Bio-nutrient formulation to enhance soil microbes and functions thereof

Abstract

The present disclosure provides a formulation comprising a bio-nutrient composition and a binder. Provided is the formation that further comprises an additive for confer additional desirable traits. The present disclosure provides a method of making the formulation in a granular and / or liquid form. Also provided is use of the formulation for improving soil health and inducing plant growth, development, and / or stress-related defense.

Description

BIO-NUTRIENT FORMULATION TO ENHANCE SOIL MICROBESAND FUNCTIONS THEREOFCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 524,422, filed on June 30, 2023, which is herein incorporated by reference in their entirety.TECHNICAL FIELD

[0002] The disclosure relates to the identification and use of bio-nutrients derived from organic and food wastes for enhancing activities and functions of soil microbes and improving plant growth and / or development.BACKGROUND

[0003] Plants require nitrogen, phosphorus, potassium, secondary and other micronutrients, for their growth and survival. Nitrogen is an essential building block of amino acids, which in turn are the building blocks of proteins. However, plants cannot use the di -nitrogen readily available in the earth's atmosphere. They require nitrate or ammonium found in soil, which can be produced by natural nitrogen fixation processes like lightning strikes, precipitation, and a symbiotic relationship with nitrogen-fixing bacteria. Alternatively, man-made fertilizers can provide ammonium to the soil.

[0004] In agriculture, plants consume the soil nitrogen as they grow, leaving the soil depleted of its available nitrogen after harvest. Farmers have several options to compensate for this nitrogen deficiency, such as crop-rotation techniques or the use of man-made fertilizers.

[0005] Overusing chemical fertilizers can have detrimental effects on the soil, including contributing to soil acidification and the formation of soil crusts. These effects can reduce the levels of organic matter and humus content in the soil, as well as negatively impact beneficial species and stunt plant growth. Additionally, excessive use of chemical fertilizers can alter the pH of the soil, encourage the growth of pests, and even result in the release of greenhouse gases.

[0006] To address these issues, organic fertilizers have been developed and utilized. Compared to conventional chemical fertilizers however, organic fertilizers typically require a larger volume of application. Unfortunately, this excess application often results in runoff that cancontaminate important sources of drinking water both above and below ground. This contamination is due to the high concentration of toxins and excess nutrients that are present in areas where they should not be, leading to harmful algae blooms that devastate aquatic life in bodies of water.

[0007] Therefore, there is a need for a safe, economical, and effective means of producing bionutrients and its formulations for agricultural application. Greenhouse propagation, turf and ornamental, and home and garden use that can combine multiple ingredients evenly and consistently and be used in hydroponics systems. Such a recyclable bio-nutrient would benefit plant growth and community's health in an environment-friendly manner.SUMMARY OF THE DISCLOSURE

[0008] The present disclosure provides a formulation comprising: formulation comprising: (a) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell; (b) a binder; and (c) optionally an additive. In some embodiments, the additive is present. In some embodiments, the additive is not present.

[0009] In some embodiments, the bio-nutrient composition comprises: i) a biosolid at a final percentage-by-weight of about 35% to about 75% of the bio-nutrient composition; ii) a grain at a final percentage-by-weight of about 10% to about 50% of the bio-nutrient composition; and iii) an algae cell at a final percentage-by-weight of about 5% to about 40% of the bionutrient composition. In some embodiments, the binder is at a percentage weight of about 1% to about 50% of said formulation.

[0010] In some embodiments, the biosolid is digested in an anaerobic digester for at least 15 days. In some embodiments, the biosolid is dewatered to a moisture content of about 10% or less. In some embodiments, the biosolid is pulverized to a mesh size between about 10 and about 300. In some embodiments, the grain is dewatered to a moisture content of about 10% or less. In some embodiments, the biosolid and / or the grain are dried at a temperature between about 155° F. and about 250° F. In some embodiments, the grain is pulverized to a mesh size of about 10 to about 300. In some embodiments, the biosolid is treated with an oil based agent, wherein the oil based agent prevents billowing. In some embodiments, the algae cell is pressed to isolate an algae cell wall. In some embodiments, the algae cell wall is pulverized to a mesh size of about 10 to about 300. In some embodiments, the algae cell wall is dried to a moisture content of about 20% or less. In some embodiments, the bio-nutrient composition is passed through a heating source, thereby being dewatered.

[0011] In some embodiments, the biosolid is a sewage sludge, a manure or a food waste. In some embodiments, the biosolid is a Class A EQ biosolid, Class A biosolid, or a Class B biosolid. In some embodiments, the grain is a brewers’ spent grain (BSG) or a distillers grain (DG). In some embodiments, the grain is a dried brewers’ spent grain (DBSG) or a dried distillers grain (DDG). In some embodiments, the algae cell class is Cyanophyceae^ Chlor ophyceae. Rhodophyceae, or Phaeophyceae . In some embodiments, the algae cell comprises a dried algae cell wall, algae process water, or a post-harvest post filtration undried algae solution.

[0012] In some embodiments, the binder is a lignin, a mineral, a plant starch, or a molasses. In some embodiments, the binder is a lignin or a lignin derivative. In some embodiments, the lignin derivative is lignosulfonate.

[0013] In some embodiments, the additive is an odor-reducing or odor-neutralizing agent. In some embodiments, the additive is selected from the group consisting of: an order neutralizer, an odor-masking agent, an essential oil, an odor absorbent, an enzyme, or a microbe. In some embodiments, the odor neutralizer is an activated charcoal, a zeolite, a baking soda, a silica gel, or a compound comprising cyclodextrin. In some embodiments, the odor-masking agent is a vanilla extract or a citrus extract. In some embodiments, the essential oil is a lavender oil, a eucalyptus oil, a peppermint oil, a lemon oil, a tea tree oil, a clove oil, an orange oil, or a cinnamon oil. In some embodiments, the odor absorbent is bentonite, kaolin clay, or peat moss. In some embodiments, the enzyme is a protease, a lipase, an amylase, a cellulase, a urease, an oxidoreductase, or a pectinase. In some embodiments, the microbe used as an additive for reducing or neutralizing odor is a species of genus Bacillus o Pseudomonas.

[0014] In some embodiments, the formulation is in a powder, granular, prill, or liquid form. In some embodiments, the liquid form of said formation is a suspension or an extract. In some embodiments, the liquid form of the formulation further comprises an enhancer. In some embodiments, the enhancer is selected from the consisting of: a preservative, a stabilizer, a buffering agent, a microbial inhibitor, a pH adjuster, and an anti-foaming agent. In some embodiments, the preservative is sodium benzoate, potassium sorbate, or citric acid. In some embodiments, the stabilizer is a xanthan gum, a chelating agent or an antioxidant. In some embodiments, the chelating agent is ethylenediaminetetraacetic acid (EDTA). In some embodiments, the antioxidant is an ascorbic acid or a tocopherol. In some embodiments, the buffering agent is phosphate, phosphate derivative, acetate, or acetate derivative. In some embodiments, the microbial inhibitor is an essential oil or a silver nanoparticle. In someembodiments, the essential oil is a thyme oil, an oregano oil, a peppermint oil, or a mint oil. In some embodiments, the pH adjuster is hydrochloric acid or sodium hydroxide. In some embodiments, the anti-foaming agent is a silicone-based antifoam.

