Compositions, methods, and systems for promoting phosphate solubilization in plants

Abstract

The present disclosure provides methods and systems for production of biostimulants that promote phosphate solubilization in plants. Embodiments described include methods of making a biostimulant composition in a bioreactor system that includes two or more containers arranged in series. The bioreactor system may include an established population of a phosphate-solubilizing microbial strain. The method may include operating the bioreactor system by transferring into the system an aqueous feedstock that comprises a microbial consortium, transferring working fluid between containers of the system, and collecting a product. The method may further include maintaining a concentration of a phosphate-solubilizing microbial strain in the system.

Description

COMPOSITIONS, METHODS, AND SYSTEMS FOR PROMOTING PHOSPHATESOLUBILIZATION IN PLANTSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application Serial No. 63 / 613,560, filed on December 21, 2023, the entire content of which is incorporated herein by reference in its entirety.BACKGROUND

[0002] The disclosure is generally related to biostimulant compositions and methods of using such biostimulant compositions to promote plant growth and enhance phosphate solubilization.

[0003] Promoting efficient production of food crops and other crops is an important goal for environmental and economic reasons. Plant growth promoting products sourced from organic materials can help to enhance crop growth, improve the efficacy of agricultural products, such as fertilizers, and reduce the environmental impacts of synthetic fertilizers and climate change. There exists a need for plant growth promoting biostimulant compositions that use abundant and available organic feedstocks.SUMMARY

[0004] Embodiments disclosed herein include a method of making a biostimulant composition, the method comprising: (a) providing a bioreactor system comprising two or more containers arranged in a series, each of the two or more containers comprising a volume of a working fluid, wherein a first container comprises a first working fluid that includes a first microbial consortium and an established population of a phosphate solubilizing bacterial strain; (b) operating the bioreactor system for a duration of time by: (i) transferring into the first container an aqueous feedstock comprising a second microbial consortium; (ii) transferring a portion of the working fluid out of each of the two or more containers into either a subsequent container of the bioreactor system or a product outflow stream; (iii) maintaining a concentration of the phosphate solubilizing bacterial strain throughout the duration of time in at least the first container at at least 80% of a concentration of the phosphate solubilizing bacterial strain at the beginning of the duration of time; and (iv) collecting at least a portion of the product outflow stream as the biostimulant composition; wherein the duration of time is at least 5 days; and wherein the phosphate solubilizing bacterial strain is not present in the aqueous feedstock or any other input into the bioreactor system during the duration of time at a concentration that ishigher than 1% of the concentration of the phosphate solubilizing microbe in the first container. In some embodiments, the duration of time is at least 7, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 200, 300, 400, or 500 days.

[0005] In some embodiments, the method further comprises adding a source of insoluble phosphate to the first container. In some embodiments, the source of insoluble phosphate comprises rock phosphate particles. In some embodiments, the aqueous feedstock and any other input into the bioreactor system does not comprise soluble phosphate or does not comprise soluble phosphate at a concentration higher than 10 mg / L.

[0006] In some embodiments, the first microbial consortium comprises other phosphate solubilizing microbes that are not the phosphate solubilizing bacterial strain, and wherein the method further comprises maintaining a concentration of soluble phosphate and a concentration of a source of insoluble phosphate in at least the first container that stimulates phosphate solubilizing activity of the phosphate solubilizing microbial strain and / or the other phosphate solubilizing microbes.

[0007] In some embodiments, the phosphate solubilizing bacterial strain is of the genus Bacillus. In some embodiments, the phosphate solubilizing bacterial strain is of the species Bacillus amyloliquefaciens o Bacillus licheniformis. In some embodiments, the phosphate solubilizing bacterial strain is one of the following: (a) a Bacillus amyloliquefaciens strain having one or more of the following: (i) a 16S rRNA gene sequence at least 95% identical to SEQ ID NO: 1; (ii) a gyrB gene sequence at least 95% identical to SEQ ID NO: 3; and (iii) a rpoB gene sequence at least 95% identical to SEQ ID NO: 5; or (b) a Bacillus licheniformis strain having one or more of the following: (i) a 16S rRNA gene sequence at least 95% identical to SEQ ID NO: 2; (ii) a gyrB gene sequence at least 95% identical to SEQ ID NO: 4; and (iii) a rpoB gene sequence at least 95% identical to SEQ ID NO: 6. In some embodiments, the phosphate solubilizing bacterial strain is the Bacillus amyloliquefaciens strain deposited under ATCC Accession No. PTA-127657 or the Bacillus licheniformis strain deposited under ATCC Accession No. PTA-127656.

[0008] In some embodiments, the phosphate solubilizing bacterial strain is not present in the aqueous feedstock or any other input into the bioreactor system during the duration of time at a concentration of greater than 100 CFU / ml. In some embodiments, the phosphate solubilizing bacterial strain is not present in the aqueous feedstock or any other input into the bioreactor system during the duration of time. In some embodiments, the maintaining of step (b)(iii) comprises maintaining the concentration of the phosphate solubilizing bacterial strain at at least IxlO3CFU / ml.

[0009] In some embodiments, before step (b), the first container further comprises an established population of other phosphate solubilizing microbes that are not the phosphate solubilizing bacterial strain, and step (b)(iii) further comprises maintaining a concentration of the other phosphate solubilizing microbes in at least the first container throughout the duration of time at at least IxlO3CFU / ml or at at least 80% of a concentration of the other phosphate solubilizing microbes at the beginning of the duration of time, wherein the other phosphate solubilizing microbes are not added to the bioreactor system during the duration of time at a concentration that is higher than 1% of the concentration of the other phosphate solubilizing microbes in the first container. In some embodiments, the other phosphate solubilizing microbes are not present in the aqueous feedstock or any other input into the bioreactor system at a concentration of greater than 104CFU / ml. In some embodiments, the population of the other phosphate solubilizing microbes in the first container is at least IxlO3CFU / ml at the beginning of the duration of time.

[0010] In some embodiments, the method comprises, before step (a), adding an inoculum of the phosphate solubilizing bacterial strain to the bioreactor system, wherein the inoculum of the phosphate solubilizing bacterial strain produces an initial population of the phosphate solubilizing bacterial strain of at least 0.5xl04CFU / ml in at least one container.

[0011] In some embodiments, before adding the inoculum of the phosphate solubilizing bacterial strain, the concentration of the phosphate solubilizing bacterial strain is less than IxlO2CFU / ml.

[0012] In some embodiments, the aqueous feedstock further comprises an organic material at least partially digestible by microbes present in at least one of the containers. In some embodiments, before the transferring of step (b)(i), the organic material had been partially digested by microbes endogenous to the organic material. In some embodiments, the method further comprises digesting the organic material in two or more serially connected containers before the transferring of step (b)(i). In some embodiments, the organic material comprises manure and / or material produced by microbial digestion of manure.

[0013] In some embodiments, the aqueous feedstock further comprises an inorganic material. In some embodiments, the rock phosphate particles had been added to a container upstream of the first container.

[0014] In some embodiments, the container upstream of the first container further comprises a hydraulic source. In some embodiments, the hydraulic source comprises a product of anaerobic digestion of manure in a serialized digestion system.

[0015] In some embodiments, the second microbial consortium comprises at least IxlO5CFU / ml. In some embodiments, the second microbial consortium comprises microbes derived from manure. In some embodiments, the operating of step (b) further comprises producing microbial metabolites that directly or indirectly promote phosphate solubilization in a plant growth medium.

[0016] In some embodiments, the transferring of step (b)(i), the transferring of step (b)(ii), and the collecting of step (b)(iv) are performed continuously throughout the duration of time.

[0017] In some embodiments, the transferring of step (b)(i), the transferring of step (b)(ii), and the collecting of step (b)(iv) are performed periodically throughout the duration of time.

[0018] In some embodiments, the method further comprises adding one or more carbon sources to at least one container of the bioreactor system. In some embodiments, the one or more carbon sources are included in the aqueous feedstock. In some embodiments, the one or more carbon sources comprise polysaccharides.

[0019] In some embodiments, the method further comprises adding one or more nitrogen sources to at least one container of the bioreactor system. In some embodiments, the one or more nitrogen sources comprise yeast.

[0020] In some embodiments, the bioreactor system comprises a clarifier container comprising a clarifier working fluid. In some embodiments, the method further comprises separating a supernatant portion of the clarifier working fluid from a floc portion of the clarifier working fluid within the clarifier container. In some embodiments, the separating comprises gravity separation. In some embodiments, the method further comprises folding the floc portion of the clarifier working fluid. In some embodiments, the folding further comprises releasing a population of the phosphate solubilizing bacterial strain into the supernatant portion without introducing floc solids into the supernatant portion. In some embodiments, the folding is performed by folding wipers in a bottom portion of the clarifier container. In some embodiments, the operating further comprises transferring the floc portion from the clarifier container to an earlier container in the bioreactor system. In some embodiments, the product outflow stream comprises the supernatant portion of the clarifier working fluid. In some embodiments, the method further comprises producing at least IxlO4CFU / ml of the phosphate solubilizing bacterial strain in the product outflow stream.

[0021] In some embodiments, the bioreactor system comprises the first container comprising a volume of a first working fluid, a second container comprising a volume of a second working fluid, and a third container comprising a volume of a third working fluid. In some embodiments, the first container comprises an outlet port fluidly connected to an inlet port of the secondcontainer and the second container comprises an outlet port fluidly connected to an input port of the third container. In some embodiments, the third container comprises an outlet port fluidly connected to a clarifier container. In some embodiments, the method further comprises maintaining the volume of each of the first working fluid, the second working fluid, and the third working fluid constant throughout the duration of time. In some embodiments, step (b) comprises operating the bioreactor system in a hydraulically balanced manner.

[0022] In some embodiments, the transferring of step (b)(i), the transferring of step (b)(ii), and the collecting of step (b)(iv) are driven by gravity. In some embodiments, the operating comprises maintaining a flow rate that results in a hydraulic retention time of the bioreactor system of at least 5 days. In some embodiments, the operating comprises maintaining the product outflow stream at a flow rate of at least 100 gallons per day. In some embodiments, the volume of working fluid in each of the two or more containers is at least 100 gallons.

[0023] In some embodiments, at least one of the two or more containers is a fluidized bed reactor. In some embodiments, at least one of the two or more containers is a packed bed reactor. In some embodiments, at least one of the two or more containers under aerobic conditions.

[0024] In some embodiments, the bioreactor system is operated continuously for at least 90 days.

[0025] In some embodiments, the method further comprises adding yeast into one of the two or more containers. In some embodiments, the yeast is Saccharomyces cerevisiae.

[0026] In some embodiments, the second microbial consortium comprises microbes endogenous to the organic material.

[0027] In some embodiments, at least one of the first working fluid, the second working fluid, or the third working fluid comprises a pH buffering system. In some embodiments, the method further comprises maintaining the pH of at least one of the first working fluid, the second working fluid, or the third working fluid between 6 and 8 throughout the duration of time.

[0028] In some embodiments, the aqueous feedstock does not include the phosphate solubilizing bacterial strain at a concentration higher than 10 CFU / ml. In some embodiments, the phosphate solubilizing bacterial strain is not added to the bioreactor system during the duration of time at a concentration that is higher than 10 CFU / ml.

[0029] In some embodiments, the bioreactor system comprises at least one container placed in the series before the first container.

[0030] In some embodiments, the method further comprises producing a population of sporulated bacteria in the product outflow stream. In some embodiments, the method furthercomprises producing a population of the phosphate solubilizing bacterial strain in the product outflow stream that is sporulated. In some embodiments, the population of the phosphate solubilizing bacterial strain that is sporulated comprises at least IxlO3CFU / ml.

[0031] In some embodiments, the method further comprises adding an additional population of the phosphate solubilizing bacterial strain to the biostimulant product.

[0032] In some embodiments, the method further comprises making at least a portion of the aqueous feedstock by a method comprising: (c) at least partially digesting manure in a series of two or more fluidly connected manure digestion containers, thereby generating at least a portion of the aqueous feedstock. In some embodiments, the series of two or more fluidly connected manure digestion containers comprise packed bed reactors. In some embodiments, the method further comprises maintaining the two or more fluidly connected manure digestion containers under anaerobic conditions. In some embodiments, step (c) comprises partially digesting the manure in only two manure digestion containers. In some embodiments, the method further comprises adding yeast to at least one of the two or manure digestion containers. In some embodiments, step (c) comprises digesting the manure by microbes endogenous to the manure. In some embodiments, step (c) comprises maintaining hydraulically balanced flow between the two or more manure digestion containers.

[0033] Also disclosed herein is a bioreactor system comprising: (a) a stream of an aqueous feedstock in fluid communication with a first container comprising a volume of a first working fluid, wherein the aqueous feedstock comprises a first microbial consortium, wherein the first working fluid comprises an established population of a phosphate solubilizing bacterial strain and a second microbial consortium, wherein a concentration of the phosphate solubilizing bacterial strain in the first working fluid is at least 100 times higher than a concentration of the phosphate solubilizing bacterial strain in the aqueous feedstock stream and in any other input into the bioreactor system; (b) one or more additional containers arranged in a series that includes the first container, wherein each of the one or more additional containers comprises a volume of a working fluid and is in fluid communication with at least one other container in the series, and wherein at least one of the one or more additional containers comprises a product outflow stream port; and (c) a product outflow stream in fluid communication with the product outflow stream port.

[0034] In some embodiments, at least the first working fluid comprises a concentration of soluble phosphate that is lower than 10 mg / L and a concentration of a source of insoluble phosphate that is at least 0.5 g / L. In some embodiments, the source of insoluble phosphate comprises 20 to 30% of insoluble phosphate by weight. In some embodiments, the source ofinsoluble phosphate comprises rock phosphate particles. In some embodiments, the rock phosphate particles are present in a container that is upstream of the first container. In some embodiments, the aqueous feedstock and any other input into the bioreactor system does not comprise soluble phosphate or does not comprise soluble phosphate at a concentration higher than 10 mg / L.

[0035] In some embodiments, the phosphate solubilizing bacterial strain is of the genus Bacillus. In some embodiments, the phosphate solubilizing bacterial strain is of the species Bacillus amyloliquefaciens o Bacillus licheniformis. In some embodiments, the phosphate solubilizing bacterial strain is one of the following: (a) a Bacillus amyloliquefaciens strain having one or more of the following: (i) a 16S rRNA gene sequence at least 95% identical to SEQ ID NO: 1; (ii) a gyrB gene sequence at least 95% identical to SEQ ID NO: 3; and (iii) a rpoB gene sequence at least 95% identical to SEQ ID NO: 5; or (b) a Bacillus licheniformis strain having one or more of the following: (i) a 16S rRNA gene sequence at least 95% identical to SEQ ID NO: 2; (ii) a gyrB gene sequence at least 95% identical to SEQ ID NO: 4; and (iii) a rpoB gene sequence at least 95% identical to SEQ ID NO: 6. In some embodiments, the phosphate solubilizing bacterial strain is the Bacillus amyloliquefaciens strain deposited under ATCC Accession No. PTA-127657 or the Bacillus licheniformis strain deposited under ATCC Accession No. PTA-127656.

[0036] In some embodiments, the bioreactor system is a continuous flow bioreactor system and the stream of the aqueous feedstock is a continuous stream. In some embodiments, each of the volume of the working fluids is constant.

[0037] In some embodiments, each of the first container and the one or more additional containers comprises a concentration of the phosphate solubilizing bacterial strain that remains at least IxlO4CFU / ml during operation of the bioreactor system. In some embodiments, the aqueous feedstock and any other input into the bioreactor system does not comprise the phosphate solubilizing bacterial strain or does not comprise a concentration of the phosphate solubilizing bacterial strain at level higher than 100 CFU / ml. In some embodiments, the first microbial consortium comprises at least IxlO4CFU / ml of microbes.

[0038] In some embodiments, the aqueous feedstock further comprises an organic material digestible by microbes present in the containers. In some embodiments, the organic material comprises manure or material derived from manure. In some embodiments, the first container further comprises yeast. In some embodiments, the first microbial consortium comprises microbes derived from manure.

[0039] In some embodiments, the container comprising the product outflow stream port is a clarifier container configured to separate a portion of a working fluid in the clarifier container into a supernatant portion and a floc portion. In some embodiments, the clarifier container comprises one or more floc folding flights configured to agitate settled floc in the clarifier container without resuspending solids in the floc portion into the supernatant portion. In some embodiments, the system further comprises a floc return stream that flows from the clarifier to an earlier container in the series. In some embodiments, the product outflow stream comprises the supernatant portion.

[0040] In some embodiments, the product outflow stream comprises at least IxlO4CFU / ml of the phosphate solubilizing bacterial strain. In some embodiments, the product outflow stream comprises at least IxlO2CFU / ml of a sporulated form of the phosphate solubilizing bacterial strain. In some embodiments, the product outflow stream comprises a total dry weight of 0.2 to 2.5 mg / ml. In some embodiments, the product outflow stream has a chemical oxygen demand between 80 to 500 mg / L. In some embodiments, the product outflow stream has an electrical conductivity between 1.3 and 3.0 mS / cm.

[0041] In some embodiments, at least the first container comprises a mixer configured to aerate the first working fluid. In some embodiments, the first working fluid and / or the working fluid in at least one of the one or more additional containers comprises aerobic conditions.

[0042] Also disclosed is a biostimulant composition made by any of the methods described above. Also disclosed is a method of promoting plant growth comprising contacting a plant, seed, or plant growth medium with the biostimulant composition. Also disclosed is a method of increasing an amount of solubilized phosphate available to a plant, the method comprising contacting a plant, seed, or plant growth medium with the biostimulant composition.

[0043] Also disclosed is a composition comprising: (a) a Bacillus amyloliquefaciens strain having one or more of the following: (i) a 16S rRNA gene sequence at least 95% identical to SEQ ID NO: 1; (ii) a gyrB gene sequence at least 95% identical to SEQ ID NO: 3; and (iii) a rpoB gene sequence at least 95% identical to SEQ ID NO: 5; and (b) a carrier. In some embodiments, the Bacillus amyloliquefaciens strain is the strain deposited under ATCC Accession No. PTA-127657, or an isolated clone thereof. In some embodiments, the composition further comprises products of digestion of an organic substrate by the Bacillus amyloliquefaciens strain. In some embodiments, the carrier comprises a fertilizer. In some embodiments, the carrier is a solid coated by the Bacillus amyloliquefaciens strain. In some embodiments, the carrier is further coated by a micronutrient. In some embodiments, the micronutrient comprises a source of insoluble phosphate. In some embodiments, the carrier is aliquid. In some embodiments, the carrier further comprises an adjuvant selected from a wetting agent, spreading agent, dispersing agent, sticking agent, dust control agent, or adhesive. In some embodiments, the concentration of the Bacillus amyloliquefaciens strain in the composition ranges from IxlO3to IxlO11cfu / ml. In some embodiments, the concentration of the Bacillus amyloliquefaciens strain in the composition ranges from IxlO4to IxlO6cfu / ml.

[0044] Also disclosed is an isolated strain of the species Bacillus amyloliquefaciens having one or more of the following: (a) a 16S rRNA gene sequence at least 95% identical to SEQ ID NO: 1; (b) a gyrB gene sequence at least 95% identical to SEQ ID NO: 3; and (c) a rpoB gene sequence at least 95% identical to SEQ ID NO: 5. In some embodiments, the A amyloliquefaciens strain is the strain deposited under ATCC Accession No. PTA-127657, or an isolated clone thereof.

[0045] Also disclosed is a composition comprising: (a) a Bacillus licheniformis strain having one or more of the following: (i) a 16S rRNA gene sequence at least 95% identical to SEQ ID NO: 2; (ii) a gyrB gene sequence at least 95% identical to SEQ ID NO: 4; and (iii) a rpoB gene sequence at least 95% identical to SEQ ID NO: 6; and (b) a carrier. In some embodiments, the Bacillus licheniformis strain is the strain deposited under ATCC Accession No. PTA-127656, or an isolated clone thereof. In some embodiments, the composition further comprises products of digestion of an organic substrate by the Bacillus licheniformis strain. In some embodiments, the carrier comprises a fertilizer. In some embodiments, the carrier is a solid coated by the Bacillus licheniformis strain. In some embodiments, the carrier is further coated by a micronutrient. In some embodiments, the micronutrient comprises a source of insoluble phosphate. In some embodiments, the carrier is a liquid. In some embodiments, the carrier further comprises an adjuvant selected from a wetting agent, spreading agent, dispersing agent, sticking agent, dust control agent, or adhesive. In some embodiments, the concentration of the Bacillus licheniformis strain in the composition ranges from IxlO3to IxlO11cfu / m In some embodiments, the concentration of the Bacillus licheniformis strain in the composition ranges from IxlO4to IxlO6.

[0046] Also disclosed is an isolated strain of the species Bacillus licheniformis having one or more of the following: (a) a 16S rRNA gene sequence at least 95% identical to SEQ ID NO: 2; (b) a gyrB gene sequence at least 95% identical to SEQ ID NO: 4; and (c) a rpoB gene sequence at least 95% identical to SEQ ID NO: 6. In some embodiments, the A licheniformis strain is the strain deposited under ATCC Accession No. PTA-127656, or an isolated clone thereof.

[0047] Also disclosed is a method for promoting growth of a plant growing in a medium, the method comprising contacting the plant or the medium with any biostimulant described above, any composition described above, or a composition comprising any of the isolated strainsdescribed above. In some embodiments, the contacting increases the amount of solubilized phosphate available to the plant by at least 5%. In some embodiments, the composition increases plant growth by at least 5% as compared to a control. In some embodiments, the composition increases uptake of a nutrient by the plant by at least 5% as compared to a control. In some embodiments, the uptake of phosphate by the plant is increased by at least 5% as compared to a control. In some embodiments, the nutrient is zinc. In some embodiments, the nutrient is sulfur, potassium, magnesium, calcium, boron, manganese, iron, and / or copper. In some embodiments, the medium is soil or a hydroponic medium.

[0048] Also disclosed is a method of remedying a phosphate deficiency in a plant growth medium, the method comprising: (a) measuring a concentration of soluble phosphate in the plant growth medium that is less than 25 ppm; and (b) after step (a), contacting the plant growth medium with any biostimulant described above, any composition described above, or a composition comprising any of the isolated strains described above.

[0049] In some aspects, provided herein are methods of promoting plant growth, comprising: contacting a plant and / or plant growth medium in which the plant is growing with a composition comprising one or more compounds, wherein the one or more compounds is selected from the group consisting of Dihydrocholesterol -trimethylsilyl-ether, Docosanoic anhydride, Quinoline, 2-monopalmitin, Beta-sitostanyl acetate, Nonane, 2,2,4,4,6,8,8-heptamethyl-, Palmitoleic acid trimethyl silyl ester, Benzen-1,3-Dicarboxylic acid, Heneicosanoic acid, Octadecanoic acid, 2- hydroxy- 1,3 -propanediyl di-ester, Fumaric acid, cyclobutyl hexadecyl ester, 1,2- diaminopropane, 2-methylpentadecanoic acid trimethylsilylester, Distearin, Myristic acid, D- (+)-Malic acid, one or more derivatives thereof, and any combination thereof.

[0050] In some embodiments, the promoting plant growth comprises promoting phosphate solubilization. In some embodiments, the concentration of the one or more compounds in the composition is at least about 1 nanomolar (nM). In some embodiments, the concentration of the one or more compounds in the composition is at least about 0.00001% of a dry weight of the composition.

[0051] In some embodiments, the contacting comprises contacting the plant with the composition. In some embodiments, the contracting comprises contacting a plant seed with the composition. In some embodiments, the contacting comprises contacting a leaf of the plant with the composition. In some embodiments, the plant growth medium comprises soil, a hydroponic medium, turface, or isolite. In some embodiments, the contacting comprises increasing an amount of solubilized phosphate available to a plant. In some embodiments, the contacting comprises increasing phosphate solubilization in the plant growth medium. In someembodiments, the contacting causes an increase in plant growth by at least 10 percent as compared to the plant and / or the medium not contacted with the one or more compounds.

[0052] In some aspects, provided herein are methods of promoting plant growth, comprising: contacting a plant and / or plant growth medium in which the plant is growing with a composition comprising one or more compounds, wherein the one or more compounds is selected from the group consisting of 1-o-h exadecylglycerol 2,3-ditrimethylsilylether, 2-monopalmitin, 3-Methyl- p-anisaldehyde, Octadecanoic acid, 2-hydroxy- 1,3 -propanediyl di-ester, Eserine, Docosanoic anhydride, trans-p-Dimethylaminocinnamonitrile, Fumaric acid, cyclobutyl hexadecyl ester, (S)- (-)-.alpha.-(l-Naphthyl)ethylamine, 2-quinolinecarboxylic acid methyl ester, Isopentyl phenyl acetate, Methyl eicosanoate, Acenaphthylene, 4-(Anisylideneamino)-cinnamic acid, one or more derivatives thereof, and any combination thereof.

[0053] In some embodiments, the composition is filter-sterilized. In some embodiments, the composition comprises phosphate-solubilizing metabolites. In some embodiments, the promoting plant growth comprises promoting phosphate solubilization. In some embodiments, the concentration of the one or more compounds in the composition is at least about 1 nanomolar (nM). In some embodiments, the concentration of the one or more compounds in the composition is at least about 0.00001% of a dry weight of the composition.

[0054] In some embodiments, the contacting comprises contacting the plant with the composition. In some embodiments, the contracting comprises contacting a plant seed with the composition. In some embodiments, the contacting comprises contacting a leaf of the plant with the composition. In some embodiments, the plant growth medium comprises soil, a hydroponic medium, turface, or isolite. In some embodiments, the contacting comprises increasing an amount of solubilized phosphate available to a plant. In some embodiments, the contacting comprises increasing phosphate solubilization in the plant growth medium. In some embodiments, the contacting causes an increase in plant growth by at least 10 percent as compared to the plant and / or the plant growth medium not contacted with the one or more compounds.

[0055] In some aspects, provided herein are compositions for promoting plant growth, comprising: (i) at least one microbial strain comprising a Bacillus amyloliquefaciens strain or a Bacillus licheniformis strain; and (ii) one or more compounds selected from the group consisting of Dihydrocholesterol-trimethylsilyl-ether, Docosanoic anhydride, Quinoline, 2-monopalmitin, Beta-sitostanyl acetate, Nonane, 2,2,4,4,6,8,8-heptamethyl, Palmitoleic acid trimethyl silyl ester, Benzen-1,3-Dicarboxylic acid, Heneicosanoic acid, Octadecanoic acid, 2-hydroxy-l, 3- propanediyl di-ester, Fumaric acid, cyclobutyl hexadecyl ester, 1,2-diaminopropane, 2-methylpentadecanoic acid trimethylsilylester, Distearin, Myristic acid, D-(+)-Malic acid, one or more derivatives thereof, and any combination thereof.

[0056] In some embodiments, the at least one microbial strain comprises Bacillus amyloliquefaciens comprising one or more of the following: (i) 16S rRNA gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 1; (ii) a gyrB gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 3; or (iii) a rpoB gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 5. In some embodiments, the at least one microbial strain comprises Bacillus licheniformis comprising one or more of the following: (i) a 16S rRNA gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 2; (ii) a gyrB gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 4; or (iii) a rpoB gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 6.

[0057] In some embodiments, Docosanoic anhydride, Heneicosanoic acid, Fumaric acid, cyclobutyl hexadecyl ester, Distearin, D-(+)-Malic acid, or any combination thereof is present in the composition at the highest relative abundance and / or Quinoline is present at the lowest relative abundance relative to a total dry weight of the composition.

[0058] In some embodiments, the composition further comprises a carrier. In some embodiments, the carrier is formulated for application to a plant and / or medium in which the plant is growing

[0059] In some aspects, provided herein are compositions for promoting plant growth, comprising: (i) at least one microbial strain comprising a Bacillus amyloliquefaciens strain or a Bacillus licheniformis strain; and (ii) one or more compounds selected from the group consisting of 1-o-hexadecylglycerol 2,3-ditrimethylsilylether, 2-monopalmitin, 3-Methyl-p-anisaldehyde, Octadecanoic acid, 2-hydroxy- 1,3 -propanediyl di-ester, Eserine, Docosanoic anhydride, trans-p- Dimethylaminocinnamonitrile, Fumaric acid, cyclobutyl hexadecyl ester, (S)-(-)-. alpha. -(1- Naphthyl)ethylamine, 2-quinolinecarboxylic acid methyl ester, Isopentyl phenyl acetate, Methyl eicosanoate, Acenaphthylene, 4-(Anisylideneamino)-cinnamic ac-id, one or more derivatives thereof, and any combination thereof.

[0060] In some embodiments, the at least one microbial strain comprises Bacillus amyloliquefaciens comprising one or more of the following: (i) 16S rRNA gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 1; (ii) a gyrB gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 3; or (iii) a rpoB gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 5. In some embodiments, the at least one microbial strain comprises Bacillus licheniformis comprising one or more of the following: (i) a 16S rRNA gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 2; (ii) agyrB gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 4; or (iii) a rpoB gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 6.

[0061] In some embodiments, Octadecanoic acid, 2-hydroxy- 1,3 -propanediyl di-ester, Eserine, Docosanoic anhydride, trans-p-Dimethylaminocinnamonitrile, Fumaric acid, cyclobutyl hexadecyl ester, (S)-(-)-. alpha. -(l-Naphthyl)ethylamine, or any combination thereof is present in the composition at the highest relative abundance and / or 3-Methyl-p-anisaldehyde is present at the lowest relative abundance relative to a total dry weight of the composition.

[0062] In some embodiments, the composition further comprises a carrier. In some embodiments, the carrier is formulated for application to a plant and / or medium in which the plant is growing.

[0063] In some aspects, provided herein are compositions for promoting plant growth, comprising: (i) two or more compounds selected from the group consisting of Dihydrocholesterol-trimethylsilyl-ether, Docosanoic anhydride, Quinoline, 2-monopalmitin, Beta-sitostanyl acetate, Nonane, 2,2,4,4,6,8,8-heptamethyl, Palmitoleic acid trimethyl silyl ester, Benzen-1,3-Dicarboxylic acid, Heneicosanoic acid, Octadecanoic acid, 2-hydroxy-l, 3- propanediyl di-ester, Fumaric acid, cyclobutyl hexadecyl ester, 1,2-diaminopropane, 2- methylpentadecanoic acid trimethylsilylester, Distearin, Myristic acid, D-(+)-Malic acid, one or more derivatives thereof, and any combination thereof; and (ii) a carrier.

[0064] In some embodiments, the carrier is formulated for application to a plant or plant growth medium in which the plant is growing. In some embodiments, the carrier comprises a fertilizer. In some embodiments, the fertilizer is a solid. In some embodiments, the carrier is a liquid.

[0065] In some embodiments, the composition is configured to increase an amount of solubilized phosphate available to a plant. In some embodiments, the composition is configured to increase phosphate solubilization in the plant growth medium.

[0066] In some embodiments, Docosanoic anhydride, Heneicosanoic acid, Fumaric acid, cyclobutyl hexadecyl ester, Distearin, D-(+)-Malic acid, or any combination thereof is present in the composition at the highest relative abundance and / or Quinoline is present at the lowest relative abundance relative to a total dry weight of the composition.

[0067] In some embodiments, the composition further comprises an adjuvant selected from the group consisting of a wetting agent, spreading agent, dispersing agent, sticking agent, dust control agent, and adhesive. In some embodiments, the composition is configured to increase or is capable of increasing an amount of solubilized phosphate available to a plant by at least 5%. In some embodiments, the composition is configured to cause or is capable of causing an increase in plant growth by at least 10 percent as compared to a control.

[0068] In some aspects, provided herein are compositions for promoting plant growth, comprising: (i) two or more compounds selected from the group consisting of l-o- hexadecylglycerol 2,3-ditrimethylsilylether, 2-monopalmitin, 3-Methyl-p-anisaldehyde, Octadecanoic acid, 2-hydroxy- 1,3 -propanediyl di-ester, Eserine, Docosanoic anhydride, trans-p- Dimethylaminocinnamonitrile, Fumaric acid, cyclobutyl hexadecyl ester, (S)-(-)-. alpha. -(1- Naphthyl)ethylamine, 2-quinolinecarboxylic acid methyl ester, Isopentyl phenyl acetate, Methyl eicosanoate, Acenaphthylene, 4-(Anisylideneamino)-cinnamic acid, one or more derivatives thereof, and any combination thereof; and (ii) a carrier.

[0069] In some embodiments, the carrier is formulated for application to a plant or plant growth medium in which the plant is growing. In some embodiments, the carrier comprises a fertilizer. In some embodiments, the fertilizer is a solid. In some embodiments, the carrier is a liquid. In some embodiments, the composition is configured to increase phosphate solubilization in the plant growth medium. In some embodiments, the composition is configured to increase phosphate solubilization in the plant growth medium.

[0070] In some embodiments, Octadecanoic acid, 2-hydroxy- 1,3 -propanediyl di-ester, Eserine, Docosanoic anhydride, trans-p-Dimethylaminocinnamonitrile, Fumaric acid, cyclobutyl hexadecyl ester, (S)-(-)-. alpha. -(l-Naphthyl)ethylamine, or any combination thereof is present in the composition at the highest relative abundance and / or 3-Methyl-p-anisaldehyde is present at the lowest relative abundance relative to a total dry weight of the composition.

[0071] In some embodiments, the composition further comprises an adjuvant selected from the group consisting of a wetting agent, spreading agent, dispersing agent, sticking agent, dust control agent, and adhesive. In some embodiments, the composition is configured to increase or is capable of increasing an amount of solubilized phosphate available to a plant by at least 5%. In some embodiments, the composition is configured to cause or is capable of causing an increase in plant growth by at least 10 percent as compared to a control.

[0072] Additional aspects and advantages of the present disclosure will become readily apparent to those skilled in this art from the following detailed description, wherein only illustrative embodiments of the present disclosure are shown and described. As will be realized, the present disclosure is capable of other and different embodiments, and its several details are capable of modifications in various obvious respects, all without departing from the disclosure. Accordingly, the drawings and description are to be regarded as illustrative in nature, and not as restrictive.INCORPORATION BY REFERENCE

[0073] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.BRIEF DESCRIPTION OF THE DRAWINGS

[0074] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings (also “Figure” and “FIG.” herein), of which:

[0075] FIG. 1 depicts a serialized PST system (PST-sIP) with a series of reactors and inputs to the digestion system.

[0076] FIG. 2 depicts field trial results for application of MS2839 in-furrow for soybean and com plants.

[0077] FIGs. 3A-3B is a series of graphs depicting the com yield and soybean yield in bushels per acre (Bu / A) after treatment with different isolates (for each pair of bars, left bar is results from Nebraska (NE) trials and right bar is results from Kentucky (KY) trials). FIG. 3A depicts the corn yield from KY and NE trials. FIG. 3B depicts the soybean yield from KY and NE trials.

[0078] FIGs. 4A-4B are a series of graphs showing corn yield across GSP (Grower Standard Practice) alone and GSP with four different bacteria strains (MS2414, MS1574, MS1835, and MS2839). FIG. 4A depicts results from North Platte, Nebraska. FIG. 4B depicts results from Brule, Nebraska.

[0079] FIGs. 5A-5B are a series of graphs showing soybean yield across GSP alone and GSP with four different bacteria strains (MS2414, MS1574, MS1835, and MS2839). FIG. 5A depicts results from North Platte, Nebraska. FIG. 5B depicts results from Brule, Nebraska.

[0080] FIG. 6 is a graph showing the results of MS2839 plant growth promotion (PGP) activity over four different tests (first corn, second corn, first soybean, and second soybean) normalized to percent over the negative control for that test.

[0081] FIG. 7 is a graph showing com greenhouse growth promotion testing between water, MS2839 alone, kelp product alone at either 0.64 ml / pot or 3.2 ml / pot, and MS2839 with kelp product at either 0.64 ml / pot or 3.2 ml / pot.

[0082] FIGs. 8A-8B depict experimental results of MS2839 and MS 1835 spore counts following seven or fourteen days of incubation. Treatments were separated based on normalfeeding e.g., N Feed) or supplemental feeding e.g., S Feed). FIG. 8A shows results of MS2839 and FIG. 8B shows results of MS 1835.

[0083] FIGs. 9A-9B depicts box plot target counts of MS 1835 (FIG. 9A) or MS2839 (FIG.9B) Treatments used either base product of the P2 digestion system or water.

[0084] FIGs. 10A-10B depicts box plot target counts of MS1835 (FIG. 10A) or MS2839 (FIG. 10B). Treatments used either liquid or powder spore preparation as the inoculation medium.

[0085] FIGs. 11A-11B depicts box plot target counts of MS1835 (FIG. 11A) or MS2839 (FIG. 11B) Treatments comprised either treatments with additional supplemented feeding or treatments that received normal feeding.

[0086] FIG. 12 is a graph depicting the amount of insoluble phosphate released into solution and made soluble (in mg / L) between untreated control (UTC) and media treated with MS2839.

[0087] FIG. 13 is a graph showing the plant growth production of Arabidopsis (measured in average leaf area, cm2) for UTC (untreated control) and PST supernatant.

[0088] FIG. 14 is a graph depicting corn shoot dry biomass (in grams) across UTC and PST 2.0 sIP. Results are from experiments with corn which was fertilized with PST 2.0 sIP coated monoammonium phosphate (MAP).

[0089] FIG. 15 is a graph depicting corn shoot dry biomass (in grams) across UTC and PST 2.0 sIP. Results are from experiments in which com was treated via in-furrow application.

