre and using environmentally friendly, environmentally produced microorganisms that pose no threat to farmland or crops.

[0025] The microorganisms described herein will reduce the level of carbon output and thus increase carbon sequestration on land by capturing CO2 in various forms. The microorganisms described herein employ various mechanisms for efficiently capturing CO2 in multiple microbial products. Notable products include bicarbonates and minerals (solids). Minerals may include calcite, aragonite, dolomite, and limestone. Limestone (CaCO3), one of the microbial products, is beneficial to agricultural land in addition to sequestrating gaseous CO2. The formation of calcite in the soil eliminates the need for farmers to use limestone treatment to maintain soil productivity. Limestone improves soil structure and increases pH.

[0026] Composition containing plant seeds and bacteria In certain embodiments, compositions comprising plant seeds and one or more microorganisms associated with the plant seeds are disclosed herein, wherein the one or more microorganisms are selected to produce or promote the formation of one or more minerals, or are derived from such microorganisms.

[0027] Biopriming and seed treatment methods: Microprime® A typical seed priming protocol involves immersing the seeds in any solution containing the necessary priming agents (inorganic and organic salts, nanoparticles, plant growth regulators, and / or plant growth-promoting bacteria), followed by refriing the seeds. This initiates the germination process other than the emergence of the radicle, as shown in Heydecker et al., 1973; Mahakham et al., 2017; McDonald, 1999; Song et al., 2017; and Wright et al., 2003 (which is incorporated herein by reference in its entirety). Seed priming using osmotic solutions (osmopriming) has been practiced for decades (Heydecker et al., 1973) and is now a common commercial practice for selected high-value horticultural seeds. The concept has also been extended to hydropriming in cereal and leguminous crops, reviving the “farm” priming technique (Harris et al., 2001). In recent years, several metal and carbon-based nanoparticles (e.g., AgNPs16, AuNPs5, CuNPs17,18, ZnNPs17,18, fullerene22, and carbon23 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 the different priming techniques (e.g., hydropriming, osmopriming, nanopriming), the internal space within the seed offers potentially ideal conditions for bacterial inoculation and colonization when this procedure is performed using microbial cells (McQuilken et al., 1998; Ashraf and Foolad, 2005; Bennett et al., 2009; Tabassum et al., 2018; Wright et al., 2003; the above references are incorporated herein by reference in their entirety).

[0028] Since the early 1990s, biopriming methods have been widely used for a wide range of crops and are undoubtedly recognized as an environmentally friendly agricultural technique (O'Callaghan, 2016; Taylor and Harman, 1990; the above references are incorporated herein by reference in their entirety). Biopriming techniques are often mistakenly defined as applying whole microorganisms, their exudates, or several biologically active compounds to the outside of seeds (provided by El-Mougy and Abdel-Kader, 2008; Muller and Berg, 2008; Song et al., 2017; Saber et al., 2012; the above references are incorporated herein by reference in their entirety). More precisely, biopriming incorporates both biological (inoculating seeds with beneficial microorganisms) and physiological (seed hydration) elements into seeds by promoting seedling emergence rate and uniformity, and also improving plant traits. Seeds treated with microorganisms differ fundamentally from other biological seed treatments in that, because the cells may be alive during the microbial treatment, the colonization and proliferation of the added microorganisms must occur within the seed. However, little of the literature to date has explained this difference in detail. Specifically, there have been no reported results or studies on: 1) the survival and / or proliferation of biological agents within the seed over a relevant timeframe (several months); 2) the shelf life and effective germination of seeds several months after treatment; 3) the effective inoculation of microbes and their interaction with plants after a relevant period of seed storage; and 4) an economically viable methodology (considering relevant factors such as the time required for seed treatment, inputs, and energy), which is scalable and therefore feasible within conventional seed business models. Furthermore, bio-osmopriming has been demonstrated to significantly enhance germination uniformity and plant growth traits when associated with bacterial coating procedures (Bennett et al., 2009; Raj et al., 2004; Sharifi, 2011; Sharifi et al., 2011; Shariffi et al., 2012).The above references are incorporated herein by reference in their entirety. 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; the above references are incorporated herein by reference in their entirety). Similarly, cell suspensions ranged widely from 10⁵ to 10⁹ cells per gram of seed and depended 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. The above references are incorporated herein by reference in their entirety).

[0029] According to the latest technology, Bacillus sp. exudates for inducing immunity in cucumber plants have been described by Song et al. (Song et al., 2017). This approach has several misleading results in both methodology and scope. This is because: 1) seeds are biostimulated by peptide-based compounds instead of using bacterial inoculants or plant growth promoters derived therefrom; 2) live microorganisms are not incorporated into the seeds to bring the microorganisms or their exudates into contact with the embryo in the early post-dormancy stage of seed germination; 3) it is not confirmed whether the biological agent (e.g., cyclodipeptide) entered the seed and stimulated the PGP effect (changes in gene expression) in the early stages of the plant embryo (before the pericarp ruptures); and 4) the stability of the plant immunity trigger elicitor over time is not reported. This last issue is particularly important because commercially viable agricultural microbial technologies must be stable over relatively long periods (e.g., more than 6 months) to fit into the current agricultural distribution system. In addition, the biological priming agents used in this referenced study are particularly unstable over time and are susceptible to changes due to biological and abiotic environmental factors (e.g., temperature, pH, biodegradation activity by other microorganisms).

[0030] The Serratia plymuthica strain HRO-C48 has been reported as a bioagent for seed inoculation procedures (Muller and Berg, 2008). This study attempted to compare three different techniques: pelletizing, 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 sector. In fact, the viability of the HRO-C48 strain was only measured over a very short storage period (30 days). Other ambiguous topics reported by the authors depend on the optimization procedure for biopriming, as 1) a high initial cell density was adjusted for seed immersion, and 2) a long incubation time of seeds in the presence of the bioagent was used (reported as 12 hours). Indeed, all such aspects are often not viable parameters for industrially and commercially implementing this method (Muller and Berg, 2008).

[0031] Several other studies that have pointed to incorporating synthetic microbial formulations into seeds have also been reported in the latest technology. For example, U.S. Patent Applications 2016 / 0338360A1 and 2016 / 0330976A1 refer to seeds containing beneficial bacteria. The methods presented in both of these referenced studies are based on directly inoculating different parts of flowers or plants to obtain seeds containing the desired microorganisms (Mitter et al, 2016a, 2016b; the above literature is incorporated herein by reference in its entirety).

[0032] Disclosed herein is a novel, effective, and reliable alternative to conventional biopriming technologies that directly addresses the problems described above. The proposed seed treatment method, named Microprime®, is a well-designed, calculated, executed, and controlled process for obtaining commercially viable seeds containing desired microorganisms. More precisely, disclosed herein is a stable microbial seed treatment method that incorporates a single microorganism and / or a synthetic consortium of microorganisms, as well as its exudates and / or its individualized biomolecules, into the seed through an industrially scalable process, taking into account process cost, time and energy, technological stability over time (both plant embryos and inoculants), adaptability to multiple soils, stability under different environmental conditions, and compatibility with conventional distribution chains for agricultural inputs. This method involves controlled, economical, and rapid hydration of seeds in an aqueous solution of a osmotic active liquid medium supplemented with specific amounts of beneficial microorganisms or a synthetic consortium of microorganisms, and / or their exudates, and / or their individualized biomolecules, in addition to surfactants that enhance permeability within the seed, and / or a group of nutrients that promote microbial colonization within the seed, and / or auxiliary reagents that promote bacterial endospore formation. The survival of the biological agents and the extension of the shelf life of the treated seeds are guaranteed by Microprime® seed technology.

[0033] The compositions and methods disclosed herein propose a novel strategy for enabling plant traits and increasing their yield. This strategy is based on two effects on plant seeds achieved by implementing a specific seed treatment method (Microprime®) described below:

[0034] 1. Functional bacterial loading of seeds: The current seed treatment method loads seeds with endospore-forming bacteria and / or endospores. As a result of Microprime® seed treatment, endospore-forming bacteria and / or endospores are allocated to the seed in the interstitial space located between the seed pericarp and its aleuron cell layer, as shown in Figures 2 and 5A and 5B. The endospore-forming bacteria and / or endospores incorporated into the seed correspond to strains that have the ability to effectively colonize the plant's rhizosphere and also have the ability to sequester CO2 by converting it to bicarbonates and ultimately carbonate minerals. The endospore conversion ability of selected bacteria and their allocation within the seed ensure stability after Microprime® seed treatment and throughout the entire commercial storage period. This process is confirmed by the bacterial cell count over time.

[0035] For the methods and compositions disclosed herein to be valuable and actually applicable on an industrial scale, they must be cost-effective and scalable. Seed treatment processes such as those disclosed herein involve multiple steps that require time, inputs, and energy. The methods and compositions disclosed herein prioritize minimizing the processing cost and time of seed treatment as much as possible. The Microprime® method aims to make the seed treatment process effective when carried out at room temperature (20-24°C) while reducing seed immersion to less than 20 minutes to 16 hours. Achieving the latter is not easy because, in addition to the desired minimum number of bacteria, endospore-forming bacteria, and / or endospores in the seeds after Microprime® seed treatment, the bacteria must remain stable and viable over time, so that the seeds (as a product) may not be affected by storage, packaging, logistics, and planting processes, just like conventional seeds that have not undergone Microprime® seed treatment.

[0036] The proposed methodology involves immersing pre-disinfected seeds in a seed treatment medium (hereinafter referred to as Microprime® solution) containing nutrients, surfactants, and salts. Figure 3 shows a process diagram illustrating the seed treatment process.

[0037] Stability of bacterial seed treatment The time-dependent stability of bacteria inside seeds is neither an easy nor a clear problem to address. When bacteria are incorporated into seeds using the seed treatment method disclosed herein (Microprime® seed treatment), the location of the bacteria inside corn seeds is shown in Figures 5A and 5B.

[0038] Figure 5A illustrates the space within a corn seed where bacteria marked with red fluorescent protein (RFP) remain after Microprime® seed treatment (pink fibers). This location is the gap between the seed pericarp and the allureon cell layer of the seed, separating the endosperm and embryo from the outer layer. This is a place where certain microorganisms can comfortably reside for a limited period before the cells inevitably die due to the depletion of available nutrients, or, in the case of certain microorganisms, it is where the process of endosporulation (bacteria from the phyla Firmicutes, Proteobacteria, and Actinomycetes) begins. The advantage of accumulating in the aforementioned location is that the microorganisms are protected from other microorganisms or external factors that could affect the immediate integrity of the microorganisms, such as dehydration.

[0039] To address the problem of long-term survival rates due to nutrient deficiencies, the methodology varies depending on the type of bacteria being treated. Some bacteria have the ability to stop growing and enter a physical state called endospores under certain conditions (primarily in scenarios where feasibility risks are perceived). As endospores, bacteria enter a dormant state that may be absent from nutrients for extended periods. For bacteria from the Firmicutes, Proteobacteria, and Actinomycetes phyla that have the ability to produce endospores, Microprime® solution is supplemented with specific salts that, when incorporated into seeds, induce the bacteria to enter this dormant state. Doing the latter ensures the long-term survival rate of bacteria in seeds. When endospores are seen again under favorable conditions with moisture and nutrients (e.g., when seeds are sown), they return to an active bacterial state (vegetative cells) and begin normal function and vegetative reproduction.

[0040] Another strategy involves directly supplementing the Microprime® solution with endospores, rather than forcing bacterial conversion during the seed treatment process. The latter demonstrated better yields in terms of endospores per seed observed after Microprime® seed treatment.

[0041] Table 1 shows several bacterial genera that are of particular interest to this proposed novel seed treatment due to their ability to convert to endospores.

[0042] [Table 1]

[0043] Bacteria of the genus Bacillus are among the most endospore-forming bacteria. To ensure sufficient bacterial growth within seeds, it is necessary to supplement the Microprime® solution with nutrients particularly compatible with the selected bacteria, or to directly add the endospores of the desired bacteria to be incorporated into the seeds to the Microprime® solution.

[0044] Biological priming of seed embryos The method of this disclosure intends to treat plant seeds with a bacterial composition designed as described above (Microprime® seed treatment). Such treatments can employ the osmotic pressure of the seed to enable bacterial incorporation, and the treatment first described by Smith et al. for introducing chemical priming agents into seeds is now called osmopriming. However, the method of this disclosure is adapted for incorporation of bacterial populations into dormant seeds, and aims to generate or promote early conditioning of the emerging plant by direct biological priming of the embryo after dormancy has ended and appropriate environmental or agricultural conditions have induced the first stage of germination. This novel approach offers unprecedented advantages over conventional bacterial formulations designed to generate or promote biological priming, because the incorporated bacteria are protected within the dormant seed and are conveniently positioned to generate or promote permanent priming of the embryo from the earliest possible developmental stage, either by themselves or by the action of their exudates. Furthermore, the treated seeds are available for normal transport, storage, coating pelletization, and sowing treatments according to standard agricultural practices, without additional requirements regarding handling, nutritional additives, preservatives, or irrigation, and without any limitations on suitability regarding pest or plant disease control agents. Thus, the method described herein (Microprime® seed treatment) also offers clear advantages over seed biopriming (Mahmood et al., 2016), because the method in the aforementioned literature involves pre-germination and dormancy arrest reactions of seeds, which reduce storage viability and limit the feasibility of handling and treatment.

[0045] In addition, the method of this disclosure differs from previously reported methods of inoculating seeds using a parent plant as a reactor for microbial growth or by inoculating the plant's sexual organs (Mitter et al., 2016a; Mitter et al., 2016b). Such methods implicitly imply an inherent bias in the types of bacteria that can ultimately be incorporated into the seeds, because successful inoculants must survive within the plant's target organs or tissues, compete with endogenous microorganisms, and access the internal space of the seed on their own. The Microprime® strategy presented herein is not hindered by endoparasitic ability or tissue survival, because the artificially incorporated bacteria do not need to be endosymbionts, do not need to face the defense response of a mature plant, or outnumber endoparasitic microbial communities, but are only required to survive long enough or have exudates that can reach the plant embryo in order to generate or promote the formation of molecular priming in the plant.

[0046] Due to the inherent advantages and differentiating features described above, the method of this disclosure is non-obvious to those skilled in the art of techniques involving bacterial plant growth stimulation. In fact, for this strategy to be successful, certain important conditions must be met by the candidate bacteria of the treatment composition, which are not necessarily considered in standard formulations of plant growth-promoting microorganisms. First, the bacterial composition must be designed using the above-mentioned efficacy and suitability criteria to avoid competition and / or antagonism within the seed. The absence of these effects must be experimentally evaluated before formulation of the composition. Seed internalization must be evaluated for each bacterium contained in the designed composition, the saturation curve determined, and the survival of the bacteria during storage time and seed treatment procedures. Furthermore, analysis of the seed endothelium must also be performed to evaluate the presence and viability of the desired bacteria and the relative abundance of each constituent strain to others (Figure 4).

[0047] Plant materials must further be conditioned before treatment with a specific bacterial composition. Seeds may be sterilized to eliminate background noise while determining the effectiveness of the Microprime® seed treatment.

[0048] After Microprime® has occurred, transcriptional analysis of marker genes related to pathogen defense, abiotic stress tolerance, and development must be determined to confirm the beneficial effects of the bacteria on the treated seeds. This analysis must be performed after the dormancy stage of the seeds, and before the rupture of the seed pericarp and endosperm and the emergence of the radicle. Evaluation of transcriptional changes in the developing embryo resulting from the previous bacterial treatment of dormant seeds is also an important step in validating the methodology, because this provides rapid confirmation of the priming effect, and the results are not affected by external factors that appear after seed rupture, including access from the seed to other microorganisms outside the developing plant tissue and / or the chemical composition of the surrounding soil or growth substrate.

[0049] The methods and compositions disclosed herein can be combined into the Microprime® Seed Treatment Method, which involves incorporating seeds into a saline solution containing a seed-compatible bacterial composition, a bacteriocompatible nutrient (when using non-endospore-forming bacteria), and a surfactant, along with minerals supplemented to increase the conversion rate of endospore-forming bacteria to endospores, in order to increase the bacterial cell load on the seeds during short-term seed immersion at room temperature.

[0050] Figure 3 illustrates this method for obtaining stable microbial seed treatment techniques.

[0051] Modified plant seeds In one embodiment, modified plant seeds comprising a microorganism or a microbial exudate incorporated into the seed are provided herein. In some embodiments, the microorganism or exudate sequesters CO2 by converting it to bicarbonates and ultimately to carbonate minerals. In some embodiments, CO2 is sequestrated by the formation of bicarbonates. In some embodiments, CO2 is sequestrated by the formation of one or more carbonate minerals. In some preferred embodiments, the microorganism is an endospore-forming bacterium or its endospores.

[0052] In some embodiments, the microorganism or exudate is incorporated into the interior of the seed. In some embodiments, the microorganism or exudate is incorporated into the seed beneath the pericarp. In some embodiments, the microorganism or exudate is incorporated into the seed between the pericarp and the aleurone cell layer. In some embodiments, the microorganism or exudate comes into contact with the embryo of the seed. In some embodiments, the microorganism or exudate does not come into contact with the embryo of the seed. In some embodiments, the microorganism or exudate comes into contact with the endosperm of the seed. In some embodiments, the microorganism or exudate does not come into contact with the endosperm of the seed. In some embodiments, the microorganism or exudate is incorporated into the seed in the space between the seed coat and the seed embryo. In some embodiments, the microorganism or exudate is incorporated in the space between the pericarp of the seed and the aleurone cell layer of the seed.

[0053] Modified plant seeds may be any type of plant seed. In some embodiments, the modified seeds are monocotyledonous plant seeds. In some embodiments, the plant seeds are maize, wheat, rice, barley, rye, sugarcane, millet, oats, or sorghum seeds. In some embodiments, the plant seeds are maize seeds. In some embodiments, the plant seeds are maize (Zea maize) seeds. In some embodiments, the seeds are modified soybean seeds. In some embodiments, the seeds are modified soybean (Glycine max) seeds. In some embodiments, the seeds are modified rice seeds. In some embodiments, the seeds are rice (Oryza sativa) seeds. In some embodiments, the modified seeds are dicotyledonous plant seeds. In some embodiments, the seeds are soybean, cotton, alfalfa, beans, quinoa, lentils, peanuts, sunflower, canola, cassava, palm oil, potato, sugar beet, cocoa, coffee, lettuce, tomato, or cabbage seeds. In some embodiments, the seeds are genetically modified organism (GMO) seeds. In some embodiments, the seeds are non-GMO seeds.

[0054] The amount of microorganisms or exudates incorporated into the seed must be sufficient to effectively sequestrate CO2 into the soil. In some embodiments, the amount of microorganisms incorporated into the seed ranges from approximately 250 colony-forming units (CFUs) to approximately 5,000 CFUs. In some embodiments, the amount of microorganisms incorporated into the seeds is approximately 250 CFU to 500 CFU, approximately 250 CFU to 750 CFU, approximately 250 CFU to 1,000 CFU, approximately 250 CFU to 2,000 CFU, approximately 250 CFU to 3,000 CFU, approximately 250 CFU to 4,000 CFU, approximately 250 CFU to 5,000 CFU, approximately 500 CFU to 750 CFU, approximately 500 CFU to 1,000 CFU, approximately 500 CFU to 2,000 CFU, approximately 500 CFU to 3,000 CFU, approximately 500 CFU to 4,000 CFU, approximately 500 CFU to 5,000 CFU, approximately 750 CFU to 1,000 CFU, and approximately 7 The CFU levels are approximately 50 CFU to 2,000 CFU, approximately 750 CFU to 3,000 CFU, approximately 750 CFU to 4,000 CFU, approximately 750 CFU to 5,000 CFU, approximately 1,000 CFU to 2,000 CFU, approximately 1,000 CFU to 3,000 CFU, approximately 1,000 CFU to 4,000 CFU, approximately 1,000 CFU to 5,000 CFU, approximately 2,000 CFU to 3,000 CFU, approximately 2,000 CFU to 4,000 CFU, approximately 2,000 CFU to 5,000 CFU, approximately 3,000 CFU to 4,000 CFU, approximately 3,000 CFU to 5,000 CFU, or approximately 4,000 CFU to 5,000 CFU. In some embodiments, the amount of microorganisms incorporated into the seed 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 seed 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 seed 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.

[0055] In some embodiments, the microorganisms or exudates incorporated into the seeds have long-term storage stability. In some embodiments, the modified seeds have storage stability for about 3 months to about 36 months. In some embodiments, the modified seeds have storage stability for about 3 months to about 6 months, about 3 months to about 9 months, about 3 months to about 12 months, about 3 months to about 15 months, about 3 months to about 18 months, about 3 months to about 21 months, about 3 months to about 24 months, about 3 months to about 30 months, about 3 months to about 36 months, about 6 months to about 9 months, about 6 months to about 12 months, Approximately 6 months to 15 months, approximately 6 months to 18 months, approximately 6 months to 21 months, approximately 6 months to 24 months, approximately 6 months to 30 months, approximately 6 months to 36 months, approximately 9 months to 12 months, approximately 9 months to 15 months, approximately 9 months to 18 months, approximately 9 months to 21 months, approximately 9 months to 24 months, approximately 9 months to 30 months, approximately 9 months Months to approximately 36 months, approximately 12 months to approximately 15 months, approximately 12 months to approximately 18 months, approximately 12 months to approximately 21 months, approximately 12 months to approximately 24 months, approximately 12 months to approximately 30 months, approximately 12 months to approximately 36 months, approximately 15 months to approximately 18 months, approximately 15 months to approximately 21 months, approximately 15 months to approximately 24 months, approximately 15 months to approximately 30 months, approximately 15 months to approximately The seeds have storage stability for 36 months, approximately 18 to 21 months, approximately 18 to 24 months, approximately 18 to 30 months, approximately 18 to 36 months, approximately 21 to 24 months, approximately 21 to 30 months, approximately 21 to 36 months, approximately 24 to 30 months, approximately 24 to 36 months, or approximately 30 to 36 months. In some embodiments, the modified seeds have storage stability for approximately 3 months, approximately 6 months, approximately 9 months, approximately 12 months, approximately 15 months, approximately 18 months, approximately 21 months, approximately 24 months, approximately 30 months, or approximately 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 approximately 6 months, 9 months, 12 months, 15 months, 18 months, 21 months, 24 months, 30 months, or 36 months.

[0056] In some embodiments, the microorganisms incorporated into the seeds are stable after incorporation. In some embodiments, the microorganisms are stable for longer than 30 days, longer than 6 months, longer than 1 year, or longer than 2 years. In some embodiments, the microorganisms are stable for longer than 30 days. In some embodiments, the microorganisms are stable for longer than 6 months. In some embodiments, the microorganisms are stable for longer than 1 year. In some embodiments, the microorganisms are stable for longer than 2 years.

[0057] The microorganisms or exudates incorporated into the plant seeds may be any of the microorganisms provided herein, or any other microorganisms. In some embodiments, the microorganism is a microbe. In some embodiments, the microorganism is an endosporic microbe. In some embodiments, the microorganism is an endosporic microbe or its endospores. In some embodiments, the microorganism is an endospore of a microorganism provided herein. In some embodiments, the microorganism is an endosporic bacterium or its endospores.

[0058] Methods for incorporating bacteria In one embodiment, a method for incorporating one or more microorganisms or their exudates into one or more plant seeds is provided herein. In some embodiments, the method includes the step of disinfecting the plant seeds. In some embodiments, the method includes the step of bringing the seeds into contact with a solution containing one or more microorganisms or their exudates. In some embodiments, the solution further includes salt. In some embodiments, the method includes the step of incubating the seeds with the solution for a certain period of time. In some embodiments, the period of time is sufficient to introduce a desired amount of the microorganisms or their exudates into the plant seeds. In some embodiments, the method incorporates a desired amount of the microorganisms or their exudates into the seeds.

[0059] In some embodiments, the method includes the step of contacting 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 contains sodium ions, lithium ions, or potassium ions. In some embodiments, the salt contains alkali metal ions. In some embodiments, the salt contains alkaline earth metal ions. In some embodiments, the salt contains halide ions. In some embodiments, the salt is an alkali or alkaline earth halide salt. In some embodiments, the salt contains chloride ions, bromide ions, or iodide ions. In some embodiments, the salt is a sulfate, phosphate, carbonate, or nitrate.