[0015] The present disclosure provides that the formulation enhances activity and / or diversity of a plurality of beneficial microbes in soil to which the formulation is applied, when comparing to the soil untreated with the formulation. In some embodiments, the beneficial microbe is Pseudomonas sp, Achromobacter sp., Bacillus sp., Nitrobacter sp., Nitrosomonas sp., Nitrospira sp., Paenibacillus sp., Rhizobium sp., Streptomyces sp., Trichoderma sp., Alkalihalobacillus hwajinpoensis, Aureobasidium pullulans, Metarhizium anisopliae, Purpureocillium lilacinum, or Pythium oligandrum. In some embodiments, the beneficial microbe is an indigenous microbe species or a genetically engineered microbe. In some embodiments, the beneficial microbe enhances production of exopolysaccharides, salicylic acid, siderophores, or 1 -aminocyclopropane- 1 -carboxylate (ACC) deaminase, thereby improving soil health and promoting plant growth and stress tolerance. In some embodiments, the formulation inhibits a plurality of soil pathogens. In some embodiments, the soil pathogen is Pythium sp., Colletotrichum sp., Fusarium sp., Curvularia sp., Sclerotinia sp., Sclerotinia homoeocarpa, Agrobacterium tumefaciens, Sclerotium rolfsii, or Leptosphaerulina americana.

[0016] In some embodiments, the formulation induces expression of a plurality of genes involved in a stress-related defense mechanism in plants to which the formulation is applied, when comparing to plants untreated with the formulation. In some embodiments, the induced stress-related defense mechanism is selected from the group consisting of: phenylpropanoid biosynthesis, glutathione metabolism, sesquiterpenoid and triterpenoid metabolism, plantpathogen interaction, ascorbate and aldarate metabolism, carotenoid biosynthesis, plant hormone signal transduction, and linoleic acid metabolism. In some embodiments, the induced stress-related defense mechanism is associated with: (i) upregulation of the expression of a plurality of gene involved in phenylpropanoid biosynthesis, which is phenylalanine ammonia lyase, trans-cinnamate 4 -monooxygenase, and / or caffeoylshikimate esterase,' (ii) upregulation of the expression of a plurality of gene involved in glutathione metabolism, which is glutathione dehydroascorbate reductase 3, glutathione peroxidase, and / or gamma- glutamy Icy clotransf erase, ' (iii) upregulation of the expression of a plurality of gene involved in sesquiterpenoid and triterpenoid metabolism, which is squalene synthase 1, squalene monooxygenase, and / or NAD+ dependent fame sal dehydrogenase,' (iv) upregulation of the expression of a plurality of gene involved in plant-pathogen interaction, which is pathogenesisrelated 1, suppressor of G2 allele of SKP1, and / or respiratory burst oxidase (v) upregulation of the expression of a plurality of gene involved in ascorbate and aldarate metabolism, which is L-gulonolactone oxidase, GDP-L-galactose phosphorylase, and / or ascorbate peroxidase, (vi) upregulation of the expression of a plurality of gene involved in carotenoid biosynthesis, which is ABA 8'-hydroxylase, lycopene beta-cyclase, and / or 9-cis-epoxycarotenoid dioxygenase, (vii) upregulation of the expression of a plurality of gene involved in plant hormone signal transduction, which is Not Responsive to PR1 protein (NPR1), jasmonic acid receptor, and / or abscisic acid receptor,' and / or (viii) upregulation of the expression of a plurality of gene involved in linoleic acid metabolism, which is linoleate 9S-lipoxygenase and / or linoleate 13 S-lipoxygenase9.

[0017] In some embodiments, the formulation induces expression of a plurality of genes involved in photosynthesis mechanism for increasing plant growth and development in plants to which the formulation is applied, when comparing to plants untreated with the formulation. In some embodiments, the induced photosynthesis mechanism is associated with upregulation of the expression of a plurality of gene, which is photosystem II subunit R, photosystem I subunit IV, plastocyanin, light-harvesting complex I chlorophyll a / b binding protein 2, and / or light-harvesting complex II chlorophyll a / b binding protein 4.

[0018] In some embodiments, the formulation induces expression of a plurality of genes involved in nutrient uptake and assimilation in plants to which the formulation is applied, when comparing to plants untreated with the formulation. In some embodiments, the induced nutrient uptake and assimilation is associated with upregulation of the expression of a plurality of gene, which is nitrate reductase, nitrite reductase, high-affinity nitrate transporter 2.1 (HAT2.1), high-affinity nitrate transporter 2.2 (HAT2.2), high-affinity nitrate transporter 2.3 (HAT2.3), glutamine synthetase, and / or mugineic acid- 3 dioxygenase .

[0019] In some embodiments, the formulation is applied with an agricultural product sequentially or simultaneously.

[0020] In some embodiments, the agricultural product is a mineral based fertilizer, synthetic fertilizer, an organic fertilizer, a micronutrient, a biostimulant, a plurality of microbes, a microbial additive, or an agricultural or landscaping chemical selected from the group consisting of a pesticide, an herbicide, a fungicide, and a surfactant.

[0021] The present disclosure provides a method of making a granular formulation comprising the steps of: (a) dry milling a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell; (b) blending the dry milled bio-nutrient composition from step (a) in a powderform; (c) adding a binder to the blended bio-nutrient composition from step (b) to produce a formulation; (d) optionally, adding an additive; and (e) granulating the formulation from step (c) or (d). In some embodiments, the additive is added to the formulation from step (d). In some embodiments, the additive is not added to the formulation from step (d).

[0022] The present disclosure provides a method of making a liquid formulation comprising the steps of: (a) dry milling a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell; (b) agitating the dry milled bio-nutrient composition from step (a) in water with a speed mixer; and (c) adding a liquid form of a binder to the wet milled bio-nutrient composition to produce a liquid formulation; (d) optionally adding an additive; and (e) filtering the formulation from step (c) or (d). In some embodiments, the additive is added to the formulation from step (d). In some embodiments, the additive is not added to the formulation from step (d).