[0090] FIG. 16 is a graph depicting uptake of macronutrients, such as nitrogen, sulfur, phosphorus, potassium, magnesium, and calcium, in com shoots (represented as %UTC, mg / shoot) after treatment with PST-sIP product at 2 qt / t rate. All results are from corn which was fertilized with MAP coated with PST-sIP product.

[0091] FIG. 17 is a graph depicting uptake of micronutrients, such as boron, zinc, manganese, copper, and iron in com shoots (represented as % UTC, pg / shoot) after treatment with PST-sIP product at 2 qt / t rate. All results are from corn which was fertilized with MAP coated with PST- sIP product.

[0092] FIG. 18 is a graph showing uptake of macronutrients in com shoots (represented as %UTC, mg / shoot) across PST-sIP. Results are from an experiment in corn in which the treatments were applied in-furrow at planting. Bars from left to right for each system are: nitrogen (N), sulfur (S), phosphorus (P), potassium (K), magnesium (Mg), and calcium (Ca). All results are from corn which was treated with PST-sIP product in-furrow at 4qt / acre application rate.

[0093] FIG. 19 is a graph showing uptake of micronutrients in corn shoots (represented as % UTC, pg / shoot) across PST-sIP. Results are from a corn experiment in which the treatmentswere applied in-furrow at planting. Bars from left to right for each system are: boron (B), zinc (Zn), manganese (Mn), iron (Fe), and copper (Cu). All results are from corn which was treated with PST-sIP product in-furrow at 4qt / acre application rate.

[0094] FIG. 20 is a graph showing dry biomass of corn over untreated control, PST, and PST- sIP conditions.

[0095] FIG. 21 is a graph showing the amount of phosphate made soluble (in mg / L) across UTC and PST-sIP system on Day 0, 3, and 7 post-inoculation. For each condition on the x-axis: left bar designates Day 0, middle bar designates Day 3, and right bar designates Day 7.

[0096] FIG. 22 is a graph showing the amount of phosphate solubilized from coated MAP fertilizer (in mg / L) across UTC, water, and PST-sIP cSPN on day 6 post-treatment.

[0097] FIG. 23 showing phosphate solubilization (in mg / L) from MAP fertilizer coated with either water, PST or PST-SIP.

[0098] FIG. 24 shows phosphate made soluble (in mg) across UTC, MS2839 alone, PBR2 feed, manure feed, and PST-sIP supernatant over 5 days.

[0099] FIG. 25 is a graph showing MS2839 spore count (cfu / ml) in supernatant for different ratios of floc. Bars from left to right for each percent floc condition are: Time Zero, 1 HR, 6 HR, 24 HR, and 48 HR post-inoculation.

[0100] FIG. 26 is a graph showing MS2839 counts (cfu / ml) from PST-sIP digestion system over time in supernatant (BP) and WB.

[0101] FIG. 27 is a graph showing counts of MS2839 (cfu / ml) from PwST-sIP digestion system over time with or without floc-folding flights. T.L. denotes trend-line for each condition.

[0102] FIG. 28 shows GC-MS chromatograms of intact samples from PwST-sIP (top 6 replicates) and PwST-non sIP (bottom 3 replicates). The numbers at the end of each label indicate respective associated batch.

[0103] FIG. 29 shows GC-MS chromatograms of filter-sterilized samples from PwST-sIP (Top 6 replicates) and PwST-non sIP (Bottom 3 replicates). The numbers at the end of each label indicate respective associated batch.

[0104] FIGs. 30A-30B show GC-MS chromatograms of intact and filter-sterilized samples of the 6 PwST-sIP batches that were extracted and analyzed. The numbers at the end of each label indicate respective associated batch. FIG. 30A shows intact samples. FIG. 30B shows filter- sterilized samples.

[0105] FIG. 31 shows GC-MS chromatograms of intact (top 3 replicates) and filter-sterilized (bottom 3 replicates) samples of the 3 PwST-non sIP batches that were extracted and analyzed. The numbers at the end of each label indicate respective associated batch.

[0106] FIGs. 32A-32B show PCA results based on GC-MS analysis of PwST-sIP and PwST- non sIP base products. FIG. 32A shows intact samples. FIG. 32B shows filter-sterilized samples.

[0107] FIG. 33 shows a Venn Diagram of the number of up-regulated compounds in the intact and filter-sterilized samples of PwST-sIP over PwST-non sIP from GC-MS analysis.

[0108] FIG. 34 shows LC-MS Chromatograms of PwST- sIP samples.

[0109] FIG. 35 shows LC-MS Chromatograms of PwST-non sIP samples.

[0110] FIGs. 36A-36B show PCA based on LC-MS analysis of PwST-sIP and PwST-non sIP base products. FIG. 36A shows intact samples. FIG. 36B shows filter-sterilized samples.

[0111] FIG. 37 shows a Venn Diagram of the number of up-regulated compounds in the intact and filter-sterilized samples of PwST-sIP over PwST-non sIP from LC-MS analysis.DETAILED DESCRIPTION

[0112] While various embodiments of the invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions may occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed.

[0113] Described herein are systems and methods that employ microbial digestion of various feedstocks. A system of the present disclosure may comprise a continuous system capable of serialized isolate production (e.g., sIP system). The isolate production of the sIP system can occur within a mixed consortium of microbes. The target isolates of a sIP digestion system may become enriched in the microbial environment and may demonstrate improved efficacy and functionality. The main targeted functionality may be phosphate solubilization of insoluble phosphate from soil or fertilizer and improved nutrient uptake in plants. A target isolate may possess commercially valuable properties and can be introduced into a continuous (e.g., serialized) reactor system comprised of a complex microbial consortia that has been modified for functionality (e.g., for phosphate solubilization). Without wishing to be bound by theory, a target isolate may provide a performance benefit to a microbial community of the digestions systems described herein, providing a chemical and / or functional synergistic relationship as it grows in the system. A digestion system described herein may comprise a phosphate solubilization technology (PST) system, with a target function to enrich a population or populations of phosphate-solubilizing microbes, metabolites, or any combination thereof.

[0114] The products of digestion methods and systems described herein can include microbes and metabolites produced by microbial digestion of feedstock substrates. In some embodiments, the products of digestion methods and systems described herein can comprise biostimulant compositions that have plant growth promoting properties when applied to plants or to a medium in which plants are growing (e.g., soil). In some embodiments, methods and systems described herein are arranged to selectively promote growth of microbes that have a desired plant growth promoting property themselves or that produce metabolites that have the desired plant growth promoting property, such that the biostimulant product has the desired plant growth promoting property. Applications of the products of the digestion systems described herein may be on dry-fertilizers, applied in conjunction with the application of fertilizers, in formulations with additional components including liquid fertilizers or micronutrient coating formulations, and / or in foliar applications. Applications of the products of the digestion systems described herein may be to a part of a plant, such as a shoot, a stem, a leaf, a lateral bud, a terminal bud, a flower, a leaf axil, a root (e.g., a primary root, a lateral root, a root hair, a root cap), or any combination thereof. These and other features of embodiments disclosed herein are described in more detail below.A. Definitions

[0115] In the following description, certain specific details are set forth in order to provide a thorough understanding of various embodiments. However, one skilled in the art will understand that the embodiments provided may be practiced without these details. Unless the context requires otherwise, throughout the specification and claims which follow, the word “comprise” and variations thereof, such as, “comprises” and “comprising” are to be construed in an open, inclusive sense, that is, as “including, but not limited to.” As used in this specification and the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the content clearly dictates otherwise. It should also be noted that the term “or” is generally employed in its sense including “and / or” unless the content clearly dictates otherwise. Further, headings provided herein are for convenience only and do not interpret the scope or meaning of the claimed embodiments.

[0116] The term “about” or “approximately” means within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, e.g., the limitations of the measurement system. For example, “about” can mean within 1 or more than 1 standard deviation, per the practice in the given value. Where particular values are described in the application and claims, unless otherwise stated the term “about” should be assumed to mean an acceptable error range for the particular value.

[0117] The term “culturing”, as used herein, can refer to the propagation of organisms on or in media of various kinds. Non-limiting examples of suitable media include tryptic soy agar (TSA), phosphate solubilizing medium which contain insoluble forms of phosphate, nutrient medium, lysogeny broth (LB medium), and / or plate count agar.

[0118] As used herein, the term “enriched culture” of an isolated microbial strain can refer to a microbial culture wherein the total microbial population of the culture contains a percentage of a target isolated strain. An enriched culture may comprise a percentage of a target isolated strain and a population of microbes enriched for a particular functionality (e.g., phosphate solubilization). In some embodiments, an enriched culture may comprise a percentage of a target isolated strain, a population of microbes enriched for a particular functionality, and metabolites enriched for a particular functionality (e.g., phosphate solubilization). The enriched culture may comprise a percentage of a total bacteria population in a container of digestion system described herein. The enriched culture may comprise a percentage of a total bacteria population in an output product (e.g., biostimulant) described herein. An enriched culture may comprise an increased amount of a target isolated strain a target population of microbes, or a combination thereof compared to a total microbial population of a culture. An enriched culture may comprise a growing population of a target isolated strain and a population of microbes enriched for a particular functionality (e.g., phosphate solubilization) over a time period. In some embodiments, an enriched culture can refer to a microbial culture wherein the total microbial population of the culture contains at least about 0.001%, at least about 0.01%, at least about 0.05%, at least about 0.1%, at least about 0.5%, at least about 1%, at least about 1.5%, at least about 2%, at least about 2.5%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 50%, or at least about 75% of a target isolated strain, a population of microbes enriched for a particular functionality, metabolites enriched for a particular functionality, or any combination thereof. In some embodiments, an enriched culture can refer to a microbial culture wherein the total microbial population of the culture contains at most about 75%, at most about 50%, at most about 25%, at most about 20%, at most about 15%, at most about 10%, at most about 5%, at most about 2.5%, at most about 2%, at most about 1.5%, at most about 1%, or at most about 0.5% of a target isolated strain, a population of microbes enriched for a particular functionality, metabolites enriched for a particular functionality, or any combination thereof. In some embodiments, an enriched culture can refer to a microbial culture wherein the total microbial population of the culture contains from about 0.5% to about 75% of a target isolated strain, a population of microbes enriched for a particular functionality, metabolites enriched for a particular functionality, or any combination thereof. In some embodiments, an enriched culture can refer toa microbial culture wherein the total microbial population of the culture contains from about 0.5% to about 1%, about 0.5% to about 2%, about 0.5% to about 3%, about 0.5% to about 5%, about 0.5% to about 10%, about 0.5% to about 15%, about 0.5% to about 20%, about 0.5% to about 25%, about 0.5% to about 50%, about 0.5% to about 60%, about 0.5% to about 75%, about 1% to about 2%, about 1% to about 3%, about 1% to about 5%, about 1% to about 10%, about 1% to about 15%, about 1% to about 20%, about 1% to about 25%, about 1% to about 50%, about 1% to about 60%, about 1% to about 75%, about 2% to about 3%, about 2% to about 5%, about 2% to about 10%, about 2% to about 15%, about 2% to about 20%, about 2% to about 25%, about 2% to about 50%, about 2% to about 60%, about 2% to about 75%, about 3% to about 5%, about 3% to about 10%, about 3% to about 15%, about 3% to about 20%, about 3% to about 25%, about 3% to about 50%, about 3% to about 60%, about 3% to about 75%, about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 50%, about 5% to about 60%, about 5% to about 75%, about 10% to about 15%, about 10% to about 20%, about 10% to about 25%, about 10% to about 50%, about 10% to about 60%, about 10% to about 75%, about 15% to about 20%, about 15% to about 25%, about 15% to about 50%, about 15% to about 60%, about 15% to about 75%, about 20% to about 25%, about 20% to about 50%, about 20% to about 60%, about 20% to about 75%, about 25% to about 50%, about 25% to about 60%, about 25% to about 75%, about 50% to about 60%, about 50% to about 75%, or about 60% to about 75% of a target isolated strain, a population of microbes enriched for a particular functionality, metabolites enriched for a particular functionality, or any combination thereof.

[0119] The term “composition” as used herein can refer to a combination of an active agent (e.g., a microbial strain described herein) and at least one other compound, carrier, or composition, which can be inert (for example, a detectable agent or liquid carrier) or active, such as, but not limited to, a fertilizer, nutrient, or pesticide. A microbial composition refers to a composition comprising at least one microbial species. A composition may comprise microbial metabolites generated in a microbial consortium of a digestion system described herein.

[0120] An “effective amount”, as used herein, can refer to an amount sufficient to effect beneficial and / or desired results. An effective amount can be administered in one or more administrations. The expression “effective microorganism” used herein in reference to a microorganism is intended to mean that the subject strain exhibits a degree of promotion of plant health, growth, yield, or a combination thereof, at a statistically significant level, that of an untreated control. In some instances, the expression “an effective amount” is used herein in reference to that quantity of microbial treatment which can be used to obtain a beneficial or desired result relative to that occurring in an untreated control under suitable conditions of treatment as described herein. Forexample, the expression “an agriculturally effective amount” is used herein in reference to that quantity of microbial treatment which can be used to obtain an agriculturally beneficial or desired result relative to that occurring in an untreated control under suitable conditions of treatment as described herein. The effective amount of an agricultural formulation or composition that may be applied for the improvement of plant health, growth, yield, or a combination thereof, can be readily determined.

[0121] A “carrier” as used herein can refer to a substance or a composition that support the survival of the microbes. Such carriers may be either organic or non-organic.

[0122] “Percentage of sequence identity”, as used herein, can be determined by comparing two optimally locally aligned sequences over a comparison window defined by the length of the local alignment between the two sequences. The amino acid sequence in the comparison window may comprise additions or deletions (e. g., gaps or overhangs) as compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two sequences.

[0123] Local alignment between two sequences may include segments of each sequence that are deemed to be sufficiently similar according to a criterion that depends on the algorithm used to perform the alignment (e. g. BLAST). The percentage of sequence identity is calculated by determining the number of positions at which the identical nucleic acid base or amino acid residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison and multiplying the result by 100. Optimal alignment of sequences for comparison may be conducted by the local homology algorithm of Smith and Waterman (Add. APL. Math. 2:482, 1981), by the global homology alignment algorithm of Needleman and Wunsch (J Mol. Biol. 48:443, 1970), by the search for similarity method of Pearson and Lipman (Proc. Natl. Acad. Sci. USA 85:2444, 1988), by heuristic implementations of these algorithms (NCBI BLAST, WU-BLAST, BLAT, SIM, BLASTZ), or by inspection. Given that two sequences have been identified for comparison, GAP and BESTFIT may be employed to determine their optimal alignment. Typically, the default values of 5.00 for gap weight and 0.30 for gap weight length are used. The term “substantial sequence identity” between polynucleotide or polypeptide sequences refers to polynucleotide or polypeptide comprising a sequence that has at least about 50% sequence identity, at least about 60% sequence identity, at least about 70% sequence identity, at least about 80% sequence identity, at least about 85% sequence identity, at least about 90% sequence identity, at least about 95% sequence identity, at least about 96% sequence identity, at least about 97% sequence identity, at least about 98% sequence identity, or at least about 99% sequence identity compared to a reference sequence using the programs. In addition, pairwise sequence homology or sequence similarity, asused, refers to the percentage of residues that are similar between two sequences aligned. Families of amino acid residues having similar side chains have been well defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Query nucleic acid and amino acid sequences can be searched against subject nucleic acid or amino acid sequences residing in public or proprietary databases. Such searches can be done using the National Center for Biotechnology Information Basic Local Alignment Search Tool (NCBI BLAST v 2.18) program. The NCBI BLAST program is available on the internet from the National Center for Biotechnology Information (blast.ncbi.nlm.nih.gov / Blast.cgi). Typically the following parameters for NCBI BLAST can be used: Filter options set to “default”, the Comparison Matrix set to “BLOSUM62”, the Gap Costs set to “Existence: 11, Extension: 1”, the Word Size set to 3, the Expect (E threshold) set to le-3, and the minimum length of the local alignment set to 50% of the query sequence length. Sequence identity and similarity may also be determined using GenomeQuest™ software (Gene-IT, Worcester Mass. USA).

[0124] The term “plant growth promotion” (e.g., “PGP”) can refer to processes that can promote plant health, growth, yield, or any combination thereof. In some embodiments, PGP can encompass a wide range of improved plant properties, including but not limited to, improved nitrogen fixation, improved phosphate uptake, improved zinc uptake, improved root development, increased leaf area, increased plant yield, increased uptake of macronutrients, increased uptake of micronutrients, increased seed germination, enhancing seed germination, enhancing early plant development, improving root growth, improving shoot growth, improving plant height, increasing nutrient uptake, mitigating transplant shock, improving plant reproduction, improving soil microbial activity, increased photosynthesis, increased abundance of functional enzymes, increased dry biomass, or an increase in accumulated biomass of the plant. In some embodiments, the microbial strains, isolates, cultures, compositions or synthetic consortia as described herein improve nutrient uptake, plant health and vigor, improve root development, increase leaf area, increase plant yield, increased uptake of macronutrients, increased uptake of micronutrients, increase seed germination, increased abundance of functional enzymes, increased dry biomass, or an increase in accumulated biomass of the plant. In some embodiments, the microbial strains, isolates, cultures, or compositions as described herein increase the size or mass of a plant or partsthereof, as compared to a control plant, or a plant that has not been treated with a substance, or parts thereof or as compared to a predetermined standard. In some embodiments, the microbial strains, isolates, cultures, compositions or synthetic consortia as described herein improve the health, vigor and yield of a plant, as compared to a control plant or a plant that has not been treated with a substance, but also can survive and multiply in microhabitats associated with the root surface.

[0125] Microbes described herein as “phosphate-solubilizing microbes” and “phosphate- solubilizing bacteria” or “phosphate solubilizers” may refer to a microbe or bacterium that either (a) has a measurable increase in phosphate made soluble relative to a control in an assay that includes suspending adding a test solution that includes the microbe or bacterium to media with insoluble, inorganic phosphate (tricalcium phosphate (Cas PCU)?), incubating the media, and measuring an amount of insoluble phosphate released into solution and made soluble using an instrument such as a SEAL AQ400 and comparing to known standards (b) makes a visible halo when grown in a colony on a plate with insoluble phosphate.

[0126] As used herein, the term “yield” can refer to the amount of harvestable plant material or plant-derived product, and is normally defined as the measurable produce of economic value of a crop.

[0127] For crop plants, “yield” can also mean the amount of harvested material per acre or unit of production. Yield may be defined in terms of quantity or quality. The harvested material may vary from crop to crop, for example, it may be seeds, above ground biomass, roots, fruits, cotton fibers, any other part of the plant, or any plant-derived product which is of economic value.

[0128] In some embodiments, microbial strains, isolates, cultures, and compositions, and biostimulant compositions described herein may be used to promote improved crop or product quality, including food quality. Improving crop quality may include improving characteristics that make a crop or product more marketable such as, for example, a desired color, size, or shape. Improving crop quality may also including making a food product with desired nutritional, nutraceutical, and / or pharmaceutical characteristics. Improving crop or product quality may include adding valuable attributes that may increase price. Improving crop or product quality may include increasing value per harvest unit, such as, for example, providing increased oil, sugar, starch, or protein yield per unit weight of harvested crop or product.

[0129] In some embodiments, the microbial strains, isolates, cultures and compositions according to the embodiments of this application can lead to plant growth promotion or plant growth improvement that is an at least 5% increase, at least 10% increase, at least 25% increase, at least 50% increase, at least 75% increase, or at least a 100% increase in the property being measured.In some embodiments, the microbial strains, isolates, cultures and compositions according to the embodiments of this application lead to plant growth promotion or plant growth improvement that is an at least 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% increase in the property being measured. In some embodiments, the microbial strains, isolates, cultures and compositions of this application may produce an above stated percentage increase in phosphate uptake, an above stated increase in phosphate solubilization capacity an above stated increase in total root weight, or in leaf area or in plant product yield (e.g., an above stated percentage increase in plant product weight).

[0130] A “control plant”, as used herein, can provide a reference point for measuring changes in phenotype of the subject plant, and may be any suitable plant cell, seed, plant component, plant tissue, plant organ or whole plant. A control plant may comprise, but is not limited to, (a) a plant which is genetically identical to the subject plant but which is not exposed to the same treatment (c.g, inoculant treatment) as the subject plant or (b) the subject plant itself, under conditions in which it has not been exposed to a particular treatment such as, for example, an inoculant or combination of inoculants and / or other chemicals. A treated plant may comprise a plant that has had an inoculum of a microbe or a biostimulant composition as described herein applied to any part of the plant (e.g., seed, stem, root, shoot, leaf, or combination thereof). A treated plant may comprise a plant that has had an inoculum of a microbe or a biostimulant composition as described herein applied using an in-furrow application. A treated plant may comprise a plant that has had an inoculum of a microbe or a biostimulant composition as described herein applied using a sidedress application, a broadcast application, a y-drop application, a tape application, or a fertigation application. A treated plant may comprise a plant that has had an inoculum of a microbe or a biostimulant composition as described herein applied to the soil. An untreated plant may comprise a plant that that has not had an inoculum of a microbe or a biostimulant composition as described herein applied directly or indirectly.

[0131] “Inoculant” as used herein can refer to any culture or preparation that comprises at least one microorganism. In some embodiments, an inoculant (sometimes as microbial inoculant, or soil inoculant) is an agricultural addition that uses beneficial microbes (including, but not limited to endophytes) to promote plant health, growth, yield, or any combination thereof. Many of the microbes suitable for use in an inoculant form symbiotic relationships with the target crops where both parties benefit (mutualism). For example, an isolated microbial strain as described herein may benefit from carbon sources from the roots of a plant and the plant may benefit from metabolites generated by metabolism of the microbe. Without wishing to be bound by theory, a plant may be colonized by the isolate and the colonization of the roots may block plant pathogensfrom accessing the roots or may provide growth and health benefits to the plant. An inoculant (e.g., inoculum of a microbe / microbial strain) can be added at one time point during an operating of a digestion system process. An inoculant (e.g., inoculum of a microbe / microbial strain) can be added at multiple time points during an operating of a digestion system process.

[0132] The term “serialized isolate production”, (e.g., sIP), can refer to specialized manipulated continuous serialized reactors that may enable the growth and enrichment of the microbes, isolates, target isolates, microorganisms, or any combination thereof, as described herein.

[0133] The term “floc” can refer to a mass formed by the aggregation of a number of fine suspended particles. Particles can comprise biological particles, non-biological particles, or a combination thereof. For example, a floc can comprise organic materials recovered from a feedstock, waste, wastewater, sludge material of a fluid, or any combination thereof used in a digestion system. A floc can comprise biosolids and / or particles from digestion products of organic materials. Floc can comprise an aggregated mass of microorganisms (e.g., bacteria).

[0134] The term “whole broth” (e.g., WB) can refer to a blend of supernatant and floc at a ratio for use in the technologies as described herein. A whole broth may comprise microbial populations (e.g., phosphate solubilizing microbes), enzymes, fungi, biosolids, or any combination thereof.

[0135] The terms “microbial consortium” or “microbial population” can refer to a group of microorganisms in an environment. Consortiums may be endosymbiotic or ectosymbiotic. Microorganisms in a microbial consortium can include, but are not limited to, bacteria, fungi, yeasts, lichens, algae, protozoa, archaea, molds, or any combination thereof.

[0136] The term “supernatant” (e.g., “base product”) can refer to the final product of the digestion system. The supernatant can be measured for amount of a microbial isolate, number of members within a microbial consortium, or types and amount of microbial metabolites with plant growth promotion capacity.

[0137] The term “load rate” can refer to a rate at which a source material is introduced into a digestion system. In some embodiments, load rate may refer to “organic load rate” or “hydraulic load rate”. Organic load rate comprises a rate at which organic feedstock is introduced into the system. Hydraulic load rate comprises a rate at which a hydraulic source is introduced into the system.

[0138] The term “working fluid” can refer to a fluid substance supporting and transporting biology and nutrients through a system of containers. For example, a working fluid may comprise organic materials, microorganisms (e.g., microbes and / or metabolites), biosolids, macronutrients, micronutrients, organic nutrients, inorganic nutrients, or any combination thereof. A working fluidcan comprise a solution that flows throughout a digestion system and may provide an enriched environment for microbes of the digestion system.

[0139] Whenever the term “at least,” “greater than,” or “greater than or equal to” precedes the first numerical value in a series of two or more numerical values, the term “at least,” “greater than” or “greater than or equal to” can apply to each of the numerical values in that series of numerical values. For example, greater than or equal to 1, 2, or 3 is equivalent to greater than or equal to 1, greater than or equal to 2, or greater than or equal to 3.

[0140] Whenever the term “no more than,” “less than,” or “less than or equal to” precedes the first numerical value in a series of two or more numerical values, the term “no more than,” “less than,” or “less than or equal to” can apply to each of the numerical values in that series of numerical values. For example, less than or equal to 3, 2, or 1 is equivalent to less than or equal to 3, less than or equal to 2, or less than or equal to 1.B. Microbial Digestion Methods and Systems

[0141] Certain embodiments disclosed herein include methods and systems in which microbes comprised in microbial consortia digest substances provided in a feedstock. The digestion systems may be comprised of a series of separate, fluidly connected containers, also referred to herein as “reactors.” In each reactor, a different microbial consortium may be established and maintained throughout continuous operation of the digestion system. The unique microbial consortia present in each reactor may provide for different physiological activities in the different reactors. Thus, different steps in digestion of a feedstock may be performed in different reactors, which may result in (1) a more complete digestion — i.e., more complete breakdown of macromolecules in the feedstock — than other types of digestion systems, and / or (2) production of a variety of microbial digestion products having plant growth promoting properties (e.g., ability to enhance or improve soil available phosphate via phosphate solubilization, phosphate uptake by plant tissues, and / or otherwise promote phosphate use efficiency).

[0142] In some embodiments, the bioreactor system (e.g., the digestion system) comprises an established population of one or more phosphate-solubilizing microbial strains in one or more containers of the system. An “established population” of a particular microbial strain is a population that remains within an operating bioreactor system without replenishing the microbial strain from outside the bioreactor system. In some embodiments, an established population is one that has not been diminished by more than about 1, 3, 5, 10, 15, 20, or 25% during continuous operation of the bioreactor system for at least about 5, 10, 15, 20, 25, 30, 60, or 90 days without adding a population of the microbial strain to the bioreactor system at a concentration higher thanabout 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 25, 50, or 100 CFU / ml. In some embodiments, an established population of a microbial strain has been established by making one or more inoculations of the microbial strain into one or more containers of the bioreactor system. In some embodiments, a measurement (e.g., a concentration or bacterial count) of an established population may be performed for a digestion system. In some embodiments, an established population is a population that is derived from a microbial population that was inoculated into the system at least about 5, 10, 20, 30, 40, 50, 60, or 90 days prior to a measurement of the established population.

[0143] In some embodiments, a bioreactor system comprises at least one microbial strain. In some embodiments, a bioreactor system comprises at least one phosphate-solubilizing microbial strain. In some embodiments, a bioreactor system comprises an established population of a first phosphate-solubilizing microbial strain and an established population of a second phosphate- solubilizing microbial strain. In some embodiments, the bioreactor system further comprises an established population of a third phosphate-solubilizing microbial strain. The established populations of the respective microbial strains may be established in individual or combined inoculations into the bioreactor system. An individual inoculation may comprise one inoculum of a microbial strain (e.g., microbe). A combined inoculation may comprise an inoculum comprising at least two microbial strains. The combined inoculation may comprise the same isolated microbial strains. The combined inoculation may comprise an isolated microbial strain and non-isolated microbial strain. The combined inoculation may comprise two or more isolated microbial strains.

[0144] In some embodiments, a reactor or a series of reactors (e.g., serialized assembly of reactors) functions to contribute to the growth of one or more microbes having desired plant growth promoting properties and / or to the production of digestion products having plant growth promoting properties. The system can comprise 1, 2, 3, 4, 5, 6, or more reactors. In some embodiments, the operation of a digestion system may lead to growth of one or more microbes having a desired plant growth promoting effect. The one or more microbes may be one or more isolated microbes added separately as an inoculum to the digestion system. The one or more microbes may also be input into the system as part of a feed material that includes a mixture of microbes. The one or more microbes may be endogenous to an organic material such as, for example, a manure, a plant, a lignocellulosic material, or an algae. The one or more microbes may also be endogenous to other types of feed materials, such as rock phosphate. In some embodiments, endogenous microbes are those microbes naturally present in feedstock material (e.g., a manure, a plant, a lignocellulosic material, or an algae). These microbes may naturally reside in a closed system and / or are present in the ecosystem of the feedstock material.

[0145] Inputs into digestion systems may include one or more of water, a microbial inoculum (e.g., inoculum of a microbe or inoculum of a microbial strain), nutrients (e.g., one or more sources of carbon, nitrogen, phosphorous, etc.), and a digestion substrate. Fluid within reactors of a digestion system may be referred to herein as a “working fluid.” In continuous operation, each reactor may have a constant volume of working fluid therein, with the rate of fluid flowing into each reactor matching the rate of fluid flowing out of each reactor. As each reactor may include a different microbial consortium and have different conditions from other reactors, the working fluid within each reactor may be considered to be distinct from working fluids within the other reactors. The total volume of working fluid within a digestion system may be referred to herein as the “total working volume” of the digestion system.

[0146] Digestion substrates included in an input stream into a digestion system may include, for example, organic materials that can be digested by microbes in the digestion system. Such organic materials may include, for example, manure, lignocellulosic material, wastewater biosolids, food waste, energy crops, yeast, guano, agricultural waste, algae, or any combination thereof. The manure may be chicken manure, cow manure, horse manure, sheep manure, alpaca manure, rabbit manure, pig manure, or any combination thereof. In some embodiments, the manure is a mixture of one, two, three, or more manures. Digestion substrates input into digestion systems may have been subject to a partial digestion before being input into the system. Thus, the input into the system may include products of digestion of an original digestion substrate by microbes endogenous to the original digestion substrate, as well as digestible materials still present in the input. In some embodiments, digestion substrates included in an input stream may include an inorganic substrate. The inorganic substrate may include, for example, sand, vermiculite, perlite, pumice, or any combination thereof. In some embodiments, the inorganic substrates comprises a mineral. In some embodiments, the inorganic substrate comprises rock phosphate.

[0147] In some embodiments, a microbial inoculum comprises a single isolated microbe. An isolated microbe may comprise a microbe generated outside of a natural environment (e.g., by plating methods or culture mediums). In some embodiments, the microbial inoculum may comprise between 1 and 5 isolated microbes (e.g., microbial strains). In some embodiments, the inoculum may comprise 1, 2, 3, 4 or 5 isolated microbes. In some embodiments, the inoculum may comprise greater than 5 isolated microbes. In some embodiments, in addition to one or more isolated microbes, a microbial inoculum input into a digestion system may include a complex mixture of microbes, which may include at least 5, 10, 20, 25, 50, 100, 200, 225, 250, 275, 300, 350, 400, or more species of microbes.

[0148] An inoculum of a microbe as described herein may have at least one plant growth promotion property (e.g., a property of plant growth). A plant growth promotion property may comprise shoot biomass, root biomass, nutrient uptake, crop yield, leaf area, chlorophyll content, photosynthetic activity, phosphate uptake, phosphate solubilization, or total biomass. A digestion system may be configured to enhance production of the inoculum of the microbe. A microbe may be a bacterial species, a fungal species, or an algal species.

[0149] In some embodiments, the inoculum of a microbe may comprise at least two isolated microbes. In some embodiments, the inoculum of a microbe may comprise at least one isolated microbe and at least one non-isolated microbe. In some embodiments, the inoculum of a microbe may comprise 1, 2, 3, 4, 5, or more microbes. An inoculum of a microbe may be transferred to a first container (e.g., reactor) of a digestion system one time, two times, three times, four times, five times, or more. An inoculum of a microbe may be transferred to a second container, a third container, a fourth container, a fifth container, a sixth container, or any container of a system described herein.

[0150] An inoculum of a microbe may have a concentration of at least about, at most about, or about 1.0 x 102cfu / ml, 1.0 x 103cfu / ml, 1.0 x 104cfu / ml, 1.0 x 105cfu / ml, 1.0 x 106cfu / ml, 1.0 x 107cfu / ml, 1.0 x 108cfu / ml, 1.0 x 109cfu / ml, 1.0 x 1010cfu / ml, 1.0 x 1011cfu / ml, or 1.0 x 1012cfu / ml, or a range between any of these two values, prior to transferring to a first container of a digestion system. An inoculum of a microbe may have a concentration of at least about, at most about, or about 1.0 x 102cfu / ml, 1.0 x 103cfu / ml, 1.0 x 104cfu / ml, 1.0 x 105cfu / ml, 1.0 x 106cfu / ml, 1.0 x 107cfu / ml, 1.0 x 108cfu / ml, 1.0 x 109cfu / ml, 1.0 x 1010cfu / ml, 1.0 x 1011cfu / ml, or 1.0 x 1012cfu / ml, or a range between any of these two values, after incubation in a digestion system described herein.

[0151] In some embodiments, the inoculum comprises a single microbe. In some embodiments, the inoculum may comprise between 1 and 5 microbes. In some embodiments, the inoculum may comprise at least 1, at least 2, at least 3, at least 4, at least 5, or more microbes. In some embodiments, the inoculum may comprise at most 5, at most 4, at most 3, at most 2, or at most 1 microbe(s).

[0152] In some embodiments, the feedstock (e.g., aqueous organic feedstock) and microbial inoculum may be transferred to the first reactor separately. In some embodiments, the feedstock (e.g., aqueous organic feedstock) is transferred to the first reactor before the microbial inoculum. In some embodiments, the microbial inoculum is transferred to the first reactor before the feedstock (e.g., aqueous organic feedstock). In some embodiments, the feedstock (e.g., aqueous organic feedstock) and microbial inoculum may be transferred to the first reactor together.

[0153] In some embodiments, the aqueous feedstock may not contain the target isolate strain (e.g., an inoculum of the microbe). For example, the aqueous feedstock may not contain the target isolate strain prior to transfer to a first container. The concentration of the target isolate microbial strain may be 0 cfu / ml. In some embodiments, the aqueous feedstock may contain the target isolate strain prior to transfer to a first container. In some embodiments, the aqueous feedstock may contain at most about 5%, at most about 4%, at most about 3%, at most about 2%, at most about 1%, at most about 0.5%, at most about 0.1%, at most about 0.05%, at most about, 0.04%, at most about 0.03%, at most about 0.02%, at most about 0.01%, at most about 0.008%, at most about 0.005%, at most about 0.004%, at most about 0.003%, at most about 0.002%, at most about 0.001%, at most about 0.0001%, or less than about 0.0001% of the target isolate strain. In some embodiments, the aqueous feedstock may not include the microbial strain (e.g., the phosphate- solubilizing microbial strain) at a concentration higher than about 1 CFU / ml, 2 CFU / ml, 3 CFU / ml, 4 CFU / ml, 5 CFU / ml, 6 CFU / ml, 7 CFU / ml, 8 CFU / ml, 9 CFU / ml, 10 CFU / ml, 11 CFU / ml, 12 CFU / ml, 13 CFU / ml, 14 CFU / ml, 15 CFU / ml, 20 CFU / ml, 25 CFU / ml, 30 CFU / ml, 40 CFU / ml, or 50 CFU / ml.

[0154] In some embodiments, the digestion system comprises a clarifier chamber or clarifier tank (CLF), which in some embodiments is a final container in a bioreactor system, as depicted in FIG. 1. The clarifier may comprise a single in-flow port and a single out-flow port. The clarifier may comprise a single in-flow port and multiple out-flow ports. In some embodiments, the clarifier comprises floc-folding wipes which rotate and release microbes that have been immobilized in the floc without introducing solids in the supernatant. The floc-folding wipers may move a working fluid in the clarifier to re-suspend microbes within the working fluid. In some embodiments, the microbes and / or an amount of target isolate strain may be re-suspended in the solution in the clarifier and transferred to the supernatant (e.g., base product). In some embodiments, the clarifier further comprises a flow line to return floc to the first reactor. The flow line may comprise a conduit from the clarifier to a container or combination of containers of the digestion system (e.g., a first container, a second container, a third container, a fourth container, a fifth container, a sixth container, or any combination thereof). The clarifier may return flow to any container (e.g., a first container, a second container, a third container, a fourth container, a fifth container, a sixth container, or any combination thereof) of a digestion system to provide a recirculation of working fluid. The working fluid recirculated from the clarifier may comprise a different microbial community (e.g., different amounts of microbes) than a working fluid of another container in the digestion system (e.g., a first working fluid, a second working fluid, a third working fluid, a fourth working fluid, a fifth working fluid, and / or a sixth working fluid).Without wishing to be bound by theory, the recirculation of flow from the clarifier to a container of the digestion system may help enrich a microbial community of a microbial consortium of a digestion system by providing working fluid from the clarifier to a different point (e.g., container) of the system. The recirculated working fluid may comprise organic materials, microbes of a microbial consortium, a target isolate, metabolites, or any combination thereof.

[0155] In the clarifier, a floc portion of a working fluid (e.g., a clarifier working fluid) may separate from a supernatant portion of a working fluid. The floc-folding wipers of the clarifier may help in separating the working fluid of the clarifier. In some embodiments, the separating may comprise gravity separation. The floc may settle on the bottom of the clarifier and the supernatant may be collected.

[0156] Biostimulant compositions produced by a digestion process as described herein may be used as-is or may be further processed before being used. For example, the outflow from the digestion system, referred to herein as “base product,” may be concentrated, sterilized, filtered, pasteurized, dehydrated before being used, or any combination of these. In some embodiments, the base product may be concentrated 2x, 3x, 4x, 5x, 6x, 7x, 8x, 9x, lOx, or more. In some embodiments, the base product may be filter sterilized to remove any bacteria or other microbes in the composition.