[0060] The salt may be present in the solution at any appropriate concentration. In some embodiments, the solution contains about 0.85% salt (w / v). In some embodiments, the solution contains about 0.1% to about 1.25% salt (w / v). In some embodiments, the solution contains about 0.1% to about 2.0% salt (w / v). In some embodiments, the solution is approximately 0.1% to approximately 0.25%, approximately 0.1% to approximately 0.5%, approximately 0.1% to approximately 0.6%, approximately 0.1% to approximately 0.7%, approximately 0.1% to approximately 0.75%, approximately 0.1% to approximately 0.8%, approximately 0.1% to approximately 0.85%, approximately 0.1% to approximately 0.9%, approximately 0.1% to approximately 0.95%, approximately 0.1% to approximately 1%, approximately 0.1% to approximately 1.25%, approximately 0.25% to approximately 0.5%, approximately 0.25% to approximately 0.6%, approximately 0.25% to approximately 0.7%, approximately 0.25% to approximately 0.75%, approximately 0.25% to approximately 0. 0.8%, approximately 0.25% to 0.85%, approximately 0.25% to 0.9%, approximately 0.25% to 0.95%, approximately 0.25% to 1%, approximately 0.25% to 1.25%, approximately 0.5% to 0.6%, approximately 0.5% to 0.7%, approximately 0.5% to 0.75%, approximately 0.5% to 0.8%, approximately 0.5% to 0.85%, approximately 0.5% to 0.9%, approximately 0.5% to 0.95%, approximately 0.5% to 1%, approximately 0.5% to 1.25%, approximately 0.6% to 0.7%, approximately 0.6% to 0.75%, approximately 0.6% to 0. 8%, approximately 0.6% to approximately 0.85%, approximately 0.6% to approximately 0.9%, approximately 0.6% to approximately 0.95%, approximately 0.6% to approximately 1%, approximately 0.6% to approximately 1.25%, approximately 0.7% to approximately 0.75%, approximately 0.7% to approximately 0.8%, approximately 0.7% to approximately 0.85%, approximately 0.7% to approximately 0.9%, approximately 0.7% to approximately 0.95%, approximately 0.7% to approximately 1%, approximately 0.7% to approximately 1.25%, approximately 0.75% to approximately 0.8%, approximately 0.75% to approximately 0.85%, approximately 0.75% to approximately 0.9%, approximately 0.75% to approximately 0.95%, approximately 0.75% to approximately 1 Contains %, approximately 0.75% to 1.25%, approximately 0.8% to 0.85%, approximately 0.8% to 0.9%, approximately 0.8% to 0.95%, approximately 0.8% to 1%, approximately 0.8% to 1.25%, approximately 0.85% to 0.9%, approximately 0.85% to 0.95%, approximately 0.85% to 1%, approximately 0.85% to 1.25%, approximately 0.9% to 0.95%, approximately 0.9% to 1%, approximately 0.9% to 1.25%, approximately 0.95% to 1%, approximately 0.95% to 1.25%, or approximately 1% to 1.25% salt (w / v).In some embodiments, the solution contains 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% of the salt (w / v). In some embodiments, the solution 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% of the 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% of salt (w / v). In some embodiments, the solution contains about 0.85% of salt (w / v). In some embodiments, the solution contains about 0.8% to about 0.9% of salt (w / v). In some embodiments, the solution contains about 0.75% to about 0.95% of salt (w / v). In some embodiments, the solution contains about 0.7% to about 1% of salt (w / v). In some embodiments, the solution contains about 0.5% to about 1.25% of salt (w / v). In some embodiments, the solution contains about 0.5% to about 2% of salt (w / v). In some embodiments, the solution contains 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% of the salt (w / v).

[0061] In some embodiments, the solution includes additional additives. In some embodiments, the solution includes dimethyl sulfoxide (DMSO), 1-dodecyl azacycloheptan-2-one, laurocaprum, 1-methyl-2-pyrrolidone (NMP), oleic acid, ethanol, methanol, polyethylene glycol (Brij 35, 58, 98), polyethylene glycol monolaurate (e.g., Tween 20), Tween 40 (sorbitol polyoxyethylene ester), Tween 60, Tween 80 (nonionic), cetylmethylammonium bromide (CTAB), urea, lecithin (solidified fatty acid derived from soybeans), 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 contains poloxamers. In some embodiments, the solution contains polyethylene glycol monolaurate (e.g., Tween 20). 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 about 0.01% to about 1% (v / v) of the solution. In some embodiments, the additional additives comprise about 0.1% (v / v) of the solution.

[0062] In some embodiments, the solution contains additional metal ions. In some embodiments, the solution contains magnesium, calcium, manganese, or a combination thereof. In some embodiments, the solution contains magnesium. In some embodiments, the solution contains calcium. In some embodiments, the solution contains manganese. In some embodiments, the solution contains magnesium and calcium. In some embodiments, the solution contains magnesium and manganese. In some embodiments, the solution contains calcium and manganese. In some embodiments, the solution contains magnesium, calcium, and manganese.

[0063] In some embodiments, the solution contains one or more nutrients for microorganisms. In some embodiments, the solution contains a bacterial growth medium. In some embodiments, the solution contains a lysogenic medium (LB), a nutrient broth, or a combination thereof. In some embodiments, the solution contains a lysogenic medium. In some embodiments, the solution contains a nutrient broth.

[0064] In some embodiments, the solution contains microorganisms. In some embodiments, the solution contains about 10 3 ~about 10 17 The solution contains microorganisms at a concentration of colony-forming units (CFU) / mL. In some embodiments, the solution is approximately 10 3 ~about 10 4 CFU / mL, approximately 10 3 ~about 10 5 CFU / mL, approximately 10 3 ~about 10 6 CFU / mL, approximately 10 3 ~about 10 7 CFU / mL, approximately 10 3 ~about 10 8 CFU / mL, approximately 10 3 ~about 10 9 CFU / mL, approximately 10 3 ~about 10 10 CFU / mL, approximately 10 3 ~about 10 12 CFU / mL, approximately 10 3 ~about 10 15 CFU / mL, approximately 10 3~about 10 17 CFU / mL, approximately 10 4 ~about 10 5 CFU / mL, approximately 10 4 ~about 10 6 CFU / mL, approximately 10 4 ~about 10 7 CFU / mL, approximately 10 4 ~about 10 8 CFU / mL, approximately 10 4 ~about 10 9 CFU / mL, approximately 10 4 ~about 10 10 CFU / mL, approximately 10 4 ~about 10 12 CFU / mL, approximately 10 4 ~about 10 15 CFU / mL, approximately 10 4 ~about 10 17 CFU / mL, approximately 10 5 ~about 10 6 CFU / mL, approximately 10 5 ~about 10 7 CFU / mL, approximately 10 5 ~about 10 8 CFU / mL, approximately 10 5 ~about 10 9 CFU / mL, approximately 10 5 ~about 10 10 CFU / mL, approximately 10 5 ~about 10 12 CFU / mL, approximately 10 5 ~about 10 15 CFU / mL, approximately 10 5 ~about 10 17 CFU / mL, approximately 10 6 ~about 10 7 CFU / mL, approximately 10 6 ~about 10 8 CFU / mL, approximately 10 6 ~about 10 9 CFU / mL, approximately 10 6 ~about 10 10 CFU / mL, approximately 10 6 ~about 10 12 CFU / mL, approximately 10 6 ~about 10 15 CFU / mL, approximately 10 6 ~about 10 17 CFU / mL, approximately 10 7~about 10 8 CFU / mL, about 10 7 ~about 10 9 CFU / mL, about 10 7 ~about 10 10 CFU / mL, about 10 7 ~about 10 12 CFU / mL, about 10 7 ~about 10 15 CFU / mL, about 10 7 ~about 10 17 CFU / mL, about 10 8 ~about 10 9 CFU / mL, about 10 8 ~about 10 10 CFU / mL, about 10 8 ~about 10 12 CFU / mL, about 10 8 ~about 10 15 CFU / mL, about 10 8 ~about 10 17 CFU / mL, about 10 9 ~about 10 10 CFU / mL, about 10 9 ~about 10 12 CFU / mL, about 10 9 ~about 10 15 CFU / mL, about 10 9 ~about 10 17 CFU / mL, about 10 10 ~about 10 12 CFU / mL, about 10 10 ~about 10 15 CFU / mL, about 10 10 ~about 10 17 CFU / mL, about 10 12 ~about 10 15 CFU / mL, about 10 12 ~about 10 17 CFU / mL, or about 10 15 ~about 10 17 CFU / mL of microorganisms. In some embodiments, the solution is 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 It contains microorganisms at a concentration of CFU / mL. 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 10 15 It contains microorganisms at a concentration of CFU / mL. In some embodiments, the solution is 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 It contains microorganisms at a concentration of CFU / mL. In some embodiments, the solution contains at least 10 6 ~10 7 The solution contains microorganisms at a concentration of CFU / mL. In some embodiments, the solution is 1 × 10⁻⁶ 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 14CFU / 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 Contains microorganisms at CFU / mL.

[0065] In some embodiments, the solution contains a desired amount of microorganisms per unit mass of seeds. In some embodiments, the solution contains about 10 3 ~about 10 17 The solution contains approximately 10 colony-forming units (CFU) per gram of seeds. In some embodiments, the solution contains approximately 10 3 ~about 10 4 CFU / gram, approximately 10 3 ~about 10 5 CFU / gram, approximately 10 3 ~about 10 6 CFU / gram, approximately 10 3 ~about 10 7 CFU / gram, approximately 10 3 ~about 10 8 CFU / gram, approximately 10 3 ~about 10 9 CFU / gram, approximately 10 3 ~about 10 10 CFU / gram, approximately 10 3 ~about 10 12 CFU / gram, approximately 10 3 ~about 10 15 CFU / gram, approximately 10 3 ~about 10 17 CFU / gram, approximately 10 4 ~about 10 5 CFU / gram, approximately 10 4 ~about 10 6 CFU / gram, approximately 10 4 ~about 10 7 CFU / gram, approximately 10 4 ~about 10 8 CFU / gram, approximately 10 4 ~about 10 9 CFU / gram, approximately 10 4 ~about 10 10 CFU / gram, approximately 10 4 ~about 10 12CFU / gram, approximately 10 4 ~about 10 15 CFU / gram, approximately 10 4 ~about 10 17 CFU / gram, approximately 10 5 ~about 10 6 CFU / gram, approximately 10 5 ~about 10 7 CFU / gram, approximately 10 5 ~about 10 8 CFU / gram, approximately 10 5 ~about 10 9 CFU / gram, approximately 10 5 ~about 10 10 CFU / gram, approximately 10 5 ~about 10 12 CFU / gram, approximately 10 5 ~about 10 15 CFU / gram, approximately 10 5 ~about 10 17 CFU / gram, approximately 10 6 ~about 10 7 CFU / gram, approximately 10 6 ~about 10 8 CFU / gram, approximately 10 6 ~about 10 9 CFU / gram, approximately 10 6 ~about 10 10 CFU / gram, approximately 10 6 ~about 10 12 CFU / gram, approximately 10 6 ~about 10 15 CFU / gram, approximately 10 6 ~about 10 17 CFU / gram, approximately 10 7 ~about 10 8 CFU / gram, approximately 10 7 ~about 10 9 CFU / gram, approximately 10 7 ~about 10 10 CFU / gram, approximately 10 7 ~about 10 12 CFU / gram, approximately 10 7 ~about 10 15 CFU / gram, approximately 10 7 ~about 10 17 CFU / gram, approximately 10 8 ~about 10 9 CFU / gram, approximately 108 ~about 10 10 CFU / gram, approximately 10 8 ~about 10 12 CFU / gram, approximately 10 8 ~about 10 15 CFU / gram, approximately 10 8 ~about 10 17 CFU / gram, approximately 10 9 ~about 10 10 CFU / gram, approximately 10 9 ~about 10 12 CFU / gram, approximately 10 9 ~about 10 15 CFU / gram, approximately 10 9 ~about 10 17 CFU / gram, approximately 10 10 ~about 10 12 CFU / gram, approximately 10 10 ~about 10 15 CFU / gram, approximately 10 10 ~about 10 17 CFU / gram, approximately 10 12 ~about 10 15 CFU / gram, approximately 10 12 ~about 10 17 CFU / gram, or about 10 15 ~about 10 17 Contains CFU / gram of seeds. In some embodiments, the solution is 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 Contains seeds CFU / gram. 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 10 15Contains seeds CFU / gram. In some embodiments, the solution is 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 Contains seeds CFU / gram. In some embodiments, the solution is 10 10 Contains less than CFU / gram of seeds. In some embodiments, the solution is 10 9 Contains less than CFU / gram of seeds. In some embodiments, the solution is 10 8 Contains less than CFU / gram of seeds. In some embodiments, the solution is 10 11 Contains less than CFU / gram of seeds in some embodiments. 5 ~about 10 9 Contains CFU / gram of seeds.

[0066] In some embodiments, the solution is about 10 3 ~about 10 17 Contains colony cells / gram of seeds. In some embodiments, the solution is about 10 3 ~about 10 4 Cells / gram, approximately 10 3 ~about 10 5 cells / gram, approximately 10 3 ~about 10 6 Cells / gram, approximately 10 3 ~about 10 7 cells / gram, approximately 10 3 ~about 10 8 Cells / gram, approximately 10 3 ~about 10 9 cells / gram, approximately 10 3 ~about 10 10 cells / gram, approximately 10 3 ~about 10 12 cells / gram, approximately 10 3 ~about 10 15 cells / gram, approximately 10 3 ~about 10 17 cells / gram, approximately 10 4~about 10 5 cells / gram, approximately 10 4 ~about 10 6 cells / gram, approximately 10 4 ~about 10 7 cells / gram, approximately 10 4 ~about 10 8 cells / gram, approximately 10 4 ~about 10 9 cells / gram, approximately 10 4 ~about 10 10 cells / gram, approximately 10 4 ~about 10 12 cells / gram, approximately 10 4 ~about 10 15 cells / gram, approximately 10 4 ~about 10 17 cells / gram, approximately 10 5 ~about 10 6 cells / gram, approximately 10 5 ~about 10 7 cells / gram, approximately 10 5 ~about 10 8 cells / gram, approximately 10 5 ~about 10 9 cells / gram, approximately 10 5 ~about 10 10 cells / gram, approximately 10 5 ~about 10 12 cells / gram, approximately 10 5 ~about 10 15 cells / gram, approximately 10 5 ~about 10 17 cells / gram, approximately 10 6 ~about 10 7 cells / gram, approximately 10 6 ~about 10 8 cells / gram, approximately 10 6 ~about 10 9 cells / gram, approximately 10 6 ~about 10 10 cells / gram, approximately 10 6 ~about 10 12 cells / gram, approximately 10 6 ~about 10 15 cells / gram, approximately 10 6 ~about 10 17 cells / gram, approximately 10 7 ~about 10 8 cells / gram, approximately 10 7~about 10 9 cells / gram, approximately 10 7 ~about 10 10 cells / gram, approximately 10 7 ~about 10 12 cells / gram, approximately 10 7 ~about 10 15 cells / gram, approximately 10 7 ~about 10 17 cells / gram, approximately 10 8 ~about 10 9 cells / gram, approximately 10 8 ~about 10 10 cells / gram, approximately 10 8 ~about 10 12 cells / gram, approximately 10 8 ~about 10 15 cells / gram, approximately 10 8 ~about 10 17 cells / gram, approximately 10 9 ~about 10 10 cells / gram, approximately 10 9 ~about 10 12 cells / gram, approximately 10 9 ~about 10 15 cells / gram, approximately 10 9 ~about 10 17 cells / gram, approximately 10 10 ~about 10 12 cells / gram, approximately 10 10 ~about 10 15 cells / gram, approximately 10 10 ~about 10 17 cells / gram, approximately 10 12 ~about 10 15 cells / gram, approximately 10 12 ~about 10 17 cells / gram, or about 10 15 ~about 10 17 Contains cells / gram of seeds. In some embodiments, the solution is 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 17Contains cells / gram of seeds. 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 10 15 Contains cells / gram of seeds. In some embodiments, the solution is 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 Contains seeds with cells / gram. In some embodiments, the solution contains fewer than 10¹⁰ cells / gram of seeds. In some embodiments, the solution contains fewer than 10⁹ cells / gram of seeds. In some embodiments, the solution contains fewer than 10⁸ cells / gram of seeds. In some embodiments, the solution contains fewer than 10¹¹ cells / gram of seeds. In some embodiments, the solution contains about 10 5 ~about 10 9 Contains seeds per cell / gram.

[0067] In some embodiments, the seeds contain a desired amount of microorganisms per seed. In some embodiments, the solution is about 10 3 ~about 10 17 It contains colony-forming units (CFU) / seeds. In some embodiments, the solution is about 10 3 ~about 10 4 CFU / seed, approximately 10 3 ~about 10 5 CFU / seed, approximately 10 3 ~about 10 6 CFU / seed, approximately 10 3 ~about 10 7 CFU / seed, approximately 10 3 ~about 10 8 CFU / seed, approximately 10 3 ~about 109 CFU / seed, approximately 10 3 ~approximately 10 10 CFU / seed, approximately 10 3 ~approximately 10 12 CFU / seed, approximately 10 3 ~approximately 10 15 CFU / seed, approximately 10 3 ~approximately 10 17 CFU / seed, approximately 10 4 ~approximately 10 5 CFU / seed, approximately 10 4 ~approximately 10 6 CFU / seed, approximately 10 4 ~approximately 10 7 CFU / seed, approximately 10 4 ~approximately 10 8 CFU / seed, approximately 10 4 ~approximately 10 9 CFU / seed, approximately 10 4 ~approximately 10 10 CFU / seed, approximately 10 4 ~approximately 10 12 CFU / seed, approximately 10 4 ~approximately 10 15 CFU / seed, approximately 10 4 ~approximately 10 17 CFU / seed, approximately 10 5 ~approximately 10 6 CFU / seed, approximately 10 5 ~approximately 10 7 CFU / seed, approximately 10 5 ~approximately 10 8 CFU / seed, approximately 10 5 ~approximately 10 9 CFU / seed, approximately 10 5 ~approximately 10 10 CFU / seed, approximately 10 5 ~approximately 10 12 CFU / seed, approximately 10 5 ~approximately 10 15 CFU / seed, approximately 10 5 ~approximately 10 17 CFU / seed, approximately 10 6 ~approximately 10 7 CFU / seed, approximately 10 6 ~approximately 10 8 CFU / seed, approximately 10 6 ~approximately 10 9 CFU / seed, approximately 10 6 ~approximately 1010 CFU / seed, approximately 10 6 ~about 10 12 CFU / seed, approximately 10 6 ~about 10 15 CFU / seed, approximately 10 6 ~about 10 17 CFU / seed, approximately 10 7 ~about 10 8 CFU / seed, approximately 10 7 ~about 10 9 CFU / seed, approximately 10 7 ~about 10 10 CFU / seed, approximately 10 7 ~about 10 12 CFU / seed, approximately 10 7 ~about 10 15 CFU / seed, approximately 10 7 ~about 10 17 CFU / seed, approximately 10 8 ~about 10 9 CFU / seed, approximately 10 8 ~about 10 10 CFU / seed, approximately 10 8 ~about 10 12 CFU / seed, approximately 10 8 ~about 10 15 CFU / seed, approximately 10 8 ~about 10 17 CFU / seed, approximately 10 9 ~about 10 10 CFU / seed, approximately 10 9 ~about 10 12 CFU / seed, approximately 10 9 ~about 10 15 CFU / seed, approximately 10 9 ~about 10 17 CFU / seed, approximately 10 10 ~about 10 12 CFU / seed, approximately 10 10 ~about 10 15 CFU / seed, approximately 10 10 ~about 10 17 CFU / seed, approximately 10 12 ~about 10 15 CFU / seed, approximately 10 12 ~about 10 17 CFU / seed, or about 10 15 ~about 10 17Contains CFU / seed. In some embodiments, the solution is 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 Contains CFU / seed. 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 10 15 Contains CFU / seed. In some embodiments, the solution is 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 Contains CFU / seed. In some embodiments, the solution is 10 10 Contains less than CFU / seed. In some embodiments, the solution is 10 9 Contains less than CFU / seed. In some embodiments, the solution is 10 8 Contains less than CFU / seed. In some embodiments, the solution is 10 11 Contains less than CFU / seed. In some embodiments, the solution contains about 10 5 ~about 10 9 Includes CFU / less than seed size.

[0068] In some embodiments, the seeds contain a desired amount of microorganisms per seed. In some embodiments, the solution is about 10 3 ~about 10 17Contains cells / seeds. In some embodiments, the solution is about 10 3 ~about 10 4 cells / seeds, about 10 3 ~about 10 5 cells / seeds, about 10 3 ~about 10 6 cells / seeds, about 10 3 ~about 10 7 cells / seeds, about 10 3 ~about 10 8 cells / seeds, about 10 3 ~about 10 9 cells / seeds, about 10 3 ~about 10 10 cells / seeds, about 10 3 ~about 10 12 cells / seeds, about 10 3 ~about 10 15 cells / seeds, about 10 3 ~about 10 17 cells / seeds, about 10 4 ~about 10 5 cells / seeds, about 10 4 ~about 10 6 cells / seeds, about 10 4 ~about 10 7 cells / seeds, about 10 4 ~about 10 8 cells / seeds, about 10 4 ~about 10 9 cells / seeds, about 10 4 ~about 10 10 cells / seeds, about 10 4 ~about 10 12 cells / seeds, about 10 4 ~about 10 15 cells / seeds, about 10 4 ~about 10 17 cells / seeds, about 10 5 ~about 10 6 cells / seeds, about 10 5 ~about 10 7 cells / seeds, about 10 5 ~about 10 8 cells / seeds, about 10 5 ~about 10 9 cells / seeds, about 10 5 ~about 10 10 cells / seeds, about 10 5 ~about 10 12Cells / seeds, approximately 10 5 ~approximately 10 15 Cells / seeds, approximately 10 5 ~approximately 10 17 Cells / seeds, approximately 10 6 ~approximately 10 7 Cells / seeds, approximately 10 6 ~approximately 10 8 Cells / seeds, approximately 10 6 ~approximately 10 9 Cells / seeds, approximately 10 6 ~approximately 10 10 Cells / seeds, approximately 10 6 ~approximately 10 12 Cells / seeds, approximately 10 6 ~approximately 10 15 Cells / seeds, approximately 10 6 ~approximately 10 17 Cells / seeds, approximately 10 7 ~approximately 10 8 Cells / seeds, approximately 10 7 ~approximately 10 9 Cells / seeds, approximately 10 7 ~approximately 10 10 Cells / seeds, approximately 10 7 ~approximately 10 12 Cells / seeds, approximately 10 7 ~approximately 10 15 Cells / seeds, approximately 10 7 ~approximately 10 17 Cells / seeds, approximately 10 8 ~approximately 10 9 Cells / seeds, approximately 10 8 ~approximately 10 10 Cells / seeds, approximately 10 8 ~approximately 10 12 Cells / seeds, approximately 10 8 ~approximately 10 15 Cells / seeds, approximately 10 8 ~approximately 10 17 Cells / seeds, approximately 10 9 ~approximately 10 10 Cells / seeds, approximately 10 9 ~approximately 10 12 Cells / seeds, approximately 10 9 ~approximately 10 15 Cells / seeds, approximately 10 9 ~approximately 10 17 Cells / seeds, approximately 10 10 ~approximately 10 12 Cells / seeds, approximately 10 10 ~approximately 1015 cells / seeds, about 10 10 ~about 10 17 cells / seeds, about 10 12 ~about 10 15 cells / seeds, about 10 12 ~about 10 17 Cells / seeds, or about 10 15 ~about 10 17 Contains cells / seeds. In some embodiments, the solution is 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 Contains cells / seeds. 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 10 15 Contains cells / seeds. In some embodiments, the solution is 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 Contains cells / seeds. In some embodiments, the solution is 10 10 Contains less than 10 cells / seeds. In some embodiments, the solution is 10 9 Contains less than 10 cells / seeds. In some embodiments, the solution is 10 8 Contains less than 10 cells / seeds. In some embodiments, the solution is 10 11 Contains less than 10 cells / seeds. In some embodiments, the solution is about 10 5 ~about 109 Includes cells / seeds smaller than a typical cell.

[0069] In some embodiments, the microorganism is a bacterium. In some embodiments, the bacterium is an endospore-forming bacterium. In some embodiments, the method includes a step of inducing endospore formation in an endospore-forming bacterium. In some embodiments, the bacteria incorporated into the seed are endospores. In some embodiments, the solution contains one or more components to induce endospore formation. In some embodiments, the solution contains potassium, ferrous sulfate, calcium, magnesium, manganese, or a combination thereof.

[0070] In some embodiments, the method includes a step of sterilizing the seeds. In some embodiments, the method includes a step of sterilizing the surface of the seeds. Any method for producing seeds having 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 immersing them 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”) indicate that there are substantially no surviving microorganisms on what has been sterilized. In some embodiments, the seeds are sterilized before incubating them in a solution containing microorganisms. In some embodiments, the seeds are sterilized after incubating them in a solution containing microorganisms. In some embodiments, a fungicide is added to the surface of the seeds.

[0071] In some embodiments, sterilized or disinfected seeds are substantially free of living microorganisms on the seed (e.g., the surface of the seed). In some embodiments, sterile or sterilized 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.