[0023] The present disclosure provides a method for improving soil health, comprising: applying the bio-nutrient formulation taught herein to soil, thereby improving soil health. In some embodiments, the formulation improves a balanced ratio of essential nutrients selected from the group consisting of: nitrogen (N), Phosphorus (P), Potassium (K), Calcium (Ca), Magnesium (Mg), Sulfur (S), Carbon (C), Hydrogen (H), Oxygen (O), and micronutrient. In some embodiments, the micronutrient is selected from the group consisting of: Iron (Fe), Manganese (Mn), Zinc (Zn), Copper (Cu), Boron (B), Molybdenum (Mo), and Chlorine (Cl). In some embodiments, the formulation increases activity and / or diversity of a plurality of beneficial microbes in soil. In some embodiments, the formulation decreases activity and / or diversity of a plurality of non-beneficial microbes in soil.

[0024] The present disclosure provides a method for altering microbiome in soil, comprising: applying the bio-nutrient formulation taught herein to soil, thereby altering microbiome in soil. In some embodiments, the formulation increases a plurality of beneficial microbes in soil. In some embodiments, the beneficial microbe enhances production of exopolysaccharides, salicylic acid, siderophores, or 1 -aminocyclopropane- 1 -carboxylate (ACC) deaminase, thereby improving soil health and promoting plant growth and stress tolerance. In some embodiments, the formulation decreases a plurality of non-beneficial microbes in soil.

[0025] The present disclosure provides a method for inducing a stress-related defense mechanism in a plant, comprising: applying the bio-nutrient formulation taught herein to a target, which is a plant, or an area around the plant, thereby improving plant defense response to biotic or abiotic stress.

[0026] The present disclosure provides a method for inducing a photosynthesis mechanism in a plant, comprising: applying the bio-nutrient formulation taught herein to a target, which is a plant, or an area around the plant, thereby improving plant growth and development.

[0027] The present disclosure provides a method for inducing a nutrient uptake and assimilation in a plant, comprising: applying the bio-nutrient formulation taught herein to a target, which is a plant, or an area around the plant, thereby improving nitrogen assimilation or uptake of a plant.BRIEF DESCRIPTION OF THE DRAWINGS

[0028] The accompanying figures, which are incorporated herein and form a part of the specification, illustrate some, but not the only or exclusive, example embodiments and / or features. It is intended that the embodiments and figures disclosed herein are to be considered illustrative rather than limiting.

[0029] For a more complete understanding of the present disclosure, the objects and advantages thereof, reference is now made to the ensuing descriptions taken in connection with the accompanying drawings briefly described as follows.

[0030] FIG. 1 is a flow chart of sustainable manufacturing of a bio-nutrient formulation.

[0031] FIG. 2 is a comparison of traditional (passive fertilizers and bio-nutrient formulations in downstream benefits of the supply chain.

[0032] FIG. 3 is a summary of the environmental benefits of the use of a bio-nutrient formulation of the present disclosure.

[0033] FIG. 4 is a set-up for a field test to compare the bio-nutrient formulation of the present disclosure with 42-0-0 polymer coated urea (synthetic fertilizer) used as a control at four different time points (TO: Prior to application; T1 = 14 days after application; T2 = 30 days after application; T3 = 60 days after application; T4 = 90 days after application). Three replicates per treatment were used in one location.

[0034] FIG. 5 presents plant and root response results of the bio-nutrient formulation in comparison to the polymer-coated urea (synthetic fertilizer).

[0035] FIG. 6 presents microbe response results of the bio-nutrient formulation in comparison to the polymer-coated urea (synthetic fertilizer).

[0036] FIG. 7 presents a summary of field test results of the bio-nutrient granular formulation of the present disclosure, in comparison to polymer-coated urea (synthetic fertilizer) tounderstand impact on root size, soil improvement, plant stress tolerance, plant disease reduction, and plant hormones produced by various microorganisms.

[0037] FIG. 8 presents modes of action (KEGG pathways) associated with plant stress and defense responses identified from enrichment analysis.

[0038] FIG. 9 presents modes of action (KEGG pathways) associated with plant growth and development identified from enrichment analysis.

[0039] FIG. 10 presents modes of action (KEGG pathways) associated with plant nutrient uptake and assimilation identified from enrichment analysis.DETAILED DESCRIPTION

[0040] The foregoing examples of the related art and limitations related therewith are intended to be illustrative and not exclusive. Other limitations of the related art will become apparent to those of skill in the art upon a reading of the specification.Definitions

[0041] While the following terms are believed to be well understood by one of ordinary skill in the art, the following definitions are set forth to facilitate explanation of the presently disclosed subject matter.

[0042] All technical and scientific terms used herein, unless otherwise defined below, are intended to have the same meaning as commonly understood by one of ordinary skill in the art. References to techniques employed herein are intended to refer to the techniques as commonly understood in the art, including variations on those techniques and / or substitutions of equivalent techniques that would be apparent to one of skill in the art.

[0043] The use of the terms “a” and “an” and “the” and similar referents in the context of describing the disclosure (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context.

[0044] The terms “comprising,” “having,” “including,” and “containing” are to be construed as open-ended terms (i.e., meaning “including, but not limited to,”) unless otherwise noted. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. For example, if the range 10-15 is disclosed, then 11, 12, 13, and 14 are also disclosed. All methods described herein can be performed in any suitable order unlessotherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as") provided herein, is intended merely to better illuminate the disclosure, and does not pose a limitation on the scope of the disclosure unless otherwise claimed. No language in the specification should be construed as indicating any nonclaimed element as essential to the practice of the disclosure.

[0045] Unless otherwise indicated, all numbers expressing quantities of ingredients, reaction conditions, and so forth used in the specification and claims are to be understood as being modified in all instances by the term “about.” The term “about,” as used herein when referring to a measurable value such as an amount of mass, weight, time, volume, concentration or percentage is meant to encompass variations of in some embodiments ±20%, in some embodiments ±10%, in some embodiments ±5%, in some embodiments ±1%, in some embodiments ±0.5%, and in some embodiments ±0.1% from the specified amount, as such variations are appropriate to perform the disclosed methods and / or employ the disclosed compositions, nucleic acids, polypeptides, etc. Accordingly, unless indicated to the contrary, the numerical parameters set forth in this specification and attached claims are approximations that can vary depending upon the desired properties sought to be obtained by the presently disclosed subject matter.

[0046] As used herein, the term “and / or” when used in the context of a list of entities, refers to the entities being present singly or in combination. Thus, for example, the phrase “A, B, C, and / or D” includes A, B, C, and D individually, but also includes any and all combinations and subcombinations of A, B, C, and D (e g., AB, AC, AD, BC, BD, CD, ABC, ABD, and BCD). In some embodiments, one or more of the elements to which the “and / or” refers can also individually be present in single or multiple occurrences in the combinations(s) and / or subcombination(s).

[0047] The term “biologically pure culture” or “substantially pure culture” refers to a culture of a bacterial species described herein containing no other bacterial species in quantities sufficient to interfere with the replication of the culture or be detected by normal bacteriological techniques.