[0157] In an aspect, provided herein is a method comprising transferring a feedstock (e.g., aqueous organic feedstock) into a first container. An inoculum of a microbe may be transferred into a first container. An aqueous feedstock (e.g., a feedstock (e.g., aqueous organic feedstock)) may be transferred into a first container. An aqueous feedstock and an inoculum of a microbe may be transferred into a first container. The first container may comprise a volume of a first working fluid. The feedstock (e.g., aqueous organic feedstock) may comprise a microbial consortium. The feedstock (e.g., aqueous organic feedstock) may comprise digestion products produced by digestion of an organic material. The feedstock (e.g., aqueous organic feedstock) may comprise a microbial consortium and digestion products produced by digestion of an organic material. The organic material may be digested by one or more microbes in the microbial consortium. The organic material may be digested by a population of microbes of the inoculum of a microbe. The digestion products described herein may comprise sugars (e.g., xylose, mannose, glucose, or any combination thereof), metabolites generated by microbes of the working fluid, fatty acids, dead microorganisms, fragments of dead microorganisms, microorganism fermentation products, enzymes, biological plant growth regulators, organic acids, chelators, or any combination thereof. The method may further comprise incubating the inoculum of a microbe under conditions that selectively promotes growth of the microbes and increases the population of the microbes. Themethod may further comprise incubating the inoculum of a microbe under conditions that selectively promotes growth of at least a portion of microbes in the microbial consortium. The terms “microbial digestion” and “digestion” can be used interchangeably.

[0158] In some embodiments, the digestion is anaerobic digestion. In some embodiments, the digestion is aerobic digestion. In some embodiments, the digestion is microaerobic digestion. In some embodiments, the digestion is aerobic digestion, microaerobic digestion, anaerobic digestion, or some combination thereof. Without wishing to be bound by theory, it is believed that during the digestion process, microbes digest the biomolecules and other nutrients present in the manure, yeast, and / or produce digestion products that include compounds that promote plant growth and soil health. In some embodiments of a digestion process, the feedstock (e.g., organic feedstock) may be mixed with water to make a feedstock (e.g., aqueous organic feedstock) for an anaerobic digestion system. The anaerobic digestion system may include a mixing tank in which the feedstock (e.g., organic feedstock) is mixed to make a fluid feed mixture or working fluid. In some embodiments, the fluid feed mixture may include manure, water, and Saccharomyces cerevisiae yeast. In some embodiments, anaerobic digestion comprises a process by which bacteria break down organic biomaterials in the absence of oxygen. The biostimulant may also contain microbes that contribute to the plant-beneficial properties of the biostimulant product. The microbes in the biostimulant product may be derived from the microbial population present in the feedstock (e.g., organic feedstock). The reactors of the bioreactor system may comprise mixers that agitate the working fluid and aerate the working fluid. The mixers may be configured to increase aerobic digestion and the increased aeration may facilitate enrichment of microbial communities of a target functionality (e.g., phosphate solubilizing microbes).

[0159] In an exemplary system of the present disclosure, a series of reactors functions to contribute to the growth of an inoculum of a microbe (e.g., isolate) having desired plant growth promoting properties. A series of reactors (e.g., serialized assembly of reactors) may also function to contribute to the production of microbial metabolites having desired plant growth promoting properties. The digestion system described herein can enrich an inoculum of a target microbe, a population of microbes within a microbial consortium with plant-growth promotion properties (e.g., phosphate solubilization), a population of metabolites with plant-growth promotion properties (e.g., phosphate solubilization), or any combination thereof. This system provides added benefits to other digestion systems in that it can target a functional community of microbes and / or metabolites with specific functionality and enrich and / or maintain the community in the digestion system. The system can comprise two, three, four, five, or more reactors chambers (e.g., containers or chambers). Without wishing to be bound by theory, the serialized reactors enablethe growth and enrichment of proprietary specialist target microbes with optimal plant growth promoting properties. The system may direct a flow of working fluid comprising an inoculum, carbon source, nutrient source, or any combination thereof, from an input feedstock (e.g., organic feedstock) to produce a base product (BP). A hydraulic source can flow into a reactor via in-flow port to comprise a first working fluid in a reactor tank. A substance providing selective pressure can flow into a reactor via in-flow port to comprise a first working fluid in a reactor tank. A hydraulic source and a substance providing selective pressure can flow into a reactor via in-flow port to comprise a first working fluid in a reactor tank. A hydraulic source may input into a first reactor or any reactor of the system. In some embodiments, a hydraulic source may input (e.g., flow) into a tank or container prior to a first reactor. In some embodiments, the container may comprise a “complete mixed reactor” (CMR). Other inputs into a system described herein may flow into any reactor of the system, including but not limited to a first reactor, a second reactor, a third reactor, or any other reactor following a first reactor.

[0160] An inoculum of a microbe (e.g., a target microbial strain) may incubate in a reactor (e.g., container) of a digestion system described herein. In some embodiments, the inoculum of a microbe may be incubated under conditions that selectively enrich and / or maintain a concentration of the microbial strain in the digestions system. These conditions to selectively enrich and / or maintain a population of the microbial strain may comprise an addition of a selective pressure source. In some cases, the population of the microbial strain may survive in the digestion system in a vegetative or sporulated state (e.g., a dormant state in the system). In some cases, the selective pressure source (e.g., selective pressure) may establish an environment to enhance the survival of the population of the microbial strain. Without wishing to be bound by theory, the selective pressure source may shift the complex microbial consortia of the digestion system to enrich at least a portion of microbes within a microbial consortium with plant growth promotion properties (e.g., phosphate solubilization). Incubation of the inoculum of the microbe and / or the portion of microbes within a microbial consortium with plant growth promotion properties may further generate metabolites with plant growth promotion properties (e.g., phosphate solubilization).

[0161] The inoculum of the microbe may comprise a phosphate-solubilizing microbe that can be maintained at a higher ion concentration in a working fluid caused by addition of the selective pressure. Survival of an inoculum of a microbe may comprise the inoculum of a microbe configured to maintain its initial amount in the environment caused by addition of the selective pressure source. Survival of an inoculum of a microbe may comprise an instance where an amount of the inoculum of the microbe is alive (e.g., maintained) at the end of a retention period of the digestion system (e.g., in a reactor or clarifier chamber). Without wishing to be bound by theory,the inoculum of a microbe may not be susceptible to the change in ionic concentration or may have solubilizing properties for the ion which may allow it to survive in the working fluid. Following addition of the selective pressure, at least a portion of microbes of the microbial consortium may enrich (e.g., grow or increase in number). These microbes of the portion of microbes in the microbial consortium may have phosphate solubilization capacities. Following addition of the selective pressure source, other microbes of the working fluid may reduce in number as the microbes of the microbial consortium are unable to survive (e.g., are susceptible) in the ionic environment. A proportion of the phosphate-solubilizing microbial strain relative to at least a portion of the microbes in a microbial consortium may be maintained in a first container of a digestion system. A proportion of the phosphate-solubilizing microbial strain relative to at least a portion of the microbes in a microbial consortium may be maintained in a second, third, fourth, fifth, sixth, seventh, or eighth container of a digestion system. A maintained phosphate- solubilizing microbial strain may change its amount in a working fluid of a digestion system less than about 0.001%, less than about 0.01%, less than about 0.1%, less than about 0.5%, less than about 1%, less than about 5%, or less than about 10% over a measured time period.

[0162] In some cases, the change in ionic concentration caused by the addition of the selective pressure may promote the growth of microbes or at least a portion of microbes in the microbial consortium. These microbes may be phosphate-solubilizing microbes. An amount of microbes or at least a portion of microbes in the microbial consortium may grow by at least about, at most about, or about 0.0001%, 0.001%, 0.01%, 0.1%, 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, or 30%, or a range between any of these values, following addition of the selective pressure. In some cases, at least a portion of phosphate-solubilizing microbes in the microbial consortium may enrich and / or grow in the system without addition of the inoculum of the microbial strain.

[0163] A selective pressure source may be added to a digestion system on a first day of a digestion process. In some embodiments, a selective pressure source may be added to the digestion system daily. In some embodiments, a selective pressure source may be added to the digestion system every 2 days, every 3 days, every 4 days, every 5 days, every 6 days, and / or weekly.

[0164] Nutrients (e.g., macronutrients, micronutrients, inorganic nutrients, or any combination thereof) can be present in the digestion system to provide an environment for bacterial growth. Macronutrients (phosphorus, carbon, nitrogen, or any combination thereof) may help enrich a population of microbes with a target functionality (e.g., phosphate solubilization). The addition of a selective pressure (e.g., an insoluble phosphate source) with inputs to facilitate microbial growth can allow for enrichment of phosphate solubilizers in the working fluid of a digestion system described herein. The phosphate solubilizers can comprise the inoculum of the microbe (e.g., apopulation of the inoculum of the microbe), phosphate-solubilizing microbes of a microbial consortium, phosphate-solubilizing metabolites produced by the inoculum of the microbe, the phosphate-solubilizing microbes of the microbial consortium, or any combination thereof.

[0165] An inoculum of a microbe described herein may contact (e.g., be applied to) a plant. In some embodiments, the contacting of an inoculum of a microbe to a plant may enhance at least one plant growth promotion property of the plant. In some embodiments, one, two, three, four, or more inoculums of a microbe may be transferred to a digestion system. An inoculum of a microbe and another inoculum of a microbe may be the same. An inoculum of a microbe and another inoculum of a microbe may be different. In some embodiments, the inoculum of the microbe and the feedstock (e.g., aqueous organic feedstock) are transferred to a container of the digestion system at the same time. In some embodiments, the inoculum of the microbe and the feedstock (e.g., aqueous organic feedstock) are not transferred to a container of the digestion system at the same time. In some embodiments, the inoculum of the microbe is transferred to a container of the digestion system prior to the feedstock (e.g., aqueous organic feedstock). In some embodiments, the inoculum of the microbe is transferred to a container of the digestion system after the feedstock (e.g., aqueous organic feedstock).

[0166] In some embodiments, the selective pressure can enrich a population of phosphate solubilizing microbes (e.g., phosphate solubilizer) in a container and / or containers of a digestion system. A population of phosphate solubilizing microbes can comprise an inoculum of a target phosphate solubilizing microbe, phosphate solubilizing microbes of a microbial consortium, phosphate solubilizing metabolites, or any combination thereof.

[0167] In some embodiments, as a working fluid flows through a digestion system, an absolute number of phosphate-solubilizing microbes may increase. In some embodiments, the absolute number of phosphate-solubilizing microbes may stay substantially the same (e.g., within 80% of a reference number) or decrease, but phosphate-solubilizing activity of microbes in the system may increase. The numbers of phosphate-solubilizing microbes in a fluid may be measured by plating a portion of the fluid on culture plates with a media containing insoluble P as the only P source and counting the number of colonies that form halos. In some embodiments, an absolute number of phosphate solubilizers can be higher, the same, or lower in a subsequent container of a bioreactor system compared to an absolute number of phosphate solubilizers in a previous container in a bioreactor system. Despite having similar absolute numbers of phosphate solubilizers in the various containers of a bioreactor system, overall phosphate solubilizing activity of microbes in the system may be stimulated by providing insoluble phosphate as the primary or only source of phosphate in the system.

[0168] As working fluid flows through a digestion system, a proportion of a particular phosphate- solubilizer or of all phosphate-solubilizers relative to a total population of bacteria may increase, stay substantially the same, or decrease. In some embodiments, a proportion of phosphate- solubilizers relative to a total population of bacteria can be higher, substantially the same, or lower in a subsequent container in a bioreactor system compared a proportion of phosphate-solubilizers relative to a total population of bacteria in a previous container in the system. Despite having similar absolute and / or relative numbers of phosphate solubilizers in the various containers of a bioreactor system, overall phosphate solubilizing activity of microbes in the system may be stimulated by providing insoluble phosphate as the primary or only source of phosphate in the system.

[0169] An inoculum of a microbe may generate metabolites in a digestion system as described herein. Microbes of the microbial consortium and / or the inoculum of the microbe may be metabolized by catalytic enzymes to produce metabolites. Metabolites may be generated by microbial metabolism. Metabolites may be generated by enzymes catalyzing biochemical reactions of the organic substrates of the feedstock (e.g., aqueous organic feedstock) in a working fluid of a digestion system as described herein. The metabolites generated by the inoculum of the microbe may have a plant growth promotion property. The metabolites generated by the inoculum of the microbe may have two or more plant growth promotion properties. The plant growth promotion properties may comprise shoot biomass, root biomass, nutrient uptake, crop yield, deaminase activity, acid production, leaf area, chlorophyll content, heat tolerance, cold tolerance, drought tolerance, or salt tolerance, or total biomass. Metabolites may be used in biostimulant compositions and / or may be applied to plants.

[0170] In some embodiments, the feedstock (e.g., aqueous organic feedstock) comprises metabolites. In some embodiments, the feedstock (e.g., aqueous organic feedstock) comprises metabolites produced by microbes endogenous to the feedstock (e.g., organic feedstock). Primary metabolites can include carbohydrates, proteins, fats, vitamins, and nucleic acid components. Metabolites can further comprise alkaloids, amino acids, biogenic amines, carboxylic acids, cresols, terpenoids, phenols (e.g., flavonoids, coumarins, tannins, lignans, stilbenes, or chromones), polyketides, eicosanoids, hormones or derivatives thereof, indoles or derivatives thereof, nucleobases, citric acid, ceramides, diglycerides, triglycerides, amides, alkanes, alcohols, stearates, sterols, organic acids or fatty acids. In some embodiments, metabolites comprise sugars and / or fatty acids. Sugars can comprise fructose, hexose, galactose, glucose, lactose, maltose, sucrose, xylose or any combination thereof. Fatty acids can comprise stearic acid, lauric acid, myristic acid, palmitic acid, octadecenoic acid, octadecadienoic acid, oleic acid, arachidic acid,behenic acid, erucic acid, adrenic acid, tricosanoic acid, lignoceric acid, nervonic acid, nonadecanoic acid, arachidic acid, myristolic acid or hydroxylated myristic acid.

[0171] Metabolites generated by the inoculum of the microbe or by at least a portion of microbes of the microbial consortium may be present in a supernatant (e.g., base product) of a digestion system. In some embodiments, the metabolites may be present by weight in a volume of solution (e.g., in mg in 100 ml). In some embodiments, a weight of metabolites per 100 ml of a base product solution can be at least about 10 mg, at least about 20 mg, at least about 30 mg, at least about 40 mg, at least about 50 mg, at least about 60 mg, at least about 70 mg, at least about 80 mg, at least about 90 mg, at least about 100 mg, at least about 120 mg, at least about 140 mg, at least about 160 mg, at least about 180 mg, at least about 200 mg, at least about 220 mg, at least about 240 mg, at least about 260 mg, at least about 280 mg, at least about 300 mg, at least about 400 mg, at least about 500 mg, or greater than about 500 mg. In some embodiments, a weight of metabolites per 100 ml of a base product solution can be at most about 500 mg, at most about 400 mg, at most about 300 mg, at most about 280 mg, at most about 260 mg, at most about 240 mg, at most about 220 mg, at most about 200 mg, at most about 180 mg, at most about 160 mg, at most about 140 mg, at most about 120 mg, at most about 100 mg, at most about 90 mg, at most about 80 mg, at most about 70 mg, at most about 60 mg, at most about 50 mg, at most about 40 mg, at most about 30 mg, at most about 20 mg, at most about 10 mg, or less than about 10 mg. In some embodiments, a weight of metabolites per 100 ml of a base product solution can be from about 10 mg to about 500 mg. In some embodiments, a weight of metabolites per 100 ml of a base product solution can be from about 10 mg to about 20 mg, about 10 mg to about 30 mg, about 10 mg to about 40 mg, about 10 mg to about 50 mg, about 10 mg to about 75 mg, about 10 mg to about 100 mg, about 10 mg to about 125 mg, about 10 mg to about 150 mg, about 10 mg to about 175 mg, about 10 mg to about 250 mg, about 10 mg to about 500 mg, about 20 mg to about 30 mg, about 20 mg to about 40 mg, about 20 mg to about 50 mg, about 20 mg to about 75 mg, about 20 mg to about 100 mg, about 20 mg to about 125 mg, about 20 mg to about 150 mg, about 20 mg to about 175 mg, about 20 mg to about 250 mg, about 20 mg to about 500 mg, about 30 mg to about 40 mg, about 30 mg to about 50 mg, about 30 mg to about 75 mg, about 30 mg to about 100 mg, about 30 mg to about 125 mg, about 30 mg to about 150 mg, about 30 mg to about 175 mg, about 30 mg to about 250 mg, about 30 mg to about 500 mg, about 40 mg to about 50 mg, about 40 mg to about 75 mg, about 40 mg to about 100 mg, about 40 mg to about 125 mg, about 40 mg to about 150 mg, about 40 mg to about 175 mg, about 40 mg to about 250 mg, about 40 mg to about 500 mg, about 50 mg to about 75 mg, about 50 mg to about 100 mg, about 50 mg to about 125 mg, about 50 mg to about 150 mg, about 50 mg to about 175 mg, about 50 mg to about 250 mg, about 50mg to about 500 mg, about 75 mg to about 100 mg, about 75 mg to about 125 mg, about 75 mg to about 150 mg, about 75 mg to about 175 mg, about 75 mg to about 250 mg, about 75 mg to about 500 mg, about 100 mg to about 125 mg, about 100 mg to about 150 mg, about 100 mg to about175 mg, about 100 mg to about 250 mg, about 100 mg to about 500 mg, about 125 mg to about150 mg, about 125 mg to about 175 mg, about 125 mg to about 250 mg, about 125 mg to about500 mg, about 150 mg to about 175 mg, about 150 mg to about 250 mg, about 150 mg to about500 mg, about 175 mg to about 250 mg, about 175 mg to about 500 mg, or about 250 mg to about 500 mg.

[0172] The term “feedstock” (e.g., “organic feedstock”) described herein can refer to raw biomaterials such as carbon compounds, proteins, and / or carbohydrates. The feedstock may be organic feedstock. The feedstock may be inorganic feedstock. The organic feedstock may be a feedstock (e.g., aqueous organic feedstock). In some cases, the organic feedstock may comprise one or more inorganic nutrients. In some embodiments, the feedstock (e.g., organic feedstock) may comprise organic substrates comprising cottonseed, algae, neem, orange seed, linseed, jojoba, kusum, rubber seed, alfalfa, sugarcane, Opuntia, coffee, Deccan hemp, or any combination thereof. In some embodiments, the feedstock can comprise an inorganic feedstock. In some embodiments, the feedstock (e.g., organic feedstock) may include, but is not limited to, manure, kelp, lignocellulose, wastewater biosolids, food waste, energy crops, glucose solution, ammonium sulfate, oils, fats, grease, or any combination thereof. In some embodiments, the feedstock is added at the beginning of the system (e.g., into a first reactor and / or a CMR). In some embodiments, the feedstock is added in a middle reactor of the system (e.g., not in the first or last reactor of the system). In some embodiments, the feedstock is added once to the system. In some embodiments, the feedstock is added two, three, four, or more times to the system. In some embodiments, the feedstock (e.g., organic feedstock) is a composition of one raw biomaterial. In some embodiments, the feedstock (e.g., organic feedstock) is a blend of two, three, four, five, six, seven, eight, nine, ten, or more biomaterials, is a blend of two, three, four, five, six, seven, eight, nine, ten, or more biomaterials.

[0173] Organic feedstock comprising carbon and nitrogen sources can flow into a reactor tank. Organic feedstock comprising carbon and nitrogen sources can flow into a reactor tank via a conduit (e.g., a pipe). The feedstock (e.g., organic feedstock) may further comprise microbial strain having a desired plant growth promoting property (e.g., a phosphate-solubilizing microbial strain). In some embodiments, the organic feedstock (e.g., aqueous feedstock or aqueous organic feedstock) can comprise a concentration of the microbial strain (e.g., the phosphate-solubilizing microbial strain) of at most about 15 CFU / ml, at most about 14 CFU / ml, at most about 13 CFU / ml,at most about 12 CFU / ml, at most about 11 CFU / ml, at most about 10 CFU / ml, at most about 9 CFU / ml, at most about 8 CFU / ml, at most about 7 CFU / ml, at most about 6 CFU / ml, at most about 5 CFU / ml, at most about 4 CFU / ml, at most about 3 CFU / ml, at most about 2 CFU / ml, at most about 1 CFU / ml, or less than about 1 CFU / ml.

[0174] In some embodiments, a reactor tank circulates working fluid within itself to recycle working fluid, wherein reactor tanks can comprise out-flow pipes to circulate and recycle working fluid within each tank. Ports and piping between tanks can assist in transferring working fluid to adjacent reactor tanks. A working fluid in a final clarifier of a system may transfer from the reactor tank to a clarifier may produce a supernatant (e.g., base product). Working fluid flows through the serialized reactor system which can aid in selective growth of the added isolate and other microbes present that have the same property as the added isolate. Base product from the clarifier can be accessed and further analyzed for microbial composition. In a digestion system described herein, a working fluid may flow from a mixing chamber through at least one reaction and to a clarifier chamber.

[0175] A feedstock (e.g., organic feedstock) can comprise digestion products from digestion of organic substrates present in the feedstock (e.g., organic feedstock). Organic substrates may improve stability of the fluid feed mixture. Organic substrates can comprise coconut coir, peat moss, hemp, wood fiber, or any combination thereof. In some embodiments, organic substrates comprise raw biomaterials present in the feedstock (e.g., aqueous organic feedstock). A digestion system may comprise a plurality of microbes and / or microorganisms derived from digestion of organic substrates in a feedstock (e.g., aqueous organic feedstock).

[0176] A feedstock (e.g., organic feedstock) described herein may comprise various organic and / or biological materials. In some embodiments, the feedstock (e.g., organic feedstock) further comprises Saccharomyces cerevisiae yeast, Saccharomyces arboricola yeast, Saccharomyces mikatae yeast, Saccharomyces jurei yeast, Saccharomyces eubayanus yeast, Saccharomyces kudriavzevii yeast, Saccharomyces uvarum yeast, or any combination thereof. In some embodiments, the feedstock (e.g., organic feedstock) may be an aqueous mixture of at least one feedstock material and water. In some embodiments, the feedstock (e.g., organic feedstock) may be an aqueous mixture of cow manure, S. cerevisiae yeast, water, or any combination thereof.

[0177] Parameters of the digestion system, such as flow rate and the solids content of the feedstock (e.g., organic feedstock), may be varied to achieve desired properties in the outflow biostimulant base product. In some embodiments, the hydraulic source is water. In some embodiments, the hydraulic source is a base product of another system. In some embodiments, the hydraulic source is a combination of water and a base product of another system. Water fromthe hydraulic source can be added to the feedstock (e.g., organic feedstock) of the digestion system to make an aqueous organic feedstock.

[0178] In some embodiments, the feedstock (e.g., aqueous organic feedstock) may further comprise a inorganic substrate. In some embodiments, the feedstock (e.g., aqueous organic feedstock) may include more than one inorganic substrate. The inorganic substrate may improve stability of the fluid feed mixture. In some embodiments, the inorganic substrate comprises sand, vermiculite, perlite, diatomaceous earth, pumice, or any combination thereof. In some embodiments, the inorganic substrates comprises a mineral. In some embodiments, the inorganic substrate comprises rock phosphate.

[0179] In some embodiments, loading inputs into a reactor can comprise a carbon source, a nitrogen source, a flour, an isolate, or any combination thereof. In some embodiments, the loading inputs comprise recycled floc from the system. In some embodiments, the loading inputs comprise a whole broth (WB). The inoculum of a microbe as described herein may metabolize the carbon source. Metabolism of carbon by the inoculum of the microbe may comprise transfer of carbonbased moieties of the carbon source to substrates in the working fluid.

[0180] The inoculum of a microbe as described herein may metabolize the nitrogen source. Metabolism of nitrogen by the inoculum of the microbe may comprise transfer of nitrogen-based moieties of the nitrogen source to substrates in the working fluid. In some embodiments, the carbon source may be transferred to a first container of the digestion system. In some embodiments, the carbon source may be transferred to a second, third, fourth, fifth, or sixth container of the digestion system. In some embodiments, the nitrogen source may be transferred to a first container of the digestion system. In some embodiments, the nitrogen source may be transferred to a second, third, fourth, fifth, or sixth container of the digestion system.

[0181] In some embodiments, the feedstock (e.g., organic feedstock) is mixed within a reactor. In some embodiments, the feedstock (e.g., organic feedstock) is mixed outside of a reactor. In some embodiments, the feedstock (e.g., organic feedstock) is mixed between one, two, three, or more reactors. In some embodiments, the feedstock (e.g., organic feedstock) is a homogenous mixture.

[0182] In some embodiments, the feedstock (e.g., organic feedstock) further comprises a microbial consortium. The terms “microbe”, “microbial strain” and “microorganism” may refer to microscopic organisms, comprising bacteria, fungi, lichens, algae, protozoa, archaea, molds, or any combination thereof. The terms “microbe” and microorganism” may be used interchangeably herein. The feedstock (e.g., organic feedstock) can comprise a microbial consortium with 2, 3, 4, 5, 6, 7, 8, 9, 10, or more microorganisms. The feedstock (e.g., organic feedstock) can comprise a microbial consortium with 2, 3, 4, 5, 6, 7, 8, 9, 10, or more groups of microorganisms. Thefeedstock (e.g., organic feedstock) can comprise a microbial consortium with 1 group of microorganisms. The microbial consortium can comprise different microorganisms. The microbial consortium can comprise the same microorganism. The microbes in the consortia may be derived from the microbes originally present within the feedstock (e.g., organic feedstock). The microbes may digest the manure, yeast, other organic raw materials, or any combination thereof to produce digestion products.

[0183] In some embodiments, microbes can be added to the start of the system (e.g., into the first reactor). In some embodiments, microbes can be added to the middle of the system (e.g., into a reactor that is not the first reactor or the final reactor of the system) or microbes can be added to the end of the system (e.g., into the final reactor). Microbes can be added concurrently with the feedstock (e.g., organic feedstock). Microbes can be added separately from the feedstock (e.g., organic feedstock). In some embodiments, microbes may be added to the system with the feedstock (e.g., organic feedstock) in the same reactor. In some embodiments, microbes may be added to the system with the feedstock (e.g., organic feedstock) in different reactors. In some embodiments, microbes may be added to the system prior to the feedstock (e.g., organic feedstock). In some embodiments, microbes may be added to the system after the feedstock (e.g., organic feedstock). In some embodiments, a period of time between addition of microbes to the system and addition of feedstock (e.g., organic feedstock) to the system can be at least about 1 minute, at least about 5 minutes, at least about 10 minutes, at least about 15 minutes, at least about 20 minutes, at least about 30 minutes, at least about 40 minutes, at least about 50 minutes, or at least about 1 hours. In some embodiments, a period of time between addition of microbes to the system and addition of feedstock (e.g., organic feedstock) to the system can be at most about 2 hours, at most about 1 hours, at most about 45 minutes, at most about 30 minutes, at most about 15 minutes, at most about 10 minutes, at most about 5 minutes, at most about 1 minute, or at most about 30 seconds.

[0184] A microbe may have nutrient solubilization properties and / or plant growth promotion properties. For example, a microbe may increase plant growth, increase shoot and / or root biomass, increase crop yield, increase soil enzymatic activity, increase photosynthesis efficiency, lower heavy metal uptake, decrease soil pH, or any combination thereof. A microbe may enhance plant growth on land with high salinity, on land with heavy metal contamination, or on land with drought conditions. A microbe (e.g., isolate) described herein may have phosphate solubilization properties.

[0185] In some embodiments, the digestion system may comprise a retention time. A retention time may comprise a time an inoculum of a microbe spends in a digestion system or a time aninoculum of a microbe spends following transfer into a first container and until collection from the digestion system. A longer retention time may be advantageous for growth or enrichment of an inoculum of a microbe of the digestion system. A shorter retention time may be advantageous for growth or enrichment of an inoculum of a microbe of the digestion system. A retention time of a digestion system may comprise at least about, at most about, or about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 6 months, 9 months, 1 year, 2 years, 3 years, 4 years, 5 years, or 10 years, or a range between any of these values.

[0186] In the first reactor, the working fluid may be agitated at a rate that allows heavier or undigested solids to settle to the bottom. An outlet at the top of the first reactors may allow the fluid to flow into the second reactor. An outlet at the bottom of the first reactor may transfer the settled solids back into a reactor tank. Each of the reactors in the series of reactors may have submerged scaffolding that provide a surface for biofilm growth. The scaffolding can be referred to as “fixed media substrates”. The flow of fluid from reactor to reactor may comprise a plug flow model, in which particles of an input fluid have the same velocity and direction of motion. In some embodiments, the flow of fluid in the digestion system is driven by gravity. In some embodiments, the flow of fluid in the digestion system is driven by a pump. The outflow from the top of the last reactor, may be used to create a product. Products of the methods and systems as described herein can be biostimulants. Biostimulants can promote plant growth or improve soil quality.C. Reactors

[0187] A digestion process to produce the biostimulant may be performed in a digestion system that includes a tank, a container, a vessel (e.g., reactor), or a series of tanks, a series of containers, or a series of vessels (e.g., reactors) through which the feedstock continuously flows. A reactor can be a fluidly connected container, system, vessel, or tank in which microbial consortia including the microbes, isolates, and / or microorganisms as described herein can be grown. A reactor can be separate or continuous. A reactor may be a physical containment system arranged in a discrete order to favor growth of particular microbes. Types of reactors can include, but are not limited to, fluidized-bed reactors (FBRs) or packed-bed reactors (PBRs).

[0188] In some embodiments, reactors may be arranged so that fluid can flow from an outflow port of a reactor into an adjacent reactor or tank. Fluid from near the top of the working fluid in a reactor may flow into the next reactor continuously. Fluid from the middle of a reactor may flow into the next reactor continuously. Fluid from the bottom of a reactor may flow into the next reactor continuously. Fluid may also be reintroduced from any outflow source into the samereactor. In some embodiments, an outflow port is between 0.1 and 35 inches below the top of the working fluid within a reactor. In some embodiments, an outflow port is at least about 1 inch, at least about 2 inches, at least about 5 inches, at least about 10 inches, at least about 20 inches, at least about 50 inches, at least about 100 inches, at least about 250 inches, at least about 500 inches, at least about 750 inches, at least about 1,000 inches, at least about 2,500 inches, at least about 5,000 inches, at least about 7,500 inches, or at least about 10,000 inches below the top of the working fluid within a reactor. In some embodiments, an outflow port is at most about 10,000 inches, at most about 7,500 inches, at most about 5,000 inches, at most about 2,500 inches, at most about 1,000 inches, at most about 750 inches, at most about 500 inches, at most about 250 inches, at most about 100 inches, at most about 50 inches, at most about 20 inches, at most about 10 inches, at most about 5 inches, at most about 2 inches, or at most about 1 inch below the top of the working fluid within a reactor. In some embodiments, the rate of outflowing product from a digestion system may match the rate of inflowing feedstock, providing for a hydraulically balanced flow throughout the system. A reactor within the system may have a unique, stable microbial consortium with distinct physiological characteristics and digestion capabilities as compared to consortia in other tanks in the system. A reactor within the system may have the same microbial consortium with similar physiological characteristics and digestion capabilities as another reactor within the system. Each reactor within the system may have the same volume capacity. Each reactor within the system may have a different volume capacity. The digestion system may comprise at least two reactors. The digestion system may comprise 2, 3, 4, 5, 6, 7, 8, 9, 10, or more reactors. Reactors of a digestion system may be arranged as a serialized assembly of reactors. A serialized assembly of reactors may have conduits (e.g., ports or outputs) connecting each reactor to an adjacent reactor and / or container. A serialized assembly of reactors may have a continuous flow of working fluid through each reactor to the adjacent reactor.

[0189] In some embodiments, a reactor may have a single in-flow port and a single out-flow port. In some embodiments, a reactor may have multiple in-flow ports and out-flow ports. In some embodiments, a reactor may have a single in-flow port and multiple out-flow ports. In some embodiments, a reactor may have multiple in-flow ports and a single out-flow port. A reactor may have another in-flow port to provide a carbon source and / or consortium inoculum. An in-flow port may be present at any location of a reactor of the digestion system. An in-flow port may be present at the top of the reactor or at the bottom of the reactor. An out-flow port may be present at the top of the reactor or at the bottom of the reactor. In some embodiments, the out-flow ports or in-flow ports described herein comprise pipes, pumps, ventilations, or other systems for transferring fluid from one vessel to another.

[0190] A reactor may have a single fluid connection. A reactor may have multiple fluid connections. The fluid connections may be present at the top of the working fluid in each reactor or near the top of the working fluid in each reactor. In some embodiments, the reactor may have flow from the bottom of the container back to the top to prevent build-up of sludge in the bottom of the reactor. In some embodiments, a reactor may have stirrers in the bottom of the container. In some embodiments, a reactor may have wipers in the bottom of the container. The wipers may stir the feedstock and prevent clogging within the reactor. The wipers may fold floc and ease floc return.

[0191] In some embodiments, the reactors may comprise packed bed reactors. In some embodiments, each of the packed bed reactors has an open cell design to allow free movement of working fluid. A fixed media (e.g., scaffolding) may be secured to the inside of each packed bed reactor. The fixed media comprises materials that increase the contact surface area for the communities of microbes with working fluid. The fixed media also provides a stable platform for anchoring biofilm. The packed bed reactors may be packed with scaffolding to increase surface area within the reactor. The scaffolding within the reactor may increase biofilm. The packed bed reactor may improve contact between the biofilm and substrates within the reactor. The fixed media can be of several types, including durable plastic, polyvinyl chloride (PVC), metal, metal alloy, glass, glass compounds, fiberglass, or any suitably robust inert material. The design and configuration of the fixed media can assume various geometric patterns that allow working fluid to freely move through each packed bed reactor and prevents fouling. Free flow supports controlled hydraulic shearing which in time promotes even distribution of working fluid. In this embodiment, the fixed media is dispersed throughout a cross sectional area of each packed bed reactor.

[0192] The scaffolding may comprise tubes, rings, or other packing materials. In some embodiments, the packed bed reactors provided herein may comprise a bundle of tubes or columns. In some embodiments, the scaffolding may comprise hexagonal, grid-like, perforated tubing, or any combination thereof. Without wishing to be bound by theory, hexagonal, grid-like, perforated scaffolding, or any combination thereof, can increase the surface area and the flow through the columns within the container. In some embodiments, the tubes or columns of the scaffolding can comprise a diameter between 0.25 and 50 inches. In some embodiments, the scaffolding can comprise a diameter of at least about 0.5 inches, at least about 0.6 inches, at least about 0.7 inches, at least about 0.8 inches, at least about 0.9 inches, at least about 1 inch, at least about 2 inches, at least about 3 inches, at least about 4 inches, at least about 5 inches, at least about 10 inches, at least about 15 inches, at least about 20 inches, at least about 25 inches, at least about30 inches, at least about 40 inches, at least about 50 inches, at least about 60 inches, or at least about 75 inches. In some embodiments, the scaffolding can comprise a diameter of at most about 75 inches, at least about 60 inches, at most about 50 inches, at most about 40 inches, at most about 30 inches, at most about 25 inches, at most about 20 inches, at most about 15 inches, at most about 10 inches, at most about 5 inches, at most about 4 inches, at most about 3 inches, at most about 2 inches, at most about 1 inches, at most about 0.9 inches, at most about 0.8 inches, at most about 0.7 inches, at most about 0.6 inches, or at most about 0.5 inches. Without wishing to be bound by theory, a system with packed bed reactors may improve production of bacterial isolates or other microbes. In some embodiments, reactors without scaffolding (i.e., reactors that are not packed bed reactors) improve production of bacterial isolates or other microbes or improves digestion of digestible substrates.

[0193] In some embodiments, the reactors may comprise fluidized bed reactors. In fluidized bed reactors, solid particles may be circulated within working fluid of the reactors, which may provide a surface for microbial colonization. Such particles may include, for example, particles of an inorganic substrate such as rock phosphate particles. In some embodiments, the fluidized bed reactors may be the same volume. In some embodiments, the fluidized bed reactors may be different volumes. In some embodiments, the fluidized bed reactors increase uniformity of particle mixing within the digestion system. The solid material of the fluidized bed reactor can have intrinsic fluid-like properties and allow for a more complete mixing. Reduction or elimination of radial and axial concentration gradients can provide for better fluid-solid contact and can achieve better uniformity of particle mixing. In some embodiments, the fluidized bed reactors increase the uniformity of temperature gradients within the digestion system. Without wishing to be bound by theory, the open container of the fluidized bed reactor can provide for a reduction in isolated hot or cold spots in the container, allowing for a uniform temperature distribution of the fluid. Hot spots can be areas of localized temperature changes during heating of a working fluid. Cold spots can be areas of localized temperature changes during cooling of a working fluid. The reduction in isolated temperature changes from the configuration of the fluidized bed reactor can provide for a more uniform temperature distribution of the fluid.

[0194] The flow rate of the digestion system may be chosen to allow for sufficient dwell time within each of the reactor for a stable and unique microbial consortium to form within each of the reactors. In some embodiments, working fluid in each reactor is continuously recycled at a rate ratio in a range of approximately 25: 1 to 35: 1, 25 to 35 gallons per minute of the recycle rate to one gallon per minute of the hydraulic feed rate. In some embodiments, working fluid in each reactor is continuously recycled at a rate ratio of at least about, at most about, or about 10: 1, atleast about 12: 1, at least about 14: 1, at least about 16: 1, at least about 18: 1, at least about 20: 1, at least about 22: 1, at least about 24: 1, at least about 26: 1, at least about 28: 1, at least about 30: 1, at least about 32: 1, at least about 34: 1, at least about 36: 1, at least about 38: 1, at least about 40: 1, at least about 45: 1, or at least about 50: 1, or a range between any of these two values. Working fluid may be recycled by a pump to prevent solids settling and to provide sufficient velocity and hydraulic shear to prevent excessive buildup and sloughing of biofilm. A digestion system provided herein may comprise a first flow rate, second flow rate, third flow rate, fourth flow rate, fifth flow rate, sixth flow rate, or seventh flow rate.