[0072] In some embodiments, plant seeds are incubated with a solution containing microorganisms for a sufficient time to incorporate the microorganisms into the seeds. In some embodiments, plant seeds are incubated with a solution containing endospore-forming bacteria or their endospores for about 1 minute to about 960 minutes. In some embodiments, plant seeds are incubated with a solution containing endospore-forming bacteria or their endospores for approximately 1 to 5 minutes, approximately 1 to 10 minutes, approximately 1 to 20 minutes, approximately 1 to 60 minutes, approximately 1 to 240 minutes, approximately 1 to 960 minutes, approximately 5 to 10 minutes, approximately 5 to 20 minutes, approximately 5 to 60 minutes, approximately 5 to 240 minutes, approximately 5 to 960 minutes, approximately 10 to 20 minutes, approximately 10 to 60 minutes, approximately 10 to 240 minutes, approximately 10 to 960 minutes, approximately 20 to 60 minutes, approximately 20 to 240 minutes, approximately 20 to 960 minutes, approximately 60 to 240 minutes, approximately 60 to 960 minutes, or approximately 240 to 960 minutes. In some embodiments, plant seeds are incubated with a solution containing endospore-forming bacteria or their endospores 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, plant seeds are incubated with a solution containing endospore-forming bacteria or their endospores 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, plant seeds are incubated with a solution containing endospore-forming bacteria or their endospores 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, plant seeds are incubated with a solution containing endospore-forming bacteria or their endospores for about 1 minute. In some embodiments, plant seeds are incubated with a solution containing endospore-forming bacteria or their endospores for about 5 minutes. In some embodiments, plant seeds are incubated with a solution containing endospore-forming bacteria or their endospores for about 10 minutes. In some embodiments, plant seeds are incubated with a solution containing endospore-forming bacteria or their endospores for about 20 minutes.In some embodiments, plant seeds are incubated with a solution containing endospore-forming bacteria or their endospores for about 60 minutes. In some embodiments, plant seeds are incubated with a solution containing endospore-forming bacteria or their endospores for about 240 minutes. In some embodiments, plant seeds are incubated with a solution containing endospore-forming bacteria or their endospores for about 960 minutes.

[0073] In some embodiments, plant seeds are incubated with a solution containing microorganisms or their exudates for about 1 minute to about 960 minutes. In some embodiments, plant seeds are incubated with a solution containing microorganisms or their exudates for approximately 1 to 5 minutes, approximately 1 to 10 minutes, approximately 1 to 20 minutes, approximately 1 to 60 minutes, approximately 1 to 240 minutes, approximately 1 to 960 minutes, approximately 5 to 10 minutes, approximately 5 to 20 minutes, approximately 5 to 60 minutes, approximately 5 to 240 minutes, approximately 5 to 960 minutes, approximately 10 to 20 minutes, approximately 10 to 60 minutes, approximately 10 to 240 minutes, approximately 10 to 960 minutes, approximately 20 to 60 minutes, approximately 20 to 240 minutes, approximately 20 to 960 minutes, approximately 60 to 240 minutes, approximately 60 to 960 minutes, or approximately 240 to 960 minutes. In some embodiments, plant seeds are incubated with a solution containing microorganisms or their exudates 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, plant seeds are incubated with a solution containing microorganisms or their exudates 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, plant seeds are incubated with a solution containing microorganisms or their exudates 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, plant seeds are incubated with a solution containing microorganisms or their exudates for about 1 minute. In some embodiments, plant seeds are incubated with a solution containing microorganisms or their exudates for about 5 minutes. In some embodiments, plant seeds are incubated with a solution containing microorganisms or their exudates for about 10 minutes. In some embodiments, plant seeds are incubated with a solution containing microorganisms or their exudates for about 20 minutes. In some embodiments, plant seeds are incubated with a solution containing microorganisms or their exudates for about 60 minutes. In some embodiments, plant seeds are incubated with a solution containing microorganisms or their exudates for about 240 minutes. In some embodiments, plant seeds are incubated with a solution containing microorganisms or their exudates for about 960 minutes.

[0074] 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 at temperatures 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 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 to The seeds are incubated with the solution at temperatures of approximately 35°C, 8-40°C, 12-16°C, 12-25°C, 12-30°C, 12-35°C, 12-40°C, 16-25°C, 16-30°C, 16-35°C, 16-40°C, 25-30°C, 25-35°C, 25-40°C, 30-35°C, 30-40°C, or 35-40°C. In some embodiments, the seeds are incubated with the solution at temperatures of approximately 2°C, 4°C, 8°C, 12°C, 16°C, 25°C, 30°C, 35°C, or 40°C. In some embodiments, the seeds are incubated with the solution at temperatures 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 temperatures of up to 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.

[0075] In some embodiments, the method includes a step of drying the seeds. In some embodiments, the seeds are dried to about 10% of their total seed moisture content. In some embodiments, the seeds are dried to about 5% to about 25% of their total seed moisture content. In some embodiments, the seeds are dried to approximately 5% to 8%, 5% to 10%, 5% to 12%, 5% to 15%, 5% to 20%, 5% to 25%, 8% to 10%, 8% to 12%, 8% to 15%, 8% to 20%, 8% to 25%, 10% to 12%, 10% to 15%, 10% to 20%, 10% to 25%, 12% to 15%, 12% to 20%, 12% to 25%, 15% to 20%, 15% to 25%, or 20% to 25% of the total seed moisture. In some embodiments, the seeds are dried to about 5%, 8%, 10%, 12%, 15%, 20%, or 25% of the total seed moisture. In some embodiments, the seeds are dried to at least about 5%, 8%, 10%, 12%, 15%, or 20% of the total seed moisture. In some embodiments, the seeds are dried to a maximum of about 8%, 10%, 12%, 15%, 20%, or 25% of the total seed moisture. In some embodiments, the seeds are dried to prevent germination. In some embodiments, the seeds are dried before planting to prevent germination.

[0076] Formulations for incorporating microorganisms In one embodiment, a formulation for incorporating a microorganism, an endospore, or its exudate into a seed is provided herein. In some embodiments, the formulation comprises one or more microorganisms or their endospores, and a salt. The one or more microorganisms may be any of the microorganisms or their endospores provided herein. In some embodiments, the one or more microorganisms comprises one or more endospore-forming bacteria or their endospores. In some embodiments, the formulation comprises a microbial exudate. The exudate may be any of the microorganisms provided herein.

[0077] In some embodiments, the formulation is a solution. In some embodiments, the formulation is an aqueous solution.

[0078] In some embodiments, the formulation contains a salt. The salt may be present in the formulation at any appropriate concentration. In some embodiments, the formulation contains about 0.85% salt (w / v). In some embodiments, the formulation contains about 0.1% to about 1.25% salt (w / v). In some embodiments, the formulation contains about 0.1% to about 2.0% salt (w / v).In some embodiments, the formulation is approximately 0.1% to approximately 0.25%, approximately 0.1% to approximately 0.5%, approximately 0.1% to approximately 0.6%, approximately 0.1% to approximately 0.7%, approximately 0.1% to approximately 0.75%, approximately 0.1% to approximately 0.8%, approximately 0.1% to approximately 0.85%, approximately 0.1% to approximately 0.9%, approximately 0.1% to approximately 0.95%, approximately 0.1% to approximately 1%, approximately 0.1% to approximately 1.25%, approximately 0.25% to approximately 0.5%, approximately 0.25% to approximately 0.6%, approximately 0.25% to approximately 0.7%, approximately 0.25% to approximately 0.75%, approximately 0.25% to approximately 0. 0.8%, approximately 0.25% to 0.85%, approximately 0.25% to 0.9%, approximately 0.25% to 0.95%, approximately 0.25% to 1%, approximately 0.25% to 1.25%, approximately 0.5% to 0.6%, approximately 0.5% to 0.7%, approximately 0.5% to 0.75%, approximately 0.5% to 0.8%, approximately 0.5% to 0.85%, approximately 0.5% to 0.9%, approximately 0.5% to 0.95%, approximately 0.5% to 1%, approximately 0.5% to 1.25%, approximately 0.6% to 0.7%, approximately 0.6% to 0.75%, approximately 0.6% to 0. 8%, approximately 0.6% to approximately 0.85%, approximately 0.6% to approximately 0.9%, approximately 0.6% to approximately 0.95%, approximately 0.6% to approximately 1%, approximately 0.6% to approximately 1.25%, approximately 0.7% to approximately 0.75%, approximately 0.7% to approximately 0.8%, approximately 0.7% to approximately 0.85%, approximately 0.7% to approximately 0.9%, approximately 0.7% to approximately 0.95%, approximately 0.7% to approximately 1%, approximately 0.7% to approximately 1.25%, approximately 0.75% to approximately 0.8%, approximately 0.75% to approximately 0.85%, approximately 0.75% to approximately 0.9%, approximately 0.75% to approximately 0.95%, approximately 0.75% to approximately 1 Contains %, approximately 0.75% to 1.25%, approximately 0.8% to 0.85%, approximately 0.8% to 0.9%, approximately 0.8% to 0.95%, approximately 0.8% to 1%, approximately 0.8% to 1.25%, approximately 0.85% to 0.9%, approximately 0.85% to 0.95%, approximately 0.85% to 1%, approximately 0.85% to 1.25%, approximately 0.9% to 0.95%, approximately 0.9% to 1%, approximately 0.9% to 1.25%, approximately 0.95% to 1%, approximately 0.95% to 1.25%, or approximately 1% to 1.25% salt (w / v). In some embodiments, the formulation contains 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% of the 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% of 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% of salt (w / v). In some embodiments, the formulation contains about 0.85% of salt (w / v). In some embodiments, the formulation contains about 0.8% to about 0.9% of salt (w / v). In some embodiments, the formulation contains about 0.75% to about 0.95% of salt (w / v). In some embodiments, the formulation contains about 0.7% to about 1% of the salt (w / v). In some embodiments, the formulation contains about 0.5% to about 1.25% of the salt (w / v). In some embodiments, the formulation contains about 0.5% to about 2% of the salt (w / v). In some embodiments, the formulation contains 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% of the salt (w / v).

[0079] 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 contains sodium ions, lithium ions, or potassium ions. In some embodiments, the salt contains alkali metal ions. In some embodiments, the salt contains alkaline earth metal ions. In some embodiments, the salt contains halide ions. In some embodiments, the salt is an alkali or alkaline earth halide salt. In some embodiments, the salt contains chloride ions, bromide ions, or iodide ions. In some embodiments, the salt is a sulfate, phosphate, carbonate, or nitrate.

[0080] In some embodiments, the formulation includes additional additives. In some embodiments, the formulation includes dimethyl sulfoxide (DMSO), 1-dodecyl azacycloheptan-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 (sorbitol polyoxyethylene ester), Tween 60, Tween 80 (nonionic), cetylmethylammonium bromide (CTAB), urea, lecithin (solidified fatty acid derived from soybeans), chitosan, poloxamer 188, poloxamer 237, poloxamer 338, poloxamer 407, or combinations 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 contains poloxamer. In some embodiments, the formulation contains polyethylene glycol monolaurate (e.g., Tween 20). 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 formulation. In some embodiments, the additional additives comprise 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.

[0081] In some embodiments, the formulation contains additional metal ions. In some embodiments, the formulation contains magnesium, calcium, manganese, or a combination thereof. In some embodiments, the formulation contains magnesium. In some embodiments, the formulation contains calcium. In some embodiments, the formulation contains manganese. In some embodiments, the formulation contains magnesium and calcium. In some embodiments, the formulation contains magnesium and manganese. In some embodiments, the formulation contains calcium and manganese. In some embodiments, the formulation contains magnesium, calcium, and manganese.

[0082] In some embodiments, the formulation comprises one or more nutrients for microorganisms. In some embodiments, the formulation comprises a bacterial growth medium. In some embodiments, the formulation comprises a lysogenic medium (LB), a nutrient broth, or a combination thereof. In some embodiments, the formulation comprises a lysogenic medium. In some embodiments, the formulation comprises a nutrient broth.

[0083] In some embodiments, the formulation includes additional components to promote endospore formation 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 contains potassium. In some embodiments, the formulation contains iron sulfide. In some embodiments, the formulation contains calcium. In some embodiments, the formulation contains magnesium. In some embodiments, the formulation contains manganese.

[0084] In some embodiments, the formulation contains microorganisms. In some embodiments, the formulation contains about 10 3 ~about 10 17 The formulation contains a number of colony-forming units (CFUs) / mL of microorganisms. In some embodiments, the formulation contains at least 1 × 10⁶ microorganisms. 6 Contains microorganisms at CFU / mL. In some embodiments, the formulation is about 10 3 ~about 10 4 CFU / mL, approximately 10 3~about 10 5 CFU / mL, approximately 10 3 ~about 10 6 CFU / mL, approximately 10 3 ~about 10 7 CFU / mL, approximately 10 3 ~about 10 8 CFU / mL, approximately 10 3 ~about 10 9 CFU / mL, approximately 10 3 ~about 10 10 CFU / mL, approximately 10 3 ~about 10 12 CFU / mL, approximately 10 3 ~about 10 15 CFU / mL, approximately 10 3 ~about 10 17 CFU / mL, approximately 10 4 ~about 10 5 CFU / mL, approximately 10 4 ~about 10 6 CFU / mL, approximately 10 4 ~about 10 7 CFU / mL, approximately 10 4 ~about 10 8 CFU / mL, approximately 10 4 ~about 10 9 CFU / mL, approximately 10 4 ~about 10 10 CFU / mL, approximately 10 4 ~about 10 12 CFU / mL, approximately 10 4 ~about 10 15 CFU / mL, approximately 10 4 ~about 10 17 CFU / mL, approximately 10 5 ~about 10 6 CFU / mL, approximately 10 5 ~about 10 7 CFU / mL, approximately 10 5 ~about 10 8 CFU / mL, approximately 10 5 ~about 10 9 CFU / mL, approximately 10 5 ~about 10 10 CFU / mL, approximately 10 5 ~about 10 12 CFU / mL, approximately 10 5 ~about 10 15 CFU / mL, approximately 10 5~about 10 17 CFU / mL, approximately 10 6 ~about 10 7 CFU / mL, approximately 10 6 ~about 10 8 CFU / mL, approximately 10 6 ~about 10 9 CFU / mL, approximately 10 6 ~about 10 10 CFU / mL, approximately 10 6 ~about 10 12 CFU / mL, approximately 10 6 ~about 10 15 CFU / mL, approximately 10 6 ~about 10 17 CFU / mL, approximately 10 7 ~about 10 8 CFU / mL, approximately 10 7 ~about 10 9 CFU / mL, approximately 10 7 ~about 10 10 CFU / mL, approximately 10 7 ~about 10 12 CFU / mL, approximately 10 7 ~about 10 15 CFU / mL, approximately 10 7 ~about 10 17 CFU / mL, approximately 10 8 ~about 10 9 CFU / mL, approximately 10 8 ~about 10 10 CFU / mL, approximately 10 8 ~about 10 12 CFU / mL, approximately 10 8 ~about 10 15 CFU / mL, approximately 10 8 ~about 10 17 CFU / mL, approximately 10 9 ~about 10 10 CFU / mL, approximately 10 9 ~about 10 12 CFU / mL, approximately 10 9 ~about 10 15 CFU / mL, approximately 10 9 ~about 10 17 CFU / mL, approximately 10 10 ~about 10 12 CFU / mL, approximately 10 10 ~about 10 15 CFU / mL, approximately 10 10~about 10 17 CFU / mL, about 10 12 ~about 10 15 CFU / mL, about 10 12 ~about 10 17 CFU / mL, or about 10 15 ~about 10 17 CFU / mL of microorganisms. In some embodiments, the formulation is 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 CFU / mL of microorganisms. In some embodiments, the formulation is 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 10 15 CFU / mL of microorganisms. In some embodiments, the formulation is at most 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 CFU / mL of microorganisms. In some embodiments, the formulation is at least 10 6 ~10 7 CFU / mL of microorganisms. In some embodiments, the formulation is 1×10 3 ~1×10 4 CFU / mL, 1×10 4 ~1×10 5 CFU / mL, 1×10 5 ~1×10 6 CFU / mL, 1×106 ~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 Contains microorganisms in CFU / mL. The microorganisms may be any of the microorganisms provided herein, or endospores of any of the microorganisms provided herein.

[0085] 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 temperatures 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 temperatures of approximately 8 to 35, 8 to 40, 12 to 16, 12 to 25, 12 to 30, 12 to 35, 12 to 40, 16 to 25, 16 to 30, 16 to 35, 16 to 40, 25 to 30, 25 to 35, 25 to 40, 30 to 35, 30 to 40, or 35 to 40°C. In some embodiments, the formulation is maintained at temperatures of approximately 2, 4, 8, 12, 16, 25, 30, 35, or 40°C. In some embodiments, the formulation is maintained at temperatures of at least 2, 4, 8, 12, 16, 25, 30, or 35°C. In some embodiments, the formulation is maintained at temperatures of up to approximately 4, 8, 12, 16, 25, 30, 35, or 40°C.

[0086] Microorganisms and exudates The microorganisms or their exudates provided herein produce or promote the formation of bicarbonates and one or more minerals. In some embodiments, the formation of bicarbonates sequesters CO2. In some embodiments, the formation of bicarbonates results in the formation of minerals. In some embodiments, the formed minerals stably sequester carbon in the soil. In some embodiments, the microorganism is a bacterium. In some embodiments, the microorganism is an endospore-forming bacterium. In some embodiments, the microorganism is the endospore of a bacterium. Whenever a microorganism (e.g., a bacterium) referred to herein is capable of forming endospores, it is intended that any endospores of that microorganism are also included. For example, if a treatment formulation of plant seeds contains a Bacillus sp., the formulation may contain endospores of a Bacillus sp.

[0087] In some embodiments, the microorganism is a microbe from the phylum Firmicutes, phylum Proteobacteria, and phylum Actinomycetes. In some embodiments, the microorganism is a microbe from the phylum Firmicutes. In some embodiments, the microorganism is a microbe from the phylum Proteobacteria. In some embodiments, the microorganism is a microbe from the phylum Actinomycetes. In some embodiments, the microorganism is an endospore of any of the microorganisms.

[0088] Rhizosphere bacteria, which fix atmospheric nitrogen, are found in plant roots. These organisms can survive in the soil and are tolerant of large amounts of CO2, which is either CO2 released from plant roots during respiration or CO2 released through respiration by nearby microorganisms and soil fauna.

[0089] Because these rhizosphere bacteria reside close to the roots, these organisms have the ability to utilize root exudates as a source of carbon and energy. Many of them have evolved to possess genes that enable them to convert CO2 into biomass or any metabolite for their own benefit. In some embodiments, the bacteria are not genetically modified. In some embodiments, the bacteria are selected for their ability to convert CO2 into bicarbonates and minerals.

[0090] Rhizobia can more actively form colonies on plant roots. Therefore, they can form stable colonies that can survive in changing soil environments, secrete antimicrobial compounds to inhibit the growth of pathogens or invaders, and form endospores that give them selective aptitudes for survival in harsh environments.

[0091] Rhizosphere bacteria have the ability to express carbonic anhydrase (CA), a well-characterized enzyme, in addition to other means of CO2 fixation. Its broad temperature tolerance (up to 50°C), wide pH range, and high levels of expression make CA an ideal candidate for CO2 sequestering in soil. This enzyme converts CO2 into bicarbonate, which is then hydrolyzed to carbonate ions. These carbonate ions react with cations present in the soil to produce minerals. Soil contains many cations (Ca... 2+ Mg 2+ Because it is rich in (Na+, K+), this allows for sustained mineralization and the formation of various minerals, which serves as a means of permanently sequestering CO2 in the soil. In some embodiments, bacteria are selected for the use of carbonic anhydrase.

[0092] In some embodiments, carbonic anhydrase is α-CA, β-CA, δ-CA, ζ-CA, η-CA, or ι-CA. In some embodiments, CA is CA-1, CA-2, CA-3, CA-4, CA-5A, CA-5B, CA-6, CA-7, CA-8, CA-9, CA-10, CA-11, CA-12, CA-13, CA-14, or CA-15.

[0093] In some embodiments, various rhizosphere bacteria are effectively loaded onto the seeds. In some embodiments, the rhizosphere bacteria include endospore-forming bacteria that enhance biological nitrogen fixation. In some embodiments, the rhizosphere bacteria include Bacillus sp, Paenibacillus sp, or both. In some embodiments, the one or more microorganisms include B. amyloliquefaciens, B. laterosporus, B. licheniformis, B. macerans, B. cereus, B. circulans, B. firmus, B. subtilis, B. sphaericus, B. megaterium, B. coagulans This includes B. coagulans, B. brevis, B. thuringiensis, B. mycoides, B. cucumis, B. endophyticus, B. pumilus, B. velezensis, B. mucilaginosus, B. tequilensis, B. methylotrophicus, or any combination thereof.In some embodiments, the one or more microorganisms include Bacillus subtilis S3C23, Bacillus subtilis MP2, Bacillus subtilis RO2C15, Bacillus subtilis RO2C22, Bacillus megaterium 6, Bacillus megaterium S3C21, Bacillus megaterium RO2C12, Bacillus cucumis S3C14, Bacillus endophyticus 5, or any combination thereof. In some embodiments, the one or more microorganisms include Bacillus subtilis S3C23. In some embodiments, the one or more microorganisms include Bacillus subtilis MP2.

[0094] CO2 sequestration by these microorganisms can be achieved by their ability to produce or promote the formation of CA. These rhizosphere bacteria can colonize roots, or otherwise replace other nearby microbial communities that could utilize nutrients from root exudates. CO2 released from roots, from soil fauna, or from microbial communities can be captured by CA via hydration to bicarbonates. Typically, cations are required to continue the mineral-producing process in order to form minerals (CaCO3, MgCO3, CaMg(CO3)2). Various cations are already present in the soil, enabling a sustained process. Ca in soil 2+ and Mg 2+ The amount can vary depending on geographical location, soil type, and irrigation pattern. These cations can be further adjusted by farmers adding limestone to maintain high soil fertility. Typical well-irrigated soil has an average of 850 kg (Ca) considering the first 15 cm depth. 2+) / acre and 218kg(Mg 2+ It has ) / acre. According to one previously published study, the amount of CO2 produced in the rhizosphere of maize is about 7000 kg / acre per maize season. Considering the amount of available CO2 and cations, a considerable amount of CO2 can be stored as minerals of Ca or Mg. Mathematically, 425 kg of CaCO3 and 114 kg of MgCO3 can be produced, but other cations (Na) in the soil + , K + Depending on the presence of ), there are many more combinations that form other minerals such as Na2CO3. Since lime treatment is carried out by farmers to maintain high soil fertility, the microorganisms disclosed herein can circumvent this requirement by biologically producing limestone (CaCO3). In addition, depending on the availability of other cations in the soil, various minerals can be formed to store gaseous CO2. These minerals include, but are not limited to, calcite, aragonite, dolomite, limestone, carbonates, magnesium carbonate, iron carbonate, magnesite, cohenite, diamond, carbonatite, ferrous carbonate, sparlite, and tyreite.