[0048] “Plant productivity” refers generally to any aspect of growth or development of a plant that is a reason for which the plant is grown. For food crops, such as grains or vegetables, “plant productivity” can refer to the yield of grain or fruit harvested from a particular crop. As used herein, improved plant productivity refers broadly to improvements in yield of grain, fruit, flowers, or other plant parts harvested for various purposes, improvements in growth of plantparts, including stems, leaves and roots, promotion of plant growth, maintenance of high chlorophyll content in leaves, increasing fruit or seed numbers, increasing fruit or seed unit weight, reducing NO2 emission due to reduced nitrogen fertilizer usage and similar improvements of the growth and development of plants.

[0049] Microbes in and around food crops can influence the traits of those crops. Plant traits that may be influenced by microbes include: yield (e.g., grain production, biomass generation, fruit development, flower set); nutrition (e.g., nitrogen, phosphorus, potassium, iron, micronutrient acquisition); abiotic stress management (e.g., drought tolerance, salt tolerance, heat tolerance); and biotic stress management (e.g., pest, weeds, insects, fungi, and bacteria). Strategies for altering crop traits include: increasing key metabolite concentrations; changing temporal dynamics of microbe influence on key metabolites; linking microbial metabolite product! on / degradati on to new environmental cues; reducing negative metabolites; and improving the balance of metabolites or underlying proteins.

[0050] As used herein, a “control sequence” refers to an operator, promoter, silencer, or terminator.

[0051] As used herein, “z z planta” may refer to in the plant, on the plant, or intimately associated with the plant, depending upon context of usage (e.g., endophytic, epiphytic, or rhizospheric associations). The plant may comprise plant parts, tissue, leaves, roots, root hairs, rhizomes, stems, seed, ovules, pollen, flowers, fruit, etc.

[0052] In some embodiments, native or endogenous control sequences of genes of the present disclosure are replaced with one or more intrageneric control sequences.

[0053] As used herein, “introduced” refers to the introduction by means of modem biotechnology, and not a naturally occurring introduction.

[0054] In some embodiments, the bacteria of the present disclosure have been modified such that they are not naturally occurring bacteria.

[0055] In some embodiments, the bacteria of the present disclosure are present in the plant in an amount of at least 103cfu, 104cfu, 105cfu, 106cfu, 107cfu, 108cfu, 109cfu, 1010cfu, 1011cfu, or 1012cfu per gram of fresh or dry weight of the plant. In some embodiments, the bacteria of the present disclosure are present in the plant in an amount of at least about 103cfu, about 104cfu, about 105cfu, about 106cfu, about 107cfu, about 108cfu, about 109cfu, about 1010cfu, about 1011cfu, or about 1012cfu per gram of fresh or dry weight of the plant. In some embodiments, the bacteria of the present disclosure are present in the plant in an amount of atleast 103to 109, 103to 107, 103to 105, 105to 109, 105to 107, 106to IO10, 106to 107cfu per gram of fresh or dry weight of the plant.

[0056] Fertilizers and exogenous nitrogen of the present disclosure may comprise the following nitrogen-containing molecules: ammonium, nitrate, nitrite, ammonia, glutamine, etc. Nitrogen sources of the present disclosure may include anhydrous ammonia, ammonia sulfate, urea, diammonium phosphate, urea-form, monoammonium phosphate, ammonium nitrate, nitrogen solutions, calcium nitrate, potassium nitrate, sodium nitrate, etc.

[0057] As used herein, “exogenous nitrogen” refers to non-atmospheric nitrogen readily available in the soil, field, or growth medium that is present under non-nitrogen limiting conditions, including ammonia, ammonium, nitrate, nitrite, urea, uric acid, ammonium acids, etc.

[0058] As used herein, “non-nitrogen limiting conditions” refers to non-atmospheric nitrogen available in the soil, field, media at concentrations greater than about 4 mM nitrogen, as disclosed by Kant et al. (2010. J. Exp. Biol. 62(4): 1499-1509), which is incorporated herein by reference.

[0059] As used herein, an “intergeneric microorganism” is a microorganism that is formed by the deliberate combination of genetic material originally isolated from organisms of different taxonomic genera. An “intergeneric mutant” can be used interchangeably with “intergeneric microorganism”. An exemplary “intergeneric microorganism” includes a microorganism containing a mobile genetic element which was first identified in a microorganism in a genus different from the recipient microorganism. Further explanation can be found, inter alia, in 40 C.F.R. § 725.3.

[0060] As used herein, an “intrageneric microorganism” is a microorganism that is formed by the deliberate combination of genetic material originally isolated from organisms of the same taxonomic genera. An “intrageneric mutant” can be used interchangeably with “intrageneric microorganism.”

[0061] As used herein, “introduced genetic material” means genetic material that is added to, and remains as a component of, the genome of the recipient.

[0062] As used herein, in the context of non-intergeneric microorganisms, the term “remodeled” is used synonymously with the term “engineered”. Consequently, a “non- intergeneric remodeled microorganism” has a synonymous meaning to “non-intergeneric engineered microorganism,” and will be utilized interchangeably. Further, the disclosure may refer to an “engineered strain” or “engineered derivative” or “engineered non-intergenericmicrobe,” these terms are used synonymously with “remodeled strain” or “remodeled derivative” or “remodeled non-intergeneric microbe.”

[0063] In some embodiments, the nitrogen fixation and assimilation genetic regulatory network comprises polynucleotides encoding genes and non-coding sequences that direct, modulate, and / or regulate microbial nitrogen fixation and / or assimilation and can comprise polynucleotide sequences of the nif cluster (e.g., nifA, niffl, nifC, nifZ), polynucleotides encoding nitrogen regulatory protein C, polynucleotides encoding nitrogen regulatory protein B, polynucleotide sequences of the gin cluster (e.g. glnA and glnD), draT, and ammonia transporters / permeases. In some cases, the Nif cluster may comprise NifB, NifH, NifD, NifK, NifE, NifN, NifX, hesa, and NifV. In some cases, the Nif cluster may comprise a subset of NifB, NifH, NifD, NifK, NifE, NifN, NifX, hesa, and NifV.

[0064] In some embodiments, the increase of nitrogen fixation and / or the production of 1% or more of the nitrogen in the plant are measured relative to control plants, which have not been exposed to the bacteria of the present disclosure. All increases or decreases in bacteria are measured relative to control bacteria. All increases or decreases in plants are measured relative to control plants.

[0065] As used herein, a “constitutive promoter” is a promoter, which is active under most conditions and / or during most development stages. There are several advantages to using constitutive promoters in expression vectors used in biotechnology, such as: high level of production of proteins used to select transgenic cells or organisms; high level of expression of reporter proteins or scorable markers, allowing easy detection and quantification; high level of production of a transcription factor that is part of a regulatory transcription system; production of compounds that requires ubiquitous activity in the organism; and production of compounds that are required during all stages of development. Non-limiting exemplary constitutive promoters include, CaMV 35S promoter, opine promoters, ubiquitin promoter, alcohol dehydrogenase promoter, etc.