[0195] Reactors can be maintained at specific temperatures which may aid in digestion and growth of microbial consortia within the system. In some embodiments, a temperature of a reactor is at least about 15°C, at least about 20°C, at least about 21°C, at least about 22°C, at least about 23°C, at least about 24°C, at least about 25°C, at least about 26°C, at least about 27°C, at least about 28°C, at least about 29°C, at least about 30°C, at least about 35°C, at least about 40°C, at least about 45°C, or at least about 50°C. In some embodiments, a temperature of a reactor is at most about 50°C, at most about 45°C, at most about 40°C, at most about 35°C, at most about 30°C, at most about 29°C, at most about 28°C, at most about 27°C, at most about 26°C, at most about 25°C, at most about 24°C, at most about 23°C, at most about 22°C, at most about 21°C, at most about 20°C, or at most about 15°C.

[0196] In some embodiments, a temperature of a reactor is about 15°C to about 45°C. In some embodiments, a temperature of a reactor is about 15°C to about 20°C, about 15°C to about 22°C, about 15°C to about 24°C, about 15°C to about 26°C, about 15°C to about 28°C, about 15°C to about 30°C, about 15°C to about 32°C, about 15°C to about 34°C, about 15°C to about 36°C, about 15°C to about 40°C, about 15°C to about 45°C, about 20°C to about 22°C, about 20°C to about 24°C, about 20°C to about 26°C, about 20°C to about 28°C, about 20°C to about 30°C, about 20°C to about 32°C, about 20°C to about 34°C, about 20°C to about 36°C, about 20°C to about 40°C, about 20°C to about 45°C, about 22°C to about 24°C, about 22°C to about 26°C, about 22°C to about 28°C, about 22°C to about 30°C, about 22°C to about 32°C, about 22°C to about 34°C, about 22°C to about 36°C, about 22°C to about 40°C, about 22°C to about 45°C, about 24°C to about 26°C, about 24°C to about 28°C, about 24°C to about 30°C, about 24°C to about 32°C, about 24°C to about 34°C, about 24°C to about 36°C, about 24°C to about 40°C, about 24°C to about 45°C, about 26°C to about 28°C, about 26°C to about 30°C, about 26°C to about 32°C, about 26°C to about 34°C, about 26°C to about 36°C, about 26°C to about 40°C, about 26°C to about 45°C, about 28°C to about 30°C, about 28°C to about 32°C, about 28°C to about 34°C, about 28°C to about 36°C, about 28°C to about 40°C, about 28°C to about 45°C,about 30°C to about 32°C, about 30°C to about 34°C, about 30°C to about 36°C, about 30°C to about 40°C, about 30°C to about 45°C, about 32°C to about 34°C, about 32°C to about 36°C, about 32°C to about 40°C, about 32°C to about 45°C, about 34°C to about 36°C, about 34°C to about 40°C, about 34°C to about 45°C, about 36°C to about 40°C, about 36°C to about 45°C, or about 40°C to about 45°C.

[0197] Reactors may be maintained under aerobic, microaerobic, or anaerobic conditions. The series of reactors in a digestion system may have different aerobic conditions. The series of reactors in a digestion system may have the same aerobic conditions. In some embodiments, a reactor may have the same aerobic condition as an adjacent reactor. In some embodiments, a reactor may have a different aerobic condition than an adjacent reactor. In some embodiments, a digestion system may have aerobic, microaerobic, anaerobic conditions, or any combination thereof.

[0198] In some embodiments, the high aeration of the working fluid of a digestion system described herein can help enrich microbes with a targeted functionality (e.g., phosphate solubilizers). In some embodiments, aerobic conditions comprise conditions with a dissolved oxygen measurement of greater than 2 mg / L. In some embodiments, aerobic conditions comprise conditions with a dissolved oxygen measurement of at least about 2 mg / L mg / L, at least about 3 mg / L mg / L, at least about 4 mg / L, at least about 5 mg / L, at least about 6 mg / L, at least about 7 mg / L, at least about 8 mg / L, at least about 9 mg / L, at least about 10 mg / L, at least about 12 mg / L, at least about 14 mg / L, at least about 15 mg / L, or greater than about 15 mg / L. In some embodiments, aerobic conditions comprise conditions with a dissolved oxygen measurement from about 2 mg / L to about 15 mg / L. In some embodiments, aerobic conditions comprise conditions with a dissolved oxygen measurement from about 2 mg / L to about 3 mg / L, about 2 mg / L to about 4 mg / L, about 2 mg / L to about 5 mg / L, about 2 mg / L to about 6 mg / L, about 2 mg / L to about 7 mg / L, about 2 mg / L to about 8 mg / L, about 2 mg / L to about 9 mg / L, about 2 mg / L to about 10 mg / L, about 2 mg / L to about 12 mg / L, about 2 mg / L to about 14 mg / L, about 2 mg / L to about 15 mg / L, about 3 mg / L to about 4 mg / L, about 3 mg / L to about 5 mg / L, about 3 mg / L to about 6 mg / L, about 3 mg / L to about 7 mg / L, about 3 mg / L to about 8 mg / L, about 3 mg / L to about 9 mg / L, about 3 mg / L to about 10 mg / L, about 3 mg / L to about 12 mg / L, about 3 mg / L to about 14 mg / L, about 3 mg / L to about 15 mg / L, about 4 mg / L to about 5 mg / L, about 4 mg / L to about 6 mg / L, about 4 mg / L to about 7 mg / L, about 4 mg / L to about 8 mg / L, about 4 mg / L to about 9 mg / L, about 4 mg / L to about 10 mg / L, about 4 mg / L to about 12 mg / L, about 4 mg / L to about 14 mg / L, about 4 mg / L to about 15 mg / L, about 5 mg / L to about 6 mg / L, about 5 mg / L to about 7 mg / L, about 5 mg / L to about 8 mg / L, about 5 mg / L to about 9 mg / L, about 5 mg / L to about 10mg / L, about 5 mg / L to about 12 mg / L, about 5 mg / L to about 14 mg / L, about 5 mg / L to about 15 mg / L, about 6 mg / L to about 7 mg / L, about 6 mg / L to about 8 mg / L, about 6 mg / L to about 9 mg / L, about 6 mg / L to about 10 mg / L, about 6 mg / L to about 12 mg / L, about 6 mg / L to about 14 mg / L, about 6 mg / L to about 15 mg / L, about 7 mg / L to about 8 mg / L, about 7 mg / L to about 9 mg / L, about 7 mg / L to about 10 mg / L, about 7 mg / L to about 12 mg / L, about 7 mg / L to about 14 mg / L, about 7 mg / L to about 15 mg / L, about 8 mg / L to about 9 mg / L, about 8 mg / L to about 10 mg / L, about 8 mg / L to about 12 mg / L, about 8 mg / L to about 14 mg / L, about 8 mg / L to about 15 mg / L, about 9 mg / L to about 10 mg / L, about 9 mg / L to about 12 mg / L, about 9 mg / L to about 14 mg / L, about 9 mg / L to about 15 mg / L, about 10 mg / L to about 12 mg / L, about 10 mg / L to about 14 mg / L, about 10 mg / L to about 15 mg / L, about 12 mg / L to about 14 mg / L, about 12 mg / L to about 15 mg / L, or about 14 mg / L to about 15 mg / L. In some embodiments, aerobic conditions comprise conditions with a dissolved oxygen measurement of between 2 mg / L and 10 mg / L. In some embodiments, microaerobic conditions comprise conditions with a dissolved oxygen measurement of less than 2 mg / L. In some embodiments, microaerobic conditions comprise conditions with a dissolved oxygen measurement of at most about 1.99 mg / L, at most about 1.8 mg / L, at most about 1.6 mg / L, at most about 1.5 mg / L, at most about 1.4 mg / L, at most about 1.3 mg / L, at most about 1.2 mg / L, at most about 1.1 mg / L, at most about 1 mg / L, at most about 0.9 mg / L, at most about 0.8 mg / L, at most about 0.7 mg / L, at most about 0.6 mg / L, at most about 0.5 mg / L, at most about 0.4 mg / L, at most about 0.3 mg / L, at most about 0.2 mg / L, at most about 0.1 mg / L, or less than about 0.1 mg / L but not 0 mg / L. In some embodiments, microaerobic conditions comprise conditions with a dissolved oxygen measurement from about 0.1 mg / L to about 1.99 mg / L. In some embodiments, microaerobic conditions comprise conditions with a dissolved oxygen measurement from about 0.1 mg / L to about 0.2 mg / L, about 0.1 mg / L to about 0.3 mg / L, about 0.1 mg / L to about 0.4 mg / L, about 0.1 mg / L to about 0.5 mg / L, about 0.1 mg / L to about 0.8 mg / L, about 0.1 mg / L to about 1 mg / L, about 0.1 mg / L to about 1.2 mg / L, about 0.1 mg / L to about 1.4 mg / L, about 0.1 mg / L to about 1.6 mg / L, about 0.1 mg / L to about 1.8 mg / L, about 0.1 mg / L to about 1.99 mg / L, about 0.2 mg / L to about 0.3 mg / L, about 0.2 mg / L to about 0.4 mg / L, about 0.2 mg / L to about 0.5 mg / L, about 0.2 mg / L to about 0.8 mg / L, about 0.2 mg / L to about 1 mg / L, about 0.2 mg / L to about 1.2 mg / L, about 0.2 mg / L to about 1.4 mg / L, about 0.2 mg / L to about 1.6 mg / L, about 0.2 mg / L to about 1.8 mg / L, about 0.2 mg / L to about 1.99 mg / L, about 0.3 mg / L to about 0.4 mg / L, about 0.3 mg / L to about 0.5 mg / L, about 0.3 mg / L to about 0.8 mg / L, about 0.3 mg / L to about 1 mg / L, about 0.3 mg / L to about 1.2 mg / L, about 0.3 mg / L to about 1.4 mg / L, about 0.3 mg / L to about 1.6 mg / L, about 0.3 mg / L to about 1.8 mg / L, about 0.3 mg / L to about 1.99 mg / L, about 0.4 mg / L to about 0.5 mg / L, about 0.4 mg / L to about 0.8 mg / L, about 0.4mg / L to about 1 mg / L, about 0.4 mg / L to about 1.2 mg / L, about 0.4 mg / L to about 1.4 mg / L, about 0.4 mg / L to about 1.6 mg / L, about 0.4 mg / L to about 1.8 mg / L, about 0.4 mg / L to about 1.99 mg / L, about 0.5 mg / L to about 0.8 mg / L, about 0.5 mg / L to about 1 mg / L, about 0.5 mg / L to about 1.2 mg / L, about 0.5 mg / L to about 1.4 mg / L, about 0.5 mg / L to about 1.6 mg / L, about 0.5 mg / L to about 1.8 mg / L, about 0.5 mg / L to about 1.99 mg / L, about 0.8 mg / L to about 1 mg / L, about 0.8 mg / L to about 1.2 mg / L, about 0.8 mg / L to about 1.4 mg / L, about 0.8 mg / L to about 1.6 mg / L, about 0.8 mg / L to about 1.8 mg / L, about 0.8 mg / L to about 1.99 mg / L, about 1 mg / L to about 1.2 mg / L, about 1 mg / L to about 1.4 mg / L, about 1 mg / L to about 1.6 mg / L, about 1 mg / L to about 1.8 mg / L, about 1 mg / L to about 1.99 mg / L, about 1.2 mg / L to about 1.4 mg / L, about 1.2 mg / L to about 1.6 mg / L, about 1.2 mg / L to about 1.8 mg / L, about 1.2 mg / L to about 1.99 mg / L, about 1.4 mg / L to about 1.6 mg / L, about 1.4 mg / L to about 1.8 mg / L, about 1.4 mg / L to about 1.99 mg / L, about 1.6 mg / L to about 1.8 mg / L, about 1.6 mg / L to about 1.99 mg / L, or about 1.8 mg / L to about 1.99 mg / L. In some embodiments, anaerobic conditions comprise conditions with a dissolved oxygen measurement of 0 mg / L.

[0199] In some embodiments, the reactors comprise high-speed mixers to move a working fluid and increase aeration. The mixers may facilitate an aerobic digestion and enrich microbial communities with a target functionality (e.g., phosphate solubilizing microbes).

[0200] Reactors may be maintained at different pH levels within a digestion system. Reactors may be maintained at the same pH levels within a digestion system. The pH of a reactor in a digestion system may be at least about 4.0, at least about 4.5, at least about 5.0, at least about 5.5, at least about 6.0, at least about 6.5, at least about 7.0, at least about 7.5, at least about 8.0, at least about 8.5, at least about 9.0, at least about 9.5, or at least about 10.0. In some embodiments, the pH of a reactor in a digestion system may be at most about 10.0, at most about 9.5, at most about 9.0, at most about 8.5, at most about 8.0, at most about 7.5, at most about 7.0, at most about 6.5, at most about 6.0, at most about 5.5, at most about 5.0, at most about 4.5, or at most about 4.0.

[0201] In some embodiments, the pH of a reactor in a digestion system may be about 3 to about 9. In some embodiments, the pH of a reactor in a digestion system may be about 3 to about 3.5, about 3 to about 4, about 3 to about 4.5, about 3 to about 5, about 3 to about 5.5, about 3 to about 6, about 3 to about 6.5, about 3 to about 7, about 3 to about 7.5, about 3 to about 8, about 3 to about 9, about 3.5 to about 4, about 3.5 to about 4.5, about 3.5 to about 5, about 3.5 to about 5.5, about 3.5 to about 6, about 3.5 to about 6.5, about 3.5 to about 7, about 3.5 to about 7.5, about 3.5 to about 8, about 3.5 to about 9, about 4 to about 4.5, about 4 to about 5, about 4 to about 5.5, about 4 to about 6, about 4 to about 6.5, about 4 to about 7, about 4 to about 7.5, about 4 to about 8, about 4 to about 9, about 4.5 to about 5, about 4.5 to about 5.5, about 4.5 to about 6, about 4.5to about 6.5, about 4.5 to about 7, about 4.5 to about 7.5, about 4.5 to about 8, about 4.5 to about 9, about 5 to about 5.5, about 5 to about 6, about 5 to about 6.5, about 5 to about 7, about 5 to about 7.5, about 5 to about 8, about 5 to about 9, about 5.5 to about 6, about 5.5 to about 6.5, about 5.5 to about 7, about 5.5 to about 7.5, about 5.5 to about 8, about 5.5 to about 9, about 6 to about 6.5, about 6 to about 7, about 6 to about 7.5, about 6 to about 8, about 6 to about 9, about 6.5 to about 7, about 6.5 to about 7.5, about 6.5 to about 8, about 6.5 to about 9, about 7 to about 7.5, about 7 to about 8, about 7 to about 9, about 7.5 to about 8, about 7.5 to about 9, or about 8 to about 9.

[0202] In some embodiments, the reactors may comprise distribution components (e.g., a distribution ring) that is subsurface of the discharge volume. The distribution component can reduce the amount of surface disruption, and / or keep the environment in the reactor anaerobic.D. Working Fluids and Microbial Consortia

[0203] A working fluid may comprise a fluidic substance that moves through a digestion system as described herein. A working fluid can comprise solid components, liquid components, gaseous components, or any combination thereof. A working fluid can comprise microbial consortia, isolated microbes or inoculum of a microbial strain (e.g., target isolates), additional organic materials, or any combination thereof. The mixture of microbial consortia, isolated microbes (e.g., target isolates), additional organic materials within a working fluid may allow for the expansion of microbes or act as a culture for an inoculum of a microbe to grow. A working fluid may comprise a pH, viscosity, temperature, surface tension, adhesion, a volume, or any combination thereof, that enhances the growth and / or functioning of microbes or microorganisms. In some embodiments, a volume of working fluid within each reactor may be continuously replenished and drawn from. In some embodiments, a volume of working fluid within each reactor is replenished and drawn from in batches (e.g., discontinuously). In some embodiments, a working fluid in a first reactor may comprise a first working fluid. In some embodiments, a working fluid in a second reactor may comprise a second working fluid. In some embodiments, a working fluid in a third reactor may comprise a third working fluid. In some embodiments, a working fluid in a fourth reactor may comprise a fourth working fluid. In some embodiments, a working fluid in a fifth reactor may comprise a fifth working fluid. In some embodiments, at least a portion of the second working fluid may be transferred to the third reactor. In some embodiments, at least a portion of the third working fluid may be transferred to the fourth reactor. In some embodiments, at least a portion of the fourth working fluid may be transferred to the fifth reactor. In some embodiments, a working fluid may be mixed in a reactor (e.g., chamber or container) prior to afirst reactor. In some embodiments, the working fluid in each reactor may be distinct from the working fluid in other reactors in the digestion system. Distinct working fluids may comprise different microbial populations. The different microbial populations may include different microbes, (e.g., bacteria, fungi, algae, or any combination thereof). Distinct working fluids may comprise different concentrations of a target isolate. Distinct working fluids may comprise different concentrations of a carbon source and / or a nitrogen source. Distinct working fluids may comprise different microbial populations, different concentrations of a target isolate, different concentrations of a carbon source, different combinations of a nitrogen source, or any combination thereof. The working fluid in a reactor of a digestion system may be similar to a working fluid of a different reactor of the digestion system.

[0204] The working fluid within each reactor may comprise different enzymes, which may be produced by microbes within the working fluid. An enzyme within a working fluid may comprise a dehydrogenase, a hydrogenase, an oxidase, a catalase, a peroxidase, a phenol o-hydroxylase, a dextransucrase, an aminotransferase, a rhodanese, a carboxylesterase, a lipase, a phosphatase, a nuclease, a phytase, an aryl sulphatase, an amylase, a cellulase, an inulase, a xylanase, a dextranase, a levanase, a poly-galacturonase, a glucosidase, a galactosidase, an invertase, a peptidase, an asparaginase, a glutaminase, an amidase, a urease, an aspartate decarboxylase, a glutamate decarboxylase an aromatic amino acid decarboxylase, or any combination thereof. An enzyme within a working fluid may comprise nitrogenase, 1 -aminocyclopropane- 1 -carboxylate deaminase (e.g., ACC-deaminase), quinoprotein glucose dehydrogenase (e.g., PQQ or quinone), gluconate 2-dehydrogenase, cellulase, endo-l,3(4)-P-glucanase, pectin lyase, or any combination thereof. The working fluid within each reactor may comprise different concentrations of enzymes. The working fluid within each reactor may comprise a different average abundance of an enzyme. An enzyme may be present at an average abundance between 0.001% to 1%. An enzyme may be present at an average abundance of less than 0.001%. An enzyme may be present at an average abundance of greater than 1%. In some embodiments, an enzyme may be present at an average abundance of at least about, at most about, or about 0.0001%, 0.001%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, or a range between any of these two values. The working fluid within each reactor may comprise enzymes with different enzymatic activity. Enzymatic activity may include, but is not limited to, nitrogen fixation, ammonia production, phosphate solubilization, cell wall lysing, or any combination thereof.

[0205] A working fluid may comprise different digestion products from working fluid within other reactors of the system. In some embodiments, a working fluid may comprise digestionproducts from a feedstock (e.g., aqueous organic feedstock) and microbial consortium at least partially derived from a previous working fluid.

[0206] The pH of a working fluid within each reactor may be different from working fluid in other reactors. The pH of a working fluid within each reactor may be the same. The pH of a working fluid may be less than 6. The pH of a working fluid may be greater than 6. The pH of a working fluid may be in a range from 2 to 11. The pH of a working fluid may be at least about, at most about, or about 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, or 11, or a range between any of these two values.

[0207] In some embodiments, a first microbial consortium may be established in a mixing chamber, in which various inputs may be mixed into a homogenous aqueous mixture to be input into a digestion reactor. In some embodiments, a digestions system described herein may comprise one or more mixing chambers in which a microbial consortium may be established. A mixing chamber may be fed an input or a series of inputs (e.g., target microbial strain, aqueous feedstock, carbon source, nitrogen source, nutrients, water, or any combination thereof) which may be mixed prior to being inputted into a first container (e.g., a first reactor). The first microbial consortium may be derived from microbes originally present in one or more digestion substrates and / or from other inputs into the mixing chamber. Inputs may include, for example, manure, digestion products of manure, and microbes derived from manure. In some embodiments, a second microbial consortium is established in a first reactor. The second microbial consortium may be derived from microbes within the mixing chamber. In some embodiments, a third microbial consortium is established in a second reactor. The third microbial consortium may be derived from the first working fluid present in the first reactor and transferred to the second reactor. In some embodiments, a fourth microbial consortium is established in a third reactor. The fourth microbial consortium may be derived from the second working fluid present in the second reactor and transferred to the third reactor. In some embodiments, a fifth microbial consortium is established in a fourth reactor. The fifth microbial consortium may be derived from the fourth working fluid present in the fourth reactor and transferred to the fifth reactor. A microbial consortium can be present in any reactor of a digestion system described herein. A microbial consortium can be derived from the working fluid of a reactor of a digestion system described herein. A first microbial consortium may be derived from inputs to a first reactor and may be present in a base product of a digestion system. A first microbial consortium may be present in a first reactor, a second reactor, a third reactor, or a clarifier. Without wishing to be bound by theory, a first microbial consortium in a working fluid may shift its microbial population and form a second microbial consortium. A second microbial consortium may be present in a first reactor, a secondreactor, a third reactor, or a clarifier. Without wishing to be bound by theory, a second microbial consortium in a working fluid may shift its microbial population and form a third microbial consortium. A third microbial consortium may be present in a first reactor, a second reactor, a third reactor, or a clarifier. Without wishing to be bound by theory, a third microbial consortium in a working fluid may shift its microbial population and form a fourth microbial consortium. A fourth microbial consortium may be present in a first reactor, a second reactor, a third reactor, or a clarifier. Without wishing to be bound by theory, a fourth microbial consortium in a working fluid may shift its microbial population and form a fifth microbial consortium. A fifth microbial consortium may be present in a first reactor, a second reactor, a third reactor, or a clarifier. Microbial consortia of the digestion system described herein may develop shifts in microbial communities based on conditions (e.g., nutrients, retention time, flow rate, pH, oxygen content, digestion products) of the reactors of the system and the working fluid. Without wishing to be bound by theory, as a working fluid incubates in a reactor of a bioreactor system described herein, microbes of a microbial consortium may become enriched, be maintained, or die which can shift a population of microbes in a microbial community in a working fluid and establish a new microbial consortium.

[0208] A portion of a first working fluid may be transferred to a second container of a serialized assembly of containers of a digestion system described herein. The working fluid of the second container may comprise a second working fluid. A portion of a second working fluid may be transferred to a third container of a serialized assembly of containers of a digestion system described herein. The working fluid of the third container may comprise a third working fluid. A portion of a third working fluid may be transferred to a fourth container of a serialized assembly of containers of a digestion system described herein. The working fluid of the fourth container may comprise a fourth working fluid. A portion of a fourth working fluid may be transferred to a fifth container of a serialized assembly of containers of a digestion system described herein. The working fluid of the fifth container may comprise a fifth working fluid. A portion of a fifth working fluid may be transferred to a sixth container of a serialized assembly of containers of a digestion system described herein. The working fluid of the sixth container may comprise a sixth working fluid.

[0209] The working fluid of a container in the digestion system may incubate in the container. A flow rate of the digestion system may increase or decrease a volume of working fluid, or the volume of the working fluid may stay the same. Adjusting a flow rate of the system may result in a longer or shorter hydraulic retention time for the system, while keeping the volume of working fluid in each container constant. The working fluid of a first, second, third, fourth, fifth, or sixthcontainer may increase in volume over a time period. The working fluid of a first, second, third, fourth, fifth, or sixth container may decrease in volume over a time period. The working fluid of a first, second, third, fourth, fifth, or sixth container may not increase or decrease in volume over a time period. The volume of working fluid in each of the containers of a digestion system may be the same. The volume of working fluid in each of the containers of a digestion system may be different. The volumes of the first working fluid, second working fluid, third working fluid, fourth working fluid, fifth working fluid, and / or sixth working fluid may be constant (e.g., unchanging over a time period). A constant volume may comprise a volume that does not increase or decrease over 1 minute, 5 minutes, 10 minutes, 30 minutes, 1 hours, 5 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, or 1 week.

[0210] As working fluid flows from each container of digestion system, the working fluid may comprise a flow rate. A first flow rate may comprise a flow rate of an aqueous feedstock inputted into a first container from a source outside the digestion system. A second flow rate may comprise a flow rate of a working fluid from a first container into a second container. A third flow rate may comprise a flow rate of a working fluid from a second container into a third container. A fourth flow rate may comprise a flow rate of a working fluid from a third container into a fourth container. A fifth flow rate may comprise a flow rate of a working fluid from a fourth container into a fifth container. A flow rate (e.g., first flow rate, second flow rate, third flow rate, fourth flow rate, fifth flow rate) may be at least about, at most about, or about 0.001, 0.003, 0.005, 0.01, 0.03, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 6.0, 7.0, 8.0, 9.0, or 10.0 gallons / min, or a range between any of these values.

[0211] A microbial consortium can comprise a population of microbes. The population of microbes can be generated from an input to the digestion system. An aqueous feedstock inputted into the digestion system may comprise a microbial consortium. Incubation of a working fluid in the digestion system may promote the growth of microbes within a microbial consortium (e.g., a first microbial consortium, a second microbial consortium, a third microbial consortium, a fourth microbial consortium, a fifth microbial consortium, a sixth microbial consortium). The microbes of a microbial consortium or at least a portion of microbes within the microbial consortium may have a desired plant growth promotion property. The plant growth promotion property may comprise shoot biomass, root biomass, nutrient uptake, crop yield, phosphate solubilization, acid production, leaf area, chlorophyll content, photosynthetic activity, or total biomass.

[0212] Reactors of a digestion system may be fluidly connected. A portion of a working fluid in a first container may be transferred to a fluidly connected second container. A portion of a working fluid in a second container may be transferred to a fluidly connected third container. A portion ofa working fluid in a third container may be transferred to a fluidly connected fourth container. A portion of a working fluid in a fourth container may be transferred to a fluidly connected fifth container. A portion of a working fluid in a fifth container may be transferred to a fluidly connected sixth container. Additional fluidly connected containers may be arranged in a similar way. In some embodiments, a transfer of working fluid between containers of a digestion system described herein may be continuous. A continuous flow of working fluid may comprise a flow of working fluid that does not stop or a flow of working fluid that stops for less than about 5 seconds, less than about 4 seconds, less than about 3 seconds, less than about 2 seconds, less than about 1 second, less than about 0.5 seconds, or less than about 0.1 seconds. A continuous flow of working fluid within a digestion system described herein (e.g., between containers of a digestion system) may have a first flow rate. A continuous flow of working fluid within a digestion system described herein (e.g., between containers of a digestion system) may have a second flow rate. In some embodiments, the first flow rate and the second flow rate are equal. A first flow rate may comprise a flow rate of fluid transferred from a source outside the digestion system into a first container. A second flow rate may comprise a rate of fluid flow from a first container to a second container. In some embodiments, an amount of working fluid and / or feedstock (e.g., aqueous organic feedstock) transferred into the first container over a time period is equal to an amount of working fluid and / or feedstock (e.g., aqueous organic feedstock) transferred into a second fluidly connected container over the same time period. In some embodiments, the first flow rate and the second flow rate are different. For example, flow rates may be different if an input stream into a first reactor is faster or slower than a stream of working fluid transferred from the first reactor to a second reactor. This difference in flow rates may result in different volumes of working fluid in containers of a digestion system.

[0213] In some embodiments, a first container of a digestion system comprises a constant volume. In some embodiments, a volume of a first container of a digestion system is different over time. In some embodiments, the flow rate between containers of the digestion system may maintain a constant volume in each container. A first container, a second container, a third container, a fourth container, a fifth container, and / or a sixth container may be maintained at a constant volume. A constant volume may be maintained by a continuous flow of fluid through a digestion system described herein. For example, similar flow rates of working fluid being transferred between containers of a digestion system may result in constant volumes of working fluid in the containers. This continuous flow of the bioreactor system may help enrich a microbial population with a targeted functionality (e.g., phosphate solubilization) by not overflowing or depleting a working fluid in a container of the system.

[0214] In some embodiments, a digestion system may be inoculated with an inoculum of a microbe (e.g., an inoculum of a microbial strain). The inoculum of a microbial strain may be an isolated microbe. An isolated microbe can comprise a microbe grown or enriched outside of a natural environment (e.g., in a culture medium or a streak plate method). In some embodiments, the inoculum of a microbe may comprise a mixture of multiple isolated microbes. The inoculum of a microbe may comprise a mixture of at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, or more isolated microbes.

[0215] In some cases, a digestion system may not be reinoculated with an isolate (e.g., microbial strain) or combination of isolates following a first inoculation. Reinoculation of a digestion system may comprise providing a microbial strain following a previous inoculation. Reinoculation of a digestion system may comprise introducing a target isolate (e.g., microbial strain) in a container of the digestion system at a time point during operating of the digestion system. In some cases, a digestion system may be reinoculated with an isolate or combination of isolates at least every 20 days, at least every 50 days, at least every 100 days, at least every 200 days, at least every 300 days, at least every 400 days, at least every 500 days, or more. In some cases, a digestion system may be inoculated with an isolate or combination of isolates on day 1 of a digestion process and reinoculated 1, 2, 3, 4, 5, or more times after day 1 of the digestion process. In some cases, a digestion system may be reinoculated with a microbial strain described herein after operating the digestion system for a time period. For example, a digestion system may be reinoculated with a microbial strain described herein after operating the digestion system for a duration of time of at least about, at most about, or about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 3 months, 4 months, 5 months, 6 months, 9 months, 12 months, 18 months, 24 months, 3 years, 4 years, 5 years, 10 years, or a range between any of these two values.

[0216] In some embodiments, the inoculation establishes an initial population of the target isolate having a concentration of at least, at most, or about IxlO2, 0.5xl03, IxlO3, 0.5xl04, IxlO4, 0.5xl05, IxlO5, 0.5xl06CFU / ml, or a range between any two of these values.

[0217] In some embodiments, the population of the microbial strain (e.g., concentration of the microbial strain) is maintained (e.g., retained) by at least about 0.00001%, at least about 0.0001%, at least about 0.001%, at least about 0.01%, at least about 0.1%, at least about 0.5%, at least about 1%, at least about 5%, at least 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, or at least about 75% from the concentration of the microbial strain added into a container (e.g., a first container) of the digestion system. Biosolids (e.g., floc)may comprise small particles from a working fluid of a digestion system. The biosolids (e.g., floc) may accumulate in a clarifier chamber over time and separate from a supernatant (e.g., base product). In some embodiments, the population of the microbial strain may be retained in the floc (e.g., biosolids) of the digestion system. In some embodiments, a majority (95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9%) of the inoculum of the microbe may be retained in the floc (e.g., biosolids) of the digestion system. Floc may be generated at any point during operating of a bioreactor system as described herein. For example, floc may be generated in a reactor of the bioreactor system (e.g., a first container, a second container, a third container, a fourth container, a fifth container, a sixth container, or any container of the system). For example, floc may be generated in a clarifier chamber of a bioreactor system. In some embodiments, floc may comprise at least a portion of phosphate-solubilizing microbes generated in a digestion system described herein.

[0218] In some embodiments, there may be at least about 1 log CFU / ml, at least about 2 logs CFU / ml, at least about 3 logs CFU / ml, at least about 4 logs CFU / ml, at least about 5 logs CFU / ml, or greater than about 5 logs CFU / ml increase in a concentration of the population of the microbial strain after inoculation in a container (e.g., first container) of the digestion system described herein. In some embodiments, there may be at most about 5 logs CFU / ml, at most about 4 logs CFU / ml, at most about 3 logs CFU / ml, at most about 2 logs CFU / ml, at most about 1 log CFU / ml, or less than about 1 log CFU / ml increase in a concentration of the population of the microbial strain after inoculation in a container (e.g., first container) of the digestion system described herein. In some embodiments, there may be from about 1 log CFU / ml to about 8 logs CFU / ml increase in a concentration of the population of the microbial strain after inoculation in a container (e.g., first container) of the digestion system described herein. In some embodiments, there may be from about 1 log CFU / ml to about 2 logs CFU / ml, about 1 log CFU / ml to about 3 logs CFU / ml, about 1 log CFU / ml to about 4 logs CFU / ml, about 1 log CFU / ml to about 5 logs CFU / ml, about 1 log CFU / ml to about 6 logs CFU / ml, about 1 log CFU / ml to about 7 logs CFU / ml, about 1 log CFU / ml to about 8 logs CFU / ml, about 2 logs CFU / ml to about 3 logs CFU / ml, about 2 logsCFU / ml to about 4 logs CFU / ml, about 2 logs CFU / ml to about 5 logs CFU / ml, about 2 logsCFU / ml to about 6 logs CFU / ml, about 2 logs CFU / ml to about 7 logs CFU / ml, about 2 logsCFU / ml to about 8 logs CFU / ml, about 3 logs CFU / ml to about 4 logs CFU / ml, about 3 logsCFU / ml to about 5 logs CFU / ml, about 3 logs CFU / ml to about 6 logs CFU / ml, about 3 logsCFU / ml to about 7 logs CFU / ml, about 3 logs CFU / ml to about 8 logs CFU / ml, about 4 logsCFU / ml to about 5 logs CFU / ml, about 4 logs CFU / ml to about 6 logs CFU / ml, about 4 logsCFU / ml to about 7 logs CFU / ml, about 4 logs CFU / ml to about 8 logs CFU / ml, about 5 logsCFU / ml to about 6 logs CFU / ml, about 5 logs CFU / ml to about 7 logs CFU / ml, about 5 logs CFU / ml to about 8 logs CFU / ml, about 6 logs CFU / ml to about 7 logs CFU / ml, about 6 logs CFU / ml to about 8 logs CFU / ml, or about 7 logs CFU / ml to about 8 logs CFU / ml increase in a concentration of the population of the microbial strain after inoculation in a container (e.g., first container) of the digestion system described herein.

[0219] In some embodiments, there may be at least about 1 log CFU / ml, at least about 2 logs CFU / ml, at least about 3 logs CFU / ml, at least about 4 logs CFU / ml, at least about 5 logs CFU / ml, or greater than about 5 logs CFU / ml decrease in a concentration of the population of the microbial strain after inoculation in a container (e.g., first container) of the digestion system described herein. In some embodiments, there may be at most about 5 logs CFU / ml, at most about 4 logs CFU / ml, at most about 3 logs CFU / ml, at most about 2 logs CFU / ml, at most about 1 log CFU / ml, or less than about 1 log CFU / ml decrease in a concentration of the population of the microbial strain after inoculation in a container (e.g., first container) of the digestion system described herein. In some embodiments, there may be from about 1 log CFU / ml to about 8 logs CFU / ml decrease in a concentration of the population of the microbial strain after inoculation in a container (e.g., first container) of the digestion system described herein. In some embodiments, there may be from about 1 log CFU / ml to about 2 logs CFU / ml, about 1 log CFU / ml to about 3 logs CFU / ml, about 1 log CFU / ml to about 4 logs CFU / ml, about 1 log CFU / ml to about 5 logs CFU / ml, about 1 log CFU / ml to about 6 logs CFU / ml, about 1 log CFU / ml to about 7 logs CFU / ml, about 1 log CFU / ml to about 8 logs CFU / ml, about 2 logs CFU / ml to about 3 logs CFU / ml, about 2 logsCFU / ml to about 4 logs CFU / ml, about 2 logs CFU / ml to about 5 logs CFU / ml, about 2 logsCFU / ml to about 6 logs CFU / ml, about 2 logs CFU / ml to about 7 logs CFU / ml, about 2 logsCFU / ml to about 8 logs CFU / ml, about 3 logs CFU / ml to about 4 logs CFU / ml, about 3 logsCFU / ml to about 5 logs CFU / ml, about 3 logs CFU / ml to about 6 logs CFU / ml, about 3 logsCFU / ml to about 7 logs CFU / ml, about 3 logs CFU / ml to about 8 logs CFU / ml, about 4 logsCFU / ml to about 5 logs CFU / ml, about 4 logs CFU / ml to about 6 logs CFU / ml, about 4 logsCFU / ml to about 7 logs CFU / ml, about 4 logs CFU / ml to about 8 logs CFU / ml, about 5 logsCFU / ml to about 6 logs CFU / ml, about 5 logs CFU / ml to about 7 logs CFU / ml, about 5 logsCFU / ml to about 8 logs CFU / ml, about 6 logs CFU / ml to about 7 logs CFU / ml, about 6 logsCFU / ml to about 8 logs CFU / ml, or about 7 logs CFU / ml to about 8 logs CFU / ml decrease in a concentration of the population of the microbial strain after inoculation in a container (e.g., first container) of the digestion system described herein.

[0220] Incubation of a microbial consortium in the digestion system under conditions of a selective pressure source (e.g., providing only insoluble phosphate source) may enrich a microbialcommunity with plant growth promotion properties (e.g., phosphate solubilizing microbes) and / or increase phosphate solubilization activity of a microbial community with or without a measurable increase in the population of phosphate solubilizers in the microbial community. Incubation of a phosphate-solubilizing target microbe and / or the phosphate-solubilizing microbes of the microbial consortium may generate metabolites with desired plant growth promotion properties (e.g., phosphate-solubilizing metabolites). The target phosphate-solubilizing microbe, phosphate- solubilizing microbes of the microbial consortium, phosphate-solubilizing metabolites, or any combination thereof may be phosphate-solubilizers in the working fluid of the system and / or in the output product (e.g., base product) of the digestion system.