[0095] In some embodiments, the amount of minerals produced may be between 50 kg / acre and a maximum of 1,000 kg / acre. In some embodiments, the amount of minerals produced may be between approximately 50 kg / acre and approximately 1,000 kg / acre. In some embodiments, the amount of minerals produced may be between approximately 50 kg / acre and approximately 100 kg / acre, approximately 50 kg / acre and approximately 200 kg / acre, approximately 50 kg / acre and approximately 300 kg / acre, approximately 50 kg / acre and approximately 400 kg / acre, approximately 50 kg / acre and approximately 500 kg / acre, approximately 50 kg / acre and approximately 600 kg / acre, approximately 50 kg / acre and approximately 700 kg / acre, and approximately 50 kg / acre and approximately 800 kg / acre. - Approximately 50 kg / acre to approximately 900 kg / acre, approximately 50 kg / acre to approximately 1,000 kg / acre, approximately 100 kg / acre to approximately 200 kg / acre, approximately 100 kg / acre to approximately 300 kg / acre, approximately 100 kg / acre to approximately 400 kg / acre, approximately 100 kg / acre to approximately 500 kg / acre, approximately 100 kg / acre to approximately 600 kg / acre, approximately 100 kg / acre to approximately 700 kg / acre, approximately 100 kg / acre to approximately 800kg / acre, approximately 100kg / acre to approximately 900kg / acre, approximately 100kg / acre to approximately 1,000kg / acre, approximately 200kg / acre to approximately 300kg / acre, approximately 200kg / acre to approximately 400kg / acre, approximately 200kg / acre to approximately 500kg / acre, approximately 200kg / acre to approximately 600kg / acre, approximately 200kg / acre to approximately 700kg / acre, approximately 200kg / acre to approximately 800kg / acre, approximately 2 00kg / acre to approximately 900kg / acre, approximately 200kg / acre to approximately 1,000kg / acre, approximately 300kg / acre to approximately 400kg / acre, approximately 300kg / acre to approximately 500kg / acre, approximately 300kg / acre to approximately 600kg / acre, approximately 300kg / acre to approximately 700kg / acre, approximately 300kg / acre to approximately 800kg / acre, approximately 300kg / acre to approximately 900kg / acre, approximately 300kg / acre to approximately 1,000kg / acre, approximately 400kg / acre to approximately 500kg / acre, approximately 400kg / acre to approximately 600kg / acre, approximately 400kg / acre to approximately 700kg / acre, approximately 400kg / acre to approximately 800kg / acre, approximately 400kg / acre to approximately 900kg / acre, approximately 400kg / acre to approximately 1,000kg / acre, approximately 500kg / acre to approximately 600kg / acre, approximately 500kg / acre to approximately 700kg / acre, approximately 500kg / acre to approximately 800kg / acre, approximately 500kg / acre to approximately 900kg / acre, approximately 500kg / acre ~ approximately 1,000 kg / acre, approximately 600 kg / acre ~ approximately 700 kg / acre, approximately 600 kg / acre ~ approximately 800 kg / acre, approximately 600 kg / acre ~ approximately 900 kg / acre, approximately 600 kg / acre ~ approximately 1,000 kg / acre, approximately 700 kg / acre ~ approximately 800 kg / acre, approximately 700 kg / acre ~ approximately 900 kg / acre, approximately 700 kg / acre ~ approximately 1,000 kg / acre, approximately 800 kg / acre ~ approximately 900 kg / acre, or approximately 900 kg / acre ~ approximately 1,It may be 000 kg / acre. In some embodiments, the amount of minerals produced may be about 50 kg / acre, about 100 kg / acre, about 200 kg / acre, about 300 kg / acre, about 400 kg / acre, about 500 kg / acre, about 600 kg / acre, about 700 kg / acre, about 800 kg / acre, about 900 kg / acre, or about 1000 kg / acre. In some embodiments, the amount of minerals produced may be at least about 50 kg / acre, about 100 kg / acre, about 200 kg / acre, about 300 kg / acre, about 400 kg / acre, about 500 kg / acre, about 600 kg / acre, about 700 kg / acre, about 800 kg / acre, or about 900 kg / acre. In some embodiments, the amount of minerals produced may be up to approximately 100 kg / acre, approximately 200 kg / acre, approximately 300 kg / acre, approximately 400 kg / acre, approximately 500 kg / acre, approximately 600 kg / acre, approximately 700 kg / acre, approximately 800 kg / acre, approximately 900 kg / acre, or approximately 1000 kg / acre.

[0096] In some embodiments, the amount of CO2 converted by microorganisms is between 0.1 tons of CO2 per acre and a maximum of 2.5 tons of CO2 per acre. In some embodiments, microorganisms convert 2.5 to 5.3 tons of CO2 / acre. In some embodiments, microorganisms convert 5.3 to 7.5 tons of CO2 / acre. In some embodiments, microorganisms convert 7.5 to 10 tons of CO2 / acre. In some embodiments, microorganisms convert 10 to 15 tons of CO2 / acre. In some embodiments, microorganisms convert 15 to 20 tons / acre. In some embodiments, the amount of CO2 converted by microorganisms is between approximately 2 tons / acre and approximately 20 tons / acre.In some embodiments, the amount of CO2 converted by microorganisms is approximately 2 tons / acre to 4 tons / acre, approximately 2 tons / acre to 6 tons / acre, approximately 2 tons / acre to 8 tons / acre, approximately 2 tons / acre to 10 tons / acre, approximately 2 tons / acre to 12 tons / acre, approximately 2 tons / acre to 14 tons / acre, approximately 2 tons / acre to 16 tons / acre, approximately 2 tons / acre to 18 tons / acre, approximately 2 tons / acre to 20 tons / acre, approximately 4 tons / acre to 6 tons / acre, approximately 4 tons / acre to approximately 8 tons / acre, approximately 4 tons / acre to approximately 10 tons / acre, approximately 4 tons / acre to approximately 12 tons / acre, approximately 4 tons / acre to approximately 14 tons / acre, approximately 4 tons / acre to approximately 16 tons / acre, approximately 4 tons / acre to approximately 18 tons / acre, approximately 4 tons / acre to approximately 20 tons / acre, approximately 6 tons / acre to approximately 8 tons / acre, approximately 6 tons / acre to approximately 10 tons / acre, approximately 6 tons / acre to approximately 12 tons / acre, approximately 6 tons / acre to approximately 14 tons / acre, approximately 6 tons / acre to approximately 16 tons / acre, approximately 6 tons / acre ~approximately 18 tons / acre, approximately 6 tons / acre to approximately 20 tons / acre, approximately 8 tons / acre to approximately 10 tons / acre, approximately 8 tons / acre to approximately 12 tons / acre, approximately 8 tons / acre to approximately 14 tons / acre, approximately 8 tons / acre to approximately 16 tons / acre, approximately 8 tons / acre to approximately 18 tons / acre, approximately 8 tons / acre to approximately 20 tons / acre, approximately 10 tons / acre to approximately 12 tons / acre, approximately 10 tons / acre to approximately 14 tons / acre, approximately 10 tons / acre to approximately 16 tons / acre, approximately 10 tons / acre to approximately 18 tons / acre, approximately The range is between 10 tons / acre and approximately 20 tons / acre, approximately 12 tons / acre and approximately 14 tons / acre, approximately 12 tons / acre and approximately 16 tons / acre, approximately 12 tons / acre and approximately 18 tons / acre, approximately 12 tons / acre and approximately 20 tons / acre, approximately 14 tons / acre and approximately 16 tons / acre, approximately 14 tons / acre and approximately 18 tons / acre, approximately 14 tons / acre and approximately 20 tons / acre, approximately 16 tons / acre and approximately 18 tons / acre, or between approximately 18 tons / acre and approximately 20 tons / acre.In some embodiments, the amount of CO2 converted by microorganisms is between approximately 2 tons / acre, approximately 4 tons / acre, approximately 6 tons / acre, approximately 8 tons / acre, approximately 10 tons / acre, approximately 12 tons / acre, approximately 14 tons / acre, approximately 16 tons / acre, approximately 18 tons / acre, or approximately 20 tons / acre. In some embodiments, the amount of CO2 converted by microorganisms is at least between approximately 2 tons / acre, approximately 4 tons / acre, approximately 6 tons / acre, approximately 8 tons / acre, approximately 10 tons / acre, approximately 12 tons / acre, approximately 14 tons / acre, approximately 16 tons / acre, or approximately 18 tons / acre. In some embodiments, the amount of CO2 converted by microorganisms is at most between approximately 4 tons / acre, approximately 6 tons / acre, approximately 8 tons / acre, approximately 10 tons / acre, approximately 12 tons / acre, approximately 14 tons / acre, approximately 16 tons / acre, approximately 18 tons / acre, or approximately 20 tons / acre.

[0097] In some embodiments, the promotion of the production of one or more minerals includes the production of ammonia and an increase in the pH of the growing medium in which the plants derived from the plant seeds are grown.

[0098] Mineralization processes can be initiated in either the cell wall or the EPS of microorganisms. The cell wall, due to its overall negative charge, has been shown to be the site of nucleation and mineralization. In some microorganisms, such as Bacillus subtilis, the EPS contains remarkably high levels of glutamic acid, aspartic acid, histidine, arginine, and lysine, which are negatively charged under alkaline conditions and can act as nucleation sites for CaCO3 precipitation. This can be beneficial because, if the microorganisms attached to the roots along with CaCO3 are unwanted, they can be removed by pulling up the plants at the end of the rich season to prevent the addition of minerals to the soil.

[0099] In some embodiments, the one or more microorganisms include a genetic modification that causes the one or more microorganisms to produce or promote the formation of more carbonic anhydrase compared to the corresponding wild-type microorganisms. In some embodiments, the genetic modification includes a nucleic acid construct, the nucleic acid construct includes one or more promoters configured to drive the expression of a carbonic anhydrase (CA) coding sequence. In some embodiments, the genetic modification includes a nucleic acid construct, the nucleic acid construct includes a carbonic anhydrase (CA) coding sequence. In some embodiments, the CA coding sequence is heterogeneous to the one or more microorganisms. In some embodiments, the CA coding sequence is endogenous to the one or more microorganisms. In some embodiments, the nucleic acid construct is codon-optimized. In some embodiments, the nucleic acid construct further includes one or more promoters configured to drive the expression of the CA coding sequence. In some embodiments, the one or more promoters drive the constitutive expression of the CA coding sequence. In some embodiments, the one or more promoters drive the inducible expression of the CA coding sequence. In some embodiments, the genetic modification includes a carbonic anhydrase gene modified by direct microbial evolution. In some embodiments, the gene modification includes a signal sequence. In some embodiments, the signal sequence includes a periplasmic signal sequence or an extracellular secretory signal. In some embodiments, the periplasmic signal sequence or the extracellular secretory signal is located at the 5' end of a carbonic anhydrase coding sequence. In some embodiments, the periplasmic signal sequence or the extracellular secretory signal is fused to the 5' end of a carbonic anhydrase coding sequence. In some embodiments, the gene modification includes one or more components of a common secretory pathway, and the gene modification results in the secretion or intracellular targeting of carbonic anhydrase by the one or more microorganisms.

[0100] In some embodiments, the carbonic anhydrase is α-CA, β-CA, δ-CA, ζ-CA, η-CA, or ι-CA. In some embodiments, the CA is CA-1, CA-2, CA-3, CA-4, CA-5A, CA-5B, CA-6, CA-7, CA-8, CA-9, CA-10, CA-11, CA-12, CA-13, CA-14, or CA-15.

[0101] In some embodiments, the one or more promoters are one or more promoters composed of Bacillus sp. or Paenibacillus sp., or are derived from the above promoters. In some embodiments, the one or more promoters are groES P, P43, P sigX P trnQ and P xylA selected from. In some embodiments, the one or more promoters are one or more housekeeping gene promoters or strongly expressed constitutive promoters, or are derived from them. In some embodiments, the one or more housekeeping gene promoters or strongly expressed constitutive promoters are P liaG P lepA P veg P gsiB P43, P trnQ P ial (bacitracin-inducible), and P xylA (xylose-inducible) selected from.

[0102] In some embodiments, the genetic modification includes the introduction of an expression vector into one or more microorganisms. In some embodiments, the genetic modification includes the modification of the chromosome of one or more microorganisms.

[0103] In some embodiments, microorganisms can fix both nitrogen and carbon dioxide. In some embodiments, nitrogen-fixing microorganisms continue to provide fixed nitrogen (ammonia) to plants through biological nitrogen fixation, which reduces the use of chemical fertilizers that pollute the environment. The final product of atmospheric nitrogen fixation (ammonia) can enhance the CO2-to-mineral process because ammonia has been reported to increase pH, which promotes mineralization. Since ammonia production from the microorganisms disclosed herein is significantly higher than that of other soil-dwelling microorganisms, even in the presence of an external nitrogen source, the mineralization process can be carried out more rapidly by the strains disclosed herein than by other soil-dwelling microorganisms.

[0104] In some embodiments, the microorganisms include genera such as Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Bradyrhizobium sp., Brevibacillus sp., Caldanerobacter sp., Caloramator sp., and 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 Gerria sp., Geobacillus sp., Geosporobacter sp., Gracilibacillus sp., Halobacillus sp.), Halonatronum sp., Heliobacterium sp., Heliophilum sp., Klebsiella sp., Laceyella sp., Lentibacillus sp., Lysinibacillus sp., Mahela sp., Metabacterium sp., Moorela sp., Natroniella sp., Oceanobacillus sp., Orenia sp., Ornithinibacillus sp., Oxalophagus sp.), Oxobacter sp., Paenibacillus sp., Paraliobacillus sp., Penicillium sp., Pelospora sp., Pelotomaculum sp., Piscibacillus sp., Planifilum sp., Pontibacillus sp., Propionispora sp., Salinibacillus sp., Salsuginibacillus sp., Seinonella sp., Shimazuella sp., Sinorhizobium Sporacetigenium sp., Sporoanaerobacter sp., Sporobacter 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 These are microbes selected from the genera Thermoanaeromonas sp., Thermobacillus sp., Thermoflavimicrobium sp., Thermovenabylum sp., Tuberibacillus sp., Virgibacillus sp., Vulcanobacillus sp., and Xanthobacter. In some embodiments, the microorganisms are selected from the genera Acetobacter sp., Actinomyces sp., Bacillus sp., Chryseobacterium sp., Coxiella sp., Ensifer sp., Glutamicibacter sp., Microbacterium sp., and Serratia sp. In some embodiments, the microorganisms are selected from the genera Acetobacter sp.In some embodiments, the microorganism is of the genus Actinomyces sp. In some embodiments, the microorganism is of the genus Bacillus sp. In some embodiments, the microorganism is of the genus Chryseobacterium sp. In some embodiments, the microorganism is of the genus Coxiella sp. In some embodiments, the microorganism is of the genus Ensifer sp. In some embodiments, the microorganism is of the genus Glutamicibacter sp. In some embodiments, the microorganism is of the genus Microbacterium sp. In some embodiments, the microorganism is of the genus Pantoea sp. In some embodiments, the microorganism is of the genus Serratia sp. In some embodiments, the microorganism is an endospore of any of the microorganisms.

[0105] In some embodiments, the microorganisms include Acetobacter cerevisiae, Bacillus cucumis, Bacillus endophyticus, Bacillus megaterium, Bacillus nakamurai, Bacillus subtilis, Chryseobacterium lactis, Ensifer adhaerens, Glutamicibacter arilaitensis, Glutamicibacter halophytocola, Microbacterium chocolatum, and Microbacterium yannicii. This includes *Serratia yannicii*, *Pantoea allii*, *Serratia marcescens*, or *Serratia ureilytica*.In some embodiments, the microorganisms include 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 includes Acetobacter cerevisiae. In some embodiments, the microorganism includes Bacillus cucumis. In some embodiments, the microorganism includes Bacillus endophyticus. In some embodiments, the microorganism includes Bacillus megaterium. In some embodiments, the microorganism includes Bacillus subtilis. In some embodiments, the microorganism includes Chryseobacterium lactis. In some embodiments, the microorganism includes Ensifer adhaerens. In some embodiments, the microorganism includes Glutamicibacter halophytocola. In some embodiments, the microorganism includes Microbacterium chocolatum. In some embodiments, the microorganism includes Pantoea allii.In some embodiments, the microorganism includes Serratia marcescens. In some embodiments, the microorganism is an endospore of any of the microorganisms.

[0106] In some embodiments, the microorganisms are endospore-forming bacteria. In some embodiments, the endospore-forming bacteria are from the genus Bacillus. In some embodiments, the endospore-forming bacteria are from the genus Bacillus sp. In some embodiments, the endospore-forming bacteria include Bacillus cucumis, Bacillus endophyticus, Bacillus megaterium, Bacillus nakamurai, and Bacillus subtilis. In some embodiments, the endospore-forming bacteria include Bacillus cucumis, Bacillus endophyticus, Bacillus megaterium, or Bacillus subtilis. In some embodiments, the endospore-forming bacteria include Bacillus cucumis. In some embodiments, the endospore-forming bacteria include Bacillus megaterium. In some embodiments, the endospore-forming bacteria include Bacillus nakamurai. In some embodiments, the endospore-forming bacteria include Bacillus subtilis. In some embodiments, the microorganism is an endospore of any of the microorganisms.

[0107] In some embodiments, the microorganism is endospore-forming. In some embodiments, the endospores are from the genus Bacillus. In some embodiments, the endospores are from the genus Bacillus (Bacillus sp.). In some embodiments, the endospores include Bacillus cucumis, Bacillus endophyticus, Bacillus megaterium, Bacillus nakamurai, or Bacillus subtilis. In some embodiments, the endospores include Bacillus cucumis, Bacillus endophyticus, Bacillus megaterium, or Bacillus subtilis. In some embodiments, the endospores include Bacillus cucumis. In some embodiments, the endospores include Bacillus megaterium. In some embodiments, the endospores include Bacillus nakamurai. In some embodiments, the endospores include Bacillus subtilis.

[0108] In some embodiments, a consortium of microorganisms is incorporated into the seeds. In some embodiments, the consortium includes genera such as Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanerobacter sp., Caloramator sp., Caminicella sp., and Cerasibacillus. 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., Gerria (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., Moorela 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 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., Thalassobacilius sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus The consortium includes two or more microorganisms selected from the genera Thermoflavimicrobium sp., Thermovenablum sp., Tuberibacillus sp., Virgibacillus sp., and Vulcanobacillus sp. In some embodiments, the consortium includes two or more microorganisms selected from the genera Acetobacter sp., Actinomyces sp., Bacillus sp., Chryseobacterium sp., Coxiella sp., Ensifer sp., Glutamicibacter sp., Microbacterium sp., or Serratia sp.The consortium comprises two or more bacteria selected from the following: In some embodiments, the consortium comprises 2, 3, 4, 5, 6, 7, 8, 9, 10, or more bacteria. In some embodiments, the consortium comprises 2 bacteria. In some embodiments, the consortium comprises 3 bacteria. In some embodiments, the consortium comprises 4 bacteria. In some embodiments, the consortium comprises 5 bacteria. In some embodiments, the consortium comprises 6 bacteria. In some embodiments, the consortium comprises endospores of any of the microorganisms.

[0109] In some embodiments, the consortium includes bacteria from the phylum Bacillus and one or more other bacteria. In some embodiments, the consortium includes bacteria from the phylum Bacillus, as well as genera such as Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanerobacter sp., Caloramator sp., and 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 Gerria 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 Sporoanaerobacter sp., Sporobacter 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 It includes one or more bacteria selected from the genera Thermobacillus sp., Thermoflavimicrobium sp., Thermovenabylum sp., Tuberibacillus sp., Virgibacillus sp., and Vulcanobacillus sp. In some embodiments, the consortium comprises two or more bacteria selected from the genera Acetobacter, Actinomyces, Bacillus, Chryseobacterium, Coxiella, Ensifer, Glutamicibacter, Microbacterium, and Serratia. In some embodiments, the consortium comprises endospores of any of the microorganisms.

[0110] In some embodiments, the consortium includes Bacillus endophyticus, Bacillus megaterium, Bacillus nakamurai, Bacillus subtilis, Chryseobacterium lactis, Ensifer adhaerens, Glutamicibacter arilaitensis, Glutamicibacter halophytocola, Microbacterium chocolatum, Microbacterium yannicii, Pantoea allii, and Serratia marcescens. It comprises a mixture of two or more bacteria selected from *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 2 bacteria. In some embodiments, the consortium comprises 3 bacteria. In some embodiments, the consortium comprises 4 bacteria. In some embodiments, the consortium comprises 5 bacteria. In some embodiments, the consortium comprises 6 bacteria. In some embodiments, the consortium comprises endospores of any of the microorganisms.

[0111] 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.

[0112] In some embodiments, the consortium comprises two or more bacteria selected from Acetobacter cereviseae, Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. In some embodiments, the consortium comprises two bacteria selected from Acetobacter cereviseae, Chryseobacterium lactis, Bacillus endophyticus, and Bacillus megaterium. In some embodiments, the consortium comprises 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 includes endospores of any of the microorganisms.

[0113] 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 cucumis. In some embodiments, the consortium comprises endospores of any of the microorganisms.

[0114] In some embodiments, exudates from any of the microorganisms provided herein are incorporated into cells. In some embodiments, the exudate contains the genera Acetonema, Actinomyces, Alkalibacillus, Ammoniphilus, Amphibacillus, Anaerobacter, Anerospora, Aneurinibacillus, Anoxybacillus, Bacillus, Brevibacillus, Caldanerobacter, Caloramator, Caminicella, and Cerasibacillus. 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., Gerria (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., Moorela 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 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., Thalassobacilius sp., Thermoacetogenium sp., Thermoactinomyces sp., Thermoalkalibacillus sp., Thermoanaerobacter sp., Thermoanaeromonas sp., Thermobacillus The exudates are from the genera Thermoflavimicrobium sp., Thermovenabylum sp., Tuberibacillus sp., Virgibacillus sp., or Vulcanobacillus sp. In some embodiments, the exudates are from the genera Acetobacter sp., Actinomyces sp., Bacillus sp., Chryseobacterium sp., Coxiella sp., Ensifer sp., Glutamicibacter sp., Microbacterium sp., and Serratia sp.It is from ). In some embodiments, the exudate contains Acetobacter cerevisiae, Bacillus cucumis, Bacillus endophyticus, Bacillus megaterium, Bacillus nakamurai, Bacillus subtilis, Chryseobacterium lactis, Ensifer adhaerens, Glutamicibacter arilaitensis, Glutamicibacter halophytocola, Microbacterium chocolatum, and Microbacterium yannicii. The exudate is from *Bacillus 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 these microorganisms.

[0115] In some embodiments, microorganisms are selected for one or more properties related to their ability to interact with plants. In some embodiments, microorganisms are selected for their compatibility. In some embodiments, microorganisms are selected so that predatory or antagonistic effects do not occur. In some embodiments, microorganisms are selected for their stability during storage. In some embodiments, microorganisms are selected for rapid colonization of plants and survival within relevant tissues. In some embodiments, microorganisms are selected for optimal incorporation into one or more seeds. In some embodiments, microorganisms remain present throughout the entire life cycle of the plant.

[0116] In some embodiments, the microorganisms incorporated into the seeds are stable after incorporation. In some embodiments, the microorganisms are stable for longer than 30 days, longer than 6 months, longer than 1 year, or longer than 2 years. In some embodiments, the microorganisms are stable for longer than 30 days. In some embodiments, the microorganisms are stable for longer than 6 months. In some embodiments, the microorganisms are stable for longer than 1 year. In some embodiments, the microorganisms are stable for longer than 2 years.

[0117] Composition containing plants and bacteria In certain embodiments, compositions comprising a plant and one or more microorganisms associated with the plant are disclosed herein, wherein the one or more microorganisms are selected to produce or promote the formation of bicarbonates and one or more minerals, or are derived from such microorganisms. In some embodiments, compositions are obtained by culturing a plant or plant seed and one or more microorganisms associated with the plant or plant seed, as described herein.

[0118] Modified plants In one embodiment, a modified plant comprising a microorganism or a microbial exudate incorporated into the plant is provided herein. In some embodiments, the microorganism or exudate produces or promotes the formation of bicarbonates and one or more minerals. In some embodiments, the formation of bicarbonates sequesters CO2. In some embodiments, the formation of bicarbonates results in the formation of minerals. In some embodiments, the formed minerals stably sequester carbon in the soil. In some preferred embodiments, the microorganism is an endospore-forming bacterium or its endospores.

[0119] In some embodiments, the microorganism or exudate is incorporated into the plant. In some embodiments, the microorganism or exudate is incorporated into the plant beneath the pericarp. In some embodiments, the microorganism or exudate is incorporated into the plant between the pericarp and the aleurone cell layer. In some embodiments, the microorganism or exudate comes into contact with the plant embryo. In some embodiments, the microorganism or exudate does not come into contact with the plant embryo. In some embodiments, the microorganism or exudate comes into contact with the plant endosperm. In some embodiments, the microorganism or exudate does not come into contact with the plant endosperm. In some embodiments, the microorganism or exudate is incorporated into the plant in the space between the plant sheath and the plant embryo. In some embodiments, the microorganism or exudate is incorporated in the space between the plant pericarp and the plant aleurone cell layer.

[0120] The modified plant may be any type of plant. In some embodiments, the modified plant is a monocotyledonous plant. In some embodiments, the plant seeds are from maize, wheat, rice, barley, rye, sugarcane, millet, oats, or sorghum. In some embodiments, the plant seeds are from maize plants. In some embodiments, the plant seeds are from maize (Zea maize) plants. In some embodiments, the modified plant is a dicotyledonous plant. In some embodiments, the plant is from soybean, cotton, alfalfa, beans, quinoa, lentils, peanuts, sunflower, canola, cassava, palm oil, potato, sugar beet, cocoa, coffee, lettuce, tomato, or cabbage plants. In some embodiments, the plant is from lettuce plants. In some embodiments, the plant is from lettuce (Lactuca sativa) plants. In some embodiments, the plant is from tomato plants. In some embodiments, the plant is from tomato (Solanum lycopersicum) plants. In some embodiments, the plant is a genetically modified organism (GMO). In some embodiments, the plant is a non-GMO plant.