[0066] As used herein, a “non-constitutive promoter” is a promoter that is active under certain conditions, in certain types of cells, and / or during certain development stages. For example, tissue specific, tissue preferred, cell type specific, cell type preferred, inducible promoters, and promoters under development control are non-constitutive promoters. Examples of promoters under developmental control include promoters that preferentially initiate transcription in certain tissues.

[0067] As used herein, “inducible” or “repressible” promoter is a promoter that is under chemical or environmental factors control. Examples of environmental conditions that may affect transcription by inducible promoters include anaerobic conditions, certain chemicals, the presence of light, acidic or basic conditions, etc.

[0068] As used herein, a “tissue specific” promoter is a promoter that initiates transcription only in certain tissues. Unlike constitutive expression of genes, tissue-specific expression is the result of several interacting levels of gene regulation. As such, in the art sometimes it is preferable to use promoters from homologous or closely related species to achieve efficient and reliable expression of transgenes in particular tissues. This is one of the main reasons for the large number of tissue-specific promoters isolated from particular tissues found in both scientific and patent literature.

[0069] As used herein, the term “operably linked” refers to the association of nucleic acid sequences on a single nucleic acid fragment so that the function of one is regulated by the other. For example, a promoter is operably linked with a coding sequence when it is capable of regulating the expression of that coding sequence (i.e., that the coding sequence is under the transcriptional control of the promoter). Coding sequences can be operably linked to regulatory sequences in a sense or antisense orientation. In another example, the complementary RNA regions of the disclosure can be operably linked, either directly or indirectly, 5' to the target mRNA, or 3' to the target mRNA, or within the target mRNA, or a first complementary region is 5' and its complement is 3' to the target mRNA.

[0070] In aspects, “applying to the plant a plurality of non-intergeneric bacteria,” includes any means by which the plant (including plant parts such as a seed, root, stem, tissue, etc.) is made to come into contact (i.e., exposed) with said bacteria at any stage of the plant’s life cycle. Consequently, “applying to the plant a plurality of non-intergeneric bacteria,” includes any of the following means of exposing the plant (including plant parts such as a seed, root, stem, tissue, etc.) to said bacteria: spraying onto plant, dripping onto plant, applying as a seed coat, applying to a field that will then be planted with seed, applying to a field already planted with seed, applying to a field with adult plants, etc. More generally, “applying to the plant a plurality of microorganisms,” such as those comprised by the consortia disclosed herein, includes any of the foregoing routes and means of application.

[0071] As used herein the term “plant” can include plant parts, tissue, leaves, roots, root hairs, rhizomes, stems, seeds, ovules, pollen, flowers, fruit, etc. Thus, when the disclosure discussesproviding a plurality of corn plants to a particular locus, it is understood that this may entail planting a com seed at a particular locus.

[0072] As used herein the terms “microorganism” or “microbe” should be taken broadly. These terms, used interchangeably, include but are not limited to, the two prokaryotic domains, Bacteria and Archaea. The term may also encompass eukaryotic fungi and protists.

[0073] As used herein, when the disclosure discuses a particular microbial deposit by accession number, it is understood that the disclosure also contemplates a microbial strain having all of the identifying characteristics of said deposited microbe, and / or a mutant thereof.

[0074] The term “microbial consortia” or “microbial consortium” refers to a subset of a microbial community of individual microbial species, or strains of a species, which can be described as carrying out a common function, or can be described as participating in, or leading to, or correlating with, a recognizable parameter, such as a phenotypic trait of interest.

[0075] The term “assemblage” or “microbial assemblage” refers to a collection or grouping of microorganisms.

[0076] The term “microbial community” means a group of microbes comprising two or more species or strains. Unlike microbial consortia, a microbial community does not have to be carrying out a common function, or does not have to be participating in, or leading to, or correlating with, a recognizable parameter, such as a phenotypic trait of interest.

[0077] As used herein, “isolate,” “isolated,” “isolated microbe,” and like terms, are intended to mean that the one or more microorganisms has been separated from at least one of the materials with which it is associated in a particular environment (for example soil, water, plant tissue, etc.). Thus, an “isolated microbe” does not exist in its naturally occurring environment; rather, it is through the various techniques described herein that the microbe has been removed from its natural setting and placed into a non-naturally occurring state of existence. Thus, the isolated strain or isolated microbe may exist as, for example, a biologically pure culture, or as spores (or other forms of the strain). In aspects, the isolated microbe may be in association with an acceptable carrier, which may be an agriculturally acceptable carrier.

[0078] In certain aspects of the disclosure, the isolated microbes exist as “isolated and biologically pure cultures.” It will be appreciated by one of skill in the art, that an isolated and biologically pure culture of a particular microbe, denotes that said culture is substantially free of other living organisms and contains only the individual microbe in question. The culture can contain varying concentrations of said microbe. The present disclosure notes that isolated and biologically pure microbes often “necessarily differ from less pure or impure materials.” See,e.g. In re Bergstrom, 427 F.2d 1394, (CCPA 1970) (discussing purified prostaglandins), see also, In re Bergy, 596 F.2d 952 (CCPA 1979) (discussing purified microbes), see also, Parke- Davis & Co. v. H.K. Mulford & Co., 189 F. 95 (S.D.N.Y. 1911) (Learned Hand discussing purified adrenaline), aff d in part, rev’d in part, 196 F. 496 (2d Cir. 1912), each of which are incorporated herein by reference. Furthermore, in some aspects, the disclosure provides for certain quantitative measures of the concentration, or purity limitations, that must be found within an isolated and biologically pure microbial culture. The presence of these purity values, in certain embodiments, is a further attribute that distinguishes the presently disclosed microbes from those microbes existing in a natural state. See, e.g., Merck & Co. v. Olin Mathieson Chemical Corp., 253 F.2d 156 (4th Cir. 1958) (discussing purity limitations for vitamin B12 produced by microbes), incorporated herein by reference.

[0079] As used herein, “individual isolates” should be taken to mean a composition, or culture, comprising a predominance of a single genera, species, or strain, of microorganism, following separation from one or more other microorganisms.

[0080] Microbes of the present disclosure may include spores and / or vegetative cells. In some embodiments, microbes of the present disclosure include microbes in a viable but non- culturable (VBNC) state.

[0081] As used herein, “spore” or “spores” refer to structures produced by bacteria and fungi that are adapted for survival and dispersal. Spores are generally characterized as dormant structures; however, spores are capable of differentiation through the process of germination. Germination is the differentiation of spores into vegetative cells that are capable of metabolic activity, growth, and reproduction. The germination of a single spore results in a single fungal or bacterial vegetative cell. Fungal spores are units of asexual reproduction, and in some cases are necessary structures in fungal life cycles. Bacterial spores are structures for surviving conditions that may ordinarily be nonconducive to the survival or growth of vegetative cells.