[0221] In some embodiments, the absolute population of the microbial inoculum of the digestion system may not decrease by more than about 0.000001%, 0.00001%, 0.0001%, 0.001%, 0.01%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, or 25% after incubation. In some embodiments, the absolute population of the microbial inoculum of the digestion system may not decrease by more than about 0.000001%, 0.00001%, 0.0001%, 0.001%, 0.01%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, or 25% after incubation for a retention period of a digestion system (e.g., 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 2 months, 3 months, 4 months, 5 months, 6 months, 1 year, 2 years, 3 years, 4 years, or 5 years). In some embodiments, the absolute population of the microbial inoculum of the digestion system may not decrease by more than about 0.000001%, 0.00001%, 0.0001%, 0.001%, 0.01%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, or 25% after incubation without inoculation of additional microbe.

[0222] In some embodiments, a proportion of a concentration of phosphate solubilizers relative to a total bacterial population count may increase across containers of a fluidly connected digestion system or may stay the same over a given period of time. In some embodiments, phosphate solubilizing activity by microbes present in a bioreactor system may be enhanced without a measurable increase in the proportion of a concentration of phosphate solubilizers relative to a total bacterial population.

[0223] Phosphate solubilization can result in the production of phosphate ions (PO43) as an insoluble source of phosphate is solubilized by microbes described herein. A measure of phosphate solubilization and capacity of microbes to solubilize phosphate can be an amount of phosphate ions (PO43) made soluble. In some embodiments, the phosphate solubilization capacity (e.g., as PO43mg / mL made soluble) of a working solution may increase in a digestion system provided herein. In some embodiments, the phosphate solubilization capacity (e.g., as PC3' mg / L made soluble) of a working solution may be at least about 1.5x higher, at least about 2x higher, atleast about 3x higher, at least about 4x higher, at least about 5x higher, at least about lOx higher, at least about 20x higher, at least about 30x higher, at least about 40x higher, at least about 50x higher, at least about 60x higher, at least about 70x higher, at least about 80x higher, at least about 90x higher, or at least about lOOx higher, or greater than about lOOx higher in a second container, third container, or fourth container, compared to that in a first container of the digestion system or compared to a similar system that lacks one or more of the features of embodiments described herein such as, for example, a population of a phosphate-solubilizing target microbe, or insoluble phosphate as the primary or only phosphate source. In some embodiments, the phosphate solubilization capacity (e.g., as PCU3' g / mL made soluble) of a working solution may be at most about lOOx higher, at most about 90x higher, at most about 80x higher, at most about 70x higher, at most about 60x higher, at most about 50x higher, at most about 40x higher, at most about 30x higher, at most about 20x higher, at most about lOx higher, at most about 5x higher, at most about 4x higher, at most about 4x higher, at most about 2x higher, or at most about 1.5x higher, or less than about 1 ,5x higher in a second container, third container, or fourth container, compared to that in a first container of the digestion system or compared to a similar system that lacks one or more of the features of embodiments described herein such as, for example, a population of a phosphate- solubilizing target microbe, or insoluble phosphate as the primary or only phosphate source.

[0224] In some embodiments, a working solution may be obtained from a reactor of a digestion system and tested for phosphate solubilizing capacity (e.g., quantified as PCU3' g / L made soluble) according to assays described herein. In some embodiments, a working solution of a digestion system described herein may have a measure of PO43' made soluble of at least about 50 mg / L, at least about 100 mg / L, at least about 150 mg / L, at least about 200 mg / L, at least about 250 mg / L, at least about 300 mg / L, at least about 350 mg / L, at least about 400 mg / L, at least about 450 mg / L, at least about 500 mg / L, at least about 600 mg / L, at least about 700 mg / L, at least about 800 mg / L, or greater than about 800 mg / L. In some embodiments, a working solution of a digestion system described herein may have a measure of PCh3' made soluble of at most about 800 mg / L, at most about 700 mg / L, at most about 600 mg / L, at most about 500 mg / L, at most about 450 mg / L, at most about 400 mg / L, at most about 350 mg / L, at most about 300 mg / L, at most about 250 mg / L, at most about 200 mg / L, at most about 150 mg / L, at most about 100 mg / L, at most about 50 mg / L, or less than about 50 mg / L. In some embodiments, a working solution of a digestion system described herein may have a measure of PO43' made soluble from about 20 mg / L to about 750 mg / L. In some embodiments, a working solution of a digestion system described herein may have a measure of PO43' made soluble from about 20 mg / L to about 50 mg / L, about 20 mg / L to about 100 mg / L, about 20 mg / L to about 150 mg / L, about 20 mg / L to about 200 mg / L, about 20 mg / Lto about 250 mg / L, about 20 mg / L to about 300 mg / L, about 20 mg / L to about 350 mg / L, about 20 mg / L to about 400 mg / L, about 20 mg / L to about 450 mg / L, about 20 mg / L to about 500 mg / L, about 20 mg / L to about 750 mg / L, about 50 mg / L to about 100 mg / L, about 50 mg / L to about 150 mg / L, about 50 mg / L to about 200 mg / L, about 50 mg / L to about 250 mg / L, about 50 mg / L to about 300 mg / L, about 50 mg / L to about 350 mg / L, about 50 mg / L to about 400 mg / L, about 50 mg / L to about 450 mg / L, about 50 mg / L to about 500 mg / L, about 50 mg / L to about 750 mg / L, about 100 mg / L to about 150 mg / L, about 100 mg / L to about 200 mg / L, about 100 mg / L to about 250 mg / L, about 100 mg / L to about 300 mg / L, about 100 mg / L to about 350 mg / L, about 100 mg / L to about 400 mg / L, about 100 mg / L to about 450 mg / L, about 100 mg / L to about 500 mg / L, about 100 mg / L to about 750 mg / L, about 150 mg / L to about 200 mg / L, about 150 mg / L to about 250 mg / L, about 150 mg / L to about 300 mg / L, about 150 mg / L to about 350 mg / L, about 150 mg / L to about 400 mg / L, about 150 mg / L to about 450 mg / L, about 150 mg / L to about 500 mg / L, about 150 mg / L to about 750 mg / L, about 200 mg / L to about 250 mg / L, about 200 mg / L to about 300 mg / L, about 200 mg / L to about 350 mg / L, about 200 mg / L to about 400 mg / L, about 200 mg / L to about 450 mg / L, about 200 mg / L to about 500 mg / L, about 200 mg / L to about 750 mg / L, about 250 mg / L to about 300 mg / L, about 250 mg / L to about 350 mg / L, about 250 mg / L to about 400 mg / L, about 250 mg / L to about 450 mg / L, about 250 mg / L to about 500 mg / L, about 250 mg / L to about 750 mg / L, about 300 mg / L to about 350 mg / L, about 300 mg / L to about 400 mg / L, about 300 mg / L to about 450 mg / L, about 300 mg / L to about 500 mg / L, about 300 mg / L to about 750 mg / L, about 350 mg / L to about 400 mg / L, about 350 mg / L to about 450 mg / L, about 350 mg / L to about 500 mg / L, about 350 mg / L to about 750 mg / L, about 400 mg / L to about 450 mg / L, about 400 mg / L to about 500 mg / L, about 400 mg / L to about 750 mg / L, about 450 mg / L to about 500 mg / L, about 450 mg / L to about 750 mg / L, or about 500 mg / L to about 750 mg / L.

[0225] Population of the microbe may be measured using methods including but not limited to spectrophotometers, cell counting, measures of turbidity, hemocytometers, electronic enumeration, measures of cell mass, and measures of cell activity.

[0226] Phosphate-solubilizing microbes may be identified using methods using agar plates or liquid testing solutions. Microbes may be tested on agar plates with insoluble phosphate and the measurement of phosphate solubilization capacity (e.g., amount of phosphate made soluble) may classify a microbe as capable of phosphate solubilization. Microbes may be tested in liquid solution containing insoluble phosphate to measure phosphate solubilization capacity. Collective phosphate solubilization activity of a community of microbes within a working fluid, product stream, or other sample from the system may be measured using assays described herein to determine whether desired stimulation of phosphate solubilizing activity is being achieved.Phosphate solubilization capacity may be measured at different time points and monitored to observe the functioning of the system.

[0227] The addition of the target isolate (e.g., the inoculum of the microbial strain / microbe) can have an added benefit in improving target functionality of working solution in a reactor of the digestion system and / or in the output base product of the digestion system. In some embodiments, addition of the inoculum of the microbial strain can improve the phosphate solubilization capacity of working fluids and base product of a digestion system compared to working fluids and base product of a digestion system without an inoculum of the microbial strain .

[0228] In some embodiments, a working solution and / or base product of a digestion system inoculated with a target isolate may have an at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, or greater than 80% increase in PCU3' concentration in comparison with that of a working solution and / or base product of a digestion system not inoculated with a target isolate. In some embodiments, a working solution and / or base product of a digestion system inoculated with a target isolate may have an at most about 80%, at most about 75%, at most about 70%, at most about 65%, at most about 60%, at most about 55%, at most about 50%, at most about 45%, at most about 40%, at most about 35%, at most about 30%, at most about 25%, at most about 20%, at most about 15% or less than about 15% increase in PCU3' concentration in comparison with that of a working solution and / or base product of a digestion system not inoculated with a target isolate.

[0229] In some embodiments, a working solution and / or base product of a digestion system inoculated with a target isolate may have from about 10% to about 80% increase in PCU3' solubilization activity in comparison with that of a working solution and / or base product of a digestion system not inoculated with a target isolate. In some embodiments, a working solution and / or base product of a digestion system inoculated with a target isolate may have from about 10% to about 15%, about 10% to about 20%, about 10% to about 25%, about 10% to about 30%, about 10% to about 35%, about 10% to about 40%, about 10% to about 45%, about 10% to about 50%, about 10% to about 60%, about 10% to about 70%, about 10% to about 80%, about 15% to about 20%, about 15% to about 25%, about 15% to about 30%, about 15% to about 35%, about 15% to about 40%, about 15% to about 45%, about 15% to about 50%, about 15% to about 60%, about 15% to about 70%, about 15% to about 80%, about 20% to about 25%, about 20% to about 30%, about 20% to about 35%, about 20% to about 40%, about 20% to about 45%, about 20% to about 50%, about 20% to about 60%, about 20% to about 70%, about 20% to about 80%, about25% to about 30%, about 25% to about 35%, about 25% to about 40%, about 25% to about 45%, about 25% to about 50%, about 25% to about 60%, about 25% to about 70%, about 25% to about 80%, about 30% to about 35%, about 30% to about 40%, about 30% to about 45%, about 30% to about 50%, about 30% to about 60%, about 30% to about 70%, about 30% to about 80%, about 35% to about 40%, about 35% to about 45%, about 35% to about 50%, about 35% to about 60%, about 35% to about 70%, about 35% to about 80%, about 40% to about 45%, about 40% to about 50%, about 40% to about 60%, about 40% to about 70%, about 40% to about 80%, about 45% to about 50%, about 45% to about 60%, about 45% to about 70%, about 45% to about 80%, about 50% to about 60%, about 50% to about 70%, about 50% to about 80%, about 60% to about 70%, about 60% to about 80%, or about 70% to about 80% increase in PC3' solubilization activity in comparison with that of a working solution and / or base product of a digestion system not inoculated with a target isolate.

[0230] A digestion system may be inoculated with a microbial strain at the start of a digestion system, which may allow for the microbial strain to flow through the system and working fluids of the reactors. Without wishing to be bound by theory, a digestion system inoculated with a phosphate solubilizing microbial strain (for example, one of the strains described herein) may increase a phosphate solubilization capacity of a working fluid and / or a base product compared to a working fluid and / or base product of an otherwise identical digestion system with no inoculated phosphate solubilizing microbial strain. Without wishing to be bound by theory, a digestion system inoculated with a phosphate solubilizing microbial strain may increase a phosphate solubilization capacity of a working fluid and / or a base product compared to a working fluid and / or base product of an otherwise identical digestion system with the phosphate solubilizing microbial strain added (e.g., spiked) at the end of the system. Without wishing to be bound by theory, a digestion system inoculated with a phosphate solubilizing microbial strain may increase a phosphate solubilization capacity of a base product compared to a base product of an otherwise identical digestion system with the phosphate solubilizing microbial strain added (e.g., spiked) at the end of the system. Incubation of the phosphate solubilizing microbial strain in the digestion system may enrich the working fluid with phosphate-solubilizing microbes, stimulate phosphate- solubilizing activity of microbes in the system with or without an increase in the population of phosphate-solubilizing microbes in the system, and / or generate phosphate-solubilizing metabolites.E. Microbial Isolates

[0231] Certain microorganisms of the present disclosure have all of the identifying characteristics of the deposited strains and, in particular, the identifying characteristics of being able to promote plant growth and / or yield as described herein. In particular, the certain microorganisms of the present disclosure can refer to the deposited microorganisms as described above, and strains derived therefrom.

[0232] Phosphorus is an essential macronutrient for plant growth and development and phosphate deficiencies can impair the physiological and biochemical functioning of plants. Phosphorus availability is important for growth promoting properties such as increasing photosynthesis, biomass of shoots and roots, strengthening the stems, increasing leafage and leaf area, formation of flowers and seeds, energy production, nitrogen fixation, resistance to infections and disease, and transformation of sugar to starch. Although most soils contain considerable amounts of phosphorus, the availability of soluble phosphorus is limited because of fixation as insoluble phosphates of aluminum, calcium, and iron, and other complexes.

[0233] Phosphate solubilizing microorganisms may be capable of hydrolyzing organic and inorganic insoluble phosphorus compounds to soluble phosphate that plants can uptake. Phosphate solubilizing microorganisms can include, but are not limited to, strains from bacteria (Bacillus, Pseudomonas, and Rhizobium), arbuscular mycorrhizal, fungi (Penicillium and Aspergillus), and actinomycetes. In some embodiments, phosphate solubilizing bacteria may comprise strains of Bacillus safensis, Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus megaterium, Commonas testosteroni, Enterobacter cloacae, Citrabacter freundii, Pseudomonas spp., Agrobacterium spp., Bacillus circulans, or any combination thereof. Mechanisms of phosphate solubilization include, but are not limited to, organic mineralization, chelation, and lowering soil pH levels. Without wishing to be bound by theory, it is believed that lower pH levels leads to a decrease in fixed phosphorus by calcium, aluminum, and iron, and hence greater phosphate solubilization. In some embodiments, the pH is between 3 and 8.5 after the addition of phosphate solubilizing bacteria. In some embodiments, the pH is lower than 5 after the addition of phosphate solubilizing bacteria.

[0234] In some embodiments, disclosed herein are phosphate solubilizing microbial strains, including compositions that include such strains and methods of using such strains to promote plant growth.

[0235] In some embodiments, the microbial strain is from a Bacillus genus. In some embodiments, the microbial strain is from a Bacillus amyloliquefaciens species. In some embodiments, the microbial strain is from a bacterial species other than Bacillusamyloliquefaciens. In some embodiments, the Bacillus amyloliquefaciens strain is deposited under Accession No. PTA-127657, or an isolated clone thereof. In some embodiments, the 16S rRNA gene of the microbial strain comprises a nucleotide sequence selected from SEQ ID NO.: 1. In some embodiments, the microbial strain comprises a 16S rRNA gene comprising a nucleotide sequence of SEQ ID NO.: 1. In some embodiments, the microbial strain comprises a 16S rRNA gene comprising a nucleotide sequence that exhibits at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, or at least 99.9% sequence identity to the nucleotide sequences as set forth in SEQ ID NO.: 1. In some embodiments, the 16S rRNA gene of the microbial strain comprises a nucleotide sequence that exhibits at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, or at least 99.9% sequence identity to any one of the nucleotide sequences as set forth in SEQ ID NO.: 1. In some embodiments, the gyrB gene of the microbial strain comprises the nucleotide sequence of SEQ ID NO: 3. In some embodiments, the microbial strain comprises a gyrB gene comprising a nucleotide sequence that exhibits at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to a nucleotide sequence as set forth in SEQ ID NO: 3. In some embodiments, the rpoB gene of the microbial strain comprises the nucleotide sequence of SEQ ID NO: 5. In some embodiments, the microbial strain comprises a rpoB gene comprising a nucleotide sequence that exhibits at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to a nucleotide sequence as set forth in SEQ ID NO: 5. Some embodiments provide a genus of plant growth-promoting microorganisms comprising any of the DNA sequences described herein and which enhances the health, growth and / or yield of a plant, as described herein.

[0236] In some embodiments, the microbial strain is from a Bacillus genus. In some embodiments, the microbial strain is from a Bacillus licheniformis species. In some embodiments, the microbial strain is from a bacterial species other Bacillus licheniformis. In some embodiments, the Bacillus licheniformis strain is deposited under Accession No. PTA- 127656, or an isolated clone thereof. In some embodiments, the 16S rRNA gene of the microbial strain comprises a nucleotide sequence selected from SEQ ID NO.: 2. In some embodiments, themicrobial strain comprises a 16S rRNA gene comprising a nucleotide sequence of SEQ ID NO.: 2. In some embodiments, the microbial strain comprises a 16S rRNA gene comprising a nucleotide sequence that exhibits at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, or at least 99.9% sequence identity to any one of the nucleotide sequences as set forth in SEQ ID NO.: 2. In some embodiments, the 16S rRNA gene of the microbial strain comprises a nucleotide sequence that exhibits at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, or at least 99.9% sequence identity to any one of the nucleotide sequences as set forth in SEQ ID NO.: 2. In some embodiments, the gyrB gene of the microbial strain comprises the nucleotide sequence of SEQ ID NO: 4. In some embodiments, the microbial strain comprises a gyrB gene comprising a nucleotide sequence that exhibits at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to a nucleotide sequence as set forth in SEQ ID NO: 4. In some embodiments, the rpoB gene of the microbial strain comprises the nucleotide sequence of SEQ ID NO: 6. In some embodiments, the microbial strain comprises a rpoB gene comprising a nucleotide sequence that exhibits at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, at least 99.6%, at least 99.7%, at least 99.8%, at least 99.9%, or 100% sequence identity to a nucleotide sequence as set forth in SEQ ID NO: 6. Some embodiments provide a genus of plant growth-promoting microorganisms comprising any of the DNA sequences described herein and which enhances the health, growth and / or yield of a plant, as described herein.

[0237] Chemical analysis of root exudates from plants treated with base product from the PST- sIP system (e.g., PST supernatant containing phosphate solubilizing bacteria) may indicate roots exudate organic acids. The metabolites of the PST base product may affect the chemistry of the root exudates. Filter-sterilized PST may affect the chemistry of the root exudates. In some embodiments, the organic acids comprise lactic acid, succinic acid, propionic acid, 2- ketogluconic acid, ketoglutaric acid, itaconic acid, isovaleric acid, isobutyric acid, acetic acid, oxalic acid, citric acid, malic acid, glyoxalic acid, fumaric acid, tartaric acid, a-ketobutyric acid, malonic acid, glycolic acid, glutamic acid, maleic acid, aspartic acid, gluconic acid, or a combination thereof. In some embodiments, the organic acids comprise lactic acid, succinicacid, malic acid, ketoglutaric acid, citric acid, or a combination thereof. In some embodiments, the released organic acids can increase solubilization of phosphates.F. Exemplary Digestion Systems for Production of Isolates

[0238] The present disclosure provides systems with conditions to produce biostimulant products with plant-growth promoting capabilities (e.g., improved phosphate solubilization capacity).

[0239] Embodiments of systems and methods described herein produce biostimulant products that may have a multi-modal way of promoting phosphate solubilization in plants. Biostimulant products produced by embodiments described herein may be used to promote plant growth by applying the products to plants and / or plant growth media (e.g., soil). One mode of action of products produced in some embodiments is enhanced phosphate solubilization included in the products. Microbes and metabolites produced a system described herein may increase the amount of soluble phosphate in soil without a plant present. Microbes and metabolites produced in a system described herein may increase the amount of soluble phosphate in soil with a plant present. Biostimulant products of the digestion systems described herein may act on a plant to create an increased amount of organic acids which may solubilize phosphates. Another mode of action may be increases in soil organic phosphate and mineralization and uptake of organic phosphate stimulated by microbes and / or microbial metabolites present in the products produced in embodiments described herein. In some cases, a microbial strain may be incubated at a phosphate (e.g., phosphorus) concentration that selectively promotes growth of the microbe. The population of the microbe may be increased. The population of the microbe may be decreased.

[0240] In an aspect, the present disclosure provides a method comprising transferring a feedstock (e.g., aqueous organic feedstock) and / or an inoculum of a phosphate-solubilizing microbe into a first container. In some embodiments, the first container may comprise a volume of a first working fluid. The feedstock (e.g., aqueous organic feedstock) may comprise a first microbial consortium, digestion products, or any combination thereof. The digestion products may be generated by digestion of organic material by microbes of the first microbial consortium. The method may further comprise incubating a population of the microbial strain in a selective pressure source. The selective pressure source may comprise insoluble phosphate. Incubating the working fluid comprising the phosphate solubilizing microbial strain and feedstock (e.g., organic feedstock) with the selective pressure may promote enrichment of phosphate- solubilizing microbes within the microbial consortium, the inoculated phosphate microbial strain, generated phosphate-solubilizing metabolites, or any combination thereof. Incubating theworking fluid comprising the phosphate solubilizing microbial strain and feedstock (e.g., organic feedstock) with the selective pressure may promote maintenance of phosphate- solubilizing microbes within the microbial consortium, the inoculated phosphate microbial strain, generated phosphate-solubilizing metabolites, or any combination thereof. Incubation of the phosphate-solubilizing microbial strain in the digestion system may increase or maintain a proportion of phosphate-solubilizing microbes (e.g., phosphate solubilizers) relative to a total bacteria count within a container of the digestion system. Incubation of the phosphate- solubilizing microbial strain in the digestion system may increase or maintain a proportion of phosphate-solubilizing microbes (e.g., phosphate solubilizers) relative to a total bacteria count across multiple containers (e.g., a first container, a second container, a third container, a fourth container, a fifth container, a sixth container, or any combination thereof) of the digestion system.

[0241] In an aspect, the present disclosure provides a method, comprising (a) transferring a feedstock (e.g., aqueous organic feedstock) and an inoculum of a microbe that is capable of promoting phosphate solubilization into a first container comprising a volume of a first working fluid, wherein the feedstock (e.g., aqueous organic feedstock) comprises: (i) a first microbial consortium; and (ii) digestion products produced by digestion of an organic material by microbes in the first microbial consortium; and (b) incubating a population of the microbial strain with a selective pressure comprising insoluble phosphate that selectively promotes growth of the microbe, thereby increasing the population of the microbe. Incubation of the inoculum of the microbe may increase or maintain a proportion of the inoculum of the microbe relative to microbe of the microbial consortium in the working fluid of the reactor of the digestion system.

[0242] In some embodiments, incubating the inoculum at a phosphate ion concentration may selectively enhance the survival of phosphate solubilizing microbes. In some embodiments, incubating the inoculum at a phosphate ion concentration may selectively inhibit the growth of microbes that may not tolerate a level of phosphate. In some embodiments, incubating the inoculum at a phosphate ion concentration may selectively reduce the number of microbes that may not tolerate a level of phosphate. In some embodiments, incubating the inoculum at a phosphate ion concentration may increase the proportion of microbes that can tolerate phosphate (e.g., phosphate solubilizers or phosphate-solubilizing microbes). Without wishing to be bound by theory, addition of a selective pressure having a phosphate ion concentration may shift a microbial population towards microbes with phosphate solubilizing properties.

[0243] In some embodiments, incubating the inoculum at a phosphate ion concentration caused by addition of a selective pressure may increase an amount of an inoculum of the microbe. Theinoculum of the microbe may not die and / or may die at a slower rate relative to a rate of death of microbes of a microbial consortium in the presence of a phosphate ion concentration caused by addition of a selective pressure.

[0244] In some embodiments, the microbe is capable of phosphate solubilization in plants or promoting soil organic phosphate content and mineralization and uptake of organic phosphate from soil. In some embodiments, the conditions of the digestion system (e.g., pH, nutrients, flow rate, retention time, selective pressure, carbon source, nitrogen source, or any combination thereof) may promote growth of one or more microbes in the first microbial consortium that may be capable of promoting plant growth or phosphate solubilization in plants, or of generating metabolites capable of promoting plant growth or phosphate solubilization in plants. In some embodiments, during the incubating the microbe or one or more microbes in the first microbial consortium, metabolites may be produced capable of promoting plant growth and phosphate solubilization. The metabolites may have phosphate solubilizing capacity and make phosphate available to plants. The metabolites may enhance the presence of bioavailable phosphate ions. An increase in phosphate ions following incubation with phosphate-solubilizing metabolites may be from phosphate solubilization or stimulation of bacteria on a source of insoluble phosphate (e.g., rock phosphate). The available phosphate for plants may promote growth, vigor, or yield of a plant. In some embodiments, the incubating increases a population of one or more microbes in the microbial consortium capable of promoting plant growth. In some embodiments, the feedstock (e.g., aqueous organic feedstock) further comprises an inorganic substrate. In some embodiments, the first microbial consortium further comprises microbes derived from the inorganic substrate. In some embodiments, the inorganic substrate comprises rock phosphate. In some embodiments, the microbe or microbes may be of the species Bacillus safensis. Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus megaterium, Commonas testosterone Enter obacter cloacae, Citrabacter freundii. Pseudomonas spp., Agrobacterium spp., Bacillus circulans, or any combination thereof. In some embodiments, the microbe is the Bacillus amyloliquefaciens strain deposited under Accession No. PTA-127657 (MS2839) or the Bacillus licheniformis strain deposited under Accession No. PTA-127656 (MS1835). In some embodiments, the first working fluid comprises (a) a second microbial consortium derived from the feedstock (e.g., aqueous organic feedstock), and / or (b) digestion products produced by digestion of substances present in the organic feedstock by the first microbial consortium and the microbe(s). In some embodiments, the method further comprises transferring a portion of the first working fluid into a second container comprising a second working fluid and incubating the second working fluid in the second container. In some embodiments, the second working fluidcomprises (a) a third microbial consortium derived from the first working fluid, and (b) digestion products produced by digestion of substances present in the first working fluid by the third microbial consortium and the microbe. In some embodiments, the amount of the feedstock (e.g., aqueous organic feedstock) transferred into the first container over a time period is equal to the amount of the first working fluid transferred into the second container over the same time period. In some embodiments, the amount of the feedstock (e.g., aqueous organic feedstock) transferred into the first container over a time period is different to the amount of the first working fluid transferred into the second container over the same time period. In some embodiments, the amount of the feedstock (e.g., aqueous organic feedstock) transferred into the first container over a time period is more compared to the amount of the first working fluid transferred into the second container over the same time period. In some embodiments, the amount of the feedstock (e.g., aqueous organic feedstock) transferred into the first container over a time period is less compared to the amount of the first working fluid transferred into the second container over the same time period. In some embodiments, the volume of the first working fluid in the first container is maintained constant. A constant volume of working fluid in the first container may comprise the same volume of working fluid over 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 10 hours, 12 hours, 18 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 1 month, 3 months, 6 months, or 1 year. In some embodiments, the volume of the first working fluid in the first container is different over time. In some embodiments, transferring the feedstock (e.g., aqueous organic feedstock) into the first container comprises continuously flowing the feedstock (e.g., aqueous organic feedstock) into the first container at a first flow rate, transferring the portion of the first working fluid into the second container comprises continuously flowing the portion of the first working fluid into the second container at a second flow rate, and the first flow rate and the second flow rate may be equal. In some embodiments, the method further comprises transferring a portion of the second working fluid to a third container comprising a third working fluid and incubating the third working fluid in the third container. In some embodiments, the method further comprises transferring a portion of the third working fluid into a fourth container comprising a fourth working fluid and incubating the fourth working fluid in the fourth container. In some embodiments, the first working fluid, the second working fluid, the third working fluid, and the fourth working fluid may be maintained at constant volumes. In some embodiments, a plant growth promoting product made by the method. In some embodiments, a method of promotingphosphate solubilization of a plant comprising contacting the plant and / or a medium in which the plant is growing with the product.

[0245] In some embodiments, a serialized set of reaction chambers may be used in a method of producing a biostimulant product, as described in more detail in the Examples below. In some embodiments, conditions within reactor chambers may be established to selectively promote the production of one or more microbes that have a specific desired plant growth promoting effect.

[0246] FIG.7 schematically illustrate examples of a digestion system 100 with conditions (e.g., microbes) that produce biostimulant products that may have a multi-modal way of promoting phosphate solubilization and / or phosphate use efficiency in plants. The system 100 can comprise a first reactor 110, a second reactor 115, a third reactor 120, and / or a clarifier chamber 125 connected sequentially in which feedstock can continuously flow and microbial consortia as described herein can be grown. The first reactor, second reactor, and / or third reactor may be fluidized bed reactors without the scaffolding or packed bed reactors with a scaffolding within the reactors. Water may act as a hydraulic source. A base product from an alternate digestion system (e.g., PST system) may act as a hydraulic source. The hydraulic source can provide continuous flow to the first reactor 110. The systems 100 may also comprise an input channel or a series of input channels that flows inputs 140, 150, 160, 170 into the digestion system. In some cases, the inputs can be added individually to the digestion systems 100. In some cases, the inputs may be added in combination with one another into the digestion system. The inputs, as described herein, may comprise one or more of water, a microbial inoculum, a microbial product or intermediate product of another digestion system, a carbon source, a nitrogen source, manure feed, selective pressure, and / or micronutrients. The inputs may comprise other organic materials. In some cases, the inputs into the digestion system may be flown into the first reactor through a tank. In other cases, the input composition can be added to the first reactor. In other cases, the input composition can be added to the second reactor, the third reactor, the fourth reactor, and / or the fifth reactor.

[0247] The system 100 may comprise a pH sensor, pH controller, or similar mechanic to monitor and / or control the pH of the reactor and / or working fluid within the reactor so that pH is maintained at a threshold. The pH sensor or controller may be automatic. A buffer addition system may also be used to control the pH of the digestion system 100. For example, if the pH of the working fluid in the first reactor is below the threshold value, an automatic base (e.g., 3M of NaOH) may be added to the first reactor until the pH reaches the threshold value (e.g., at least about 7). In some embodiments, a pH in a reactor of digestion system 100 may be between 4.0- 9.0. In some embodiments, the pH in the reactor may be at least about 3.0, at least about 3.5, atleast about 4.0, at least about 4.5, at least about 5.0, at least about 5.5, at least about 6.0, at least about 6.5, at least about 7.0, at least about 7.5, at least about 8.0, at least about 8.5, at least about 9.0, at least about 9.5, or at least about 10.0.

[0248] In some cases, fluid (e.g., working fluid) can flow in a hydraulically balanced manner. The clarifier chamber 125 produces biostimulant products or digestion products (e.g., base products) 130.

[0249] In some cases, the first reactor 110, second reactor 115, and / or third reactor 120 of the system comprise high-speed mixers 180 to provide a continuous aerobic system. Fluid from an outflow port of the first reactor 110 can flow into the second reactor 115 continuously. Fluid from an outflow port of the second reactor 115 can flow into the third reactor 120 continuously. Fluid from an outflow port of the third reactor 120 can flow into the fourth reactor 125 continuously.

[0250] The outflow port may be positioned on the top, middle, and / or bottom of a reactor. Additionally, each reactor or clarifier chamber may comprise another outflow port for reintroducing a working fluid back into the same reactor or clarifier chamber, and may be pumped back to just below the surface of the same reactor to maintain homogeneous conditions within the working solutions. In some cases, the working fluid from each reactor is recirculated within each reactor from the bottom of the reactor back to just below the surface of the working solution to maintain a homogeneous environment for fermentation. Working fluid may be recirculated via a conduit or output of a reactor and reintroduced into the same reactor. Without wishing to be bound by theory, recirculation of a working fluid within a reactor may support enrichment and / or growth of an inoculum of a microbe or another component of a working fluid as described herein. For example, fluid from the first reactor may be reintroduced back into the first reactor. Biosolids (e.g., floc) may be generated through the process. Biosolids may comprise organic materials recovered from the feedstock, waste, wastewater, sludge material, or any combination thereof, of a working fluid of a digestion system described herein. In some embodiments, floc can comprise biosolids. In some embodiments, floc may comprise biosolids and / or other solid particles from digestion products of organic materials. Suspended particles from the organic substrates of the working fluid may aggregate to form floc. Floc may be produced throughout the fermentation process in the digestion system. Floc can be collected in a clarifier chamber and reintroduced to a container (e.g., reactor) of the system to serve as a seed of microbes.

[0251] In some cases, the clarifier chamber 125 may comprise an outflow port for reintroducing fluid back to the first reactor 110. In some embodiments, the supernatant (or base product) 130from the clarifier chamber may be continuously collected, and a portion of the floc at the bottom of the clarifier can be returned to the first reactor.

[0252] In some embodiments, the supernatant (or base product) from the clarifier chamber 125 may be continuously collected, and a portion of the floc at the bottom of the clarifier can be returned to the first reactor at a concentration of at least about 0.5 L / day, at least about 1.0 L / day, at least about 1.5 L / day, at least about 2.0 L / day, at least about 2.5 L / day, at least about 3.0 L / day, at least about 3.5 L / day, at least about 4.0 L / day, at least about 4.1 L / day, at least about 4.2 L / day, at least about 4.3 L / day, at least about 4.4 L / day, at least about 4.5 L / day, at least about 4.51 L / day, at least about 4.52 L / day, at least about 4.53 L / day, at least about 4.54 L / day, at least about 4.55 L / day, at least about 4.56 L / day, at least about 4.57 L / day, at least about 4.58 L / day, at least about 4.59 L / day, at least about 4.6 L / day, at least about 4.7 L / day, at least about 4.8 L / day, at least about 4.9 L / day, at least about 5.0 L / day, at least about 6.0 L / day, at least about 7.0 L / day, at least about 8.0 L / day, at least about 9.0 L / day, or at least about 10.0 L / day.

[0253] In some embodiments, the supernatant (or base product) from the clarifier chamber 125 may be continuously collected, and a portion of the floc at the bottom of the clarifier can be returned to the first reactor at a concentration of at most about 10.0 L / day, at most about 9.0 L / day, at most about 8.0 L / day, at most about 7.0 L / day, at most about 6.0 L / day, at most about5.5 L / day, at most about 5.0 L / day, at most about 4.9 L / day, at most about 4.8 L / day, at most about 4.7 L / day, at most about 4.6 L / day, at most about 4.59 L / day, at most about 4.58 L / day, at most about 4.57 L / day, at most about 4.56 L / day, at most about 4.55 L / day, at most about 4.54 L / day, at most about 4.53 L / day, at most about 4.52 L / day, at most about 4.51 L / day, at most about 4.5 L / day, at most about 4.4 L / day, at most about 4.3 L / day, at most about 4.2 L / day, at most about 4.1 L / day, at most about 4.0 L / day, at most about 3.5 L / day, at most about 3.0 L / day, at most about 2.5 L / day, at most about 2.0 L / day, at most about 1.5 L / day, at most about 1.0 L / day, or at most about 0.5 L / day.

[0254] In some embodiments, the supernatant (or base product) from the clarifier chamber 125 may be continuously collected, and a portion of the floc at the bottom of the clarifier can be returned to the first reactor at a concentration of about 0.5 L / day to about 15 L / day. In some embodiments, the supernatant (or base product) from the clarifier chamber 125 may be continuously collected, and a portion of the floc at the bottom of the clarifier can be returned to the first reactor at a concentration of about 0.5 L / day to about 1 L / day, about 0.5 L / day to about1.5 L / day, about 0.5 L / day to about 2 L / day, about 0.5 L / day to about 3 L / day, about 0.5 L / day to about 4 L / day, about 0.5 L / day to about 4.5 L / day, about 0.5 L / day to about 5 L / day, about0.5 L / day to about 7.5 L / day, about 0.5 L / day to about 10 L / day, about 0.5 L / day to about 12 L / day, about 0.5 L / day to about 15 L / day, about 1 L / day to about 1.5 L / day, about 1 L / day to about 2 L / day, about 1 L / day to about 3 L / day, about 1 L / day to about 4 L / day, about 1 L / day to about 4.5 L / day, about 1 L / day to about 5 L / day, about 1 L / day to about 7.5 L / day, about 1 L / day to about 10 L / day, about 1 L / day to about 12 L / day, about 1 L / day to about 15 L / day, about 1.5 L / day to about 2 L / day, about 1.5 L / day to about 3 L / day, about 1.5 L / day to about 4 L / day, about 1.5 L / day to about 4.5 L / day, about 1.5 L / day to about 5 L / day, about 1.5 L / day to about 7.5 L / day, about 1.5 L / day to about 10 L / day, about 1.5 L / day to about 12 L / day, about 1.5 L / day to about 15 L / day, about 2 L / day to about 3 L / day, about 2 L / day to about 4 L / day, about 2 L / day to about 4.5 L / day, about 2 L / day to about 5 L / day, about 2 L / day to about 7.5 L / day, about 2 L / day to about 10 L / day, about 2 L / day to about 12 L / day, about 2 L / day to about 15 L / day, about 3 L / day to about 4 L / day, about 3 L / day to about 4.5 L / day, about 3 L / day to about 5 L / day, about 3 L / day to about 7.5 L / day, about 3 L / day to about 10 L / day, about 3 L / day to about 12 L / day, about 3 L / day to about 15 L / day, about 4 L / day to about 4.5 L / day, about 4 L / day to about 5 L / day, about 4 L / day to about 7.5 L / day, about 4 L / day to about 10 L / day, about 4 L / day to about 12 L / day, about 4 L / day to about 15 L / day, about 4.5 L / day to about 5 L / day, about 4.5 L / day to about 7.5 L / day, about 4.5 L / day to about 10 L / day, about 4.5 L / day to about 12 L / day, about 4.5 L / day to about 15 L / day, about 5 L / day to about 7.5 L / day, about 5 L / day to about 10 L / day, about 5 L / day to about 12 L / day, about 5 L / day to about 15 L / day, about 7.5 L / day to about 10 L / day, about 7.5 L / day to about 12 L / day, about 7.5 L / day to about 15 L / day, about 10 L / day to about 12 L / day, about 10 L / day to about 15 L / day, or about 12 L / day to about 15 L / day.