[0121] The amount of microorganisms or exudates incorporated into the plant must be at a level sufficient to effectively sequestrate CO2. In some embodiments, the amount of microorganisms incorporated into the plant is approximately 250 colony-forming units (CFU) to approximately 5,000 CFU. In some embodiments, the amount of microorganisms incorporated into the plant is approximately 250 CFU to 500 CFU, approximately 250 CFU to 750 CFU, approximately 250 CFU to 1,000 CFU, approximately 250 CFU to 2,000 CFU, approximately 250 CFU to 3,000 CFU, approximately 250 CFU to 4,000 CFU, approximately 250 CFU to 5,000 CFU, approximately 500 CFU to 750 CFU, approximately 500 CFU to 1,000 CFU, approximately 500 CFU to 2,000 CFU, approximately 500 CFU to 3,000 CFU, approximately 500 CFU to 4,000 CFU, approximately 500 CFU to 5,000 CFU, approximately 750 CFU to 1,000 CFU, and approximately 7 The CFU levels are approximately 50 CFU to 2,000 CFU, approximately 750 CFU to 3,000 CFU, approximately 750 CFU to 4,000 CFU, approximately 750 CFU to 5,000 CFU, approximately 1,000 CFU to 2,000 CFU, approximately 1,000 CFU to 3,000 CFU, approximately 1,000 CFU to 4,000 CFU, approximately 1,000 CFU to 5,000 CFU, approximately 2,000 CFU to 3,000 CFU, approximately 2,000 CFU to 4,000 CFU, approximately 2,000 CFU to 5,000 CFU, approximately 3,000 CFU to 4,000 CFU, approximately 3,000 CFU to 5,000 CFU, or approximately 4,000 CFU to 5,000 CFU. In some embodiments, the amount of microorganisms incorporated into the plant 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 plant 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 plant is up to 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 is incorporated into the plant. In some embodiments, at least about 1000 CFU is incorporated into the plant.

[0122] In some embodiments, the microorganisms or exudates incorporated into the plant have long-term storage stability. In some embodiments, the modified plant has storage stability for about 3 months to about 36 months. In some embodiments, the modified plant has storage stability for about 3 months to about 6 months, about 3 months to about 9 months, about 3 months to about 12 months, about 3 months to about 15 months, about 3 months to about 18 months, about 3 months to about 21 months, about 3 months to about 24 months, about 3 months to about 30 months, about 3 months to about 36 months, about 6 months to about 9 months, about 6 months to about 12 months, Approximately 6 months to 15 months, approximately 6 months to 18 months, approximately 6 months to 21 months, approximately 6 months to 24 months, approximately 6 months to 30 months, approximately 6 months to 36 months, approximately 9 months to 12 months, approximately 9 months to 15 months, approximately 9 months to 18 months, approximately 9 months to 21 months, approximately 9 months to 24 months, approximately 9 months to 30 months, approximately 9 months Months to approximately 36 months, approximately 12 months to approximately 15 months, approximately 12 months to approximately 18 months, approximately 12 months to approximately 21 months, approximately 12 months to approximately 24 months, approximately 12 months to approximately 30 months, approximately 12 months to approximately 36 months, approximately 15 months to approximately 18 months, approximately 15 months to approximately 21 months, approximately 15 months to approximately 24 months, approximately 15 months to approximately 30 months, approximately 15 months to approximately The plants have storage stability for 36 months, approximately 18 to 21 months, approximately 18 to 24 months, approximately 18 to 30 months, approximately 18 to 36 months, approximately 21 to 24 months, approximately 21 to 30 months, approximately 21 to 36 months, approximately 24 to 30 months, approximately 24 to 36 months, or approximately 30 to 36 months. In some embodiments, the modified plants have storage stability for approximately 3 months, approximately 6 months, approximately 9 months, approximately 12 months, approximately 15 months, approximately 18 months, approximately 21 months, approximately 24 months, approximately 30 months, or approximately 36 months. In some embodiments, the modified plants 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 plants have storage stability for up to approximately 6 months, 9 months, 12 months, 15 months, 18 months, 21 months, 24 months, 30 months, or 36 months.

[0123] In some embodiments, the microorganisms incorporated into the plant are stable after incorporation. In some embodiments, the microorganisms are stable for longer than 30 days, longer than 6 months, longer than 1 year, or longer than 2 years. In some embodiments, the microorganisms are stable for longer than 30 days. In some embodiments, the microorganisms are stable for longer than 6 months. In some embodiments, the microorganisms are stable for longer than 1 year. In some embodiments, the microorganisms are stable for longer than 2 years.

[0124] The microorganisms incorporated into the plant or their exudates may be any of the microorganisms provided herein or any other microorganisms. In some embodiments, the microorganism is a microbe. In some embodiments, the microorganism is an endospore-forming microbe. In some embodiments, the microorganism is an endospore-forming microbe or its endospores. In some embodiments, the microorganism is an endospore of a microorganism provided herein. In some embodiments, the microorganism is an endospore-forming bacterium or its endospores.

[0125] Methods for incorporating bacteria In one embodiment, a method for incorporating one or more microorganisms or their exudates into one or more plants is provided herein. In some embodiments, the method includes the step of disinfecting the plants. In some embodiments, the method includes the step of bringing the plants into contact with a solution containing one or more microorganisms or their exudates. In some embodiments, the solution further includes salt. In some embodiments, the method includes the step of incubating the plants with the solution for a certain period of time. In some embodiments, the period of time is sufficient to introduce a desired amount of the microorganisms or their exudates into the plants. In some embodiments, the method incorporates a desired amount of the microorganisms or their exudates into the plants.

[0126] In some embodiments, the above method includes the step of contacting a plant with a solution containing a salt as described herein.

[0127] In some embodiments, the solution includes additional additives as described herein.

[0128] In some embodiments, the solution contains additional metal ions as described herein.

[0129] In some embodiments, the solution comprises one or more nutrients for microorganisms as described herein. In some embodiments, the solution comprises a bacterial growth medium. In some embodiments, the solution comprises a lysogenic medium (LB), a nutrient broth, or a combination thereof. In some embodiments, the solution comprises a lysogenic medium. In some embodiments, the solution comprises a nutrient broth.

[0130] In some embodiments, the solution contains microorganisms such as those described herein.

[0131] In some embodiments, the solution contains a desired amount of microorganisms per unit of plant mass as described herein.

[0132] In some embodiments, the plant contains a desired amount of microorganisms per plant, as described herein. In some embodiments, the microorganisms are bacteria. In some embodiments, the bacteria are endosporic bacteria. In some embodiments, the method includes a step of inducing endosporulation of endosporic bacteria. In some embodiments, the bacteria incorporated into the plant are endospores. In some embodiments, the solution contains one or more components to induce endosporulation. In some embodiments, the solution contains potassium, ferrous sulfate, calcium, magnesium, manganese, or a combination thereof.

[0133] In some embodiments, the method includes a step of sterilizing the plant. In some embodiments, the method includes a step of sterilizing the surface of the plant. Any method for producing a plant having a sterilized surface may be employed. In some embodiments, the plant is sterilized with a bleach solution. In some embodiments, the plant is sterilized before immersing it in a solution containing one or more microorganisms. In some embodiments, the plant is a sterilized plant. In some embodiments, the plant has a sterilized surface. As used herein, “sterilizing,” “sterilized,” and related terms (e.g., “disinfecting”) indicate that there are substantially no surviving microorganisms on what has been sterilized. In some embodiments, the plant is sterilized before incubating it in a solution containing microorganisms. In some embodiments, the plant is sterilized after incubating it in a solution containing microorganisms. In some embodiments, a fungicide is added to the surface of the plant.

[0134] In some embodiments, sterilized or disinfected plants are substantially free of living microorganisms on the plant (e.g., on the plant surface). In some embodiments, sterile or sterilized plants 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 plant.

[0135] In some embodiments, the plants are incubated with a solution containing microorganisms for a sufficient time to incorporate the microorganisms into the plants.

[0136] Microorganisms and exudates The microorganisms or their exudates provided herein produce or promote the formation of bicarbonates and one or more minerals. In some embodiments, the formation of bicarbonates sequesters CO2. In some embodiments, the formation of bicarbonates results in the formation of minerals. In some embodiments, the formed minerals stably sequester carbon in the soil. In some embodiments, the microorganisms are bacteria as described herein. In some embodiments, the microorganisms are endospore-forming bacteria. In some embodiments, the microorganisms are endospores of bacteria. Whenever a microorganism (e.g., bacteria) referred herein can form endospores, it is intended that any endospores of that microorganism are also included. For example, if a plant treatment formulation contains Bacillus sp., the formulation may contain endospores of Bacillus sp.

[0137] In some embodiments, the microorganism is a microbe from the phylum Firmicutes, phylum Proteobacteria, and phylum Actinomycetes. In some embodiments, the microorganism is a microbe from the phylum Firmicutes. In some embodiments, the microorganism is a microbe from the phylum Proteobacteria. In some embodiments, the microorganism is a microorganism from the phylum Actinomycetes. In some embodiments, the microorganism is an endospore of any of the microorganisms.

[0138] Rhizosphere bacteria, which fix atmospheric nitrogen, are found in plant roots. These organisms can survive in the soil and are tolerant of large amounts of CO2, which is either CO2 released from plant roots during respiration or CO2 released through respiration by nearby microorganisms and soil fauna.

[0139] In some embodiments, the bacteria are not genetically modified. In some embodiments, the bacteria are selected for their ability to convert CO2 into bicarbonates and ultimately minerals. In some embodiments, the bacteria are selected for their use of carbonic anhydrase.

[0140] In some embodiments, various rhizosphere bacteria are effectively loaded onto the seeds. In some embodiments, the rhizosphere bacteria include endospore-forming bacteria that enhance biological nitrogen fixation. In some embodiments, the rhizosphere bacteria include Bacillus sp, Paenibacillus sp, or both. In some embodiments, the one or more microorganisms include B. amyloliquefaciens, B. laterosporus, B. licheniformis, B. macerans, B. cereus, B. circulans, B. firmus, B. subtilis, B. sphaericus, B. megaterium, B. coagulans This includes B. coagulans, B. brevis, B. thuringiensis, B. mycoides, B. cucumis, B. endophyticus, B. pumilus, B. velezensis, B. mucilaginosus, B. tequilensis, B. methylotrophicus, or any combination thereof.In some embodiments, the one or more microorganisms include Bacillus subtilis S3C23, Bacillus subtilis MP2, Bacillus subtilis RO2C15, Bacillus subtilis RO2C22, Bacillus megaterium 6, Bacillus megaterium S3C21, Bacillus megaterium RO2C12, Bacillus cucumis S3C14, Bacillus endophyticus 5, or any combination thereof. In some embodiments, the one or more microorganisms include Bacillus subtilis S3C23. In some embodiments, the one or more microorganisms include Bacillus subtilis MP2.

[0141] In some embodiments, the one or more microorganisms include a genetic modification that causes the one or more microorganisms to produce or promote the formation of more carbonic anhydrase compared to the corresponding wild-type microorganisms. In some embodiments, the genetic modification includes a nucleic acid construct, the nucleic acid construct includes one or more promoters configured to drive the expression of a carbonic anhydrase (CA) coding sequence. In some embodiments, the genetic modification includes a nucleic acid construct, the nucleic acid construct includes a carbonic anhydrase (CA) coding sequence. In some embodiments, the CA coding sequence is heterogeneous to the one or more microorganisms. In some embodiments, the CA coding sequence is endogenous to the one or more microorganisms. In some embodiments, the nucleic acid construct is codon-optimized. In some embodiments, the nucleic acid construct further includes one or more promoters configured to drive the expression of the CA coding sequence. In some embodiments, the one or more promoters drive the constitutive expression of the CA coding sequence. In some embodiments, the one or more promoters drive the inducible expression of the CA coding sequence. In some embodiments, the genetic modification includes a carbonic anhydrase gene modified by direct microbial evolution. In some embodiments, the gene modification includes a signal sequence. In some embodiments, the signal sequence includes a periplasmic signal sequence or an extracellular secretory signal. In some embodiments, the periplasmic signal sequence or the extracellular secretory signal is located at the 5' end of a carbonic anhydrase coding sequence. In some embodiments, the periplasmic signal sequence or the extracellular secretory signal is fused to the 5' end of a carbonic anhydrase coding sequence. In some embodiments, the gene modification includes one or more components of a common secretory pathway, and the gene modification results in the secretion or intracellular targeting of carbonic anhydrase by the one or more microorganisms.

[0142] In some embodiments, the one or more promoters are composed of or derived from the Bacillus sp. or Paenibacillus sp. genera. In some embodiments, the one or more promoters are P groES P43, P sigX , P trnQ , and P xylA Selected from. In some embodiments, the one or more promoters are one or more housekeeping gene promoters or strongly expressed constitutive promoters, or derived therefrom. In some embodiments, the one or more housekeeping gene promoters or strongly expressed constitutive promoters are P liaG , P lepA , P veg , P gsiB P43, P trnQ , P ial (Bacitracin-induced), and P xylA Selected from (xylose-inducible).

[0143] In some embodiments, the gene modification includes the introduction of an expression vector into one or more microorganisms. In some embodiments, the gene modification includes modifications to the chromosomes of one or more microorganisms.

[0144] In some embodiments, microorganisms can fix both nitrogen and carbon dioxide. In some embodiments, nitrogen-fixing microorganisms continue to provide fixed nitrogen (ammonia) to plants through biological nitrogen fixation, which reduces the use of chemical fertilizers that pollute the environment. The final product of atmospheric nitrogen fixation (ammonia) can enhance the CO2-to-mineral process because ammonia has been reported to increase pH, which promotes mineralization. Since ammonia production from the microorganisms disclosed herein is significantly higher than that of other soil-dwelling microorganisms, even in the presence of an external nitrogen source, the mineralization process can be carried out more rapidly by the strains disclosed herein than by other soil-dwelling microorganisms.

[0145] In some embodiments, the microorganisms include genera such as Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanerobacter sp., Caloramator sp., Caminicella sp., and Cerasibacillus. 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., Gerria 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., Moorela 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 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 The microbes are selected from the genera Thermovenablum sp., Tuberibacillus sp., Virgibacillus sp., and Vulcanobacillus sp. In some embodiments, the microorganisms are selected from the genera 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 of the genus Acetobacter sp. In some embodiments, the microorganism is of the genus Actinomyces sp. In some embodiments, the microorganism is of the genus Bacillus sp.In some embodiments, the microorganism is Chryseobacterium sp. In some embodiments, the microorganism is Coxiella sp. In some embodiments, the microorganism is Ensifer sp. In some embodiments, the microorganism is Glutamicibacter sp. In some embodiments, the microorganism is Microbacterium sp. In some embodiments, the microorganism is Pantoea sp. In some embodiments, the microorganism is Serratia sp. In some embodiments, the microorganism is an endospore of any of the microorganisms.

[0146] In some embodiments, microorganisms are selected for one or more properties related to their ability to interact with plants. In some embodiments, microorganisms are selected for compatibility. In some embodiments, microorganisms are selected so that phagocytic or antagonistic effects do not occur. In some embodiments, microorganisms are selected for stability during storage. In some embodiments, microorganisms are selected for rapid colonization of plants and survival within relevant tissues. In some embodiments, microorganisms are selected for optimal integration into one or more plants. In some embodiments, microorganisms remain present throughout the entire life cycle of the plant.

[0147] In some embodiments, the microorganisms incorporated into the plant are stable after incorporation. In some embodiments, the microorganisms are stable for longer than 30 days, longer than 6 months, longer than 1 year, or longer than 2 years. In some embodiments, the microorganisms are stable for longer than 30 days. In some embodiments, the microorganisms are stable for longer than 6 months. In some embodiments, the microorganisms are stable for longer than 1 year. In some embodiments, the microorganisms are stable for longer than 2 years.

[0148] Methods for producing bicarbonates and minerals In certain embodiments, methods for promoting the formation and mineralization of bicarbonates are disclosed herein, the methods comprising (a) culturing a plant and one or more microorganisms associated with the plant, the plant's roots, and / or the rhizosphere of the roots, wherein the one or more microorganisms are selected to produce or promote the formation of bicarbonates and one or more minerals, or are derived from such microorganisms.

[0149] Modified plants In some embodiments, the one or more microorganisms associated with the plant are located in the roots or rhizosphere of the plant. In some embodiments, the one or more microorganisms associated with the plant are located in the roots or rhizosphere of the plant by irrigation.

[0150] In some embodiments, the plant is derived from a seedling, which is irrigated with microorganisms disclosed herein, selected to stimulate the production of the one or more minerals disclosed herein by the plant. In some embodiments, the plant is obtained by culturing a plant seed and one or more microorganisms associated with the plant seed, as described herein. In some embodiments, the seed is stimulated by a method described herein.

[0151] In one embodiment, a method is provided herein that includes the step of cultivating a plant containing a microorganism or an exudate of a microorganism incorporated into the plant. In some preferred embodiments, the microorganism is an endospore-forming bacterium or its endospores.

[0152] In some embodiments, the amount of minerals produced may be between 50 kg / acre and a maximum of 1,000 kg / acre. In some embodiments, the amount of minerals produced may be between approximately 50 kg / acre and approximately 1,000 kg / acre. In some embodiments, the amount of minerals produced may be between approximately 50 kg / acre and approximately 100 kg / acre, approximately 50 kg / acre and approximately 200 kg / acre, approximately 50 kg / acre and approximately 300 kg / acre, approximately 50 kg / acre and approximately 400 kg / acre, approximately 50 kg / acre and approximately 500 kg / acre, approximately 50 kg / acre and approximately 600 kg / acre, approximately 50 kg / acre and approximately 700 kg / acre, and approximately 50 kg / acre and approximately 800 kg / acre. - Approximately 50 kg / acre to approximately 900 kg / acre, approximately 50 kg / acre to approximately 1,000 kg / acre, approximately 100 kg / acre to approximately 200 kg / acre, approximately 100 kg / acre to approximately 300 kg / acre, approximately 100 kg / acre to approximately 400 kg / acre, approximately 100 kg / acre to approximately 500 kg / acre, approximately 100 kg / acre to approximately 600 kg / acre, approximately 100 kg / acre to approximately 700 kg / acre, approximately 100 kg / acre to approximately 800kg / acre, approximately 100kg / acre to approximately 900kg / acre, approximately 100kg / acre to approximately 1,000kg / acre, approximately 200kg / acre to approximately 300kg / acre, approximately 200kg / acre to approximately 400kg / acre, approximately 200kg / acre to approximately 500kg / acre, approximately 200kg / acre to approximately 600kg / acre, approximately 200kg / acre to approximately 700kg / acre, approximately 200kg / acre to approximately 800kg / acre, approximately 2 00kg / acre to approximately 900kg / acre, approximately 200kg / acre to approximately 1,000kg / acre, approximately 300kg / acre to approximately 400kg / acre, approximately 300kg / acre to approximately 500kg / acre, approximately 300kg / acre to approximately 600kg / acre, approximately 300kg / acre to approximately 700kg / acre, approximately 300kg / acre to approximately 800kg / acre, approximately 300kg / acre to approximately 900kg / acre, approximately 300kg / acre to approximately 1,000kg / acre, approximately 400kg / acre to approximately 500kg / acre, approximately 400kg / acre to approximately 600kg / acre, approximately 400kg / acre to approximately 700kg / acre, approximately 400kg / acre to approximately 800kg / acre, approximately 400kg / acre to approximately 900kg / acre, approximately 400kg / acre to approximately 1,000kg / acre, approximately 500kg / acre to approximately 600kg / acre, approximately 500kg / acre to approximately 700kg / acre, approximately 500kg / acre to approximately 800kg / acre, approximately 500kg / acre to approximately 900kg / acre, approximately 500kg / acre ~ approximately 1,000 kg / acre, approximately 600 kg / acre ~ approximately 700 kg / acre, approximately 600 kg / acre ~ approximately 800 kg / acre, approximately 600 kg / acre ~ approximately 900 kg / acre, approximately 600 kg / acre ~ approximately 1,000 kg / acre, approximately 700 kg / acre ~ approximately 800 kg / acre, approximately 700 kg / acre ~ approximately 900 kg / acre, approximately 700 kg / acre ~ approximately 1,000 kg / acre, approximately 800 kg / acre ~ approximately 900 kg / acre, or approximately 900 kg / acre ~ approximately 1,It may be 000 kg / acre. In some embodiments, the amount of minerals produced may be about 50 kg / acre, about 100 kg / acre, about 200 kg / acre, about 300 kg / acre, about 400 kg / acre, about 500 kg / acre, about 600 kg / acre, about 700 kg / acre, about 800 kg / acre, about 900 kg / acre, or about 1000 kg / acre. In some embodiments, the amount of minerals produced may be at least about 50 kg / acre, about 100 kg / acre, about 200 kg / acre, about 300 kg / acre, about 400 kg / acre, about 500 kg / acre, about 600 kg / acre, about 700 kg / acre, about 800 kg / acre, or about 900 kg / acre. In some embodiments, the amount of minerals produced may be up to approximately 100 kg / acre, approximately 200 kg / acre, approximately 300 kg / acre, approximately 400 kg / acre, approximately 500 kg / acre, approximately 600 kg / acre, approximately 700 kg / acre, approximately 800 kg / acre, approximately 900 kg / acre, or approximately 1000 kg / acre.

[0153] In some embodiments, the microorganism or exudate is incorporated into the plant. In some embodiments, the microorganism or exudate is incorporated into the plant beneath the pericarp. In some embodiments, the microorganism or exudate is incorporated into the plant between the pericarp and the aleurone cell layer. In some embodiments, the microorganism or exudate comes into contact with the plant embryo. In some embodiments, the microorganism or exudate does not come into contact with the plant embryo. In some embodiments, the microorganism or exudate comes into contact with the plant endosperm. In some embodiments, the microorganism or exudate does not come into contact with the plant endosperm. In some embodiments, the microorganism or exudate is incorporated into the plant in the space between the plant sheath and the plant embryo. In some embodiments, the microorganism or exudate is incorporated in the space between the plant pericarp and the plant aleurone cell layer.

[0154] The modified plant may be any type of plant. In some embodiments, the modified plant is a monocotyledonous plant. In some embodiments, the plant seeds are from maize, wheat, rice, barley, rye, sugarcane, millet, oats, or sorghum. In some embodiments, the plant seeds are from maize plants. In some embodiments, the plant seeds are from maize (Zea maize) plants. In some embodiments, the modified plant is a dicotyledonous plant. In some embodiments, the plant is from soybean, cotton, alfalfa, beans, quinoa, lentils, peanuts, sunflower, canola, cassava, palm oil, potato, sugar beet, cocoa, coffee, lettuce, tomato, or cabbage plants. In some embodiments, the plant is from lettuce plants. In some embodiments, the plant is from lettuce (Lactuca sativa) plants. In some embodiments, the plant is from tomato plants. In some embodiments, the plant is from tomato (Solanum lycopersicum) plants. In some embodiments, the plant is a genetically modified organism (GMO). In some embodiments, the plant is a non-GMO plant. In some embodiments, the plant is a monocotyledonous or dicotyledonous plant. In some embodiments, the plant is corn, wheat, rice, sorghum, barley, rye, sugarcane, millet, oats, soybeans, cotton, alfalfa, beans, quinoa, lentils, peanuts, sunflowers, canola, cassava, palm oil, potatoes, sugar beets, cocoa, coffee, lettuce, tomatoes, peas, or cabbage.

[0155] The amount of microorganisms or exudates incorporated into the plant must be at a level sufficient to effectively sequestrate CO2. In some embodiments, the amount of microorganisms incorporated into the plant is approximately 250 colony-forming units (CFU) to approximately 5,000 CFU. In some embodiments, the amount of microorganisms incorporated into the plant is approximately 250 CFU to 500 CFU, approximately 250 CFU to 750 CFU, approximately 250 CFU to 1,000 CFU, approximately 250 CFU to 2,000 CFU, approximately 250 CFU to 3,000 CFU, approximately 250 CFU to 4,000 CFU, approximately 250 CFU to 5,000 CFU, approximately 500 CFU to 750 CFU, approximately 500 CFU to 1,000 CFU, approximately 500 CFU to 2,000 CFU, approximately 500 CFU to 3,000 CFU, approximately 500 CFU to 4,000 CFU, approximately 500 CFU to 5,000 CFU, approximately 750 CFU to 1,000 CFU, and approximately 7 The CFU levels are approximately 50 CFU to 2,000 CFU, approximately 750 CFU to 3,000 CFU, approximately 750 CFU to 4,000 CFU, approximately 750 CFU to 5,000 CFU, approximately 1,000 CFU to 2,000 CFU, approximately 1,000 CFU to 3,000 CFU, approximately 1,000 CFU to 4,000 CFU, approximately 1,000 CFU to 5,000 CFU, approximately 2,000 CFU to 3,000 CFU, approximately 2,000 CFU to 4,000 CFU, approximately 2,000 CFU to 5,000 CFU, approximately 3,000 CFU to 4,000 CFU, approximately 3,000 CFU to 5,000 CFU, or approximately 4,000 CFU to 5,000 CFU. In some embodiments, the amount of microorganisms incorporated into the plant 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 plant 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 plant is up to 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 is incorporated into the plant. In some embodiments, at least about 1000 CFU is incorporated into the plant.