[0082] As used herein, the phrase “synthetic composition” refers to a composition that includes at least one microbe that has been cultured and / or genetically engineered.

[0083] As used herein, a monocotyledonous plant or monocot refers to a flowering plant having embryos with one cotyledon or seed leaf, parallel leaf veins, and flower parts in multiples of three. Examples of monocots taught in the disclosure include, but are not limited to turfgrass, com / maize, rice, wheat, barley, sorghum, rye, millet, oat, annual ryegrass, orchid, iris, lily, onion, and palm. Examples of turfgrass include, but are not limited to Agrostis spp. (bentgrass species including colonial bentgrass and creeping bentgrasses), Poa pratensis(Kentucky bluegrass), Lolium spp. (ryegrass species including annual ryegrass and perennial ryegrass), Festuca arundinacea (tall fescue) Festuca rubra commutata (Chewings fescue), Cynodon dactylon (bermudagrass, Pennisetum clandestinum (kikuyu grass), Stenotaphrum secundatum (St. Augustine grass), Zoysia japonica (zoysia grass), and Dichondra micrantha.

[0084] The monocotyledonous and dicotyledonous plants, such as crops including grain crops (e.g., wheat, maize, rice, millet, barley), fruit crops (e.g., tomato, apple, grape, peach, pear, plum, raspberry, black raspberry, blackberry, cane berry, cherry, avocado, strawberry, wild strawberry, orange), forage crops (e.g., alfalfa), root vegetable crops (e.g., carrot, potato, sugar beets, yam), leafy vegetable crops (e.g., lettuce, spinach); flowering plants (e.g., petunia, rose, chrysanthemum), conifers and pine trees (e.g., pine fir, spruce); plants used in phytoremediation (e.g., heavy metal accumulating plants); oil crops (e.g., sunflower, rape seed) and plants used for experimental purposes (e.g., Arabidopsis).

[0085] In the present disclosure, the plants are intended to comprise without limitation angiosperm and gymnosperm plants such as acacia, alfalfa, amaranth, apple, apricot, artichoke, ash tree, asparagus, avocado, banana, barley, beans, beet, birch, beech, blackberry, black raspberry, blueberry, broccoli, Brussel's sprouts, cabbage, cane berry, canola, cantaloupe, carrot, cassava, cauliflower, cedar, a cereal, celery, chestnut, cherry, Chinese cabbage, citrus, Clementine, clover, coffee, corn, cotton, cowpea, cucumber, cypress, eggplant, elm, endive, eucalyptus, fennel, figs, fir, geranium, grape, grapefruit, groundnuts, ground cherry, gum hemlock, hickory, kale, kiwifruit, kohlrabi, larch, lettuce, leek, lemon, lime, locust, pine, maidenhair, maize, mango, maple, melon, millet, mushroom, mustard, nuts, oak, oats, oil palm, okra, onion, orange, an ornamental plant or flower or tree, papaya, palm, parsley, parsnip, pea, peach, peanut, pear, peat, pepper, persimmon, pigeon pea, peach, pine, pineapple, plantain, plum, pomegranate, potato, pumpkin, radicchio, radish, rapeseed, raspberry, rice, rye, sorghum, safflower, sallow, soybean, spinach, spruce, squash, strawberry, sugar beet, sugarcane, sunflower, sweet potato, sweet corn, tangerine, tea, tobacco, tomato, trees, triticale, turf grasses, turnips, vine, walnut, watercress, watermelon, wheat, wild strawberry, yams, yew, and zucchini.

[0086] As used herein, the term “bio-nutrient”, “bio nutrient”, or “biologically active nutrient” refers to a biologically active compound or compound that influences, signals and proliferates microbes existing naturally within the soil or modified and added into the soil. The bio-nutrient composition of the present disclosure refers to the array of essential nutrients and beneficial compounds that are present in biological materials, such as soil amendments, fertilizers, andplant or microbial products. These nutrients and compounds play crucial roles in promoting plant growth, improving soil health, enhancing crop productivity, and promoting sustainable agricultural practices. The bio-nutrient composition typically includes macro-nutrients, micronutrients, and other beneficial bioactive compounds, which support plant growth and soil health.

[0087] As used herein, the term “macronutrients” include (i) Nitrogen (N), which is essential for protein synthesis, chlorophyll formation, and overall plant growth; (ii) Phosphorus (P), which is crucial for energy transfer, DNA / RNA synthesis, and root development; (iii) Potassium (K), which is important for enzyme activation, water regulation, and stress resistance; (iv) Calcium (Ca), which is involved in cell wall structure, cell division, and signalling; (v) Magnesium (Mg), which is central component of chlorophyll and involved in photosynthesis; and / or (vi) Sulfur (S), which is important for amino acids and protein synthesis.

[0088] As used herein, the term “micronutrients” include (i) Iron (Fe), which is vital for chlorophyll synthesis and electron transport in photosynthesis; (ii) Manganese (Mn), which is involved in enzyme activation and photosynthesis; (iii) Zinc (Zn), which is essential for enzyme function, protein synthesis, and growth regulation; (iv) Copper (Cu), which is important for photosynthesis, respiration, and lignin synthesis; (v) Boron (B): Required for cell wall formation and reproductive development; (vi) Molybdenum (Mo), which is necessary for nitrogen fixation and enzyme function; and / or (vii) Chlorine (Cl), which is involved in osmotic and ionic balance.

[0089] As used herein, the term “beneficial bioactive compounds” include (i) amino acids which serve as building blocks for proteins and precursors for growth regulators; (ii) vitamins which function as coenzymes in metabolic processes; (iii) phytohormones which includes auxins, cytokinins, gibberellins, and other hormones that regulate plant growth and development; (iv) humic and fulvic acids which improve soil structure, nutrient availability, and microbial activity; and / or (v) organic matter that enhances soil fertility, water retention, and microbial diversity.

[0090] In some embodiments, the bio-nutrients comprise three main macro nutrients: nitrogen (N), phosphorus (P), and potassium (K), which is applied to soil to supply nutrients to a plant or a microbe. In some embodiments, the bio-nutrients further comprises micronutrients. In some embodiments, the term further comprises elements of a soil amendment including but not limited to organic matter, organic carbon, humic acid, fulvic acid, carbohydrates, amino acids, lignins and other elements commonly known to improve soil tilth (overall soil quality). In someembodiments, bio-nutrients may comprise active or live microorganisms that improve the chemical and biological aspects of soils, restore soil fertility, and encourage plant growth. In some embodiments, a bio-nutrient composition may comprise inactive, inert, or dead microorganisms that contribute to the increase of organic matter and carbon in soil.