[0255] As shown in FIG. 7, the clarifier 125 may comprise a floc flight system (e.g., flocfolding flights) that can improve the concentration of the added specific isolate(s) in the base product. In some embodiments, the floc may return back to the system at least about a 0.1% v / v rate per day, at least about a 0.5% v / v rate per day, at least about a 1.0% v / v rate per day, at least about a 1.1% v / v rate per day, at least about a 1.2% v / v rate per day, at least about a 1.3% v / v rate per day, at least about a 1.4% v / v rate per day, at least about a 1.5% v / v rate per day, at least about a 1.6% v / v rate per day, at least about a 1.7% v / v rate per day, at least about a 1.8% v / v rate per day, at least about a 1.9% v / v rate per day, at least about a 2.0% v / v rate per day, at least about a 2.1% v / v rate per day, at least about a 2.2% v / v rate per day, at least about a 2.3% v / v rate per day, at least about a 2.4% v / v rate per day, at least about a 2.5% v / v rate per day, at least about a 2.6% v / v rate per day, at least about a 2.7% v / v rate per day, at least about a 2.8% v / v rate per day, at least about a 2.9% v / v rate per day, at least about a 3.0% v / v rate per day, at leastabout a 4.0% v / v rate per day, at least about a 5.0% v / v rate per day, at least about a 6.0% v / v rate per day, at least about a 7.0% v / v rate per day, at least about a 8.0% v / v rate per day, at least about a 9.0% v / v rate per day, or at least about a 10.0% v / v rate per day.

[0256] In some embodiments, the floc may return back to the system at most about a 10.0% v / v rate per day, 9.0% v / v rate per day, 8.0% v / v rate per day, 7.0% v / v rate per day, 6.0% v / v rate per day, 5.0% v / v rate per day, 4.0% v / v rate per day, 3.0% v / v rate per day, 2.9% v / v rate per day, 2.8% v / v rate per day, 2.7% v / v rate per day, 2.6% v / v rate per day, 2.5% v / v rate per day, 2.4% v / v rate per day, 2.3% v / v rate per day, 2.2% v / v rate per day, 2.1% v / v rate per day, 2.0% v / v rate per day, 1.9% v / v rate per day, 1.8% v / v rate per day, 1.7% v / v rate per day, 1.6% v / v rate per day, 1.5% v / v rate per day, 1.4% v / v rate per day, 1.3% v / v rate per day, 1.2% v / v rate per day, 1.1% v / v rate per day, 1.0% v / v rate per day, 0.5% v / v rate per day, or 0.1% v / v rate per day.

[0257] In some embodiments, the floc may return back to the system at about a about 0.1 %v / v rate per day to about 15 %v / v rate per day. In some embodiments, the floc may return back to the system at about a about 0.1 %v / v rate per day to about 0.5 %v / v rate per day, about 0.1 %v / v rate per day to about 1 %v / v rate per day, about 0.1 %v / v rate per day to about 1.5 %v / v rate per day, about 0.1 %v / v rate per day to about 2 %v / v rate per day, about 0.1 %v / v rate per day to about 3 %v / v rate per day, about 0.1 %v / v rate per day to about 4 %v / v rate per day, about 0.1 %v / v rate per day to about 5 %v / v rate per day, about 0.1 %v / v rate per day to about 7 %v / v rate per day, about 0.1 %v / v rate per day to about 10 %v / v rate per day, about 0. 1 %v / v rate per day to about 12 %v / v rate per day, about 0.1 %v / v rate per day to about 15 %v / v rate per day, about 0.5 %v / v rate per day to about 1 %v / v rate per day, about 0.5 %v / v rate per day to about 1.5 %v / v rate per day, about 0.5 %v / v rate per day to about 2 %v / v rate per day, about 0.5 %v / v rate per day to about 3 %v / v rate per day, about 0.5 %v / v rate per day to about 4 %v / v rate per day, about 0.5 %v / v rate per day to about 5 %v / v rate per day, about 0.5 %v / v rate per day to about 7 %v / v rate per day, about 0.5 %v / v rate per day to about 10 %v / v rate per day, about 0.5 %v / v rate per day to about 12 %v / v rate per day, about 0.5 %v / v rate per day to about 15 %v / v rate per day, about 1 %v / v rate per day to about 1.5 %v / v rate per day, about 1 %v / v rate per day to about 2 %v / v rate per day, about 1 %v / v rate per day to about 3 %v / v rate per day, about 1 %v / v rate per day to about 4 %v / v rate per day, about 1 %v / v rate per day to about 5 %v / v rate per day, about 1 %v / v rate per day to about 7 %v / v rate per day, about 1 %v / v rate per day to about 10 %v / v rate per day, about 1 %v / v rate per day to about 12 %v / v rate per day, about 1 %v / v rate per day to about 15 %v / v rate per day, about 1.5 %v / v rate per day to about 2 %v / v rate per day, about 1.5 %v / v rate per day to about 3 %v / v rate per day, about 1.5 %v / v rate per day to about 4%v / v rate per day, about 1.5 %v / v rate per day to about 5 %v / v rate per day, about 1.5 %v / v rate per day to about 7 %v / v rate per day, about 1.5 %v / v rate per day to about 10 %v / v rate per day, about 1.5 %v / v rate per day to about 12 %v / v rate per day, about 1.5 %v / v rate per day to about 15 %v / v rate per day, about 2 %v / v rate per day to about 3 %v / v rate per day, about 2 %v / v rate per day to about 4 %v / v rate per day, about 2 %v / v rate per day to about 5 %v / v rate per day, about 2 %v / v rate per day to about 7 %v / v rate per day, about 2 %v / v rate per day to about 10 %v / v rate per day, about 2 %v / v rate per day to about 12 %v / v rate per day, about 2 %v / v rate per day to about 15 %v / v rate per day, about 3 %v / v rate per day to about 4 %v / v rate per day, about 3 %v / v rate per day to about 5 %v / v rate per day, about 3 %v / v rate per day to about 7 %v / v rate per day, about 3 %v / v rate per day to about 10 %v / v rate per day, about 3 %v / v rate per day to about 12 %v / v rate per day, about 3 %v / v rate per day to about 15 %v / v rate per day, about 4 %v / v rate per day to about 5 %v / v rate per day, about 4 %v / v rate per day to about 7 %v / v rate per day, about 4 %v / v rate per day to about 10 %v / v rate per day, about 4 %v / v rate per day to about 12 %v / v rate per day, about 4 %v / v rate per day to about 15 %v / v rate per day, about 5 %v / v rate per day to about 7 %v / v rate per day, about 5 %v / v rate per day to about 10 %v / v rate per day, about 5 %v / v rate per day to about 12 %v / v rate per day, about 5 %v / v rate per day to about 15 %v / v rate per day, about 7 %v / v rate per day to about 10 %v / v rate per day, about 7 %v / v rate per day to about 12 %v / v rate per day, about 7 %v / v rate per day to about 15 %v / v rate per day, about 10 %v / v rate per day to about 12 %v / v rate per day, about 10 %v / v rate per day to about 15 %v / v rate per day, or about 12 %v / v rate per day to about 15 %v / v rate per day.

[0258] In some embodiments, the digestion system can comprise a hydraulic source 150. In some embodiments, base product feed from a digestion system (e.g., P2) may act as a hydraulic source 150. In some embodiments, water can act as a hydraulic source 150. In some embodiments, when coupled to the first reactor 110, the hydraulic source 150 can provide continuous flow of water to the complete mixed reactor 110. In some embodiments, a water feed may act as a hydraulic source 150 and, when coupled to the first reactor 110, provides a continuous flow of water to the complete mixed reactor 110.

[0259] In some embodiments, as solids accumulate over time in the clarifier chamber 125, a range of at least about 20-25% solids v / v may be maintained in the digestion system. In some embodiments, additional floc may be harvested from the digestion system and removed. In some embodiments, at least about 5% solids v / v, at least about 10% solids v / v, at least about 15% solids v / v, at least about 16% solids v / v, at least about 17% solids v / v, at least about 18% solids v / v, at least about 19% solids v / v, at least about 20% solids v / v, at least about 21% solids v / v, at-n-least about 22% solids v / v, at least about 23% solids v / v, at least about 24% solids v / v, at least about 25% solids v / v, at least about 26% solids v / v, at least about 27% solids v / v, at least about 28% solids v / v, at least about 29% solids v / v, at least about 30% solids v / v, at least about 35% solids v / v, at least about 40% solids v / v, at least about 45% solids v / v, or at least about 50% solids v / v may be maintained in the digestion system.

[0260] In some embodiments, at most about 50% solids v / v, at most about 45% solids v / v, at most about 40% solids v / v, at most about 35% solids v / v, at most about 30% solids v / v, at most about 29% solids v / v, at most about 28% solids v / v, at most about 27% solids v / v, at most about 26% solids v / v, at most about 25% solids v / v, at most about 24% solids v / v, at most about 23% solids v / v, at most about 22% solids v / v, at most about 21% solids v / v, at most about 20% solids v / v, at most about 19% solids v / v, at most about 18% solids v / v, at most about 17% solids v / v, at most about 16% solids v / v, at most about 15% solids v / v, at most about 14% solids v / v, at most about 13% solids v / v, at most about 12% solids v / v, at most about 11% solids v / v, at most about 10% solids v / v, or at most about 5% solids v / v may be maintained in the digestion system.

[0261] In some embodiments, about 0.1% solids v / v to about 60% solids v / v may be maintained in the digestion system. In some embodiments, about 0.1% solids v / v to about 1% solids v / v, about 0.1% solids v / v to about 5% solids v / v, about 0.1% solids v / v to about 10% solids v / v, about 0.1% solids v / v to about 15% solids v / v, about 0.1% solids v / v to about 20% solids v / v, about 0.1% solids v / v to about 25% solids v / v, about 0.1% solids v / v to about 30% solids v / v, about 0.1% solids v / v to about 35% solids v / v, about 0.1% solids v / v to about 40% solids v / v, about 0.1% solids v / v to about 50% solids v / v, about 0.1% solids v / v to about 60% solids v / v, about 1% solids v / v to about 5% solids v / v, about 1% solids v / v to about 10% solids v / v, about 1% solids v / v to about 15% solids v / v, about 1% solids v / v to about 20% solids v / v, about 1% solids v / v to about 25% solids v / v, about 1% solids v / v to about 30% solids v / v, about 1% solids v / v to about 35% solids v / v, about 1% solids v / v to about 40% solids v / v, about 1% solids v / v to about 50% solids v / v, about 1% solids v / v to about 60% solids v / v, about 5% solids v / v to about 10% solids v / v, about 5% solids v / v to about 15% solids v / v, about 5% solids v / v to about 20% solids v / v, about 5% solids v / v to about 25% solids v / v, about 5% solids v / v to about 30% solids v / v, about 5% solids v / v to about 35% solids v / v, about 5% solids v / v to about 40% solids v / v, about 5% solids v / v to about 50% solids v / v, about 5% solids v / v to about 60% solids v / v, about 10% solids v / v to about 15% solids v / v, about 10% solids v / v to about 20% solids v / v, about 10% solids v / v to about 25% solids v / v, about 10% solids v / v to about 30% solids v / v, about 10% solids v / v to about 35% solids v / v, about 10% solids v / v to about 40% solids v / v, about 10% solids v / v to about 50% solids v / v, about 10% solids v / v to about 60% solids v / v, about15% solids v / v to about 20% solids v / v, about 15% solids v / v to about 25% solids v / v, about15% solids v / v to about 30% solids v / v, about 15% solids v / v to about 35% solids v / v, about15% solids v / v to about 40% solids v / v, about 15% solids v / v to about 50% solids v / v, about15% solids v / v to about 60% solids v / v, about 20% solids v / v to about 25% solids v / v, about20% solids v / v to about 30% solids v / v, about 20% solids v / v to about 35% solids v / v, about20% solids v / v to about 40% solids v / v, about 20% solids v / v to about 50% solids v / v, about20% solids v / v to about 60% solids v / v, about 25% solids v / v to about 30% solids v / v, about25% solids v / v to about 35% solids v / v, about 25% solids v / v to about 40% solids v / v, about25% solids v / v to about 50% solids v / v, about 25% solids v / v to about 60% solids v / v, about30% solids v / v to about 35% solids v / v, about 30% solids v / v to about 40% solids v / v, about30% solids v / v to about 50% solids v / v, about 30% solids v / v to about 60% solids v / v, about35% solids v / v to about 40% solids v / v, about 35% solids v / v to about 50% solids v / v, about35% solids v / v to about 60% solids v / v, about 40% solids v / v to about 50% solids v / v, about40% solids v / v to about 60% solids v / v, or about 50% solids v / v to about 60% solids v / v.

[0262] In some embodiments, the input composition, as described herein, may comprise a selective pressure or a phosphate source (e.g., phosphate ion source) 140. Without wishing to be bound by theory, the selective pressure source (e.g., phosphate source) may shift the complex microbial consortia of the digestion system 100 to increased phosphate solubilizing microbes. The selective pressure source may change the ionic concentration of the working fluid. Microbes of the microbial consortia may not be able to live in the new ionic environment and a shift in the complex microbial consortia may occur. In some embodiments, the selective pressure may create a high ionic concentration which selectively inhibits the growth of some microbes within the working fluid, while promoting the growth or survival of other microbes (e.g., an inoculum of a microbe) in the working fluid. The phosphate solubilizing microbes may comprise microbes that can survive a higher phosphate ion concentration in a working fluid caused by addition of the selective pressure. In some embodiments, the inoculum of a microbe may comprise a phosphate solubilizing microbe. Addition of the selective pressure to the digestion system 100 may promote survival of the inoculum of the microbe. The terms “selective pressure source” and “selective pressure” may be used interchangeably. Addition of a selective pressure source (e.g., phosphate source) may increase a concentration of phosphate within the working fluid of the digestion system 100.

[0263] In some embodiments, the sole or primary source of phosphate in a bioreactor system may comprise rock phosphate. In some embodiments, the sole or primary source of phosphate may comprise different rock phosphates from different sources that may contain differentcompositions of phosphorous and / or other bound micronutrients, macronutrients, or metals. In some embodiments, the sole or primary source of phosphate may comprise rock phosphate mined from different geographies. A sole or primary source of phosphate may be added to enrich for a phosphate-solubilizing community of microbes and / or metabolites and / or to stimulate phosphate-solubilizing activity by microbes within the system. The sole or primary source of phosphate (e.g., rock phosphate) may be added to the digestion system 100 on a first day of a digestion process. The sole or primary source of phosphate (e.g., rock phosphate) may be added to the first reactor 110, the second reactor 115, or another reactor of the digestion system. A source of insoluble phosphate(e.g., rock phosphate) may be added into the first reactor at a range of 10.0-50.0 grams per day, based on the span of the hydraulic retention time of the system. A hydraulic retention time may comprise an amount of time a working fluid is maintained in a container (e.g., is not transferred out) of a digestion system as described herein. In some embodiments, a selective pressure (e.g., rock phosphate) may be added to the digestion system at a concentration of at least about 1 gram / day, at least about 2 grams / day, at least about 5 grams / day, at least about 7 grams / day, at least about 10 grams / day, at least about 15 grams / day, at least about 20 grams / day, at least about 25 grams / day, at least about 30 grams / day, at least about 35 grams / day, at least about 40 grams / day, at least about 45 grams / day, at least about 50 grams / day, at least about 55 grams / day, at least about 60 grams / day, at least about 70 grams / day, at least about 80 grams / day, at least about 90 grams / day, or at least about 100 grams / day. In some embodiments, rock phosphate may be added to the digestion system at a concentration of at most about 100 grams / day, at most about 90 grams / day, at most about 80 grams / day, at most about 70 grams / day, at most about 60 grams / day, at most about 55 grams / day, at most about 50 grams / day, at most about 45 grams / day, at most about 40 grams / day, at most about 35 grams / day, at most about 30 grams / day, at most about 25 grams / day, at most about 20 grams / day, at most about 15 grams / day, at most about 10 grams / day, at most about 7 grams / day, at most about 5 grams / day, at most about 2 grams / day, or at most about 1 gram / day.

[0264] In some embodiments, a source of insoluble phosphate (e.g., rock phosphate) may be added to the digestion system at a concentration of about 5 grams / day to about 100 grams / day. In some embodiments, a source of insoluble phosphate (e.g., rock phosphate) may be added to the digestion system at a concentration of about 5 grams / day to about 10 grams / day, about 5 grams / day to about 15 grams / day, about 5 grams / day to about 20 grams / day, about 5 grams / day to about 25 grams / day, about 5 grams / day to about 30 grams / day, about 5 grams / day to about 35 grams / day, about 5 grams / day to about 40 grams / day, about 5 grams / day to about 50 grams / day,about 5 grams / day to about 60 grams / day, about 5 grams / day to about 75 grams / day, about 5 grams / day to about 100 grams / day, about 10 grams / day to about 15 grams / day, about 10 grams / day to about 20 grams / day, about 10 grams / day to about 25 grams / day, about 10 grams / day to about 30 grams / day, about 10 grams / day to about 35 grams / day, about 10 grams / day to about 40 grams / day, about 10 grams / day to about 50 grams / day, about 10 grams / day to about 60 grams / day, about 10 grams / day to about 75 grams / day, about 10 grams / day to about 100 grams / day, about 15 grams / day to about 20 grams / day, about 15 grams / day to about 25 grams / day, about 15 grams / day to about 30 grams / day, about 15 grams / day to about 35 grams / day, about 15 grams / day to about 40 grams / day, about 15 grams / day to about 50 grams / day, about 15 grams / day to about 60 grams / day, about 15 grams / day to about 75 grams / day, about 15 grams / day to about 100 grams / day, about 20 grams / day to about 25 grams / day, about 20 grams / day to about 30 grams / day, about 20 grams / day to about 35 grams / day, about 20 grams / day to about 40 grams / day, about 20 grams / day to about 50 grams / day, about 20 grams / day to about 60 grams / day, about 20 grams / day to about 75 grams / day, about 20 grams / day to about 100 grams / day, about 25 grams / day to about 30 grams / day, about 25 grams / day to about 35 grams / day, about 25 grams / day to about 40 grams / day, about 25 grams / day to about 50 grams / day, about 25 grams / day to about 60 grams / day, about 25 grams / day to about 75 grams / day, about 25 grams / day to about 100 grams / day, about 30 grams / day to about 35 grams / day, about 30 grams / day to about 40 grams / day, about 30 grams / day to about 50 grams / day, about 30 grams / day to about 60 grams / day, about 30 grams / day to about 75 grams / day, about 30 grams / day to about 100 grams / day, about 35 grams / day to about 40 grams / day, about 35 grams / day to about 50 grams / day, about 35 grams / day to about 60 grams / day, about 35 grams / day to about 75 grams / day, about 35 grams / day to about 100 grams / day, about 40 grams / day to about 50 grams / day, about 40 grams / day to about 60 grams / day, about 40 grams / day to about 75 grams / day, about 40 grams / day to about 100 grams / day, about 50 grams / day to about 60 grams / day, about 50 grams / day to about 75 grams / day, about 50 grams / day to about 100 grams / day, about 60 grams / day to about 75 grams / day, about 60 grams / day to about 100 grams / day, or about 75 grams / day to about 100 grams / day.

[0265] In some embodiments, the input composition to the digestion system 100 may comprise a carbon source. The carbon source may be glucose, gluconic acid, lactose, sucrose, pyruvate, simple sugars, a filtered manure, or a combination thereof 160. The manure may be cow manure, chicken manure, horse manure, sheep manure, alpaca manure, rabbit manure, pig manure, or a combination thereof. In some cases, the manure may be added to the digestion system on a firstday of a digestion process. In some embodiments, the carbon source may be added to the second reactor 115 of the digestion system. In some embodiments, the carbon source may be added to a first reactor 110 or a third reactor 120. In some embodiments, the filtered manure comprises a chemical oxygen demand of 1200-3000 mg / L. In some embodiments, the filtered manure may be added to the second reactor 115 to maintain a concentration range of 250-2000 mL of carbon per day based on the span of the hydraulic retention time of the system. In some embodiments, the concentration of carbon that may be maintained can be based on the total concentration of all components of the working fluid in the at a point during the digestion process of the digestion system.

[0266] In some embodiments, the carbon source (e.g., glucose, manure) may be added to a reactor to maintain a concentration of at least about, at most about, or about 50 mL / day, 100 mL / day, 150 mL / day, 200 mL / day, 225 mL / day, 250 mL / day, 275 mL / day, 300 mL / day, 350 mL / day, 400 mL / day, 500 mL / day, 600 mL / day, 700 mL / day, 800 mL / day, 900 mL / day, 1000 mL / day, 1250 mL / day, 1500 mL / day, 1750 mL / day, 2000 mL / day, 2250 mL / day, 2500 mL / day, 2750 mL / day, 3000 mL / day, 4000 mL / day, 5000 mL / day, 7500 mL / day, or 10000 mL / day, or a range between any of these two values.

[0267] An intermediate product of a digestion system (e.g., a P2 digestion system) may be used as an input. In some embodiments, a solution from a packed bed digestion system 170 (e.g., PBR-2 feed) may be added to a reactor of the digestion system described herein. In some embodiments, a mixture of water and raw manure (e.g., manure that has not been incubated in a water slurry for more than 5, 10, 15, 20, or 24 hours) may be used as an input. In some embodiments, a PBR-2 feed may be added to the second reactor 115. In some embodiments, a PBR-2 feed may be added to a first reactor 110 or a third reactor 120. In some embodiments, the PBR-2 feed may be added to the second reactor 115 to maintain a concentration range of 500.0- 4000.0 mL per day based on the span of the hydraulic retention time of the system. In some embodiments, the PBR-2 feed of an external digestion system may be added to a reactor to maintain a concentration of at least about, at most about, or about 200 mL / day, 225 mL / day, 250 mL / day, 275 mL / day, 300 mL / day, 350 mL / day, 400 mL / day, 500 mL / day, 600 mL / day, 700 mL / day, 800 mL / day, 900 mL / day, 1000 mL / day, 1250 mL / day, 1500 mL / day, 1750 mL / day, 2000 mL / day, 2250 mL / day, 2500 mL / day, 2750 mL / day, 3000 mL / day, 4000 mL / day, 5000 mL / day, 7500 mL / day, or 10000 mL / day, or a range between any of these two values.

[0268] In some embodiments, the input composition to the digestion system 100 may comprise a nitrogen source. The nitrogen source may be ammonium sulfate, ammonium chloride, ammonium nitrate, sodium nitrate, yeast extract, yeast, or a combination thereof. In some cases,the nitrogen source may be added to the digestion system on a first day of a digestion process. In some cases, the nitrogen source may be added to the digestion system on a second day, third day, fourth day, or any day following a first day of a digestion process. In some embodiments, the nitrogen source may be added to the second reactor 115 of the digestion system. In some embodiments, the nitrogen source may be added to a first reactor 110, or a third reactor 120 of the system. The yeast may be Saccharomyces cerevisiae yeast. The yeast may be added into a reactor of the system to maintain a concentration range of 0.50-5.0 grams of nitrogen per day based on the span of the hydraulic retention time of the system. In some embodiments, the concentration of nitrogen that may be maintained can be based on the total concentration of all components of the working fluid in the at a point during the digestion process of the digestion system.

[0269] In some embodiments, the yeast may be added to a reactor to maintain a concentration of at least about, at most about, or about 0.1 grams / day, 0.2 grams / day, 0.3 grams / day, 0.4 grams / day, 0.5 grams / day, 0.6 grams / day, 0.7 grams / day, 0.8 grams / day, 0.9 grams / day, 1.0 grams / day, 1.25 grams / day, 1.5 grams / day, 1.75 grams / day, 2.0 grams / day, 2.5 grams / day, 3.0 grams / day, 3.5 grams / day, 4.0 grams / day, 4.5 grams / day, 5.0 grams / day, 5.5 grams / day, 6.0 grams / day, 6.5 grams / day, 7.0 grams / day, 7.5 grams / day, 8.0 grams / day, 9.0 grams / day, or 10.0 grams / day, or a range between any of these two values.

[0270] In some embodiments, the input composition to the digestion system 100 may comprise micronutrients. Micronutrients may comprise important elements for microorganisms to support physiological functions. The micronutrients may comprise less than about 1%, less than about 2%, less than about 3%, less than about 4%, or less than about 5% of a dry weight of a plant. Elemental micronutrients may comprise boron, zinc, manganese, copper, chlorine, molybdenum, or a combination thereof. The addition of micronutrients or compounds that provide micronutrients may enrich an inoculum of a microbe and / or microbes of a microbial consortium of a working fluid of a digestion system as described herein.

[0271] In some embodiments, the digestion system 100 may be inoculated with a Bacillus amyloliquefaciens strain (e.g., the Bacillus amyloliquefaciens strain deposited under ATCC Accession No. PTA-127657, or a Bacillus amyloliquefaciens strain having a 16S rRNA gene sequence at least 98% identical to SEQ ID NO: 1) and / or a Bacillus licheniformis strain (e.g., a Bacillus licheniformis strain having a 16S rRNA gene sequence at least 98% identical to SEQ ID NO: 2). A microbial inoculum may be added to the first reactor 110. The microbial inoculum may be added to a second reactor 115 or a third reactor 120. A microbial inoculum may be present in a P2 base product feed, a filtered manure feed, or a PBR-2 feed as described herein. Insome cases, a digestion 100 system may be inoculated with an isolate or combination of isolates once. In some cases, a digestion system 100 may not be reinoculated with an isolate or combination of isolates following a first inoculation. In some cases, a digestion system 100 may be reinoculated with an isolate or combination of isolates at least every 20 days, at least every 50 days, at least every 100 days, at least every 200 days, at least every 300 days, at least every 400 days, at least every 500 days, or more. In some cases, a digestion system 300 may be reinoculated with an isolate or combination of isolates at most every 500 days, at most every 400 days, at most every 300 days, at most every 200 days, at most every 100 days, at most every 50 days, at most every 20 days, or less. In some cases, a digestion system 100 may be inoculated with an isolate or combination of isolates on day 1 of a digestion process and reinoculated 1, 2, 3, 4, 5, or more times after day 1 of the digestion process. In some embodiments, the population of the microbe does not decrease by more than 60%, by more than 50%, by more than 45%, by more than 40%, by more than 35%, by more than 30%, by more than 25%, by more than 20%, by more than 15%, by more than 10%, or by more than 5% between a first and a second transferring of the isolate (e.g., microbe). In some embodiments, the population of the microbe is maintained by at least about 5%, at least 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, or at least about 75%.

[0272] The inoculum of a microbe may be an isolated microbe. An isolated microbe can comprise a microbe grown or enriched outside of a natural environment (e.g., in a culture medium or a streak plate method). In some embodiments, the inoculum of a microbe may comprise a mixture of multiple isolated microbes. The inoculum of a microbe may comprise a mixture of at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, or more isolated microbes.

[0273] In some embodiments, the input composition, as described herein, may comprise plantbased materials (e.g., soluble plant-based materials). The plant-based material may be com flour, cereal flour, com gluten, soy protein hydrolysate, or any combination thereof. The plant-based material may be added to the reactor to at any concentration range of 0.2-3% w / v based on the span of the hydraulic retention time of the digestion system. In some embodiments, the plantbased material may be added to the reactor to at a concentration range of at least about, at most about, or about 0.05% w / v, 0.075% w / v, 0.1% w / v, 0.125% w / v, 0.15% w / v, 0.175% w / v, 0.2% w / v, 0.25% w / v, 0.3% w / v, 0.5% w / v, 0.75% w / v, 1.0% w / v, 1.5% w / v, 2.0% w / v, 2.5% w / v, 3.0% w / v, 3.5% w / v, 4.0% w / v, 5.0% w / v, 6.0% w / v, 7.0% w / v, 8.0% w / v, 9.0% w / v, 10.0% w / v, or a range between any two of these values.

[0274] A working fluid may be maintained within a reactor 110. A maintained working fluid may be a working fluid as it rests in a reactor 110 of a digestion system. Working fluid may be maintained within each reactor of a digestion system. A volume of a working fluid may be maintained by changing a flow rate of the digestion system. A volume of a working fluid may be maintained by changing a rate at which working fluid is recycled from a clarifier chamber to a reactor of the digestion system. Without wishing to be bound by theory, maintenance of a working fluid may support enrichment and / or growth of an inoculum of a microbe or another component of a working fluid as described herein. In some embodiments, the first reactor 110, the second reactor 115, and / or the third reactor 120 may be at least about a 1 gallon, at least about a 2 gallon, at least about a 3 gallon, at least about a 4 gallon, at least about a 5 gallon, at least about a 6 gallon, at least about a 7 gallon, at least about a 8 gallon, at least about a 9 gallon, at least about a 10 gallon, at least about a l l gallon, at least about a 12 gallon, at least about a 13 gallon, at least about a 14 gallon, at least about a 15 gallon, at least about a 16 gallon, at least about a 17 gallon, at least about a 18 gallon, at least about a 19 gallon, at least about a 20 gallon, at least about a 25 gallon, at least about a 30 gallon, at least about a 40 gallon, at least about a 50 gallon reactor, at least about a 100 gallon reactor, at least about a 500 gallon reactor, at least about a 1,000 gallon reactor, at least about a 2,000 gallon reactor, at least about a 3,000 gallon reactor, at least about a 4,000 gallon reactor, at least about a 5,000 gallon reactor, at least about a 10,000 gallon reactor, or at least about a 12,000 gallon reactor.

[0275] In some embodiments, the first reactor 110, the second reactor 115, and / or the third reactor 120 may be at most about a 50 gallon, at most about a 100 gallon, at most about a 500 gallon at most about a 1,000 gallon, at most about a 10,000 gallon, at most about a 12,000 gallon, at most about a 40 gallon, at most about a 30 gallon, at most about a 25 gallon, at most about a 20 gallon, at most about a 19 gallon, at most about a 18 gallon, at most about a 17 gallon, at most about a 16 gallon, at most about a 15 gallon, at most about a 14 gallon, at most about a 13 gallon, at most about a 12 gallon, at most about a l l gallon, at most about a 10 gallon, at most about a 9 gallon, at most about a 8 gallon, at most about a 7 gallon, at most about a 6 gallon, at most about a 5 gallon, at most about a 4 gallon, at most about a 3 gallon, at most about a 2 gallon, or at most about 1 gallon reactor.

[0276] In some embodiments, the first reactor 110, the second reactor 115, and / or the third reactor 120 may be about a 1 gallon to 75 gallon reactor. In some embodiments, the first reactor 110, the second reactor 115, and / or the third reactor 120 may be about a 1 gallon to 3 gallon, 1 gallon to 5 gallon, 1 gallon to 7 gallon, 1 gallon to 10 gallon, 1 gallon to 12 gallon, 1 gallon to 15 gallon, 1 gallon to 17 gallon, 1 gallon to 20 gallon, 1 gallon to 25 gallon, 1 gallon to 50gallon, 1 gallon to 75 gallon, 3 gallon to 5 gallon, 3 gallon to 7 gallon, 3 gallon to 10 gallon, 3 gallon to 12 gallon, 3 gallon to 15 gallon, 3 gallon to 17 gallon, 3 gallon to 20 gallon, 3 gallon to 25 gallon, 3 gallon to 50 gallon, 3 gallon to 75 gallon, 5 gallon to 7 gallon, 5 gallon to 10 gallon, 5 gallon to 12 gallon, 5 gallon to 15 gallon, 5 gallon to 17 gallon, 5 gallon to 20 gallon, 5 gallon to 25 gallon, 5 gallon to 50 gallon, 5 gallon to 75 gallon, 7 gallon to 10 gallon, 7 gallon to 12 gallon, 7 gallon to 15 gallon, 7 gallon to 17 gallon, 7 gallon to 20 gallon, 7 gallon to 25 gallon, 7 gallon to 50 gallon, 7 gallon to 75 gallon, 10 gallon to 12 gallon, 10 gallon to 15 gallon, 10 gallon to 17 gallon, 10 gallon to 20 gallon, 10 gallon to 25 gallon, 10 gallon to 50 gallon, 10 gallon to 75 gallon, 12 gallon to 15 gallon, 12 gallon to 17 gallon, 12 gallon to 20 gallon, 12 gallon to 25 gallon, 12 gallon to 50 gallon, 12 gallon to 75 gallon, 15 gallon to 17 gallon, 15 gallon to 20 gallon, 15 gallon to 25 gallon, 15 gallon to 50 gallon, 15 gallon to 75 gallon, 17 gallon to 20 gallon, 17 gallon to 25 gallon, 17 gallon to 50 gallon, 17 gallon to 75 gallon, 20 gallon to 25 gallon, 20 gallon to 50 gallon, 20 gallon to 75 gallon, 25 gallon to 50 gallon, 25 gallon to 75 gallon, or 50 gallon to 75 gallon reactor.

[0277] In some embodiments, the total volume of working fluid within all of the reactors and clarifier may be maintained at 60 gallons. In some embodiments, the total volume of working fluid within all of the reactors and clarifier may be maintained at least about, at most about, or about 30 gallons, 40 gallons, 45 gallons, 50 gallons, 55 gallons, 60 gallons, 65 gallons, 70 gallons, 75 gallons, 80 gallons, 85 gallons, 90 gallons, 95 gallons, 100 gallons, or a range between any of these two values.

[0278] A hydraulic rate of flow (e.g., hydraulic flow rate) can help transfer working fluid between reactors of a digestion system. The hydraulic rate of flow may be 12 mL / min to maintain a retention time of about 13 days and may be varied accordingly for a retention time from 7 to 21 days depending on production volume requirements. In some embodiments, the hydraulic rate of flow through the digestion system may be at least about, at most about, or about 5 ml / min, 5.5 ml / min, 6 ml / min, 6.5 ml / min, 7 ml / min, 7.5 ml / min, 8 ml / min, 9 ml / min, 10 ml / min, 11 ml / min, 12 ml / min, 13 ml / min, 14 ml / min, 14.5 ml / min, 15 ml / min, 15.5 ml / min, 16 ml / min, 16.5 ml / min, 17 ml / min, 17.5 ml / min, 20 ml / min, 22.5 ml / min, 25 ml / min, 30 ml / min, 35 ml / min, 40 ml / min, 50 ml / min, 60 ml / min, or 75 ml / min, or a range between any two of these values.

[0279] In some embodiments, the hydraulic rate of flow through the digestion system 100 may be at least about 2 ml / min, at least about 5 ml / min, at least about 10 ml / min, at least about 20 ml / min, at least about 25 ml / min, at least about 50 ml / min, at least about 75 ml / min, at least about 100 ml / min, at least about 150 ml / min, at least about 200 ml / min, at least about 250ml / min, at least about 500 ml / min, at least about 750 ml / min, at least about 1,000 ml / min, at least about 1,500 ml / min, at least about 2,000 ml / min, at least about 2,500 ml / min, at least about 3,000 ml / min, at least about 4,000 ml / min, at least about 5,000 ml / min, at least about 10,000 ml / min, at least about 15,000 ml / min, or at least about 20,000 ml / min. In some embodiments, the hydraulic rate of flow through the digestion system 100 may be at most about 5,000 ml / min, at most about 4,000 ml / min, at most about 3,000 ml / min, at most about 2,500 ml / min, at most about 2,000 ml / min, at most about 1,500 ml / min, at most about 1,000 ml / min, at most about 750 ml / min, at most about 500 ml / min, at most about 250 ml / min, at most about 200 ml / min, at most about 150 ml / min, at most about 100 ml / min, at most about 75 ml / min, at most about 50 ml / min, at most about 25 ml / min, at most about 20 ml / min, at most about 10 ml / min, at most about 5 ml / min, or at most about 2 ml / min.