[0156] In some embodiments, the microorganisms or exudates incorporated into the plant have long-term storage stability. In some embodiments, the modified plant has storage stability for about 3 months to about 36 months. In some embodiments, the modified plant has storage stability for about 3 months to about 6 months, about 3 months to about 9 months, about 3 months to about 12 months, about 3 months to about 15 months, about 3 months to about 18 months, about 3 months to about 21 months, about 3 months to about 24 months, about 3 months to about 30 months, about 3 months to about 36 months, about 6 months to about 9 months, about 6 months to about 12 months, Approximately 6 months to 15 months, approximately 6 months to 18 months, approximately 6 months to 21 months, approximately 6 months to 24 months, approximately 6 months to 30 months, approximately 6 months to 36 months, approximately 9 months to 12 months, approximately 9 months to 15 months, approximately 9 months to 18 months, approximately 9 months to 21 months, approximately 9 months to 24 months, approximately 9 months to 30 months, approximately 9 months Months to approximately 36 months, approximately 12 months to approximately 15 months, approximately 12 months to approximately 18 months, approximately 12 months to approximately 21 months, approximately 12 months to approximately 24 months, approximately 12 months to approximately 30 months, approximately 12 months to approximately 36 months, approximately 15 months to approximately 18 months, approximately 15 months to approximately 21 months, approximately 15 months to approximately 24 months, approximately 15 months to approximately 30 months, approximately 15 months to approximately The plants have storage stability for 36 months, approximately 18 to 21 months, approximately 18 to 24 months, approximately 18 to 30 months, approximately 18 to 36 months, approximately 21 to 24 months, approximately 21 to 30 months, approximately 21 to 36 months, approximately 24 to 30 months, approximately 24 to 36 months, or approximately 30 to 36 months. In some embodiments, the modified plants have storage stability for approximately 3 months, approximately 6 months, approximately 9 months, approximately 12 months, approximately 15 months, approximately 18 months, approximately 21 months, approximately 24 months, approximately 30 months, or approximately 36 months. In some embodiments, the modified plants 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 plants have storage stability for up to approximately 6 months, 9 months, 12 months, 15 months, 18 months, 21 months, 24 months, 30 months, or 36 months.

[0157] In some embodiments, the microorganisms incorporated into the plant are stable after incorporation. In some embodiments, the microorganisms are stable for longer than 30 days, longer than 6 months, longer than 1 year, or longer than 2 years. In some embodiments, the microorganisms are stable for longer than 30 days. In some embodiments, the microorganisms are stable for longer than 6 months. In some embodiments, the microorganisms are stable for longer than 1 year. In some embodiments, the microorganisms are stable for longer than 2 years.

[0158] The microorganisms incorporated into the plant or their exudates may be any of the microorganisms provided herein or any other microorganisms. In some embodiments, the microorganism is a microbe. In some embodiments, the microorganism is an endospore-forming microbe. In some embodiments, the microorganism is an endospore-forming microbe or its endospores. In some embodiments, the microorganism is an endospore of a microorganism provided herein. In some embodiments, the microorganism is an endospore-forming bacterium or its endospores.

[0159] Microorganisms and exudates The microorganisms or their exudates provided herein can produce or promote the formation of bicarbonates and one or more minerals. In some embodiments, the formation of bicarbonates sequesters CO2. In some embodiments, the formation of bicarbonates results in the formation of minerals. In some embodiments, the formed minerals stably sequester carbon in the soil. In some embodiments, the microorganisms are endospore-forming bacteria. In some embodiments, the microorganisms are bacterial endospores. Whenever a microorganism (e.g., bacteria) referred herein can form endospores, it is intended that any endospores of that microorganism are also included. For example, if a plant treatment formulation contains Bacillus sp., the formulation may contain Bacillus sp. endospores.

[0160] In some embodiments, the microorganism is a microbe from the phylum Firmicutes, phylum Proteobacteria, and phylum Actinomycetes. In some embodiments, the microorganism is a microbe from the phylum Firmicutes. In some embodiments, the microorganism is a microbe from the phylum Proteobacteria. In some embodiments, the microorganism is a microbe from the phylum Actinomycetes. In some embodiments, the microorganism is an endospore of any of the microorganisms.

[0161] Rhizosphere bacteria, which fix atmospheric nitrogen, are found in plant roots. These organisms can survive in the soil and are tolerant of large amounts of CO2, which is either CO2 released from plant roots during respiration or CO2 released through respiration by nearby microorganisms and soil fauna.

[0162] In some embodiments, the bacteria are not genetically modified. In some embodiments, the bacteria are selected for their ability to convert CO2 into bicarbonates and ultimately minerals. In some embodiments, the bacteria are selected for the use of carbonic anhydrase. In some embodiments, the bacteria are selected for the use of carbonic anhydrase. In some embodiments, various rhizosphere bacteria are effectively loaded onto the seeds. In some embodiments, the rhizosphere bacteria include Bacillus sp., Paenibacillus sp., or both. In some embodiments, the one or more microorganisms are B. amyloliquefaciens, B. laterosporus, B. licheniformis, B. macerans, B. cereus, B. circulans, B. firmus, B. subtilis, B. sphaericus, B. megaterium, B. coagulans ( This includes B. coagulans, B. brevis, B. thuringiensis, B. mycoides, B. cucumis, B. endophyticus, B. pumilus, B. velezensis, B. mucilaginosus, B. tequilensis, B. methylotrophicus, or any combination thereof.In some embodiments, the one or more microorganisms include Bacillus subtilis S3C23, Bacillus subtilis MP2, Bacillus subtilis RO2C15, Bacillus subtilis RO2C22, Bacillus megaterium 6, Bacillus megaterium S3C21, Bacillus megaterium RO2C12, Bacillus cucumis S3C14, Bacillus endophyticus 5, or any combination thereof. In some embodiments, the one or more microorganisms include Bacillus subtilis S3C23. In some embodiments, the one or more microorganisms include Bacillus subtilis MP2.

[0163] In some embodiments, the one or more microorganisms include a genetic modification that causes the one or more microorganisms to produce or promote the formation of more carbonic anhydrase compared to the corresponding wild-type microorganisms. In some embodiments, the genetic modification includes a nucleic acid construct, the nucleic acid construct includes one or more promoters configured to drive the expression of a carbonic anhydrase (CA) coding sequence. In some embodiments, the genetic modification includes a nucleic acid construct, the nucleic acid construct includes a carbonic anhydrase (CA) coding sequence. In some embodiments, the CA coding sequence is heterogeneous to the one or more microorganisms. In some embodiments, the CA coding sequence is endogenous to the one or more microorganisms. In some embodiments, the nucleic acid construct is codon-optimized. In some embodiments, the nucleic acid construct further includes one or more promoters configured to drive the expression of the CA coding sequence. In some embodiments, the one or more promoters drive the constitutive expression of the CA coding sequence. In some embodiments, the one or more promoters drive the inducible expression of the CA coding sequence. In some embodiments, the genetic modification includes a carbonic anhydrase gene modified by direct microbial evolution. In some embodiments, the gene modification includes a signal sequence. In some embodiments, the signal sequence includes a periplasmic signal sequence or an extracellular secretory signal. In some embodiments, the periplasmic signal sequence or the extracellular secretory signal is located at the 5' end of a carbonic anhydrase coding sequence. In some embodiments, the periplasmic signal sequence or the extracellular secretory signal is fused to the 5' end of a carbonic anhydrase coding sequence. In some embodiments, the gene modification includes one or more components of a common secretory pathway, and the gene modification results in the secretion or intracellular targeting of carbonic anhydrase by the one or more microorganisms.

[0164] In some embodiments, the one or more promoters are composed of or derived from the Bacillus sp. or Paenibacillus sp. genera. In some embodiments, the one or more promoters are P groES P43, P sigX , P trnQ , and P xylA Selected from. In some embodiments, the one or more promoters are one or more housekeeping gene promoters or strongly expressed constitutive promoters, or derived therefrom. In some embodiments, the one or more housekeeping gene promoters or strongly expressed constitutive promoters are P liaG , P lepA , P veg , P gsiB P43, P trnQ , P ial (Bacitracin-induced), and P xylA Selected from (xylose-inducible).

[0165] In some embodiments, the gene modification includes the introduction of an expression vector into one or more microorganisms. In some embodiments, the gene modification includes modifications to the chromosomes of one or more microorganisms.

[0166] In some embodiments, microorganisms can fix both nitrogen and carbon dioxide. In some embodiments, nitrogen-fixing microorganisms continue to provide fixed nitrogen (ammonia) to plants through biological nitrogen fixation, which reduces the use of chemical fertilizers that pollute the environment. The final product of atmospheric nitrogen fixation (ammonia) can enhance the CO2-to-mineral process because ammonia has been reported to increase pH, which promotes mineralization. Since ammonia production from the microorganisms disclosed herein is significantly higher than that of other soil-dwelling microorganisms, even in the presence of an external nitrogen source, the mineralization process can be carried out more rapidly by the strains disclosed herein than by other soil-dwelling microorganisms.

[0167] Methods for sequestering or converting carbon into bicarbonates and minerals In certain embodiments, a method for sequestering carbon is disclosed herein, the method comprising the steps of a. culturing a plant and one or more microorganisms associated with the plant, wherein the one or more microorganisms are selected to produce or promote the formation of bicarbonates and one or more minerals, or are derived from such microorganisms.

[0168] Modified plants In one embodiment, a method for carbon sequestration is provided herein, comprising the step of cultivating a modified plant containing a microorganism or a microbial exudate incorporated into the plant. In some embodiments, the microorganism or exudate produces or promotes the formation of bicarbonates and one or more minerals. In some embodiments, the formation of bicarbonates sequesters CO2. In some embodiments, the formation of bicarbonates results in the formation of minerals. In some embodiments, the formed minerals stably sequester carbon in the soil. In some preferred embodiments, the microorganism is an endospore-forming bacterium or its endospores.

[0169] In some embodiments, the amount of CO2 converted by microorganisms is between 0.1 tons of CO2 per acre and a maximum of 2.5 tons of CO2 per acre. In some embodiments, microorganisms convert 2.5 to 5.3 tons of CO2 / acre. In some embodiments, microorganisms convert 5.3 to 7.5 tons of CO2 / acre. In some embodiments, microorganisms convert 7.5 to 10 tons of CO2 / acre. In some embodiments, microorganisms convert 10 to 15 tons of CO2 / acre. In some embodiments, microorganisms convert 15 to 20 tons / acre. In some embodiments, the amount of CO2 converted by microorganisms is between approximately 2 tons / acre and approximately 20 tons / acre.In some embodiments, the amount of CO2 converted by microorganisms is approximately 2 tons / acre to 4 tons / acre, approximately 2 tons / acre to 6 tons / acre, approximately 2 tons / acre to 8 tons / acre, approximately 2 tons / acre to 10 tons / acre, approximately 2 tons / acre to 12 tons / acre, approximately 2 tons / acre to 14 tons / acre, approximately 2 tons / acre to 16 tons / acre, approximately 2 tons / acre to 18 tons / acre, approximately 2 tons / acre to 20 tons / acre, approximately 4 tons / acre to 6 tons / acre, approximately 4 tons / acre to approximately 8 tons / acre, approximately 4 tons / acre to approximately 10 tons / acre, approximately 4 tons / acre to approximately 12 tons / acre, approximately 4 tons / acre to approximately 14 tons / acre, approximately 4 tons / acre to approximately 16 tons / acre, approximately 4 tons / acre to approximately 18 tons / acre, approximately 4 tons / acre to approximately 20 tons / acre, approximately 6 tons / acre to approximately 8 tons / acre, approximately 6 tons / acre to approximately 10 tons / acre, approximately 6 tons / acre to approximately 12 tons / acre, approximately 6 tons / acre to approximately 14 tons / acre, approximately 6 tons / acre to approximately 16 tons / acre, approximately 6 tons / acre ~approximately 18 tons / acre, approximately 6 tons / acre to approximately 20 tons / acre, approximately 8 tons / acre to approximately 10 tons / acre, approximately 8 tons / acre to approximately 12 tons / acre, approximately 8 tons / acre to approximately 14 tons / acre, approximately 8 tons / acre to approximately 16 tons / acre, approximately 8 tons / acre to approximately 18 tons / acre, approximately 8 tons / acre to approximately 20 tons / acre, approximately 10 tons / acre to approximately 12 tons / acre, approximately 10 tons / acre to approximately 14 tons / acre, approximately 10 tons / acre to approximately 16 tons / acre, approximately 10 tons / acre to approximately 18 tons / acre, approximately The range is between 10 tons / acre and approximately 20 tons / acre, approximately 12 tons / acre and approximately 14 tons / acre, approximately 12 tons / acre and approximately 16 tons / acre, approximately 12 tons / acre and approximately 18 tons / acre, approximately 12 tons / acre and approximately 20 tons / acre, approximately 14 tons / acre and approximately 16 tons / acre, approximately 14 tons / acre and approximately 18 tons / acre, approximately 14 tons / acre and approximately 20 tons / acre, approximately 16 tons / acre and approximately 18 tons / acre, or between approximately 18 tons / acre and approximately 20 tons / acre.In some embodiments, the amount of CO2 converted by microbe is between approximately 2 tons / acre, approximately 4 tons / acre, approximately 6 tons / acre, approximately 8 tons / acre, approximately 10 tons / acre, approximately 12 tons / acre, approximately 14 tons / acre, approximately 16 tons / acre, approximately 18 tons / acre, or approximately 20 tons / acre. In some embodiments, the amount of CO2 converted by microorganisms is at least between approximately 2 tons / acre, approximately 4 tons / acre, approximately 6 tons / acre, approximately 8 tons / acre, approximately 10 tons / acre, approximately 12 tons / acre, approximately 14 tons / acre, approximately 16 tons / acre, or approximately 18 tons / acre. In some embodiments, the amount of CO2 converted by microorganisms is at most between approximately 4 tons / acre, approximately 6 tons / acre, approximately 8 tons / acre, approximately 10 tons / acre, approximately 12 tons / acre, approximately 14 tons / acre, approximately 16 tons / acre, approximately 18 tons / acre, or approximately 20 tons / acre.

[0170] In some embodiments, the microorganism or exudate is incorporated into the plant. In some embodiments, the microorganism or exudate is incorporated into the plant beneath the pericarp. In some embodiments, the microorganism or exudate is incorporated into the plant between the pericarp and the aleurone cell layer. In some embodiments, the microorganism or exudate comes into contact with the plant embryo. In some embodiments, the microorganism or exudate does not come into contact with the plant embryo. In some embodiments, the microorganism or exudate comes into contact with the plant endosperm. In some embodiments, the microorganism or exudate does not come into contact with the plant endosperm. In some embodiments, the microorganism or exudate is incorporated into the plant in the space between the plant sheath and the plant embryo. In some embodiments, the microorganism or exudate is incorporated in the space between the plant pericarp and the plant aleurone cell layer.

[0171] The modified plant may be any type of plant. In some embodiments, the modified plant is a monocotyledonous plant. In some embodiments, the plant seeds are from maize, wheat, rice, barley, rye, sugarcane, millet, oats, or sorghum. In some embodiments, the plant seeds are from maize plants. In some embodiments, the plant seeds are from maize (Zea maize) plants. In some embodiments, the modified plant is a dicotyledonous plant. In some embodiments, the plant is from soybean, cotton, alfalfa, beans, quinoa, lentils, peanuts, sunflower, canola, cassava, palm oil, potato, sugar beet, cocoa, coffee, lettuce, tomato, or cabbage plants. In some embodiments, the plant is from lettuce plants. In some embodiments, the plant is from lettuce (Lactuca sativa) plants. In some embodiments, the plant is from tomato plants. In some embodiments, the plant is from tomato (Solanum lycopersicum) plants. In some embodiments, the plant is a genetically modified organism (GMO). In some embodiments, the plant is a non-GMO plant.

[0172] The amount of microorganisms or exudates incorporated into the plant must be at a level sufficient to effectively sequestrate CO2. In some embodiments, the amount of microorganisms incorporated into the plant is approximately 250 colony-forming units (CFU) to approximately 5,000 CFU. In some embodiments, the amount of microorganisms incorporated into the plant is approximately 250 CFU to 500 CFU, approximately 250 CFU to 750 CFU, approximately 250 CFU to 1,000 CFU, approximately 250 CFU to 2,000 CFU, approximately 250 CFU to 3,000 CFU, approximately 250 CFU to 4,000 CFU, approximately 250 CFU to 5,000 CFU, approximately 500 CFU to 750 CFU, approximately 500 CFU to 1,000 CFU, approximately 500 CFU to 2,000 CFU, approximately 500 CFU to 3,000 CFU, approximately 500 CFU to 4,000 CFU, approximately 500 CFU to 5,000 CFU, approximately 750 CFU to 1,000 CFU, and approximately 7 The CFU levels are approximately 50 CFU to 2,000 CFU, approximately 750 CFU to 3,000 CFU, approximately 750 CFU to 4,000 CFU, approximately 750 CFU to 5,000 CFU, approximately 1,000 CFU to 2,000 CFU, approximately 1,000 CFU to 3,000 CFU, approximately 1,000 CFU to 4,000 CFU, approximately 1,000 CFU to 5,000 CFU, approximately 2,000 CFU to 3,000 CFU, approximately 2,000 CFU to 4,000 CFU, approximately 2,000 CFU to 5,000 CFU, approximately 3,000 CFU to 4,000 CFU, approximately 3,000 CFU to 5,000 CFU, or approximately 4,000 CFU to 5,000 CFU. In some embodiments, the amount of microorganisms incorporated into the plant 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 plant 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 plant is up to 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 is incorporated into the plant. In some embodiments, at least about 1000 CFU is incorporated into the plant.

[0173] In some embodiments, the microorganisms or exudates incorporated into the plant have long-term storage stability. In some embodiments, the modified plant has storage stability for about 3 months to about 36 months. In some embodiments, the modified plant has storage stability for about 3 months to about 6 months, about 3 months to about 9 months, about 3 months to about 12 months, about 3 months to about 15 months, about 3 months to about 18 months, about 3 months to about 21 months, about 3 months to about 24 months, about 3 months to about 30 months, about 3 months to about 36 months, about 6 months to about 9 months, about 6 months to about 12 months, Approximately 6 months to 15 months, approximately 6 months to 18 months, approximately 6 months to 21 months, approximately 6 months to 24 months, approximately 6 months to 30 months, approximately 6 months to 36 months, approximately 9 months to 12 months, approximately 9 months to 15 months, approximately 9 months to 18 months, approximately 9 months to 21 months, approximately 9 months to 24 months, approximately 9 months to 30 months, approximately 9 months Months to approximately 36 months, approximately 12 months to approximately 15 months, approximately 12 months to approximately 18 months, approximately 12 months to approximately 21 months, approximately 12 months to approximately 24 months, approximately 12 months to approximately 30 months, approximately 12 months to approximately 36 months, approximately 15 months to approximately 18 months, approximately 15 months to approximately 21 months, approximately 15 months to approximately 24 months, approximately 15 months to approximately 30 months, approximately 15 months to approximately The plants have storage stability for 36 months, approximately 18 to 21 months, approximately 18 to 24 months, approximately 18 to 30 months, approximately 18 to 36 months, approximately 21 to 24 months, approximately 21 to 30 months, approximately 21 to 36 months, approximately 24 to 30 months, approximately 24 to 36 months, or approximately 30 to 36 months. In some embodiments, the modified plants have storage stability for approximately 3 months, approximately 6 months, approximately 9 months, approximately 12 months, approximately 15 months, approximately 18 months, approximately 21 months, approximately 24 months, approximately 30 months, or approximately 36 months. In some embodiments, the modified plants 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 plants have storage stability for up to approximately 6 months, 9 months, 12 months, 15 months, 18 months, 21 months, 24 months, 30 months, or 36 months.

[0174] In some embodiments, the microorganisms incorporated into the plant are stable after incorporation. In some embodiments, the microorganisms are stable for longer than 30 days, longer than 6 months, longer than 1 year, or longer than 2 years. In some embodiments, the microorganisms are stable for longer than 30 days. In some embodiments, the microorganisms are stable for longer than 6 months. In some embodiments, the microorganisms are stable for longer than 1 year. In some embodiments, the microorganisms are stable for longer than 2 years.

[0175] The microorganisms incorporated into the plant or their exudates may be any of the microorganisms provided herein or any other microorganisms. In some embodiments, the microorganism is a microbe. In some embodiments, the microorganism is an endospore-forming microbe. In some embodiments, the microorganism is an endospore-forming microbe or its endospores. In some embodiments, the microorganism is an endospore of a microorganism provided herein. In some embodiments, the microorganism is an endospore-forming bacterium or its endospores.

[0176] Methods for incorporating bacteria In one embodiment, a method for incorporating one or more microorganisms or their exudates into one or more plants is provided herein. In some embodiments, the method includes the step of disinfecting the plants. In some embodiments, the method includes the step of bringing the plants into contact with a solution containing one or more microorganisms or their exudates. In some embodiments, the solution further includes salt. In some embodiments, the method includes the step of incubating the plants with the solution for a certain period of time. In some embodiments, the period of time is sufficient to introduce a desired amount of the microorganisms or their exudates into the plants. In some embodiments, the method incorporates a desired amount of the microorganisms or their exudates into the plants.

[0177] In some embodiments, the above method includes the step of contacting a plant with a solution containing a salt as described herein.

[0178] In some embodiments, the solution includes additional additives as described herein.

[0179] In some embodiments, the solution contains additional metal ions as described herein.

[0180] In some embodiments, the solution comprises one or more nutrients for microorganisms as described herein. In some embodiments, the solution comprises a bacterial growth medium. In some embodiments, the solution comprises a lysogenic medium (LB), a nutrient broth, or a combination thereof. In some embodiments, the solution comprises a lysogenic medium. In some embodiments, the solution comprises a nutrient broth.

[0181] In some embodiments, the solution contains microorganisms such as those described herein.

[0182] In some embodiments, the solution contains a desired amount of microorganisms per unit of plant mass as described herein.

[0183] In some embodiments, the plant contains a desired amount of microorganisms per plant, as described herein. In some embodiments, the microorganisms are bacteria. In some embodiments, the bacteria are endosporic bacteria. In some embodiments, the method includes a step of inducing endosporulation of endosporic bacteria. In some embodiments, the bacteria incorporated into the plant are endospores. In some embodiments, the solution contains one or more components to induce endosporulation. In some embodiments, the solution contains potassium, ferrous sulfate, calcium, magnesium, manganese, or a combination thereof.

[0184] In some embodiments, the method includes a step of sterilizing the plant. In some embodiments, the method includes a step of sterilizing the surface of the plant. Any method for producing a plant having a sterilized surface may be employed. In some embodiments, the plant is sterilized with a bleach solution. In some embodiments, the plant is sterilized before immersing it in a solution containing one or more microorganisms. In some embodiments, the plant is a sterilized plant. In some embodiments, the plant has a sterilized surface. As used herein, “sterilizing,” “sterilized,” and related terms (e.g., “disinfecting”) indicate that there are substantially no surviving microorganisms on what has been sterilized. In some embodiments, the plant is sterilized before incubating it in a solution containing microorganisms. In some embodiments, the plant is sterilized after incubating it in a solution containing microorganisms. In some embodiments, a fungicide is added to the surface of the plant.

[0185] In some embodiments, sterilized or disinfected plants are substantially free of living microorganisms on the plant (e.g., on the plant surface). In some embodiments, sterile or sterilized plants 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 plant.

[0186] In some embodiments, the plants are incubated with a solution containing microorganisms for a sufficient time to incorporate the microorganisms into the plants.

[0187] Microorganisms and exudates The microorganisms or their exudates provided herein produce or promote the formation of bicarbonates and one or more minerals. In some embodiments, the formation of bicarbonates sequesters CO2. In some embodiments, the formation of bicarbonates results in the formation of minerals. In some embodiments, the formed minerals stably sequester carbon in the soil. In some embodiments, the microorganisms are bacteria as described herein. In some embodiments, the microorganisms are endospore-forming bacteria. In some embodiments, the microorganisms are endospores of bacteria. Whenever a microorganism (e.g., bacteria) referred herein can form endospores, it is intended that any endospores of that microorganism are also included. For example, if a plant treatment formulation contains Bacillus sp., the formulation may contain endospores of Bacillus sp.

[0188] In some embodiments, the microorganism is a microbe from the phylum Firmicutes, phylum Proteobacteria, and phylum Actinomycetes. In some embodiments, the microorganism is a microbe from the phylum Firmicutes. In some embodiments, the microorganism is a microbe from the phylum Proteobacteria. In some embodiments, the microorganism is a microbe from the phylum Actinomycetes. In some embodiments, the microorganism is an endospore of any of the microorganisms.

[0189] Rhizosphere bacteria, which fix atmospheric nitrogen, are found in plant roots. These microorganisms can survive in the soil and are tolerant of large amounts of CO2, which is either CO2 released from plant roots during respiration or CO2 released through respiration by nearby microorganisms and soil fauna.