[0091] In some embodiments, “a bio-nutrient composition” comprises a biosolid, a grain (such as a dried brewers spent grain or a dried distillers grain), and an algae cell or cell wall. In further embodiments, “a bio-nutrient formulation” refers to a bio-nutrient composition (comprising a biosolid, a grain, and an algae cell) and a binder.Overview

[0092] The present disclosure provides methods of making a dry, granular and liquid form of the bio-nutrient formulation that enhances the biological activities and functions of microbes native / indigenous to soil, as well as microbes that are enhanced or genetically edited and applied to plants, without respect to specific plant or crop genus family and species.

[0093] In some embodiments, this bio-nutrient formulation is plant agnostic, not focusing on the crop or plant but rather on the underlying / core microbial functions that manage plant functions including but not limited to drought, salt, heat stress, disease pressure, promoting immune system mechanisms, macro, secondary and micronutrient uptake / regulation, carbon sequestration, metals solubilization, salt tolerance and mineralization / volatilization. In some embodiments, the formulation of the present disclosure can enhance soil health by maintaining healthy soil with balanced nutrient composition, which ensures sustained plant growth and resilience. In some embodiments, the formulation of the present disclosure can improve fertilization in soil, which can optimize nutrient availability and uptake. In some embodiments, the formulation of the present disclosure can improve soil structure and nutrient-holding capacity.

[0094] In other embodiments, this bio-nutrient formulation directly influences the physiological and biochemical processes in plants, affecting their growth, development, and ability to tolerate various stresses. Plants absorb nutrients from the soil through their roots. The availability of bio-nutrients of the present disclosure in the soil directly affects the plant's ability to uptake these nutrients. For example, nitrogen is absorbed as nitrate (NCh ) or ammonium (NH4+), phosphorus as phosphate (PCri3), and potassium as K+. Nitrogen can be utilized for the synthesis of amino acids, proteins, and chlorophyll, which are fundamental for growth andphotosynthesis. Adequate phosphorus levels promote healthy root development, and potassium influences turgor pressure and cell elongation, supporting overall plant growth.

[0095] The bio-nutrient formulation of the present disclosure can robust growth by providing the necessary building blocks for cell division, enlargement, and differentiation. The bionutrient formulation of the present disclosure can ensure timely flowering, fruiting, and seed development by regulating hormonal balances and energy allocation. The bio-nutrient formulation of the present disclosure can enhance the plant's ability to cope with abiotic and biotic stresses through improved structural integrity, metabolic resilience, and activation of defense mechanisms. Thus, proper management and supply of these bio-nutrients is crucial for achieving optimal plant health and productivity.

[0096] The present disclosure also provides methods of utilizing the bio-nutrient formulation including various forms of organic waste and food waste / residual materials; (i) forms of biosolids such as treated sewage sludge or manure, (ii) forms of grain wastes from ethanol, alcohol and beer production, and (iii) forms of algae and kelp derived from fresh water sources, salt or brackish water sources, or environmental cleanup sites and other cleanup and carbon capture technologies. In some embodiments, these forms of organic waste integrate a variety of nutrients including macro, secondary, and micronutrients, organic carbon, organic matter, humic acid, fulvic acid, carbohydrates, amino acids, and inert or dead microbiology. These components function as feedstock for a diversity of microbes that govern plant functions.

[0097] The present disclosure further provides methods that may be used to combine this bionutrient formulation product with existing synthetic or organic soil inputs products (including fertilizer blends, chemical, microbe, and soil amendment products) to enhance the effects of partner products.

[0098] The present disclosure then provides bio-nutrient compositions or formulations produced from methods taught herein. In some embodiments, a bio-nutrient composition comprises (i) biosolids (ii) dried distillers grains or brewers grains, (iii) dried algae biomass. In order to form a bio-nutrient formulation, the bio-nutrient composition taught herein is added with a binder, such as lignin or lignosulfonate, which enhances the function, performance, handling, and application of the bio-nutrient fertilizer. In some embodiments, this bio-nutrient formulation can be used with existing synthetic or organic soil inputs products (including raw minerals, bulk fertilizer ingredients, blended fertilizers, chemical, microbe, and soil amendment products) to enhance the effects of products.Bio-nutrient compounds

[0099] The widespread use of fertilizers comes with a multitude of inherent problems. First, if the fertilizer contains hazardous chemicals or additives, these can be washed away by irrigation, and sent to downstream collection sites. Second, organic fertilizers often require significantly greater volume application of fertilizers as compared to conventional chemicalbased fertilizers. This excessive application often leads to runoff and pollution of important sources of downstream surface and subsurface drinking water. This causes large concentrations of toxins and excess nutrients to be present in areas where they should not, as well as cause algae blooms in bodies of water that decimate aquatic life. Third, synthetic fertilizers often have damaging side effects to soil including harming microbes, reducing soil fertility, changing soil pH (too acidic or too alkaline), nitrogen leaching, soil compaction, reduction in soil organic matter, loss of soil carbon. Further synthetic fertilizers contain high levels of soluble salts, causing root damage, leaf and tissue damage, damaging soil structure, reducing water infiltration and therefore contributing to erosion and plant loss.

[0100] The present disclosure teaches the recycling of organic waste from sewage and industry. With treated properly, these waste materials can be used to address the metabolic and nutritional needs of plants and soil microbiology as well as ensure the safety of the public.

[0101] This present disclosure provides a bio-nutrient composition that effects a meaningful and measurable change in activity of soil microbiology. In some embodiments, the bio-nutrient composition reduces the need for traditional fertilizer, pesticides, and fungicides. In some embodiments, the bio-nutrient composition confers improved growth to the point where many weeds were out-crowded. In some embodiments, it aids microbes that solubilize and mineralize a variety of nutrients, effects a reduction of salts, and enhances sequestration of carbon. In addition, the improved nutritional status and growth rendered the turf more resilient to both biotic and abiotic challenges.

[0102] The present disclosure provides a bio-nutrient compound, interchangeably used as a bio-nutrient composition, that comprises a biosolid, a grain (such as a dried brewers spent grain or a dried distillers grain), and an algae cell (or an algae cell wall). This bio-nutrient compound is used as a non-toxic, slow-release fertilizer to be used in the agricultural and home and garden industries. This slow-release fertilizer composition and preparation method is well described in US Patent No. 10,351,483, which is incorporated by reference herein.