[0280] In some embodiments, the hydraulic rate of flow through the digestion system 100 may be from about 5 ml / min to about 20,000 ml / min. In some embodiments, the hydraulic rate of flow through the digestion system 100 may be from about 5 ml / min to about 10 ml / min, about 5 ml / min to about 15 ml / min, about 5 ml / min to about 20 ml / min, about 5 ml / min to about 25 ml / min, about 5 ml / min to about 50 ml / min, about 5 ml / min to about 100 ml / min, about 5 ml / min to about 250 ml / min, about 5 ml / min to about 500 ml / min, about 5 ml / min to about 1,000 ml / min, about 5 ml / min to about 2,500 ml / min, about 5 ml / min to about 5,000 ml / min, about 5,000 ml / min to about 10,000 ml / min, about 5,000 ml / min to about 15,000 ml / min, about 10,000 ml / min to about 20,000 ml / min, about 10 ml / min to about 15 ml / min, about 10 ml / min to about 20 ml / min, about 10 ml / min to about 25 ml / min, about 10 ml / min to about 50 ml / min, about 10 ml / min to about 100 ml / min, about 10 ml / min to about 250 ml / min, about 10 ml / min to about 500 ml / min, about 10 ml / min to about 1,000 ml / min, about 10 ml / min to about 2,500 ml / min, about 10 ml / min to about 5,000 ml / min, about 15 ml / min to about 20 ml / min, about 15 ml / min to about 25 ml / min, about 15 ml / min to about 50 ml / min, about 15 ml / min to about 100 ml / min, about 15 ml / min to about 250 ml / min, about 15 ml / min to about 500 ml / min, about 15 ml / min to about 1,000 ml / min, about 15 ml / min to about 2,500 ml / min, about 15 ml / min to about 5,000 ml / min, about 20 ml / min to about 25 ml / min, about 20 ml / min to about 50 ml / min, about 20 ml / min to about 100 ml / min, about 20 ml / min to about 250 ml / min, about 20 ml / min to about 500 ml / min, about 20 ml / min to about 1,000 ml / min, about 20 ml / min to about 2,500 ml / min, about 20 ml / min to about 5,000 ml / min, about 25 ml / min to about 50 ml / min, about 25 ml / min to about 100 ml / min, about 25 ml / min to about 250 ml / min, about 25 ml / min to about 500 ml / min, about 25 ml / min to about 1,000 ml / min, about 25 ml / min to about 2,500 ml / min, about 25 ml / min to about 5,000 ml / min, about 50 ml / min to about 100 ml / min, about 50 ml / minto about 250 ml / min, about 50 ml / min to about 500 ml / min, about 50 ml / min to about 1,000 ml / min, about 50 ml / min to about 2,500 ml / min, about 50 ml / min to about 5,000 ml / min, about 100 ml / min to about 250 ml / min, about 100 ml / min to about 500 ml / min, about 100 ml / min to about 1,000 ml / min, about 100 ml / min to about 2,500 ml / min, about 100 ml / min to about 5,000 ml / min, about 250 ml / min to about 500 ml / min, about 250 ml / min to about 1,000 ml / min, about 250 ml / min to about 2,500 ml / min, about 250 ml / min to about 5,000 ml / min, about 500 ml / min to about 1,000 ml / min, about 500 ml / min to about 2,500 ml / min, about 500 ml / min to about 5,000 ml / min, about 1,000 ml / min to about 2,500 ml / min, about 1,000 ml / min to about 5,000 ml / min, or about 2,500 ml / min to about 5,000 ml / min.

[0281] A retention time may comprise a time an inoculum of a microbe spends in a digestion system or a time an inoculum of a microbe spends following transfer into a first container and until collection from the digestion system. A longer retention time may be advantageous for growth or enrichment of an inoculum of a microbe of the digestion system. A shorter retention time may be advantageous for growth or enrichment of an inoculum of a microbe of the digestion system. In some embodiments, the retention time of digestion system 100 may be 13 days. In some embodiments, the retention time of the digestion system may be at least about 1 day, at least about 2 days, at least about 3 days, at least about 4 days, at least about 5 days, at least about 6 days, at least about 7 days, at least about 8 days, at least about 9 days, at least about 10 days, at least about 11 days, at least about 12 days, at least about 13 days, at least about 14 days, at least about 15 days, at least about 16 days, at least about 17 days, at least about 18 days, at least about 19 days, at least about 20 days, at least about 25 days, at least about 30 days, or at least about 50 days. In some embodiments, the retention time of the digestion system 100 may be at most about 50 days, at most about 30 days, at most about 25 days, at most about 20 days, at most about 19 days, at most about 18 days, at most about 17 days, at most about 16 days, at most about 15 days, at most about 14 days, at most about 13 days, at most about 12 days, at most about 11 days, at most about 10 days, at most about 9 days, at most about 8 days, at most about 7 days, at most about 6 days, at most about 5 days, at most about 4 days, at most about 3 days, at most about 2 days, or at most about 1 day.

[0282] In some embodiments, the retention time of the digestion system 100 may be about 1 day to about 30 days. In some embodiments, the retention time of the digestion system may be about 1 day to about 3 days, about 1 day to about 5 days, about 1 day to about 8 days, about 1 day to about 10 days, about 1 day to about 12 days, about 1 day to about 14 days, about 1 day to about 16 days, about 1 day to about 18 days, about 1 day to about 20 days, about 1 day to about 25 days, about 1 day to about 30 days, about 3 days to about 5 days, about 3 days to about 8 days,about 3 days to about 10 days, about 3 days to about 12 days, about 3 days to about 14 days, about 3 days to about 16 days, about 3 days to about 18 days, about 3 days to about 20 days, about 3 days to about 25 days, about 3 days to about 30 days, about 5 days to about 8 days, about 5 days to about 10 days, about 5 days to about 12 days, about 5 days to about 14 days, about 5 days to about 16 days, about 5 days to about 18 days, about 5 days to about 20 days, about 5 days to about 25 days, about 5 days to about 30 days, about 8 days to about 10 days, about 8 days to about 12 days, about 8 days to about 14 days, about 8 days to about 16 days, about 8 days to about 18 days, about 8 days to about 20 days, about 8 days to about 25 days, about 8 days to about 30 days, about 10 days to about 12 days, about 10 days to about 14 days, about 10 days to about 16 days, about 10 days to about 18 days, about 10 days to about 20 days, about 10 days to about 25 days, about 10 days to about 30 days, about 12 days to about 14 days, about 12 days to about 16 days, about 12 days to about 18 days, about 12 days to about 20 days, about 12 days to about 25 days, about 12 days to about 30 days, about 14 days to about 16 days, about 14 days to about 18 days, about 14 days to about 20 days, about 14 days to about 25 days, about 14 days to about 30 days, about 16 days to about 18 days, about 16 days to about 20 days, about 16 days to about 25 days, about 16 days to about 30 days, about 18 days to about 20 days, about 18 days to about 25 days, about 18 days to about 30 days, about 20 days to about 25 days, about 20 days to about 30 days, or about 25 days to about 30 days.

[0283] In some embodiments, the digestion system 100 may produce between 2.85 to 8.56 gallons of base product flowing from the clarifiers per day. In some embodiments, the digestion system may produce at least about 0.5 gallons, at least about 1 gallon, at least about 2 gallons, at least about 2.5 gallons, at least about 3.0 gallons, at least about 3.5 gallons, at least about 4.0 gallons, at least about 4.5 gallons, at least about 5.0 gallons, at least about 6.0 gallons, at least about 7.0 gallons, at least about 8.0 gallons, at least about 8.5 gallons, at least about 9.0 gallons, at least about 10.0 gallons, at least about 12.5 gallons, at least about 15.0 gallons, or at least about 20.0 gallons. In some embodiments, the digestion system may produce at most about 20.0 gallons, at most about 15.0 gallons, at most about 12.5 gallons, at most about 10.0 gallons, at most about 9.0 gallons, at most about 8.5 gallons, at most about 8.0 gallons, at most about 7.0 gallons, at most about 6.0 gallons, at most about 5.0 gallons, at most about 4.5 gallons, at most about 4.0 gallons, at most about 3.5 gallons, at most about 3.0 gallons, at most about 2.5 gallons, at most about 2.0 gallons, at most about 1.0 gallons, or at most about 0.5 gallon per day.

[0284] In some embodiments, the digestion system may produce about 0.1 gallons to about 15 gallons per day. In some embodiments, the digestion system may produce about 0.1 gallons to about 1 gallon, about 0.1 gallons to about 2 gallons, about 0.1 gallons to about 3 gallons, about0.1 gallons to about 4 gallons, about 0.1 gallons to about 5 gallons, about 0.1 gallons to about 7.5 gallons, about 0.1 gallons to about 8 gallons, about 0.1 gallons to about 8.5 gallons, about 0.1 gallons to about 9 gallons, about 0.1 gallons to about 10 gallons, about 0.1 gallons to about 15 gallons, about 1 gallon to about 2 gallons, about 1 gallon to about 3 gallons, about 1 gallon to about 4 gallons, about 1 gallon to about 5 gallons, about 1 gallon to about 7.5 gallons, about 1 gallon to about 8 gallons, about 1 gallon to about 8.5 gallons, about 1 gallon to about 9 gallons, about 1 gallon to about 10 gallons, about 1 gallon to about 15 gallons, about 2 gallons to about 3 gallons, about 2 gallons to about 4 gallons, about 2 gallons to about 5 gallons, about 2 gallons to about 7.5 gallons, about 2 gallons to about 8 gallons, about 2 gallons to about 8.5 gallons, about2 gallons to about 9 gallons, about 2 gallons to about 10 gallons, about 2 gallons to about 15 gallons, about 3 gallons to about 4 gallons, about 3 gallons to about 5 gallons, about 3 gallons to about 7.5 gallons, about 3 gallons to about 8 gallons, about 3 gallons to about 8.5 gallons, about3 gallons to about 9 gallons, about 3 gallons to about 10 gallons, about 3 gallons to about 15 gallons, about 4 gallons to about 5 gallons, about 4 gallons to about 7.5 gallons, about 4 gallons to about 8 gallons, about 4 gallons to about 8.5 gallons, about 4 gallons to about 9 gallons, about4 gallons to about 10 gallons, about 4 gallons to about 15 gallons, about 5 gallons to about 7.5 gallons, about 5 gallons to about 8 gallons, about 5 gallons to about 8.5 gallons, about 5 gallons to about 9 gallons, about 5 gallons to about 10 gallons, about 5 gallons to about 15 gallons, about 7.5 gallons to about 8 gallons, about 7.5 gallons to about 8.5 gallons, about 7.5 gallons to about 9 gallons, about 7.5 gallons to about 10 gallons, about 7.5 gallons to about 15 gallons, about 8 gallons to about 8.5 gallons, about 8 gallons to about 9 gallons, about 8 gallons to about 10 gallons, about 8 gallons to about 15 gallons, about 8.5 gallons to about 9 gallons, about 8.5 gallons to about 10 gallons, about 8.5 gallons to about 15 gallons, about 9 gallons to about 10 gallons, about 9 gallons to about 15 gallons, or about 10 gallons to about 15 gallons per day.

[0285] In some embodiments, the digestion system 100 may produce about 100 gallons, about 250 gallons, about 500 gallons, about 1,000 gallons, about 1,500 gallons, about 2,000 gallons, about 3,000 gallons, about 4,000 gallons, about 5,000 gallons, about 6,000 gallons, about 7,000 gallons, about 8,000 gallons, about 9,000 gallons, or about 10,000 gallons or more per day. In some embodiments, the digestion system 100 may produce from about 10 gallons to about 2,500 gallons per day. In some embodiments, the digestion system 100 may produce from about 10 gallons to about 25 gallons, about 10 gallons to about 50 gallons, about 10 gallons to about 100 gallons, about 10 gallons to about 150 gallons, about 10 gallons to about 200 gallons, about 10 gallons to about 250 gallons, about 10 gallons to about 500 gallons, about 10 gallons to about 1,000 gallons, about 10 gallons to about 1,500 gallons, about 10 gallons to about 2,000 gallons,about 10 gallons to about 2,500 gallons, about 25 gallons to about 50 gallons, about 25 gallons to about 100 gallons, about 25 gallons to about 150 gallons, about 25 gallons to about 200 gallons, about 25 gallons to about 250 gallons, about 25 gallons to about 500 gallons, about 25 gallons to about 1,000 gallons, about 25 gallons to about 1,500 gallons, about 25 gallons to about 2,000 gallons, about 25 gallons to about 2,500 gallons, about 50 gallons to about 100 gallons, about 50 gallons to about 150 gallons, about 50 gallons to about 200 gallons, about 50 gallons to about 250 gallons, about 50 gallons to about 500 gallons, about 50 gallons to about 1,000 gallons, about 50 gallons to about 1,500 gallons, about 50 gallons to about 2,000 gallons, about 50 gallons to about 2,500 gallons, about 100 gallons to about 150 gallons, about 100 gallons to about 200 gallons, about 100 gallons to about 250 gallons, about 100 gallons to about 500 gallons, about 100 gallons to about 1,000 gallons, about 100 gallons to about 1,500 gallons, about 100 gallons to about 2,000 gallons, about 100 gallons to about 2,500 gallons, about 150 gallons to about 200 gallons, about 150 gallons to about 250 gallons, about 150 gallons to about 500 gallons, about 150 gallons to about 1,000 gallons, about 150 gallons to about 1,500 gallons, about 150 gallons to about 2,000 gallons, about 150 gallons to about 2,500 gallons, about 200 gallons to about 250 gallons, about 200 gallons to about 500 gallons, about 200 gallons to about 1,000 gallons, about 200 gallons to about 1,500 gallons, about 200 gallons to about 2,000 gallons, about 200 gallons to about 2,500 gallons, about 250 gallons to about 500 gallons, about 250 gallons to about 1,000 gallons, about 250 gallons to about 1,500 gallons, about 250 gallons to about 2,000 gallons, about 250 gallons to about 2,500 gallons, about 500 gallons to about 1,000 gallons, about 500 gallons to about 1,500 gallons, about 500 gallons to about 2,000 gallons, about 500 gallons to about 2,500 gallons, about 1,000 gallons to about 1,500 gallons, about 1,000 gallons to about 2,000 gallons, about 1,000 gallons to about 2,500 gallons, about 1,500 gallons to about 2,000 gallons, about 1,500 gallons to about 2,500 gallons, or about 2,000 gallons to about 2,500 gallons per day.

[0286] In some embodiments, the digestion system 100 may produce between 19.97 to 59.91 gallons of base product per week. In some embodiments, the digestion system may produce at least about, at most about, or about 5 gallons, 10 gallons, 15 gallons, 18 gallons, 20 gallons, 25 gallons, 30 gallons, 35 gallons, 40 gallons, 45 gallons, 50 gallons, 55 gallons, 60 gallons, 65 gallons, 70 gallons, 75 gallons, 85 gallons, or 100 gallons of base product per week, or a range between any of these two values.

[0287] In some embodiments, the digestion system 100 may produce about 500 gallons, about 1,000 gallons, about 2,000 gallons, about 2,500 gallons, about 5,000 gallons, about 7,000 gallons, about 10,000 gallons, about 15,000 gallons, about 20,000 gallons, about 25,000 gallons,about 30,000 gallons, about 40,000 gallons, about 50,000 gallons, about 60,000 gallons, about 70,000 gallons or more per week. In some embodiments, the digestion system 100 may produce from about 100 gallons to about 10,000 gallons per week. In some embodiments, the digestion system 100 may produce from about 100 gallons to about 250 gallons, about 100 gallons to about 500 gallons, about 100 gallons to about 1,000 gallons, about 100 gallons to about 1,500 gallons, about 100 gallons to about 2,000 gallons, about 100 gallons to about 2,500 gallons, about 100 gallons to about 5,000 gallons, about 100 gallons to about 7,000 gallons, about 100 gallons to about 7,500 gallons, about 100 gallons to about 8,000 gallons, about 100 gallons to about 10,000 gallons, about 250 gallons to about 500 gallons, about 250 gallons to about 1,000 gallons, about 250 gallons to about 1,500 gallons, about 250 gallons to about 2,000 gallons, about 250 gallons to about 2,500 gallons, about 250 gallons to about 5,000 gallons, about 250 gallons to about 7,000 gallons, about 250 gallons to about 7,500 gallons, about 250 gallons to about 8,000 gallons, about 250 gallons to about 10,000 gallons, about 500 gallons to about 1,000 gallons, about 500 gallons to about 1,500 gallons, about 500 gallons to about 2,000 gallons, about 500 gallons to about 2,500 gallons, about 500 gallons to about 5,000 gallons, about 500 gallons to about 7,000 gallons, about 500 gallons to about 7,500 gallons, about 500 gallons to about 8,000 gallons, about 500 gallons to about 10,000 gallons, about 1,000 gallons to about1.500 gallons, about 1,000 gallons to about 2,000 gallons, about 1,000 gallons to about 2,500 gallons, about 1,000 gallons to about 5,000 gallons, about 1,000 gallons to about 7,000 gallons, about 1,000 gallons to about 7,500 gallons, about 1,000 gallons to about 8,000 gallons, about 1,000 gallons to about 10,000 gallons, about 1,500 gallons to about 2,000 gallons, about 1,500 gallons to about 2,500 gallons, about 1,500 gallons to about 5,000 gallons, about 1,500 gallons to about 7,000 gallons, about 1,500 gallons to about 7,500 gallons, about 1,500 gallons to about 8,000 gallons, about 1,500 gallons to about 10,000 gallons, about 2,000 gallons to about 2,500 gallons, about 2,000 gallons to about 5,000 gallons, about 2,000 gallons to about 7,000 gallons, about 2,000 gallons to about 7,500 gallons, about 2,000 gallons to about 8,000 gallons, about 2,000 gallons to about 10,000 gallons, about 2,500 gallons to about 5,000 gallons, about 2,500 gallons to about 7,000 gallons, about 2,500 gallons to about 7,500 gallons, about 2,500 gallons to about 8,000 gallons, about 2,500 gallons to about 10,000 gallons, about 5,000 gallons to about 7,000 gallons, about 5,000 gallons to about 7,500 gallons, about 5,000 gallons to about 8,000 gallons, about 5,000 gallons to about 10,000 gallons, about 7,000 gallons to about 7,500 gallons, about 7,000 gallons to about 8,000 gallons, about 7,000 gallons to about 10,000 gallons, about7.500 gallons to about 8,000 gallons, about 7,500 gallons to about 10,000 gallons, or about 8,000 gallons to about 10,000 gallons per week.

[0288] In some embodiments, the base product collected from the digestion system 100 may have a pH of at least about 3.0, at least about 4.0, at least about 5.0, at least about 5.5, at least about 6.0, at least about 6.2, at least about 6.4, at least about 6.6, at least about 6.8, at least about 7.0, at least about 7.2, at least about 7.4, at least about 7.6, at least about 7.8, at least about 8.0, at least about 8.5, at least about 9.0, at least about 9.5, or at least about 10.0. In some embodiments, the base product collected from the digestion system 100 may have a pH of at most about 10.0, at most about 9.5, at most about 9.0, at most about 8.5, at most about 8.0, at most about 7.8, at most about 7.6, at most about 7.4, at most about 7.2, at most about 7.0, at most about 6.8, at most about 6.6, at most about 6.4, at most about 6.2, at most about 6.0, at most about 5.5, at most about 5.0, at most about 4.0, or at most about 3.0.

[0289] In some embodiments, the base product collected from the digestion system 100 may have a pH from about 3 to about 9. In some embodiments, the base product collected from the digestion system 100 may have a pH from about 3 to about 4, about 3 to about 5, about 3 to about 6, about 3 to about 6.5, about 3 to about 7, about 3 to about 7.2, about 3 to about 7.4, about 3 to about 7.6, about 3 to about 7.8, about 3 to about 8, about 3 to about 9, about 4 to about 5, about 4 to about 6, about 4 to about 6.5, about 4 to about 7, about 4 to about 7.2, about 4 to about 7.4, about 4 to about 7.6, about 4 to about 7.8, about 4 to about 8, about 4 to about 9, about 5 to about 6, about 5 to about 6.5, about 5 to about 7, about 5 to about 7.2, about 5 to about 7.4, about 5 to about 7.6, about 5 to about 7.8, about 5 to about 8, about 5 to about 9, about 6 to about 6.5, about 6 to about 7, about 6 to about 7.2, about 6 to about 7.4, about 6 to about 7.6, about 6 to about 7.8, about 6 to about 8, about 6 to about 9, about 6.5 to about 7, about 6.5 to about 7.2, about 6.5 to about 7.4, about 6.5 to about 7.6, about 6.5 to about 7.8, about 6.5 to about 8, about 6.5 to about 9, about 7 to about 7.2, about 7 to about 7.4, about 7 to about 7.6, about 7 to about 7.8, about 7 to about 8, about 7 to about 9, about 7.2 to about 7.4, about 7.2 to about 7.6, about 7.2 to about 7.8, about 7.2 to about 8, about 7.2 to about 9, about 7.4 to about 7.6, about 7.4 to about 7.8, about 7.4 to about 8, about 7.4 to about 9, about 7.6 to about 7.8, about 7.6 to about 8, about 7.6 to about 9, about 7.8 to about 8, about 7.8 to about 9, or about 8 to about 9.

[0290] The digestion process of a digestion system 100 may be an aerobic fermentation process. The working fluid in each reactor described herein may be circulated within the reactor in such a way as to that highly aerates the system using high-speed mixers 180. The dissolved oxygen within the first reactor 110, the second reactor 115, and / or the third reactor 120 may be at least about, at most about, or about 2.0, 2.5, 3.0, 4.0, 5.0, 6.0, 7.5, 8.0, 9.0, 10.0 mg / L, or a range between any of these two values.G. Biostimulant Compositions

[0291] A biostimulant may enhance water uptake and / or nutrient utilization in a plant and improves soil quality. A biostimulant and / or application of a biostimulant may enhance a yield of a crop. A biostimulant can comprise a microbe and / or microorganism described herein. A biostimulant may be a product of a digestion system as described herein. The inoculum of a microbe described herein may comprise a plant growth promotion property for a biostimulant product. The inoculum of a microbe described herein may be added to a biostimulant comprising plant extracts, protein hydrolysates, chemical biostimulants, humic or fulvic acids, seaweed extracts, liquid manure, other beneficial bacteria and / or fungi (e.g., Bacillus or rhizobiiim). or any combination thereof. The biostimulant composition may comprise a population of phosphate- solubilizing microbes enriched in the digestion system. In some embodiments, the biostimulant composition comprises a population of the inoculum of the phosphate-solubilizing microbe, at least a portion of phosphate-solubilizing microbes of the microbial consortium, phosphate- solubilizing metabolites generated in the digestion system, or any combination thereof. The phosphate-solubilizers enriched in the digestion system described herein may be added to plant extracts, protein hydrolysates, chemical biostimulants, humic or fulvic acids, seaweed extracts, liquid manure, other beneficial bacteria and / or fungi (e.g., Bacillus or rhizobiiim). or any combination thereof. In some embodiments, the present disclosure provides a biostimulant composition comprising chemical species and / or microbes that promote plant growth, including increasing a plant’s or crop’s phosphate use efficiency. Biostimulant compositions described herein may include dead microorganisms, sporulated microorganisms, fragments of dead microorganisms, viable microorganisms, microorganism digestion products, microbial metabolites and secondary metabolites, enzymes, biological plant growth regulators, organic acids, chelators, or any combination thereof.

[0292] In some embodiments, biostimulant compositions described herein may be characterized by mass spectrometry or NMR spectroscopy. In some embodiments, the biostimulant composition has an associated LC-MS chromatogram. In some embodiments, the biostimulant composition has an associated GC-MS chromatogram. In some embodiments, the biostimulant composition has an associated 'H-NMR spectrum. In some embodiments, the biostimulant composition has an associated13C-NMR spectrum.

[0293] In some embodiments, biostimulant compositions include viable microbes. In some embodiments, the microbes include bacteria that may be derived from the bacterial population present in manure, rock phosphate, or other digestion substrates. In some embodiments, themicrobes include bacteria that may be added to a digestion system as isolated microbial strains. In some embodiments, the bacteria include one or more bacteria of the species Bacillus safensis. Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus megaterium, Pseudomonas spp., Agrobacterium spp., Bacillus circulans, or any combination thereof. In some embodiments, the bacteria include one or more of microbial strains MS2839 (ATCC Accession No. PTA 127657) or MS1835 (ATCC Accession No. PTA-127656). In some embodiments, any one or any combination of these bacterial species comprises at least about 0.000001%, at least about 0.00001%, at least about 0.00005%, at least about 0.0001%, at least about 0.0005%, at least about 0.001%, at least about 0.0015%, at least about 0.002%, at least about 0.003%, at least about 0.004%, at least about 0.005%, at least about 0.01%, at least about 0.05%, at least about 0.1%, at least about 0.5%, at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 11%, at least about 12%, at least about 13%, at least about 14%, at least about 15%, at least about 20%, or more than about 20% of the total bacterial species present in the biostimulant, as determined by metagenomic sequencing or semi-quantitative PCR sequencing. In some embodiments, any one of these bacterial species comprises at most about 20%, at most about 15%, at most about 14%, at most about 13%, at most about 12%, at most about 11%, at most about 10%, at most about 9%, at most about 8%, at most about 7%, at most about 6%, at most about 5%, at most about 4%, at most about 3%, at most about 2%, at most about 1%, at most about 0.5%, at most about 0.1%, at most about 0.05%, at most about 0.01%, at most about 0.005%, at most about 0.004%, at most about 0.003%, at most about 0.002%, at most about 0.0015%, at most about 0.0005%, at most about 0.0001%, at most about 0.00005%, at most about 0.00001%, at most about 0.000001%, or less than about 0.000001% of the bacterial species present in the biostimulant, as determined by metagenomic sequencing or semi-quantitative PCR sequencing.

[0294] In some embodiments, the biostimulant is filter sterilized and does not comprise viable microbes. In some embodiments, the dry weight of the microbial biomass is less than 0.0001% in relation to the total dry weight of the biostimulant composition. In some embodiments, the biostimulant compositions may comprise metabolites from the microbial communities of the digestion system described herein. As working fluid is transferred in a digestion system, metabolites may be generated by metabolism of organic substrates. The metabolites may be produced with or without introduction of an isolate or target isolate. The metabolites may have positive effects on plant growth and plant growth promotion properties. In some embodiments, metabolites produced by the digestion systems described herein may enhance nutrient availability and / or nutrient use efficiency.

[0295] In some embodiments, the biostimulant comprises to IxlO8CFU / ml of bacteria. In some embodiments, the biostimulant comprises at least about, at most about, or about 100 CFU / ml, 500 CFU / ml, IxlO3CFU / ml, IxlO4CFU / ml, IxlO5CFU / ml, IxlO6CFU / ml, 5xl06CFU / ml, IxlO7CFU / ml, 5xl07CFU / ml, IxlO8CFU / ml, 5xl08CFU / ml, IxlO9CFU / ml, 5xl09CFU / ml, IxlO10CFU / ml, IxlO11CFU / ml IxlO12CFU / ml IxlO13CFU / ml IxlO14CFU / ml, or IxlO15CFU / ml of bacteria, or a range between any two of these values.

[0296] In some embodiments, the biostimulant has a pH of from 7 to 8. In some embodiments, the biostimulant has a pH of at least about, at most about, or about 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, or 11, or a range between any of these two values. In some embodiments, the density of the biostimulant is about 0.997 to 0.999 g / cm3or is about 0.998 g / cm3. In some embodiments, the density of the biostimulant is at least about, at most about, or about 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, or 1.5, or a range between any of these two values. In some embodiments, the biostimulant has a solids content of 0.01 to 2%. In some embodiments, the biostimulant has a solids content of at least about, at most about, or about 0.001%, 0.01%, 0.05%, 0.1%, 0.5%, 1%, 1.1%, 1.2%< 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 3%, 4%, or 5%, or a range between any of these two values. In some embodiments, the solids content is at least about, at most about, or about 0.01%, 0.05%, 0.1%, 0.15%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, or 2.0% w / w, or a range between any two of these values. In some embodiments, the chemical oxygen demand (COD) of the biostimulant is from 100 to 250 mg / L. In some embodiments, the COD is at least about, at most about, or about 10, 25, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 400, or 500 mg / L, or a range between any two of these values. COD values may vary with the concentration rate of the biostimulant, and may increase as concentration increases. In some embodiments, the biostimulant has a total nitrogen content of about 0.0002% w / w or between about 0.0001% and 0.0003% w / w. In some embodiments, the biostimulant has a total nitrogen content of at least about, at most about, or about 0.00001%, 0.00005%, 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.001%, or 0.01%, or a range between any of these two values. In some embodiments, the biostimulant has a total phosphorous content of about 0.00001% w / w or between about 0.000005% and 0.000015% w / w. In some embodiments, the biostimulant has a total phosphorus content of at least about, at most about, or about 0.00001%, 0.00002%, 0.00003%, 0.00004%, 0.00005%, 0.00006%, 0.00007%, 0.00008%, 0.00009%, 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.001%, or 0.01%, or a range between any of these two values. In some embodiments, the biostimulant has a total potassium content of about 0.0001% w / w or between about 0.00005% and 0.00015% w / w. In someembodiments, the biostimulant has a total potassium content of at least about, at most about, or about 0.00001, 0.00005%, 0.00006%, 0.00007%, 0.00008%, 0.00009%, 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.001%, or 0.01%, or a range between any of these two values.

[0297] In some embodiments, the biostimulant compositions described herein can comprise one or more compounds. The one or more compounds may comprise chemical compounds or derivatives thereof. The derivatives of any compound described herein can comprise stereoisomers, tautomers, solvates, isomers, pharmaceutically acceptable salts, or any combination thereof.

[0298] A bioproduct of a bioreactor system described herein can comprise one or more compounds. The PST and / or PwST product can comprise one or more compounds. The one or more compounds may be bioactive. The one or more compounds may have at least one plant growth promoting property as described herein. In some embodiments, a product (e.g., bioproduct or biostimulant composition) can comprise phenolic acids, fatty acyls (e.g., acylcarnitines or free fatty acids), organic acids and derivatives thereof, glycerophospholipids (e.g., phosphatidylcholines, p-glycoproteins, or phosphatidylinositols), alkaloids, carbohydrates and derivatives thereof, amino acids and metabolites thereof (e.g., small peptides), aldehydes, ketones, esters, benzenes and substituted derivatives thereof (e.g., phenolics), alcohols, amines, heterocyclic compounds, flavonoids (e.g., isoflavones), or any combination thereof. In some embodiments, the one or more compounds can comprise phenolic acids (e.g., phenols), acylcarnitines, organic acids and derivatives thereof (e.g., oxo dicarboxylic acids, carboxylic acids, or phosphonoacetic acids), phosphatidylcholines, amino acids and derivatives thereof (e.g., isoleucine derivatives), small peptides, phosphatidylglycerophosphate, ketones (e.g., resorcinols, chaicones, or aromatic ketones), free fatty acids (e.g., endocannibinoid analogues), alcohols (e.g., triterpenoids or 3Beta-sterol), aldehydes (e.g., acetal), phosphatidylinositols (e.g., glycerophosphocholines), benzenes and substituted derivatives thereof (e.g., dibenzofurans), heterocyclic compounds (e.g., steroids), isoflavones (e.g., 7-hydroxyisoflavones), polyamines (e.g., ureas), amines (e.g., benzamides), carotenoids (e.g., apo carotenoids), or any combination thereof.

[0299] In some embodiments, a product (e.g., bioproduct or biostimulant composition) can comprise 9,11-methane-epoxy Prostaglandin, Flalpha, Arg-Ser-Tyr, Ser-Tyr-Arg, 8-[(lr,2r,5r)- 2-(2-Carboxyethyl)-5-hydroxy-3-oxocyclopentyl]-6-oxooctanoic acid, 15(S)-Fluprostenol, Arg- Cys-Tyr, Catharanthine, Ochrolifuanine A, Sorbitol-6-phosphate, all-trans-Carophyll yellow, Arg-Ser-Val, D-Mannose 6-phosphate, Asn-Phe-Ala-Arg, FFA(16:0), 4- Hydroxybenzeneacetonitrile, 7-[(2R)-3-hydroxy-2-(3-hydroxy-5-methylnon-l-enyl)-5-oxocyclopentyl]-3-methoxyheptanoic acid, Ala-Pro- Asp, Gln-Val-Ile-Asp, H-Trp-ser-OH, 3,8- Dihydroxy-l-pentanoyl-6-pentyl-l lH-dibenzo(b,e)(l,4)dioxepin-l 1-one, tetranor-PGFM, Proleu, Pro-Gly-Ile, Oxolinic acid, 2-(4-hydroxy-3-methoxyphenyl)-3,4-dihydro-2H-l-benzopyran- 3,7-diol, Iriflophenone trimethyl ether, l,2-di-(5Z-hexadecenoyl)-sn-glycero-3- phosphoethanolamine, {4-[3-(5-hydroxy-2,2-dimethyl-2H-chromen-6- yl)propanoyl]phenyl}oxidanesulfonic acid, one or more derivatives thereof, or any combination thereof. In some embodiments, a product (e.g., bioproduct or biostimulant composition) can comprise Val-val, Prostaglandin D3, l-(3,4-Dimethoxyphenyl)-2-(2-methoxyphenoxy)propane-1.3-diol, Ser-Lys-Val, Tert-butyl 4-((3-(pyridin-4-yl)-l,2,4-oxadiazol-5-yl)methoxy)piperidine-1 -carboxylate, Actinonin, [(2S,6S,9R)-4,4,11,1 l-tetramethyl-3,5,7,10,12- pentaoxatricyclo[7.3.0.02,6]dodecan-6-yl]methyl sulfamate, prostaglandin B2, Haplopine, Asn- Tyr-Arg, 2-Pyrocatechuic acid, Monocrotaline, Glu-Tyr-Ile-Glu, Psoralidin, Lophocerine, Dodecyl glucopyranoside, (R)-Warfarin, Docosahexaenoic acid, Pro-Tyr, (2R)-3-(5-acetamido-2-hydroxyphenyl)sulfanyl-2-azaniumylpropanoate, His-Val-Ile, , Acetylphosphate, Lumichrome, prostaglandin Bl, Tyr-Asp-Ser, 4-Pregnen-17alpha,20beta-diol-3-one-20-sulfate, Osthole, Kojic acid, Didymin, , Protoheme, Homolanthionine dizwitterion, 7,14,15,16- tetrahydroxy-3-methyl-3,4,5,6,7,8,9,10,l l,12-decahydro-lH-2-benzoxacyclotetradecin-l-one, alpha-Ketoisovaleric acid, 2-(4-Morpholinyl)-8-phenyl-4H-l-benzopyran-4-one, Tetrabutylammonium, DL-Carbidopa, L-Tyrosine, 24, 5-benzyl-5-(pyridin-3-yl)imidazolidine-2.4-dione, Adrenoyl ethanolamidem, one or more derivatives thereof, or any combination thereof. In some embodiments, a product (e.g., bioproduct or biostimulant composition) can comprise Diosmin, Phenol glucuronide, Salsolinol, phlorisobutyrophenone, 3-methyl-N- naphthalen-2-ylbutanamide, (+)-Eudesmin, Bis(2-methylpropanoyloxy)-9, 10-epoxy-p-mentha- 1,3,5-triene, FFA(17:0), 3,5-dimethyl PIT-1, Isoleucylcysteine, Gly-Gly-Leu, 4'-Apo-beta- carotenal, 6-[(4Ar,8as)-octahydroquinolin-l(2h)-ylsulfonyl]-l,2,3,4-tetrahydroquinoline, Tryptamine, Arg-Arg-Lys, Metanephrine, Oleic Acid-biotin, Hydroxycotinine, Pseudoyohimbine, Methyl 3,12-dihydroxy-l l-ketoisoallospirostan-3-hemisuccinate, Phenacetin, Ala-Leu- Ala-Pro-Lys, L-Tyrosine ethyl ester, 4-Methoxyestrone, Pro-Phe-Gly, 3'-N'- Acetylfusarochromanone, Allopurinol riboside, Lys-Leu-Thr-Arg, Butyl linoleate, Glyceryl arachidonate, Thr-Leu-Val-Arg, 3 -Thiatetradecanoic Acid, 8-iso Prostaglandin F2d4, 2- Propylglutaric acid, Gln-Gly-Tyr, Mellein, 4-(Methylsulfanyl)butan-2-ol, Pro-Ile-Gly, (S)-(S)-1- ((2S,3 S)-3-Hexyl-4-oxooxetan-2-yl)tridecan-2-yl 2-formamido-4-methylpentanoate, Indole-3- acetamide, N-nonanoyl-L-Homoserine lactone, Carbaprostacyclin-biotin, Phe-Cys-Leu-Phe- Arg, Methyl carb amyl PAF C-8, Carnitine C6:0, 4,4'-Methylenedianiline, 2 -Phenoxy ethanol,Val-trp, Vicianose, 2-Phenylethynesulfonamide, one or more derivatives thereof, or any combination thereof.

[0300] The product of a bioreactor system described herein may be intact (e.g., not filter- sterilized) or it may be filter-sterilized. The filter-sterilized product can comprise metabolites. The metabolites can help phosphate solubilization. The metabolites can improve phosphate solubilization. The product can be from a sIP system described herein (e.g., a PST-sIP or PwST- sIP system described herein). In some embodiments, the composition can comprise at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 100, 1000, 5000, 10000, 25000, 50000, or greater than about 50000 compounds (e.g., different compounds or different classes of compounds) described herein.

[0301] In some embodiments, the intact sample from the bioreactor system described herein may be tested for one or more compounds. The bioreactor system can comprise a sIP system, in which the system can be inoculated with one or more isolate (e.g., microbial strains) described herein. The bioreactor system can comprise a non-sIP system, in which the system is not inoculated with one or more isolate (e.g., microbial strains) described herein. Without wishing to be bound by theory, the addition of the isolate to the bioreactor system can influence the composition of the working fluid and / or product of the system. Thus, the product (e.g., biostimulant composition) of a sIP system can comprise different compounds, different abundances of compounds, or different concentrations of compounds compared to those in a product (e.g., biostimulant composition) of a non-sIP system.