[0190] In some embodiments, the bacteria are not genetically modified. In some embodiments, the bacteria are selected for their ability to convert CO2 into bicarbonates and ultimately minerals. In some embodiments, the bacteria are selected for the use of carbonic anhydrase. In some embodiments, the bacteria are selected for the use of carbonic anhydrase. In some embodiments, various rhizosphere bacteria are effectively loaded onto the seeds. In some embodiments, the rhizosphere bacteria include endospore-forming bacteria that enhance biological nitrogen fixation. In some embodiments, the rhizosphere bacteria include Bacillus sp., Paenibacillus sp., or both. In some embodiments, the one or more microorganisms are B. amyloliquefaciens, B. laterosporus, B. licheniformis, B. macerans, B. cereus, B. circulans, B. firmus, B. subtilis, B. sphaericus, B. megaterium, B. coagulans ( This includes B. coagulans, B. brevis, B. thuringiensis, B. mycoides, B. cucumis, B. endophyticus, B. pumilus, B. velezensis, B. mucilaginosus, B. tequilensis, B. methylotrophicus, or any combination thereof.In some embodiments, the one or more microorganisms include Bacillus subtilis S3C23, Bacillus subtilis MP2, Bacillus subtilis RO2C15, Bacillus subtilis RO2C22, Bacillus megaterium 6, Bacillus megaterium S3C21, Bacillus megaterium RO2C12, Bacillus cucumis S3C14, Bacillus endophyticus 5, or any combination thereof. In some embodiments, the one or more microorganisms include Bacillus subtilis S3C23. In some embodiments, the one or more microorganisms include Bacillus subtilis MP2.

[0191] In some embodiments, the one or more microorganisms include a genetic modification that causes the one or more microorganisms to produce or promote the formation of more carbonic anhydrase compared to the corresponding wild-type microorganisms. In some embodiments, the genetic modification includes a nucleic acid construct, the nucleic acid construct includes one or more promoters configured to drive the expression of a carbonic anhydrase (CA) coding sequence. In some embodiments, the genetic modification includes a nucleic acid construct, the nucleic acid construct includes a carbonic anhydrase (CA) coding sequence. In some embodiments, the CA coding sequence is heterogeneous to the one or more microorganisms. In some embodiments, the CA coding sequence is endogenous to the one or more microorganisms. In some embodiments, the nucleic acid construct is codon-optimized. In some embodiments, the nucleic acid construct further includes one or more promoters configured to drive the expression of the CA coding sequence. In some embodiments, the one or more promoters drive the constitutive expression of the CA coding sequence. In some embodiments, the one or more promoters drive the inducible expression of the CA coding sequence. In some embodiments, the genetic modification includes a carbonic anhydrase gene modified by direct microbial evolution. In some embodiments, the gene modification includes a signal sequence. In some embodiments, the signal sequence includes a periplasmic signal sequence or an extracellular secretory signal. In some embodiments, the periplasmic signal sequence or the extracellular secretory signal is located at the 5' end of a carbonic anhydrase coding sequence. In some embodiments, the periplasmic signal sequence or the extracellular secretory signal is fused to the 5' end of a carbonic anhydrase coding sequence. In some embodiments, the gene modification includes one or more components of a common secretory pathway, and the gene modification results in the secretion or intracellular targeting of carbonic anhydrase by the one or more microorganisms.

[0192] In some embodiments, the one or more promoters are composed of or derived from the Bacillus sp. or Paenibacillus sp. genera. In some embodiments, the one or more promoters are P groES P43, P sigX , P trnQ , and P xylA Selected from. In some embodiments, the one or more promoters are one or more housekeeping gene promoters or strongly expressed constitutive promoters, or derived therefrom. In some embodiments, the one or more housekeeping gene promoters or strongly expressed constitutive promoters are P liaG , P lepA , P veg , P gsiB P43, P trnQ , P ial (Bacitracin-induced), and P xylA Selected from (xylose-inducible).

[0193] In some embodiments, the gene modification includes the introduction of an expression vector into one or more microorganisms. In some embodiments, the gene modification includes modifications to the chromosomes of one or more microorganisms.

[0194] In some embodiments, microorganisms can fix both nitrogen and carbon dioxide. In some embodiments, nitrogen-fixing microorganisms continue to provide fixed nitrogen (ammonia) to plants through biological nitrogen fixation, which reduces the use of chemical fertilizers that pollute the environment. The final product of atmospheric nitrogen fixation (ammonia) can enhance the CO2-to-mineral process because ammonia has been reported to increase pH, which promotes mineralization. Since ammonia production from the microorganisms disclosed herein is significantly higher than that of other soil-dwelling microorganisms, even in the presence of an external nitrogen source, the mineralization process can be carried out more rapidly by the strains disclosed herein than by other soil-dwelling microorganisms.

[0195] In some embodiments, the microorganisms include genera such as Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanerobacter sp., Caloramator sp., Caminicella sp., and Cerasibacillus. 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., Gerria 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., Moorela 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 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 The microbes are selected from the genera Thermovenablum sp., Tuberibacillus sp., Virgibacillus sp., and Vulcanobacillus sp. In some embodiments, the microorganisms are selected from the genera 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 of the genus Acetobacter sp. In some embodiments, the microorganism is of the genus Actinomyces sp. In some embodiments, the microorganism is of the genus Bacillus sp.In some embodiments, the microorganism is Chryseobacterium sp. In some embodiments, the microorganism is Coxiella sp. In some embodiments, the microorganism is Ensifer sp. In some embodiments, the microorganism is Glutamicibacter sp. In some embodiments, the microorganism is Microbacterium sp. In some embodiments, the microorganism is Pantoea sp. In some embodiments, the microorganism is Serratia sp. In some embodiments, the microorganism is an endospore of any of the microorganisms.

[0196] In some embodiments, microorganisms are selected for one or more properties related to their ability to interact with plants. In some embodiments, microorganisms are selected for compatibility. In some embodiments, microorganisms are selected so that phagocytic or antagonistic effects do not occur. In some embodiments, microorganisms are selected for stability during storage. In some embodiments, microorganisms are selected for rapid colonization of plants and survival within relevant tissues. In some embodiments, microorganisms are selected for optimal integration into one or more plants. In some embodiments, microorganisms remain present throughout the entire life cycle of the plant.

[0197] In some embodiments, the microorganisms incorporated into the plant are stable after incorporation. In some embodiments, the microorganisms are stable for longer than 30 days, longer than 6 months, longer than 1 year, or longer than 2 years. In some embodiments, the microorganisms are stable for longer than 30 days. In some embodiments, the microorganisms are stable for longer than 6 months. In some embodiments, the microorganisms are stable for longer than 1 year. In some embodiments, the microorganisms are stable for longer than 2 years.

[0198] composition containing bacteria In certain embodiments, compositions comprising one or more microorganisms are disclosed herein, wherein the one or more microorganisms are selected to produce or promote the formation of bicarbonates and one or more minerals, or are derived from such microorganisms.

[0199] The microorganisms or their exudates provided herein produce or promote the formation of bicarbonates and one or more minerals. In some embodiments, the formation of bicarbonates sequesters CO2. In some embodiments, the formation of bicarbonates results in the formation of minerals. In some embodiments, the formed minerals stably sequester carbon in the soil. In some embodiments, the microorganisms are bacteria as described herein. In some embodiments, the microorganisms are endospore-forming bacteria. In some embodiments, the microorganisms are endospores of bacteria. Whenever a microorganism (e.g., bacteria) referred herein can form endospores, it is intended that any endospores of that microorganism are also included. For example, if a plant treatment formulation contains Bacillus sp., the formulation may contain endospores of Bacillus sp.

[0200] In some embodiments, the microorganism is a microbe from the phylum Firmicutes, phylum Proteobacteria, and phylum Actinomycetes. In some embodiments, the microorganism is a microbe from the phylum Firmicutes. In some embodiments, the microorganism is a microbe from the phylum Proteobacteria. In some embodiments, the microorganism is a microbe from the phylum Actinomycetes. In some embodiments, the microorganism is an endospore of any of the microorganisms.

[0201] Rhizosphere bacteria, which fix atmospheric nitrogen, are found in plant roots. These organisms can survive in the soil and are tolerant of large amounts of CO2, which is either CO2 released from plant roots during respiration or CO2 released through respiration by nearby microorganisms and soil fauna.

[0202] In some embodiments, the bacteria are not genetically modified. In some embodiments, the bacteria are selected for their ability to convert CO2 into bicarbonates and ultimately minerals. In some embodiments, the bacteria are selected for the use of carbonic anhydrase. In some embodiments, the bacteria are not genetically modified. In some embodiments, the bacteria are selected for their ability to convert CO2 into bicarbonates and minerals. In some embodiments, the bacteria are selected for the use of carbonic anhydrase. In some embodiments, the bacteria are selected for the use of carbonic anhydrase. In some embodiments, various rhizosphere bacteria are effectively loaded onto the seeds. In some embodiments, the rhizosphere bacteria include endospore-forming bacteria. In some embodiments, the rhizosphere bacteria include Bacillus sp., Paenibacillus sp., or both. In some embodiments, the one or more microorganisms are B. amyloliquefaciens, B. laterosporus, B. licheniformis, B. macerans, B. cereus, B. circulans, B. firmus, B. subtilis, B. sphaericus, B. megaterium, B. coagulans ( This includes B. coagulans, B. brevis, B. thuringiensis, B. mycoides, B. cucumis, B. endophyticus, B. pumilus, B. velezensis, B. mucilaginosus, B. tequilensis, B. methylotrophicus, or any combination thereof.In some embodiments, the one or more microorganisms include Bacillus subtilis S3C23, Bacillus subtilis MP2, Bacillus subtilis RO2C15, Bacillus subtilis RO2C22, Bacillus megaterium 6, Bacillus megaterium S3C21, Bacillus megaterium RO2C12, Bacillus cucumis S3C14, Bacillus endophyticus 5, or any combination thereof. In some embodiments, the one or more microorganisms include Bacillus subtilis S3C23. In some embodiments, the one or more microorganisms include Bacillus subtilis MP2.

[0203] In some embodiments, microorganisms can fix both nitrogen and carbon dioxide. In some embodiments, nitrogen-fixing microorganisms continue to provide fixed nitrogen (ammonia) to plants through biological nitrogen fixation, which reduces the use of chemical fertilizers that pollute the environment. The final product of atmospheric nitrogen fixation (ammonia) can enhance the process from CO2 to bicarbonates and minerals, as ammonia has been reported to increase pH, which promotes mineralization. Since ammonia production from the microorganisms disclosed herein is significantly higher than that of other soil-dwelling microorganisms, even in the presence of an external nitrogen source, the mineralization process can be carried out more rapidly by the strains disclosed herein than by other soil-dwelling microorganisms.

[0204] In some embodiments, the microorganisms include genera such as Acetonema sp., Actinomyces sp., Alkalibacillus sp., Ammoniphilus sp., Amphibacillus sp., Anaerobacter sp., Anaerospora sp., Aneurinibacillus sp., Anoxybacillus sp., Bacillus sp., Brevibacillus sp., Caldanerobacter sp., Caloramator sp., Caminicella sp., and Cerasibacillus. 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., Gerria 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., Moorela 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 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 The microbes are selected from the genera Thermovenablum sp., Tuberibacillus sp., Virgibacillus sp., and Vulcanobacillus sp. In some embodiments, the microorganisms are selected from the genera 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 of the genus Acetobacter sp. In some embodiments, the microorganism is of the genus Actinomyces sp. In some embodiments, the microorganism is of the genus Bacillus sp.In some embodiments, the microorganism is Chryseobacterium sp. In some embodiments, the microorganism is Coxiella sp. In some embodiments, the microorganism is Ensifer sp. In some embodiments, the microorganism is Glutamicibacter sp. In some embodiments, the microorganism is Microbacterium sp. In some embodiments, the microorganism is Pantoea sp. In some embodiments, the microorganism is Serratia sp. In some embodiments, the microorganism is an endospore of any of the microorganisms.

[0205] In some embodiments, microorganisms are selected for one or more properties related to their ability to interact with plants. In some embodiments, microorganisms are selected for compatibility. In some embodiments, microorganisms are selected so that phagocytic or antagonistic effects do not occur. In some embodiments, microorganisms are selected for stability during storage. In some embodiments, microorganisms are selected for rapid colonization of plants and survival within relevant tissues. In some embodiments, microorganisms are selected for optimal integration into one or more plants. In some embodiments, microorganisms remain present throughout the entire life cycle of the plant.

[0206] In some embodiments, the microorganisms incorporated into the plant are stable after incorporation. In some embodiments, the microorganisms are stable for longer than 30 days, longer than 6 months, longer than 1 year, or longer than 2 years. In some embodiments, the microorganisms are stable for longer than 30 days. In some embodiments, the microorganisms are stable for longer than 6 months. In some embodiments, the microorganisms are stable for longer than 1 year. In some embodiments, the microorganisms are stable for longer than 2 years.

[0207] definition Whenever the terms “at least,” “greater than,” or “greater than or equal to” precede the first number in a set of two or more numbers, those terms apply to each of the numbers in that set. For example, 1, 2, or 3 or more is equivalent to 1 or more, 2 or more, or 3 or more.

[0208] Whenever the terms "no more than," "less than," or "less than or equal to" are placed before the first number in a set of two or more numbers, the terms "no more than," "less than," or "less than or equal to" apply to each of the numbers in that set. For example, 3, 2, or 1 or less is equivalent to 3 or less, 2 or less, or 1 or less.

[0209] The use of absolute or sequential terms, such as “will,” “will not,” “shall,” “shall not,” “must,” “must not,” “first,” “initially,” “next,” “sequently,” “before,” “after,” “lastly,” and “final,” is intended as illustrative examples and does not limit the scope of the embodiments disclosed herein.

[0210] As used herein, the singular forms “a,” “an,” and “the” are intended to include the plural forms similarly, unless the context otherwise expressly indicates. Furthermore, the terms “including,” “includes,” “having,” “has,” and “with,” or their variations thereof, are intended to include in a manner similar to the term “comprising,” to the extent that they are used in the detailed description and / or in any of the claims.

[0211] The term “irrigation system” may also refer to an artificial process that applies a controlled amount of water to nourish plants, as well as to support crop production, as described herein, and may be known here as “sprinkling.” In some embodiments, the term “irrigation system” may include foliar spraying, furrow fertilization, sprinkler systems, humidifiers, or misting systems.

[0212] As used herein, the expressions “at least one,” “one or more,” and “and / or” are open-ended expressions that are both conjunctive and disjunctive when in operation. For example, each of the expressions “at least one of A, B, and C,” “at least one of A, B, or C,” “one or more of A, B, and C,” “one or more of A, B, or C,” and “A, B, and / or C” means A alone, B alone, C alone, A and B together, A and C together, B and C together, or A, B, and C together.

[0213] As used herein, “or” may refer to “and,” “or,” or “and / or,” and may be used exclusively or inclusively. For example, the term “A or B” may refer to “A or B,” “A but not B,” “B but not A,” and “A and B.” In some cases, the context may indicate a particular meaning.

[0214] Any systems, methods, software, and platforms described herein are modular. Therefore, terms such as “first” and “second” do not necessarily imply priority, order of importance, or order of actions.

[0215] The term "about" when referring to a number or range means that the number or range mentioned is an approximation within the experimental variability (or statistical experimental error), and that the number or range may vary, for example, from 1% to 15%, from the stated number or range. In examples, the term "about" refers to ±10% of the stated number or value.

[0216] The terms “increased,” “increasing,” or “increase” are used herein in general to mean an increase of a statistically significant amount. In some aspects, the terms “increased” or “increase” mean an increase of at least 10% compared to a baseline level, for example, an increase of at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or up to 100%, or any increase between 10% and 100%, compared to a baseline level, standard, or control. Other examples of “increase” include an increase of at least 2 times, at least 5 times, at least 10 times, at least 20 times, at least 50 times, at least 100 times, at least 1000 times, or more compared to a baseline level.

[0217] The terms “decreased,” “decreasing,” or “decrease” are used herein to generally mean a statistically significant reduction. In some embodiments, “decreased” or “decrease” means a reduction of at least 10% compared to a baseline level, for example, at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90%, or up to 100% (absence or undetectable level compared to a baseline level), or any reduction between 10% and 100%. In the context of markers or symptoms, these terms mean a statistically significant reduction of such a level. The reduction may be, for example, at least 10%, at least 20%, at least 30%, at least 40%, or more, and is preferably reduced to a level that is considered to be within the normal range for an individual without the given disease.

[0218] While preferred 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 only as examples. The present invention is not intended to be limited by any specific examples provided herein. Although the present invention is described in relation to the foregoing specification, the descriptions and examples of embodiments herein are not intended to be constrained. Those skilled in the art will be able to conceive of many modifications, changes, and substitutions without departing from the present invention. Furthermore, it will be understood that all aspects of the present invention are not limited to any specific descriptions, configurations, or relative proportions described herein, depending on various conditions and variables. It should be understood that various alternatives to the embodiments of the present invention described herein may be used in carrying out the present invention. Therefore, the present invention is intended to extend to any such alternatives, modifications, variations, or equivalents. The following claims define the scope of the present invention, and it is intended that methods and structures within the scope of these claims and their equivalents are encompassed thereby.