[0302] In some embodiments, one or more compounds can be found at a greater abundance in a product (e.g., intact product) of a sIP system compared to an abundance of the one or more compounds in a product (e.g., intact product) of a non-sIP system. In some embodiments, one or more compounds of a product of a system (e.g., sIP system) can comprise Dihydrocholesteroltrimethyl silyl-ether, Docosanoic anhydride, Quinoline, 2-monopalmitin, Beta-sitostanyl acetate, Nonane, 2,2,4,4,6,8,8-heptamethyl-, Palmitoleic acid trimethylsilyl ester, Benzen-1,3- Dicarboxylic acid, Heneicosanoic acid, Octadecanoic acid, 2 -hydroxy- 1,3 -propanediyl di-ester, Fumaric acid, cyclobutyl hexadecyl ester, 1,2-diaminopropane, 2-methylpentadecanoic acid trimethyl silylester, Distearin, Myristic acid, D-(+)-Malic acid, one or more derivatives thereof, or any combination thereof. In some embodiments, an abundance of the one or more compounds in a product (e.g., intact sample) from a sIP bioreactor system described herein may be at least about 2-fold, at least about 5-fold, at least about 10-fold, at least about 20-fold, at least about 30- fold, at least about 40-fold, at least about 50-fold, at least about 100-fold, at least about 200-fold, at least about 300-fold, at least about 400-fold, at least about 500-fold, at least about 1000-fold,or greater than about 1000-fold higher than an abundance of the one or more compounds in a product (e.g., sample) from a non-sIP bioreactor system described herein. In some embodiments, an abundance of the one or more compounds in a product (e.g., intact sample) from a sIP bioreactor system describ...

Claims

CLAIMSWHAT IS CLAIMED IS:

1. A method of making a biostimulant composition, the method comprising:(a) providing a bioreactor system comprising two or more containers arranged in a series, each of the two or more containers comprising a volume of a working fluid, wherein a first container comprises a first working fluid that includes a first microbial consortium and an established population of a phosphate solubilizing bacterial strain;(b) operating the bioreactor system for a duration of time by:(i) transferring into the first container an aqueous feedstock comprising a second microbial consortium;(ii) transferring a portion of the working fluid out of each of the two or more containers into either a subsequent container of the bioreactor system or a product outflow stream;(iii) maintaining a concentration of the phosphate solubilizing bacterial strain throughout the duration of time in at least the first container at at least 80% of a concentration of the phosphate solubilizing bacterial strain at the beginning of the duration of time; and(iv) collecting at least a portion of the product outflow stream as the biostimulant composition; wherein the duration of time is at least 5 days; and wherein the phosphate solubilizing bacterial strain is not present in the aqueous feedstock or any other input into the bioreactor system during the duration of time at a concentration that is higher than 1% of the concentration of the phosphate solubilizing microbe in the first container.

2. The method of claim 1, further comprising adding a source of insoluble phosphate to the first container.

3. The method of claim 2, wherein the source of insoluble phosphate comprises rock phosphate particles.

4. The method of claim 1 or 2, wherein the aqueous feedstock and any other input into the bioreactor system does not comprise soluble phosphate or does not comprise soluble phosphate at a concentration higher than 10 mg / L.

5. The method of claim 1, wherein the first microbial consortium comprises other phosphate solubilizing microbes that are not the phosphate solubilizing bacterial strain, and wherein the method further comprises maintaining a concentration of soluble phosphate and a concentration of a source of insoluble phosphate in at least the first container that stimulates phosphate solubilizing activity of the phosphate solubilizing microbial strain and / or the other phosphate solubilizing microbes.

6. The method of any one of claims 1 to 5, wherein the phosphate solubilizing bacterial strain is of the genus Bacillus.

7. The method of any one of claims 1 to 6, wherein the phosphate solubilizing bacterial strain is of the species Bacillus amyloliquefaciens o Bacillus licheniformis.

8. The method of any one of claims 1 to 7, wherein the phosphate solubilizing bacterial strain is one of the following:(a) a Bacillus amyloliquefaciens strain having one or more of the following:(i) a 16S rRNA gene sequence at least 95% identical to SEQ ID NO: 1;(ii) a gyrB gene sequence at least 95% identical to SEQ ID NO: 3; and(iii) a rpoB gene sequence at least 95% identical to SEQ ID NO: 5; or(b) a Bacillus licheniformis strain having one or more of the following:(i) a 16S rRNA gene sequence at least 95% identical to SEQ ID NO: 2;(ii) a gyrB gene sequence at least 95% identical to SEQ ID NO: 4; and(iii) a rpoB gene sequence at least 95% identical to SEQ ID NO: 6.

9. The method of any one of claims 1 to 8, wherein the phosphate solubilizing bacterial strain is the Bacillus amyloliquefaciens strain deposited under ATCC Accession No. PTA-127657 or the Bacillus licheniformis strain deposited under ATCC Accession No. PTA-127656.

10. The method of any one of claims 1 to 9, wherein the phosphate solubilizing bacterial strain is not present in the aqueous feedstock or any other input into the bioreactor system during the duration of time at a concentration of greater than 100 CFU / ml.

11. The method of any one of claims 1 to 10, wherein the phosphate solubilizing bacterial strain is not present in the aqueous feedstock or any other input into the bioreactor system during the duration of time.

12. The method of any one of claims 1 to 11, wherein the maintaining of step (b)(iii) comprises maintaining the concentration of the phosphate solubilizing bacterial strain at at least IxlO3CFU / ml.

13. The method of any one of claims 1 to 12, wherein, before step (b), the first container further comprises an established population of other phosphate solubilizing microbes that are not the phosphate solubilizing bacterial strain, and wherein step (b)(iii) further comprises maintaining a concentration of the other phosphate solubilizing microbes in at least the first container throughout the duration of time at at least IxlO3CFU / ml or at at least 80% of a concentration of the other phosphate solubilizing microbes at the beginning of the duration of time, wherein the other phosphate solubilizing microbes are not added to the bioreactor system during the duration of time at a concentration that is higher than 1% of the concentration of the other phosphate solubilizing microbes in the first container.

14. The method of claim 13, wherein the other phosphate solubilizing microbes are not present in the aqueous feedstock or any other input into the bioreactor system at a concentration of greater than 104CFU / ml.

15. The method of claim 13 or 14, wherein the population of the other phosphate solubilizing microbes in the first container is at least IxlO3CFU / ml at the beginning of the duration of time.

16. The method of any one of claims 1 to 15, further comprising, before step (a), adding an inoculum of the phosphate solubilizing bacterial strain to the bioreactor system, wherein the inoculum of the phosphate solubilizing bacterial strain produces an initial population of the phosphate solubilizing bacterial strain of at least 0.5xl04CFU / ml in at least one container.

17. The method of claim 16, wherein, before adding the inoculum of the phosphate solubilizing bacterial strain, the concentration of the phosphate solubilizing bacterial strain is less than IxlO2CFU / ml.

18. The method of any one of claims 1 to 17, wherein the aqueous feedstock further comprises an organic material at least partially digestible by microbes present in at least one of the containers.

19. The method of claim 18, wherein, before the transferring of step (b)(i), the organic material had been partially digested by microbes endogenous to the organic material.

20. The method of claim 18, further comprising digesting the organic material in two or more serially connected containers before the transferring of step (b)(i).

21. The method of any one of claims 18 to 20, wherein the organic material comprises manure and / or material produced by microbial digestion of manure.

22. The method of any one of claims 1 to 21, wherein the aqueous feedstock further comprises an inorganic material.

23. The method of any one of claims 2 to 22, wherein the rock phosphate particles had been added to a container upstream of the first container.

24. The method of claim 23, wherein the container upstream of the first container further comprises a hydraulic source.

25. The method of claim 24, wherein the hydraulic source comprises a product of anaerobic digestion of manure in a serialized digestion system.

26. The method of any one of claims 1 to 25, wherein the second microbial consortium comprises at least IxlO5CFU / ml.

27. The method of claim 26, wherein the second microbial consortium comprises microbes derived from manure.

28. The method of any one of claims 1 to 27, wherein the operating of step (b) further comprises producing microbial metabolites that directly or indirectly promote phosphate solubilization in a plant growth medium.

29. The method of any one of claims 1 to 28, wherein the transferring of step (b)(i), the transferring of step (b)(ii), and the collecting of step (b)(iv) are performed continuously throughout the duration of time.

30. The method of any one of claims 1 to 29, wherein the transferring of step (b)(i), the transferring of step (b)(ii), and the collecting of step (b)(iv) are performed periodically throughout the duration of time.

31. The method of any one of claims 1 to 30, further comprising adding one or more carbon sources to at least one container of the bioreactor system.

32. The method of claim 31, wherein the one or more carbon sources are included in the aqueous feedstock.

33. The method of claim 31 or 32, wherein the one or more carbon sources comprise polysaccharides.

34. The method of any one of claims 1 to 33, further comprising adding one or more nitrogen sources to at least one container of the bioreactor system.

35. The method of claim 34, wherein the one or more nitrogen sources comprise yeast.

36. The method of any one of claims 1 to 35, wherein the bioreactor system comprises a clarifier container comprising a clarifier working fluid.

37. The method of claim 36, further comprising separating a supernatant portion of the clarifier working fluid from a floc portion of the clarifier working fluid within the clarifier container.

38. The method of claim 37, wherein the separating comprises gravity separation.

39. The method of claim 37 or 38, further comprising folding the floc portion of the clarifier working fluid.

40. The method of claim 39, wherein the folding further comprises releasing a population of the phosphate solubilizing bacterial strain into the supernatant portion without introducing floc solids into the supernatant portion.

41. The method of claim 39 or 40, wherein the folding is performed by folding wipers in a bottom portion of the clarifier container.

42. The method of any one of claims 37 to 41, wherein the operating further comprises transferring the floc portion from the clarifier container to an earlier container in the bioreactor system.

43. The method of any one of claims 37 to 42, wherein the product outflow stream comprises the supernatant portion of the clarifier working fluid.

44. The method of any one of claims 1 to 43, wherein the method further comprises producing at least IxlO4CFU / ml of the phosphate solubilizing bacterial strain in the product outflow stream.

45. The method of any one of claims 1 to 44, wherein the bioreactor system comprises the first container comprising a volume of a first working fluid, a second container comprising a volume of a second working fluid, and a third container comprising a volume of a third working fluid.

46. The method of claim 45, wherein the first container comprises an outlet port fluidly connected to an inlet port of the second container and the second container comprises an outlet port fluidly connected to an input port of the third container.

47. The method of claim 46, wherein the third container comprises an outlet port fluidly connected to a clarifier container.

48. The method of any one of claims 45 to 47, further comprising maintaining the volume of each of the first working fluid, the second working fluid, and the third working fluid constant throughout the duration of time.

49. The method of any one of claims 1 to 48, wherein step (b) comprises operating the bioreactor system in a hydraulically balanced manner.

50. The method of any one of claims 1 to 49, wherein the transferring of step (b)(i), the transferring of step (b)(ii), and the collecting of step (b)(iv) are driven by gravity.

51. The method of any one of claims 1 to 50, wherein the operating comprises maintaining a flow rate that results in a hydraulic retention time of the bioreactor system of at least 5 days.

52. The method of any one of claims 1 to 51, wherein the operating comprises maintaining the product outflow stream at a flow rate of at least 100 gallons per day.

53. The method of any one of claims 1 to 52, wherein the volume of working fluid in each of the two or more containers is at least 100 gallons.

54. The method of any one of claims 1 to 53, wherein at least one of the two or more containers is a fluidized bed reactor.

55. The method of any one of claims 1 to 54, wherein at least one of the two or more containers is a packed bed reactor.

56. The method of any one of claims 1 to 55, further comprising maintaining at least one of the two or more containers under aerobic conditions.

57. The method of any one of claims 1 to 56, wherein the bioreactor system is operated continuously for at least 90 days.

58. The method of any one of claims 1 to 57, further comprising adding yeast into one of the two or more containers.

59. The method of claim 58, wherein the yeast is Saccharomyces cerevisiae.

60. The method of any one of claims 18 to 59, wherein the second microbial consortium comprises microbes endogenous to the organic material.

61. The method of any one of claims 45 to 60, wherein at least one of the first working fluid, the second working fluid, or the third working fluid comprises a pH buffering system.

62. The method of any one of claims 45 to 61, further comprising maintaining the pH of at least one of the first working fluid, the second working fluid, or the third working fluid between 6 and 8 throughout the duration of time.

63. The method of any one of claims 1 to 62, wherein the aqueous feedstock does not include the phosphate solubilizing bacterial strain at a concentration higher than 10 CFU / ml.

64. The method of any one of claims 1 to 63, wherein the phosphate solubilizing bacterial strain is not added to the bioreactor system during the duration of time at a concentration that is higher than 10 CFU / ml.

65. The method of any one of claims 1 to 64, wherein the bioreactor system comprises at least one container placed in the series before the first container.

66. The method of any one of claims 1 to 65, further comprising producing a population of sporulated bacteria in the product outflow stream.

67. The method of any one of claims 1 to 66, further comprising producing a population of the phosphate solubilizing bacterial strain in the product outflow stream that is sporulated.

68. The method of claim 67, wherein the population of the phosphate solubilizing bacterial strain that is sporulated comprises at least IxlO3CFU / ml.

69. The method of any one of claims 1 to 68, further comprising adding an additional population of the phosphate solubilizing bacterial strain to the biostimulant product.

70. The method of any one of claims 1 to 69, wherein the method further comprises making at least a portion of the aqueous feedstock by a method comprising:(c) at least partially digesting manure in a series of two or more fluidly connected manure digestion containers, thereby generating at least a portion of the aqueous feedstock.

71. The method of claim 70, wherein the series of two or more fluidly connected manure digestion containers comprise packed bed reactors.

72. The method of claim 70 or 71, further comprising maintaining the two or more fluidly connected manure digestion containers under anaerobic conditions.

73. The method of any one of claims 70 to 72, wherein step (c) comprises partially digesting the manure in only two manure digestion containers.

74. The method of any one of claims 70 to 73, further comprising adding yeast to at least one of the two or manure digestion containers.

75. The method of any one of claims 70 to 74, wherein step (c) comprises digesting the manure by microbes endogenous to the manure.

76. The method of any one of claims 70 to 75, wherein step (c) comprises maintaining hydraulically balanced flow between the two or more manure digestion containers.

77. A bioreactor system comprising:(a) a stream of an aqueous feedstock in fluid communication with a first container comprising a volume of a first working fluid, wherein the aqueous feedstock comprises a first microbial consortium, wherein the first working fluid comprises an established population of a phosphate solubilizing bacterial strain and a second microbial consortium, wherein a concentration of the phosphate solubilizing bacterial strain in the first working fluid is at least 100 times higher than a concentration of thephosphate solubilizing bacterial strain in the aqueous feedstock stream and in any other input into the bioreactor system;(b) one or more additional containers arranged in a series that includes the first container, wherein each of the one or more additional containers comprises a volume of a working fluid and is in fluid communication with at least one other container in the series, and wherein at least one of the one or more additional containers comprises a product outflow stream port; and(c) a product outflow stream in fluid communication with the product outflow stream port.

78. The system of claim 77, wherein at least the first working fluid comprises a concentration of soluble phosphate that is lower than 10 mg / L and a concentration of a source of insoluble phosphate that is at least 0.5 g / L.

79. The system of claim 78, wherein the source of insoluble phosphate comprises 20 to 30% of insoluble phosphate by weight.

80. The system of claim 78 or 79, wherein the source of insoluble phosphate comprises rock phosphate particles.

81. The system of claim 80, wherein the rock phosphate particles are present in a container that is upstream of the first container.

82. The system of any one of claims 77 to 80, wherein the aqueous feedstock and any other input into the bioreactor system does not comprise soluble phosphate or does not comprise soluble phosphate at a concentration higher than 10 mg / L.

83. The system of any one of claims 77 to 82, wherein the phosphate solubilizing bacterial strain is of the genus Bacillus.

84. The system of any one of claims 77 to 83, wherein the phosphate solubilizing bacterial strain is of the species Bacillus amyloliquefaciens o Bacillus licheniformis .

85. The system of any one of claims 77 to 84, wherein the phosphate solubilizing bacterial strain is one of the following:(a) a Bacillus amyloliquefaciens strain having one or more of the following:(i) a 16S rRNA gene sequence at least 95% identical to SEQ ID NO: 1;(ii) a gyrB gene sequence at least 95% identical to SEQ ID NO: 3; and(iii) a rpoB gene sequence at least 95% identical to SEQ ID NO: 5; or(b) a Bacillus licheniformis strain having one or more of the following:(i) a 16S rRNA gene sequence at least 95% identical to SEQ ID NO: 2;(ii) a gyrB gene sequence at least 95% identical to SEQ ID NO: 4; and(iii) a rpoB gene sequence at least 95% identical to SEQ ID NO: 6.

86. The system of any one of claims 77 to 85, wherein the phosphate solubilizing bacterial strain is the Bacillus amyloliquefaciens strain deposited under ATCC Accession No. PTA- 127657 or the Bacillus licheniformis strain deposited under ATCC Accession No. PTA- 127656.

87. The system of any one of claims 77 to 86, wherein the bioreactor system is a continuous flow bioreactor system and the stream of the aqueous feedstock is a continuous stream.

88. The system of any one of claims 77 to 87, wherein each of the volume of the working fluids is constant.

89. The system of any one of claims 77 to 88, wherein each of the first container and the one or more additional containers comprises a concentration of the phosphate solubilizing bacterial strain that remains at least IxlO4CFU / ml during operation of the bioreactor system.

90. The system of any one of claims 77 to 89, wherein the aqueous feedstock and any other input into the bioreactor system does not comprise the phosphate solubilizing bacterial strain or does not comprise a concentration of the phosphate solubilizing bacterial strain at level higher than 100 CFU / ml.

91. The system of any one of claims 77 to 90, wherein the first microbial consortium comprises at least IxlO4CFU / ml of microbes.

92. The system of any one of claims 77 to 91, wherein the aqueous feedstock further comprises an organic material digestible by microbes present in the containers.

93. The system of claim 92, wherein the organic material comprises manure or material derived from manure.

94. The system of any one of claims 77 to 93, wherein the first container further comprises yeast.

95. The system of claim 94, wherein the first microbial consortium comprises microbes derived from manure.

96. The system of any one of claims 77 to 95, wherein the container comprising the product outflow stream port is a clarifier container configured to separate a portion of a working fluid in the clarifier container into a supernatant portion and a floc portion.

97. The system of claim 96, wherein the clarifier container comprises one or more floc folding flights configured to agitate settled floc in the clarifier container without resuspending solids in the floc portion into the supernatant portion.

98. The system of claim 96 or 97, further comprising a floc return stream that flows from the clarifier to an earlier container in the series.

99. The system of any one of claims 96 to 98, wherein the product outflow stream comprises the supernatant portion.

100. The system of claim 99, wherein the product outflow stream comprises at least IxlO4CFU / ml of the phosphate solubilizing bacterial strain.

101. The system of claim 99 or 100, wherein the product outflow stream comprises at least IxlO2CFU / ml of a sporulated form of the phosphate solubilizing bacterial strain.

102. The system of any one of claims 99 to 101, wherein the product outflow stream comprises a total dry weight of 0.2 to 2.5 mg / ml.

103. The system of any one of claims 99 to 102, wherein the product outflow stream has a chemical oxygen demand between 80 to 500 mg / L.

104. The system of any one of claims 99 to 103, wherein the product outflow stream has an electrical conductivity between 1.3 and 3.0 mS / cm.

105. The system of any one of claims 77 to 104, wherein at least the first container comprises a mixer configured to aerate the first working fluid.

106. The system of any one of claims 77 to 105, wherein the first working fluid and / or the working fluid in at least one of the one or more additional containers comprises aerobic conditions.

107. A biostimulant composition made by the method of any one of claims 1 to 76 or the system of any one of claims 77 to 106.

108. A method of promoting plant growth comprising contacting a plant, seed, or plant growth medium with the biostimulant composition of claim 107.

109. A method of increasing an amount of solubilized phosphate available to a plant, the method comprising contacting a plant, seed, or plant growth medium with the biostimulant composition of claim 107.

110. A composition comprising:(a) a Bacillus amyloliquefaciens strain having one or more of the following:(i) a 16S rRNA gene sequence at least 95% identical to SEQ ID NO: 1;(ii) a gyrB gene sequence at least 95% identical to SEQ ID NO: 3; and(iii) a rpoB gene sequence at least 95% identical to SEQ ID NO: 5; and(b) a carrier.

111. The composition of claim 110, wherein the Bacillus amyloliquefaciens strain is the strain deposited under ATCC Accession No. PTA-127657, or an isolated clone thereof.

112. The composition of claim 110 or 111, further comprising products of digestion of an organic substrate by the Bacillus amyloliquefaciens strain.

113. The composition of any one of claims 110 to 112, wherein the carrier comprises a fertilizer.

114. The composition of any one of claims 110 to 113, wherein the carrier is a solid coated by the Bacillus amyloliquefaciens strain.

115. The composition of claim 114, wherein the carrier is further coated by a micronutrient.

116. The composition of claim 115, wherein the micronutrient comprises a source of insoluble phosphate.

117. The composition of any one of claims 110 to 113, wherein the carrier is a liquid.

118. The composition of any one of claims 110 to 117, wherein the carrier further comprises an adjuvant selected from a wetting agent, spreading agent, dispersing agent, sticking agent, dust control agent, or adhesive.

119. The composition of any one of claims 110 to 118, wherein the concentration of the Bacillus amyloliquefaciens strain in the composition ranges from IxlO3to IxlO11cfu / ml.

120. The composition of any one of claims 110 to 119, wherein the concentration of the Bacillus amyloliquefaciens strain in the composition ranges from IxlO4to IxlO6cfu / ml.

121. An isolated strain of the species Bacillus amyloliquefaciens having one or more of the following:(a) a 16S rRNA gene sequence at least 95% identical to SEQ ID NO: 1;(b) a gyrB gene sequence at least 95% identical to SEQ ID NO: 3; and(c) a rpoB gene sequence at least 95% identical to SEQ ID NO: 5.

122. The isolated strain of claim 121, wherein the B. amyloliquefaciens strain is the strain deposited under ATCC Accession No. PTA-127657, or an isolated clone thereof.

123. A composition comprising:(a) a Bacillus licheniformis strain having one or more of the following:(i) a 16S rRNA gene sequence at least 95% identical to SEQ ID NO: 2;(ii) a gyrB gene sequence at least 95% identical to SEQ ID NO: 4; and(iii) a rpoB gene sequence at least 95% identical to SEQ ID NO: 6; and(b) a carrier.

124. The composition of claim 123, wherein the Bacillus licheniformis strain is the strain deposited under ATCC Accession No. PTA-127656, or an isolated clone thereof.

125. The composition of claim 123 or 124, further comprising products of digestion of an organic substrate by the Bacillus licheniformis strain.

126. The composition of any one of claims 123 to 125, wherein the carrier comprises a fertilizer.

127. The composition of any one of claims 123 to 126, wherein the carrier is a solid coated by the Bacillus licheniformis strain.

128. The composition of claim 127, wherein the carrier is further coated by a micronutrient.

129. The composition of claim 128, wherein the micronutrient comprises a source of insoluble phosphate.

130. The composition of any one of claims 123 to 126, wherein the carrier is a liquid.

131. The composition of any one of claims 123 to 130, wherein the carrier further comprises an adjuvant selected from a wetting agent, spreading agent, dispersing agent, sticking agent, dust control agent, or adhesive.

132. The composition of any one of claims 123 to 131, wherein the concentration of the Bacillus licheniformis strain in the composition ranges from IxlO3to IxlO11cfu / ml.

133. The composition of any one of claims 123 to 132, wherein the concentration of the Bacillus licheniformis strain in the composition ranges from IxlO4to IxlO6.

134. An isolated strain of the species Bacillus licheniformis having one or more of the following:(a) a 16S rRNA gene sequence at least 95% identical to SEQ ID NO: 2;(b) a gyrB gene sequence at least 95% identical to SEQ ID NO: 4; and(c) a rpoB gene sequence at least 95% identical to SEQ ID NO: 6.

135. The isolated strain of claim 134, wherein the B. licheniformis strain is the strain deposited under ATCC Accession No. PTA-127656, or an isolated clone thereof.

136. A method for promoting growth of a plant growing in a medium, the method comprising contacting the plant or the medium with the biostimulant of claim 107, the composition of any one of claims 110 to 120 or 123 to 133, or a composition comprising the isolated strain of any one of claims 121, 122, 134, and 135.

137. The method of claim 136, wherein the contacting increases the amount of solubilized phosphate available to the plant by at least 5%.

138. The method of claim 136 or 137, wherein the composition increases plant growth by at least 5% as compared to a control.

139. The method of any one of claims 136 to 138, wherein the composition increases uptake of a nutrient by the plant by at least 5% as compared to a control.

140. The method of claim 139, wherein the nutrient is phosphate, and wherein the uptake of phosphate by the plant is increased by at least 5% as compared to a control.

141. The method of claim 139, wherein the nutrient is zinc.

142. The method of claim 139, wherein the nutrient is sulfur, potassium, magnesium, calcium, boron, manganese, iron, and / or copper.

143. The method of any one of claims 136 to 142, wherein the medium is soil or a hydroponic medium.

144. A method of remedying a phosphate deficiency in a plant growth medium, the method comprising:(a) measuring a concentration of soluble phosphate in the plant growth medium that is less than 25 ppm; and(b) after step (a), contacting the plant growth medium with the biostimulant of claim 107, the composition of any one of claims 110 to 120 or 123 to 133, or a composition comprising the isolated strain of any one of claims 121, 122, 134, and 135.

145. A method of promoting plant growth, comprising:(a) contacting a plant and / or plant growth medium in which the plant is growing with a composition comprising one or more compounds, wherein the one or more compounds is selected from the group consisting of Dihydrocholesterol-trimethylsilyl- ether, Docosanoic anhydride, Quinoline, 2-monopalmitin, Beta-sitostanyl acetate, Nonane, 2,2,4,4,6,8,8-heptamethyl-, Palmitoleic acid trimethyl silyl ester, Benzen-1,3- Dicarboxylic acid, Heneicosanoic acid, Octadecanoic acid, 2-hydroxy-l,3- propanediyl di-ester, Fumaric acid, cyclobutyl hexadecyl ester, 1,2-diaminopropane, 2-methyl pentadecanoic acid tri methyl silyl ester, Distearin, Myristic acid, D-(+)-Malic acid, one or more derivatives thereof, and any combination thereof.

146. The method of claim 145, wherein the promoting plant growth comprises promoting phosphate solubilization.

147. The method of claim 145 or 146, wherein the concentration of the one or more compounds in the composition is at least about 1 nanomolar (nM).

148. The method of any one of claims 145-147, wherein the concentration of the one or more compounds in the composition is at least about 0.00001% of a dry weight of the composition.

149. The method of any one of claims 145-148, wherein the contacting comprises contacting the plant with the composition.

150. The method of any one of claims 145-149, wherein the contracting comprises contacting a plant seed with the composition.

151. The method of any one of claims 145-150, wherein the contacting comprises contacting a leaf of the plant with the composition.

152. The method of any one of claims 145-151, wherein the plant growth, medium comprises soil, a hydroponic medium, turface, or isolite.

153. The method of any one of claims 145-152, wherein the contacting comprises increasing an amount of solubilized phosphate available to a plant.

154. The method of any one of claims 145-153, wherein the contacting comprises increasing phosphate solubilization in the plant growth medium.

155. The method of any one of claims 145-154, wherein the contacting causes an increase in plant growth by at least 10 percent as compared to the plant and / or the medium not contacted with the one or more compounds.

156. A method of promoting plant growth, comprising:(a) contacting a plant and / or plant growth medium in which the plant is growing with a composition comprising one or more compounds, wherein the one or more compounds is selected from the group consisting of 1 -o-hexadecylglycerol 2,3- ditrimethylsilylether, 2 -monopalmitin, 3-Methyl-p-anisaldehyde, Octadecanoic acid, 2-hydroxy-l,3-propanediyl di-ester, Eserine, Docosanoic anhydride, trans-p- Diniethylaminocinnamonitrile, Fumaric acid, cyclobutyl hexadecyl ester, (S)-(-)- . alpha. -(l-Naphthyl)ethylamine, 2-quinolinecarboxylic acid methyl ester, Isopentyl phenyl acetate, Methyl eicosanoate, Acenaphthylene, 4-(Anisylideneamino)-cinnamic acid, one or more derivatives thereof, and any combination thereof.

157. The method of claim 156, wherein the composition is filter-sterilized.

158. The method of claim 156 or 157, wherein the composition comprises phosphate- solubilizing metabolites.

159. The method of any one of claims 156-158, wherein the promoting plant growth comprises promoting phosphate solubilization.

160. The method of any one of claims 156-159, wherein the concentration of the one or more compounds in the composition is at least about 1 nanomolar (nM).

161. The method of any one of claims 156-160, wherein the concentration of the one or more compounds in the composition is at least about 0.00001% of a dry weight of the composition.

162. The method of any one of claims 156-161, wherein the contacting comprises contacting the plant with the composition.

163. The method of any one of claims 156-162, wherein the contracting comprises contacting a plant seed with the composition.

164. The method of any one of claims 156-163, wherein the contacting comprises contacting a leaf of the plant with the composition.

165. The method of any one of claims 156-164, wherein the plant growth medium comprises soil, a hydroponic medium, turface, or isolite.

166. The method of any one of claims 156-165, wherein the contacting comprises increasing an amount of solubilized phosphate available to a plant.

167. The method of any one of claims 156-166, wherein the contacting comprises increasing phosphate solubilization in the plant growth medium.

168. The method of any one of claims 156-167, wherein the contacting causes an increase in plant growth by at least 10 percent as compared to the plant and / or the plant growth medium not contacted with the one or more compounds.

169. A composition for promoting plant growth, comprising:(a) at least one microbial strain comprising a Bacillus amyloliquefaciens strain or a Bacillus licheniformis strain; and(b) one or more compounds selected from the group consisting of Dihydrocholesterol-trimethylsilyl-ether, Docosanoic anhydride, Quinoline, 2- monopalmitin, Beta-sitostanyl acetate, Nonane, 2,2,4,4,6,8,8-heptamethyl, Palmitoleic acid trimethylsilyl ester, Benzen-1,3-Dicarboxylic acid, Heneicosanoic acid, Octadecanoic acid, 2 -hydroxy- 1,3 -propanediyl di-ester, Fumaric acid, cyclobutyl hexadecyl ester, 1,2-diaminopropane, 2- methylpentadecanoic acid trimethylsilylester, Distearin, Myristic acid, D-(+)- Malic acid, one or more derivatives thereof, and any combination thereof.

170. The composition of claim 169, wherein the at least one microbial strain comprises Bacillus amyloliquefaciens comprising one or more of the following:(a) 16S rRNA gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 1;(b) a gyrB gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 3; or(c) a rpoB gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 5.

171. The composition of claim 169, wherein the at least one microbial strain comprises Bacillus licheniformis comprising one or more of the following:(a) a 16S rRNA gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 2;(b) a gyrB gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 4; or(c) a rpoB gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 6.

172. The composition of any one of claims 169-171, wherein Docosanoic anhydride, Heneicosanoic acid, Fumaric acid, cyclobutyl hexadecyl ester, Distearin, D-(+)-Malic acid,or any combination thereof is present in the composition at the highest relative abundance and / or Quinoline is present at the lowest relative abundance relative to a total dry weight of the composition.

173. The composition of any one of claims 169-172, further comprising a carrier.

174. The composition of claim 173, wherein the carrier is formulated for application to a plant and / or medium in which the plant is growing.

175. A composition for promoting plant growth, comprising:(a) at least one microbial strain comprising a Bacillus amyloliquefaciens strain or a Bacillus licheniformis strain; and(b) one or more compounds selected from the group consisting of l-o- hexadecylglycerol 2,3-ditrimethylsilylether, 2-monopalmitin, 3-Methyl-p- anisaldehyde, Octadecanoic acid, 2 -hydroxy- 1,3 -propanediyl di-ester, Eserine, Docosanoic anhydride, trans-p-Dimethylaminocinnamonitrile, Fumaric acid, cyclobutyl hexadecyl ester, (S)-(-)-. alpha. -(l-Naphthyl)ethylamine, 2- quinolinecarboxylic acid methyl ester, Isopentyl phenyl acetate, Methyl eicosanoate, Acenaphthylene, 4-(Anisylideneamino)-cinnamic acid, one or more derivatives thereof, and any combination thereof.

176. The composition of claim 175, wherein the at least one microbial strain comprises Bacillus amyloliquefaciens comprising one or more of the following:(a) 16S rRNA gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 1;(b) a gyrB gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 3; or(c) a rpoB gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 5.

177. The composition of claim 175, wherein the at least one microbial strain comprises Bacillus licheniformis comprising one or more of the following:(a) a 16S rRNA gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 2;(b) a gyrB gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 4; or(c) a rpoB gene sequence at least 95% identical to the sequence set forth in SEQ ID NO: 6.

178. The composition of any one of claims 175-177, wherein Octadecanoic acid, 2 -hydroxy- 1,3- propanediyl di-ester, Eserine, Docosanoic anhydride, trans-p-Dimethylaminocinnamonitrile, Fumaric acid, cyclobutyl hexadecyl ester, (S)-(-)-.alpha.-(l-Naphthyl)ethylamine, or any combination thereof is present in the composition at the highest relative abundance and / or 3-Methyl-p-anisaldehyde is present at the lowest relative abundance relative to a total dry weight of the composition.

179. The composition of any one of claims 175-178, further comprising a carrier.

180. The composition of claim 179, wherein the carrier is formulated for application to a plant and / or medium in which the plant is growing.

181. A composition for promoting plant growth, comprising:(a) two or more compounds selected from the group consisting of Dihydrocholesterol-trimethylsilyl-ether, Docosanoic anhydride, Quinoline, 2- monopalmitin, Beta-sitostanyl acetate, Nonane, 2,2,4,4,6,8,8-heptamethyl, Palmitoleic acid trimethylsilyl ester, Benzen-1,3-Dicarboxylic acid, Heneicosanoic acid, Octadecanoic acid, 2-hydroxy- 1,3 -propanediyl di-ester, Fumaric acid, cyclobutyl hexadecyl ester, 1,2-diaminopropane, 2- methylpentadecanoic acid trimethylsilylester, Distearin, Myristic acid, D-(+)~ Malic acid, one or more derivatives thereof, and any combination thereof; and(b) a carrier.

182. The composition of claim 181, wherein the carrier is formulated for application to a plant or plant growth medium in which the plant is growing.

183. The composition of claim 181 or 182, wherein the carrier comprises a fertilizer.

184. The composition of claim 183, wherein the fertilizer is a solid.

185. The composition of any one of claims 181-184, wherein the carrier is a liquid.

186. The composition of any one of claims 181-185, wherein the composition is configured to increase an amount of solubilized phosphate available to a plant.

187. The composition of any one of claims 181-186, wherein the composition is configured to increase phosphate solubilization in the plant growth medium.

188. The composition of any one of claims 181-187, wherein Docosanoic anhydride, Heneicosanoic acid, Fumaric acid, cyclobutyl hexadecyl ester, Distearin, D-(+)-Malic acid, or any combination thereof is present in the composition at the highest relative abundance and / or Quinoline is present at the lowest relative abundance relative to a total dry weight of the composition.

189. The composition of any one of claims 181-188, wherein the composition further compri ses an adjuvant selected from the group consisting of a wetting agent, spreading agent, dispersing agent, sticking agent, dust control agent, and adhesive.

190. The composition of any one of claims 181-189, wherein the composition is configured to increase or is capable of increasing an amount of solubilized phosphate available to a plant by at least 5%.

191. The composition of any one of claims 181-190, wherein the composition is configured to cause or is capable of causing an increase in plant growth by at least 10 percent as compared to a control.

192. A composition for promoting plant growth, comprising:(a) two or more compounds selected from the group consisting of l-o- hexadecylglycerol 2,3-ditrimethylsilylether, 2-monopalmitin, 3-Methyl-p- anisaldehyde, Octadecanoic acid, 2 -hydroxy- 1,3 -propanediyl di-ester, Eserine, Docosanoic anhydride, trans-p-Dimethylaminocinnamonitrile, Fumaric acid, cyclobutyl hexadecyl ester, (S)-(-)-.alpha.-(l-Naphthyl)ethylamine, 2- quinolinecarboxylic acid methyl ester, Isopentyl phenylacetate, Methyleicosanoate, Acenaphthylene, 4-(Anisylideneamino)-cinnamic acid, one or more derivatives thereof, and any combination thereof; and(b) a carrier.

193. The composition of claim 192, wherein the carrier is formulated for application to a plant or plant growth medium in which the plant is growing.

194. The composition of claim 192 or 193, wherein the carrier comprises a fertilizer.

195. The composition of claim 194, wherein the fertilizer is a solid.

196. The composition of any one of claims 192-195, wherein the carrier is a liquid.

197. The composition of any one of claims 192-196, wherein the composition is configured to increase phosphate solubilization in the plant growth medium.

198. The composition of any one of claims 192-197, wherein the composition is configured to increase phosphate solubilization in the plant growth medium.

199. The composition of any one of claims 192-198, wherein Octadecanoic acid, 2 -hydroxy- 1,3- propanediyl di-ester, Eserine, Docosanoic anhydride, trans-p-Dimethylaminocinnamonitrile, Fumaric acid, cyclobutyl hexadecyl ester, (S)-(-)-.alpha.-(l-Naphthyl)ethylamine, or any combination thereof is present in the composition at the highest relative abundance and / or 3-Methyl-p-anisaldehyde is present at the lowest relative abundance relative to a total dry weight of the composition.

200. The composition of any one of claims 192-199, wherein the composition further comprises an adjuvant selected from the group consisting of a wetting agent, spreading agent, dispersing agent, sticking agent, dust control agent, and adhesive.

201. The composition of any one of claims 192-200, wherein the composition is configured to increase or is capable of increasing an amount of solubilized phosphate available to a plant by at least 5%.

202. The composition of any one of claims 192-201, wherein the composition is configured to cause or is capable of causing an increase in plant growth by at least 10 percent as compared to a control.

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