[0219] Numbered Embodiments 1. A composition comprising one or more microorganisms, wherein the one or more microorganisms are located in the space between the membrane and cell layer of a plant or part thereof, and are selected to produce or promote the formation of one or more bicarbonates, carbonates, or one or more minerals, or are derived from the one or more microorganisms. 2. The composition according to any one of Embodiment 1, wherein the plant or part thereof is a plant root, plant stem, plant leaf, plant seed, plant fruit, plant tuber, or plant root nodule. 3. The composition according to any one of Embodiments 1 to 2, wherein the plant or part thereof includes a commercial plant or a part thereof. 4. The composition according to any one of Embodiments 1 to 3, wherein the commercial plant or a part thereof is corn, wheat, rice, sorghum, barley, rye, sugarcane, millet, oats, soybeans, cotton, alfalfa, beans, quinoa, lentils, peanuts, sunflowers, canola, cassava, palm oil, potatoes, sugar beets, cocoa, coffee, lettuce, tomatoes, peas, cabbage, fruit trees, nut trees, forest trees, grasslands, or turfgrass. 5. The composition according to any one of Embodiments 1 to 4, wherein a portion thereof is a plant seed, and one or more microorganisms related to the plant seed are arranged in the space between the seed coat and the seed embryo of the plant seed. 6. The composition according to any one of Embodiments 1 to 5, wherein the one or more microorganisms related to the plant or a part thereof are arranged as a coating on the plant or a part thereof. 7. The composition according to any one of Embodiments 1 to 6, wherein the one or more microorganisms relating to the plant or a part thereof are applied to the plant seeds through an irrigation system. 8. The composition according to any one of Embodiments 1 to 7, wherein the irrigation system includes in-furrow treatment technology. 9. The composition according to any one of embodiments 1 to 8, wherein the irrigation system includes a spraying method. 10. The composition according to any one of Embodiments 1 to 9, wherein the bicarbonate sequesters carbon. 11. The composition according to any one of Embodiments 1 to 10, wherein the carbon is gaseous carbon. 12. The composition according to any one of Embodiments 1 to 11, wherein the gaseous carbon is carbon dioxide. 13. The composition according to any one of Embodiments 1 to 12, wherein the carbonate sequesters carbon. 14. The composition according to any one of Embodiments 1 to 13, wherein the carbon is gaseous carbon. 15. The composition according to any one of Embodiments 1 to 14, wherein the gaseous carbon is carbon dioxide. 16. The composition according to any one of embodiments 1 to 15, wherein the one or more minerals sequester carbon. 17. The composition according to any one of Embodiments 1 to 16, wherein the carbon is gaseous carbon. 18. The composition according to any one of Embodiments 1 to 17, wherein the gaseous carbon is carbon dioxide. 19. The composition according to any one of Embodiments 1 to 18, wherein a portion thereof is a plant seed, and one or more microorganisms associated with the plant seed are arranged in the space between the seed pericarp and the seed allureon cell layer of the plant seed. 20. The composition according to any one of Embodiments 1 to 19, wherein the one or more microorganisms comprises one or more carbonic anhydrases. 21. The composition according to any one of Embodiments 1 to 20, wherein the one or more carbonic anhydrases include carbonic anhydrase alpha class. 22. The composition according to any one of Embodiments 1 to 21, wherein the one or more carbonic anhydrases include carbonic anhydrase beta class. 23. The composition according to any one of Embodiments 1 to 22, wherein the one or more carbonic anhydrases comprise carbonic anhydrase gamma class. 24. The composition according to any one of Embodiments 1 to 23, wherein the one or more carbonic anhydrases include carbonic anhydrase delta classes. 25. The composition according to any one of Embodiments 1 to 24, wherein the one or more carbonic anhydrases comprise carbonic anhydrase zeta classes. 26. The composition according to any one of Embodiments 1 to 25, wherein the one or more carbonic anhydrases comprise carbonic anhydrase etaclass. 27. The composition according to any one of Embodiments 1 to 26, wherein the one or more carbonic anhydrases include carbonic anhydrase iotaclass. 28. The composition according to any one of Embodiments 1 to 27, wherein the one or more microorganisms include bacteria, archaea, fungi, or viruses. 29. The composition according to any one of Embodiments 1 to 28, wherein the one or more microorganisms include the bacteria. 30. The composition according to any one of Embodiments 1 to 29, wherein the bacteria include endospore-forming bacteria. 31. The aforementioned bacteria include the genera Acetonema, Actinomyces, Alkalibacillus, Ammoniphilus, Amphibacillus, Anaerobacter, Anerospora, Anneurinibacillus, Anoxybacillus, Bacillus, Brevibacillus, Caldanerobacter, Caloramator, Caminicella, and Cerasibacillus. 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., Gerria (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., Moorela 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., Sporobacter 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., Thermovenablum A composition according to any one of Embodiments 1 to 30, comprising bacteria from the genera *Tuberibacillus sp.*, *Virgibacillus sp.*, *Vulcanobacillus sp.*, or a combination thereof. 32. The composition according to any one of Embodiments 1 to 31, wherein the bacteria include bacteria belonging to the phylum Firmicutes. 33. The composition according to any one of Embodiments 1 to 32, wherein the bacteria include rhizosphere bacteria. 34. The composition according to any one of Embodiments 1 to 33, wherein the rhizosphere bacteria include the genus Bacillus, the genus Paenibacillus, or both. 35. The aforementioned bacteria are B. amyloliquefaciens, B. laterosporus, B. licheniformis, B. macerans, B. cereus, B. circulans, B. firmus, B. subtilis, B. megaterium, B. coagulans, B. brevis, and B. sphericus. A composition according to any one of Embodiments 1 to 34, comprising B. phaericus, B. thuringiensis, B. mycoides, B. cucumis, B. endophyticus, B. pumilus, B. velezensis, B. mucilaginosus, B. tequilensis, B. methylotrophicus, or any combination thereof. 36. The composition according to any one of Embodiments 1 to 35, wherein the bacteria include Bacillus subtilis S3C23, Bacillus subtilis MP2, Bacillus subtilis RO2C15, Bacillus subtilis RO2C22, Bacillus megaterium 6, Bacillus megaterium S3C21, Bacillus megaterium RO2C12, Bacillus cucumis S3C14, Bacillus endophyticus 5, or any combination thereof. 37. The composition according to any one of Embodiments 1 to 36, wherein the bacterium comprises Bacillus subtilis S3C23. 38. The composition according to any one of Embodiments 1 to 37, wherein the bacterium comprises Bacillus subtilis MP2. 39. The composition according to any one of Embodiments 1 to 38, wherein the bacterium comprises Ensifer adhaerens S3C10. 40. The composition according to any one of Embodiments 1 to 39, wherein the bacteria include Paenibacillus polymyxa, Paenibacillus taohuashanense, Paenibacillus pocheonensis, Paenibacillus aceris, Paenibacillus catalpa, Paenibacillus rigui, Paenibacillus pabuli, Paenibacillus brasiliensis, or any combination thereof. 41. The composition according to any one of Embodiments 1 to 40, wherein the bacteria include Paenibacillus polymyxa RO3C16, Paenibacillus taohuashanense TY4D5, Paenibacillus pocheonensis S2C3, Paenibacillus aceris VF2D2, Paenibacillus catalpa TY2B5, Paenibacillus rigui TY2D5, Paenibacillus pabuli PG2A8, or any combination thereof. 42. The composition according to any one of Embodiments 1 to 41, wherein the bacteria include non-endospore-forming bacteria. 43. The composition according to any one of Embodiments 1 to 42, wherein the bacteria include bacteria belonging to the phylum Proteobacteria. 44. The aforementioned bacteria include the genera Klebsiella, Rhizobium, Bradyrhizobium, Ochrobactrum, Sinorhizobium, Xanthobacter, Methylobacterium, Actinomyces, Kosakonia, Azotobacter, Acetobacter, Herbaspirillum, Pseudomonas, Paraburkholderia, and Ralstonia. A composition according to any one of Embodiments 1 to 43, comprising Geobacter sp., Serratia sp., Pantoea sp., Ensifer sp., Enterobacter sp., or any combination thereof. 45. The composition according to any one of Embodiments 1 to 44, wherein the bacteria include bacteria belonging to the phylum Actinomycetes. 46. ​​The composition according to any one of Embodiments 1 to 45, wherein the bacteria include the genera Streptomyces sp., Coxiella sp., and Frankia sp. 47. The composition according to any one of Embodiments 1 to 46, wherein the bacteria include bacteria belonging to the phylum Cyanobacteria. 48. The composition according to any one of Embodiments 1 to 47, wherein the bacteria include the genus Cyanobacteria sp. 49. The composition according to any one of Embodiments 1 to 48, wherein the bacteria include bacteria belonging to the phylum Chloroflexus. 50. The composition according to any one of Embodiments 1 to 49, wherein the one or more microorganisms include one or more fungi related to the plant or a part thereof. 51. The composition according to any one of Embodiments 1 to 50, wherein the one or more fungi associated with the plant or a part thereof are placed in the space between the seed coat and the seed embryo of the plant or a part thereof. 52. The composition according to any one of Embodiments 1 to 51, wherein the one or more fungi associated with the plant or a part thereof are arranged as a coating on the plant or a part thereof. 53. The composition according to any one of Embodiments 1 to 52, wherein the one or more fungi associated with a plant or a part thereof are applied to the plant or a part thereof by an in-furrow treatment technique. 54. The composition according to any one of Embodiments 1 to 53, wherein the one or more fungi associated with the plant or a part thereof are applied to the plant or a part thereof by a spraying method. 55. The composition according to any one of Embodiments 1 to 54, wherein the one or more fungi associated with the plant or a part thereof are applied to the plant or a part thereof via the irrigation system. 56. The composition according to any one of Embodiments 1 to 55, wherein the one or more fungi associated with the plant or a part thereof are arranged in the space between the seed pericarp and the seed allureon cell layer of the plant or a part thereof. 57. The composition according to any one of Embodiments 1 to 56, wherein the one or more fungi include arbuscular mycorrhizal fungi. 58. A composition according to any one of Embodiments 1 to 57, wherein one or more fungi include ectomycorrhizal fungi. 59. A composition according to any one of Embodiments 1 to 58, wherein one or more fungi include fungi from the genus Trichoderma. 60. A composition according to any one of Embodiments 1 to 59, wherein one or more fungi include fungi from the genus Penicillium. 61. The composition according to any one of Embodiments 1 to 60, wherein the one or more minerals include calcite, aragonite, dolomite, limestone, or any combination thereof. 62. The composition according to any one of Embodiments 1 to 61, wherein the one or more minerals include CaCO3, MgCO3, CaMg(CO3)2, or any combination thereof. 63. The composition according to any one of Embodiments 1 to 62, wherein the promotion of the production of one or more minerals includes the production of ammonia and an increase in the pH of a growing medium in which a plant or a plant derived from the plant or a part thereof is grown. 64. The composition according to any one of Embodiments 1 to 63, wherein a portion thereof is a plant seed, and the one or more microorganisms are not naturally present in the space between the seed pericarp and the seed allureon cell layer of the plant or a portion thereof. 65. The composition according to any one of Embodiments 1 to 64, wherein the plant or part thereof is a monocotyledonous plant or a dicotyledonous plant. 66. A method for promoting mineralization, the method comprising a step of culturing a plant or a part thereof and one or more microorganisms associated with the plant or the part thereof, wherein the one or more microorganisms are selected to produce or promote the formation of bicarbonates, carbonates, or one or more minerals, or are derived from the above microorganisms. 67. The method according to any one of Embodiment 66, wherein the plant or part thereof is a commercial plant, plant root, plant stem, plant leaf, plant seed, plant fruit, plant tuber, or plant root nodule. 68. The method according to any one of embodiments 66 to 67, wherein the one or more microorganisms relating to the plant or a part thereof are located in the plant roots or rhizosphere of the plant or a part thereof. 69. The method according to any one of embodiments 66 to 68, wherein the one or more microorganisms associated with the plant or part thereof are placed in the plant roots or rhizosphere of the plant or part thereof by an irrigation system. 70. The method according to any one of embodiments 66 to 69, wherein the irrigation system includes in-furrow treatment technology. 71. The method according to any one of embodiments 66 to 70, wherein the irrigation system includes a spraying method. 72. The method according to any one of embodiments 66 to 71, wherein the plant or a part thereof is derived from a seedling that is integrated with the microorganisms via the irrigation system in order to stimulate the production of the one or more minerals by the plant or a part thereof. 73. The method according to any one of embodiments 66 to 72, wherein the bicarbonate sequesters carbon. 74. The method according to any one of embodiments 66 to 73, wherein the carbon is gaseous carbon. 75. The method according to any one of embodiments 66 to 74, wherein the gaseous carbon is carbon dioxide. 76. The method according to any one of embodiments 66 to 75, wherein the carbonate sequesters carbon. 77. The method according to any one of embodiments 66 to 76, wherein the carbon is gaseous carbon. 78. The method according to any one of embodiments 66 to 77, wherein the gaseous carbon is carbon dioxide. 79. The method according to any one of embodiments 66 to 78, wherein the one or more minerals sequester carbon. 80. The method according to any one of embodiments 66 to 79, wherein the carbon is gaseous carbon. 81. The method according to any one of embodiments 66 to 80, wherein the gaseous carbon is carbon dioxide. 82. The method according to any one of embodiments 66 to 81, wherein the one or more microorganisms include bacteria, archaea, fungi, or viruses. 83. The method according to any one of embodiments 66 to 82, wherein the one or more microorganisms include the bacteria. 84. The method according to any one of embodiments 66 to 83, wherein the bacteria include endospore-forming bacteria. 85. The method according to any one of embodiments 66 to 84, wherein the bacteria include rhizosphere bacteria. 86. The method according to any one of embodiments 66 to 85, wherein the rhizosphere bacteria include the genera Bacillus sp, Paenibacillus sp, or both. 87. The bacteria mentioned above are B. amyloliquefaciens, B. laterosporus, B. licheniformis, B. macerans, B. cereus, B. circulans, B. firmus, B. subtilis, B. sphaericus, B. megaterium, B. coagulans, and B. brevis. The method according to any one of embodiments 66 to 86, comprising (B. brevis), B. thuringiensis, B. mycoides, B. cucumis, B. endophyticus, B. pumilus, B. velezensis, B. mucilaginosus, B. tequilensis, B. methylotrophicus, or any combination thereof. 88. The method according to any one of embodiments 66 to 87, wherein the bacteria include Bacillus subtilis S3C23, Bacillus subtilis MP2, Bacillus subtilis RO2C15, Bacillus subtilis RO2C22, Bacillus megaterium 6, Bacillus megaterium S3C21, Bacillus megaterium RO2C12, Bacillus cucumis S3C14, Bacillus endophyticus 5, or any combination thereof. 89. The method according to any one of embodiments 66 to 88, wherein the bacterium comprises Bacillus subtilis S3C23. 90. The method according to any one of embodiments 66 to 89, wherein the bacterium comprises Bacillus subtilis MP2. 91. The method according to any one of embodiments 66 to 90, wherein the bacteria comprises Ensifer adhaerens S3C10. 92. The method according to any one of embodiments 66 to 91, wherein the one or more microorganisms include one or more fungi associated with the plant or a part thereof. 93. The method according to any one of embodiments 66 to 92, wherein the one or more fungi associated with the plant or part thereof are placed in the plant roots or rhizosphere of the plant or part thereof by the irrigation system. 94. The method according to any one of embodiments 66 to 93, wherein the one or more fungi include arbuscular mycorrhizal fungi. 95. The method according to any one of embodiments 66 to 94, wherein one or more fungi include ectomycorrhizal fungi. 96. The method according to any one of embodiments 66 to 95, wherein one or more fungi include fungi from the genus Trichoderma. 97. The method according to any one of embodiments 66 to 96, wherein one or more fungi include fungi from the genus Penicillium. 98. The method according to any one of embodiments 66 to 97, wherein the one or more microorganisms result in the formation of one or more carbonic anhydrases. 99. The method according to any one of embodiments 66 to 98, wherein the one or more microorganisms result in the formation of one or more carbonic anhydrases belonging to the alpha class. 100. The method according to any one of embodiments 66 to 99, wherein the one or more microorganisms result in the formation of one or more carbonic anhydrases belonging to the beta class. 101. The method according to any one of embodiments 66 to 100, wherein the one or more microorganisms result in the formation of one or more carbonic anhydrases belonging to the gamma class. 102. The method according to any one of embodiments 66 to 101, wherein the one or more microorganisms result in the formation of one or more carbonic anhydrases belonging to the delta class. 103. The method according to any one of embodiments 66 to 102, wherein the one or more microorganisms result in the formation of one or more carbonic anhydrases belonging to the zeta class. 104. The method according to any one of embodiments 66 to 103, wherein the one or more microorganisms result in the formation of one or more carbonic anhydrases belonging to the etaclass. 105. The method according to any one of embodiments 66 to 104, wherein the one or more microorganisms result in the formation of one or more carbonic anhydrases belonging to the iotaclass. 106. The method according to any one of embodiments 66 to 105, wherein the one or more minerals include calcite, aragonite, dolomite, limestone, or any combination thereof. 107. The method according to any one of embodiments 66 to 106, wherein the promotion of the production of one or more minerals includes the production of ammonia and an increase in the pH of the medium in which the plant or a part thereof is growing. 108. The method according to any one of embodiments 66 to 107, wherein the one or more microorganisms are not naturally present in the one or more roots. 109. The method according to any one of embodiments 66 to 108, wherein the plant or part thereof is a monocotyledonous plant or a dicotyledonous plant. 110. The method according to any one of embodiments 66 to 109, wherein the plant or part thereof includes a commercial plant or part thereof. 111. The method according to any one of embodiments 66 to 110, wherein the commercial plant or a portion thereof consists essentially of the group comprising corn, wheat, rice, sorghum, barley, rye, sugarcane, millet, oats, soybeans, cotton, alfalfa, beans, quinoa, lentils, peanuts, sunflowers, canola, cassava, palm oil, potatoes, sugar beets, cocoa, coffee, lettuce, tomatoes, peas, cabbage, fruit trees, nut trees, forest trees, grasslands, or turfgrass. 112. A composition comprising one or more microorganisms, wherein the one or more microorganisms are selected to produce or promote the formation of bicarbonates, carbonates, or one or more minerals, or are derived from such microorganisms. 113. The composition according to any one of Embodiments 112, wherein the plant or part thereof is a plant root, plant stem, plant leaf, plant seed, plant fruit, plant tuber, or plant root nodule. 114. The composition according to any one of embodiments 112 to 113, wherein the plant or part thereof includes a commercial plant or a part thereof. 115. The composition according to any one of Embodiments 112 to 114, wherein the commercial plant or a portion thereof is corn, wheat, rice, sorghum, barley, rye, sugarcane, millet, oats, soybeans, cotton, alfalfa, beans, quinoa, lentils, peanuts, sunflowers, canola, cassava, palm oil, potatoes, sugar beets, cocoa, coffee, lettuce, tomatoes, peas, cabbage, fruit trees, nut trees, forest trees, grasslands, or turfgrass. 116. The composition according to any one of embodiments 112 to 115, wherein the bicarbonate sequesters carbon. 117. The composition according to any one of embodiments 112 to 116, wherein the carbon is gaseous carbon. 118. The composition according to any one of embodiments 112 to 117, wherein the gaseous carbon is carbon dioxide. 119. The composition according to any one of embodiments 112 to 118, wherein the carbonate sequesters carbon. 120. The composition according to any one of embodiments 112 to 119, wherein the carbon is gaseous carbon. 121. The composition according to any one of embodiments 112 to 120, wherein the gaseous carbon is carbon dioxide. 122. The composition according to any one of embodiments 112 to 121, wherein the one or more minerals sequester carbon. 123. The composition according to any one of embodiments 112 to 122, wherein the carbon is gaseous carbon. 124. The composition according to any one of embodiments 112 to 123, wherein the gaseous carbon is carbon dioxide. 125. The composition according to any one of embodiments 112 to 124, wherein the one or more microorganisms include bacteria, archaea, fungi, or viruses. 126. The composition according to any one of embodiments 112 to 125, wherein the one or more microorganisms include the bacteria. 127. The composition according to any one of Embodiments 112 to 126, wherein the bacteria include endospore-forming bacteria. 128. The composition according to any one of embodiments 112 to 127, wherein the bacteria include rhizosphere bacteria. 129. The composition according to any one of Embodiments 112 to 128, wherein the rhizosphere bacteria include the genus Bacillus, the genus Paenibacillus, or both. 130. The aforementioned bacteria include B. amyloliquefaciens, B. laterosporus, B. licheniformis, B. macerans, B. cereus, B. circulans, B. firmus, B. subtilis, B. sphaericus, B. megaterium, B. coagulans, B. brevis ( A composition according to any one of Embodiments 112 to 129, comprising B. brevis, B. thuringiensis, B. mycoides, B. cucumis, B. endophyticus, B. pumilus, B. velezensis, B. mucilaginosus, B. tequilensis, B. methylotrophicus, or any combination thereof. 131. The composition according to any one of Embodiments 112 to 130, wherein the bacteria include Bacillus subtilis S3C23, Bacillus subtilis MP2, Bacillus subtilis RO2C15, Bacillus subtilis RO2C22, Bacillus megaterium 6, Bacillus megaterium S3C21, Bacillus megaterium RO2C12, Bacillus cucumis S3C14, Bacillus endophyticus 5, or any combination thereof. 132. The composition according to any one of Embodiments 112 to 131, wherein the bacterium comprises Bacillus subtilis S3C23. 133. The composition according to any one of Embodiments 112 to 132, wherein the bacterium comprises Bacillus subtilis MP2. 134. The composition according to any one of embodiments 112 to 133, wherein the bacterium comprises Ensifer adhaerens S3C10. 135. The composition according to any one of Embodiments 112 to 134, wherein the one or more microorganisms include one or more fungi related to the plant or a part thereof. 136. The composition according to any one of embodiments 112 to 135, wherein the one or more fungi associated with the plant or part thereof are placed in the plant roots or rhizosphere of the plant or part thereof by the irrigation system. 137. The composition according to any one of embodiments 112 to 136, wherein the one or more fungi include arbuscular mycorrhizal fungi. 138. A composition according to any one of Embodiments 112 to 137, wherein one or more fungi include ectomycorrhizal fungi. 139. A composition according to any one of embodiments 112 to 138, wherein one or more fungi include fungi from the genus Trichoderma. 140. A composition according to any one of Embodiments 112 to 139, wherein one or more fungi include fungi from the genus Penicillium. 141. The composition according to any one of embodiments 112 to 140, wherein the one or more minerals include calcite, aragonite, dolomite, limestone, or any combination thereof. 142. The composition according to any one of Embodiments 112 to 141, wherein the promotion of the production of one or more minerals includes the production of ammonia and an increase in the pH of the medium in which the plant or a part thereof is growing. 143. The composition according to any one of Embodiments 112 to 142, wherein the one or more microorganisms comprises one or more carbonic anhydrases. 144. The composition according to any one of embodiments 112 to 143, wherein the one or more carbonic anhydrases include carbonic anhydrase α class. 145. The composition according to any one of embodiments 112 to 144, wherein the one or more carbonic anhydrases comprise carbonic anhydrase beta class. 146. The composition according to any one of embodiments 112 to 145, wherein the one or more carbonic anhydrases comprise carbonic anhydrase gamma class. 147. The composition according to any one of embodiments 112 to 146, wherein the one or more carbonic anhydrases comprise carbonic anhydrase delta classes. 148. The composition according to any one of embodiments 112 to 147, wherein the one or more carbonic anhydrases comprise carbonic anhydrase zeta classes. 149. The composition according to any one of embodiments 112 to 148, wherein the one or more carbonic anhydrases comprise carbonic anhydrase etaclass. 150. The composition according to any one of embodiments 112 to 149, wherein the one or more carbonic anhydrases include carbonic anhydrase iotaclass. [Examples]

[0220] The methods and compositions disclosed herein are designed, in connection with plants or plant seeds, to microbiologically sequestrate carbon by the formation of bicarbonates and one or more minerals.

[0221] Definition of Microbial Formulation: To achieve early conditioning, this disclosure uses seed treatment compositions comprising a synthetic consortium or a single isolated bacterial strain or endospores in a suspension medium. Typically, plant cultivation compositions and methods include diverse and environmentally adapted plant-associated bacteria belonging to a wide variety of bacterial genera distributed in different taxa within the Proteobacteria phylum (alpha, beta, gamma, and delta proteobacteria classes), as well as the Firmicutes, Bacteroidetes, and Actinomycetes phyla. The inventors have isolated and characterized rhizosphere bacteria belonging to various genera, typically plant-associated microorganisms, within these large taxa, which can be applied to seeds using the methods of this disclosure to effectively colonize roots and ultimately sequester CO2. The compositions include one, two, three, or more different bacterial strains that have been cultured separately and mixed for Microprime® seed treatment.

[0222] Example 1. Seed treatment to increase the load of non-endospore-forming bacteria, endospore-forming bacteria, and / or bacterial endospores: Microprime® technology. While CO2-fixing microorganisms exist in the soil, CO2 sequestration is ineffective for the following reasons: Many microorganisms cannot utilize CO2 as an energy source. Many microorganisms are not located near the roots where CO2 is released, and the...

Claims

1. A composition for carbon sequestration comprising a plant or a part thereof, or a plant seed, and one or more bacteria applied to the plant or a part thereof, or the plant seed, The one or more bacteria are either one or more bacteria selected to produce one or more compounds or to promote the production of one or more compounds, or derived from one or more bacteria selected to produce one or more compounds or to promote the production of one or more compounds, and the bacteria include bacteria selected from the genera Bacillus and Encifer, or any combination thereof. The one or more compounds described above contain bicarbonate ions, carbonate ions, or a combination thereof, thereby sequestering carbon. composition.

2. The composition according to claim 1, wherein the one or more bacteria applied to the plant seed are arranged in the space between the seed coat and the seed embryo of the plant seed.

3. The composition according to claim 1, wherein the one or more bacteria applied to the plant seed are arranged as a coating on the plant seed.

4. The composition according to any one of claims 1 to 3, wherein the bicarbonate ion or the carbonate ion sequesters carbon.

5. The composition according to any one of claims 1 to 4, wherein the one or more bacteria applied to the plant seed are arranged in the space between the seed pericarp and the seed allureon cell layer of the plant seed.

6. The composition according to any one of claims 1 to 5, wherein the plant seeds are monocotyledonous plant seeds or dicotyledonous plant seeds.

7. The composition according to claim 1, wherein the plant seeds are corn seeds, wheat seeds, rice seeds, sorghum seeds, barley seeds, rye seeds, sugarcane seeds, millet seeds, oat seeds, soybean seeds, cotton seeds, alfalfa seeds, legume seeds, quinoa seeds, lentil seeds, peanut seeds, sunflower seeds, canola seeds, cassava seeds, oil palm seeds, potato seeds, sugar beet seeds, cocoa seeds, coffee seeds, lettuce seeds, tomato seeds, pea seeds, or cabbage seeds.

8. The composition according to claim 1, comprising the plant or a part thereof, and one or more bacteria applied to the plant or a part thereof.

9. The composition according to claim 8, wherein the plant or part thereof is a plant root, plant stem, plant leaf, plant fruit, plant tuber, or plant root nodule.

10. The composition according to any one of claims 1 to 9, wherein the bacteria include endospore-forming bacteria.

11. The composition according to claim 10, wherein the bacteria include the genus Bacillus (Bacillus sp.).

12. The aforementioned bacteria include B. amyloliquefaciens, B. laterosporus, B. licheniformis, B. macerans, B. cereus, B. circulars, B. firmus, B. subtilis, B. megatherium, B. coagulans, B. brevis, B. sphaericus, B. thuringiensis, and B. A composition according to any one of claims 1 to 8, comprising B. mycoides, B. cucumis, B. endophyticus, B. pumilus, B. velezensis, B. mucilaginosus, B. tequilensis, B. methylotropicus, or any combination thereof.

13. The composition according to any one of claims 1 to 9, wherein the bacterium comprises Bacillus subtilis.

14. The composition according to any one of claims 1 to 8, wherein the bacteria include non-endospore-forming bacteria.

15. The composition according to any one of claims 1 to 13, wherein the one or more compounds include calcite, aragonite, dolomite, limestone, or any combination thereof.

16. The one or more compounds mentioned above are CaCO ３ , MgCO ３ CaMg(CO ３ ) ２ The composition according to any one of claims 1 to 15, comprising, or any combination thereof.

17. The composition according to any one of claims 1 to 16, wherein the promotion of the generation of one or more compounds includes the generation of ammonia and an increase in the pH of the culture medium in which the plant or a part thereof is growing.

18. The composition according to any one of claims 1 to 17, wherein one or more of the compounds include minerals.

19. The composition according to claim 18, wherein the mineral comprises a carbonate mineral.

20. The composition according to claim 19, wherein the carbonate mineral contains carbonate ions.

21. The composition according to claim 20, wherein the carbonate mineral sequesters carbon.

22. A method for sequestering carbon, wherein the method is (a) A step of culturing a plant or a part thereof, or a plant seed, and one or more bacteria applied to the plant or a part thereof, or the plant seed, Here, the one or more bacteria are one or more bacteria selected to produce one or more compounds or to promote the production of one or more compounds, or are derived from one or more bacteria selected to produce one or more compounds or to promote the production of one or more compounds, and the bacteria include bacteria selected from the genera Bacillus and Encifer, or any combination thereof. The one or more compounds described above contain carbonate ions, bicarbonate ions, or a combination thereof, thereby sequestering carbon. method.

23. The method according to claim 22, wherein the plant or part thereof is a plant root, plant stem, plant leaf, plant fruit, plant tuber, or plant root nodule.

24. The method according to claim 22, wherein the one or more bacteria applied to the plant seed are arranged in the space between the seed coat and the seed embryo of the plant seed.

25. The method according to claim 22, wherein the one or more bacteria applied to the plant seed are arranged as a coating on the plant seed.

26. The method according to claim 22, wherein the plant or part thereof is selected from the group essentially consisting of corn, wheat, rice, sorghum, barley, rye, sugarcane, millet, oats, soybeans, cotton, alfalfa, beans, quinoa, lentils, peanuts, sunflowers, canola, cassava, oil palm, potatoes, sugar beets, cocoa, coffee, lettuce, tomatoes, peas, cabbage, fruit trees, nut trees, forest trees, grasslands, or turfgrass.

27. The method according to any one of claims 22 to 26, wherein the one or more bacteria applied to the plant are located in the roots or rhizosphere of the plant or a part thereof.

28. The method according to any one of claims 22 to 27, wherein the one or more bacteria applied to the plant are placed in the plant roots or rhizosphere of the plant or a part thereof by an irrigation system.

29. The method according to claim 28, wherein the irrigation system includes furrow treatment technology.

30. The method according to claim 28, wherein the irrigation system includes a spraying method.

31. The method according to any one of claims 22 to 30, wherein the plant or part thereof is derived from a seedling that is integrated with the bacteria via an irrigation system in order to stimulate the production of one or more compounds by the plant or part thereof.

32. The method according to any one of claims 22 to 31, wherein the bacteria include endospore-forming bacteria.

33. The method according to any one of claims 22 to 31, wherein the bacteria include rhizosphere bacteria.

34. The method according to claim 33, wherein the rhizosphere bacteria include the genus Bacillus (Bacillus sp.).

35. The aforementioned bacteria include B. amyloliquefaciens, B. laterosporus, B. licheniformis, B. macerans, B. cereus, B. circulars, B. firmus, B. subtilis, B. megatherium, B. coagulans, B. brevis, B. sphaericus, B. thuringiensis, and B. The method according to any one of claims 22 to 31, comprising B. mycoides, B. cucumis, B. endophyticus, B. pumilus, B. velezensis, B. mucilaginosus, B. tequilensis, B. methylotropicus, or any combination thereof.

36. The method according to any one of claims 22 to 31, wherein the bacterium comprises Bacillus subtilis.

37. The method according to any one of claims 22 to 36, wherein the plant or a part thereof grows from the plant seed.

38. The method according to any one of claims 22 to 37, wherein the one or more compounds include minerals.

39. The method according to claim 38, wherein the mineral includes a carbonate mineral.

40. The method according to claim 39, wherein the carbonate mineral contains carbonate ions.

41. The method according to claim 40, wherein the carbonate mineral sequesters carbon.

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