[0103] (i) Biosolids

[0104] The present disclosure teaches that a bio-nutrient composition comprises a biosolid. In some embodiments, the bio-nutrient composition comprises the biosolid at a percentage by weight of 0% to 100%, by weight of about 1% to about 99%, by weight of about 2% to about 98%, by weight of about 3% to about 97%, by weight of about 4% to about 96%, by weight of about 5% to about 95%, by weight of about 6% to about 94%, by weight of about 7% to about 93%, by weight of about 8% to about 92%, by weight of about 9% to about 91%, by weight of about 10% to about 90%, by weight of about 11% to about 89%, by weight of about 12% to about 88%, by weight of about 13% to about 87%, by weight of about 14% to about 86%, about 15% to about 85%, by weight of about 16% to about 84%, by weight of about 17% to about 83%, by weight of about 18% to about 82%, by weight of about 19% to about 81%, by weight of about 20% to about 80%, by weight of about 22% to about 79%, by weight of about 24% to about 78%, by weight of about 26% to about 77%, by weight of about 28% to about 76%, by weight of about 30% to about 75%, by weight of about 31% to about 75%, by weight of about 32% to about 75%, by weight of about 33% to about 75%, by weight of about 34% to about 75%, or by weight of about 35% to about 75%.

[0105] The present disclosure teaches that a bio-nutrient composition comprises a biosolid. In some embodiments, the bio-nutrient composition comprises the biosolid at a percentage by weight of 0%, by weight of about 1%, by weight of about 2%, by weight of about 3%, by weight of about 4%, by weight of about 5%, by weight of about 6%, by weight of about 7%, by weight of about 8%, by weight of about 9%, by weight of about 10%, by weight of about 11%, by weight of about 12%, by weight of about 13%, by weight of about 14%, by weight of about 15%, by weight of about 16%, by weight of about 17%, by weight of about 18%, by weight of about 19%, by weight of about 20%, by weight of about 21%, by weight of about 22%, by weight of about 23%, by weight of about 24%, by weight of about 25%, by weight of about 26%, by weight of about 27%, by weight of about 28%, by weight of about 29%, by weight of about 30%, by weight of about 31%, by weight of about 32%, by weight of about 33%, by weight of about 34%, by weight of about 35%, by weight of about 36%, by weight of about 37%, by weight of about 38%, by weight of about 39%, by weight of about 40%, by weight of about 41%, by weight of about 42%, by weight of about 43%, by weight of about 44%, by weight of about 45%, by weight of about 46%, by weight of about 47%, by weight of about 48%, by weight of about 49%, by weight of about 50%, by weight of about 51%, by weight of about 52%, by weight of about 53%, by weight of about 54%, by weight of about 55%, by weight of about 56%, by weight of about 57%, by weight of about 58%, by weight ofabout 59%, by weight of about 60%, by weight of about 61%, by weight of about 62%, by weight of about 63%, by weight of about 64%, by weight of about 65%, by weight of about 66%, by weight of about 67%, by weight of about 68%, by weight of about 69%, by weight of about 70%, by weight of about 71%, by weight of about 72%, by weight of about 73%, by weight of about 74%, by weight of about 75%, by weight of about 76%, by weight of about 77%, by weight of about 78%, by weight of about 79%, by weight of about 80%, by weight of about 81%, by weight of about 82%, by weight of about 83%, by weight of about 84%, by weight of about 85%, by weight of about 86%, by weight of about 87%, by weight of about 88%, by weight of about 89%, by weight of about 90%, by weight of about 91%, by weight of about 92%, by weight of about 93%, by weight of about 94%, by weight of about 95%, by weight of about 96%, by weight of about 97%, by weight of about 98%, by weight of about 99%, or by weight of 100%.

[0106] The biosolids are a by-product of the wastewater treatment process, which involves separating the liquids from the solids. The resulting solids are then subjected to physical and chemical treatment to produce a semi-solid, nutrient-rich substance known as biosolids.

[0107] The terms "biosolids" and "sewage sludge" are often used interchangeably. The term “sewage sludge” refers to the solids separated during the treatment of municipal wastewater. The definition includes domestic septage. In some embodiments, biosolids refers to treated sewage sludge that meets the EPA pollutant and pathogen requirements for land application and surface disposal.

[0108] To be used in beneficial ways, biosolids must meet state and federal requirements. For instance, they can be applied to agricultural land and reclamation sites such as mining sites. When applied to land at the appropriate rate, biosolids provide numerous benefits, including adding nutrients, improving soil structure, and facilitating water reuse. Additionally, land application of biosolids can offer economic and waste management benefits, such as conserving landfill space, reducing the demand for non-renewable resources like phosphorus, and decreasing the need for synthetic fertilizers. However, biosolids can also be disposed of through incineration, landfilling, or other forms of surface disposal.

[0109] To ensure that biosolids are handled, processed, and land-applied in a way that minimizes the potential risks to human health, there are existing requirements and guidance. These requirements divide biosolids into two categories: "Class A" and "Class B," which are determined based on their treatment methods. Each class has specified requirements for reducing pollutants, pathogens, and vector attraction, as well as general management practices.

[0110] For Class A biosolids, the treatment processes outlined in 40 CFR Part 503 eliminate all pathogens, including viruses. However, Class B biosolids may contain pathogens even when the requirements outlined in 40 CFR Part 503 are met. It is important to follow the EPA's site restrictions, which allow time for pathogen degradation, when harvesting crops and turf, grazing animals, and allowing for public contact. Overall, these guidelines help ensure that biosolids are processed and handled in a way that is safe for human health and the environment.

[0111] Facultative bacteria are used under anaerobic conditions to decompose the organic and bacterial matter within the biosolids. These are typically classified as “Class B” as they may contain pathogens after digestion by the bacteria. In contrast, “Class A” biosolids are heat dried and are more favorable in their usage for animal and human crop growth. “Class A Exceptional Quality (EQ)” have undergone additional treatment and are granted the classification of “unrestricted use” as a soil amendment and fertilizer. To acquire this certification, the process must be proven to remove enteric viruses, bacteria, and other pathogens. Drying the biosolids wherein the total composition of moisture is less than 10% has the negative of lower fertilization value because nitrogen content is only 2% to 5%. As market value for fertilizer is principally based on nitrogen content, raising the nitrogen concentration is favorable for the plants as well as the business. Methods disclosed herein function to benefit the growth of plants, as well as the health of the environment and community.

[0112] In some embodiments, biosolids are heat-treated between about 155° F. and about 250° F to break down pathogens, viruses, bacteria and / or fungi, as well as to help break down Per- and Polyfluoroalkyl substances (PF AS) and other chemicals that may be present in biosolids. In some embodiments, the Class A Exceptional Quality process involves the range of this high heat.

[0113] In some embodiments, the biosolid is dewatered to a moisture content of about 15% or less, or about 10% or less. In some embodiments, the biosolid is dried at a temperature between about 100°F. and about 300°F, between about 125° F. and about 275°F., between about 150°F and about 250°F, or between about 155° F. and about 250° F.

[0114] The present disclosure teaches that the biosolids is passed through a heating source such as drum drier, spray drier, rotary drum, or dehydration / dewatering pad. The drying biosolids has the total composition of moisture about 30% or less, about 25% or less, about 20% or less, about 15% or less, about 14% or less, about 13% or less, about 12% or less, about 11% or less, about 10% or less, about 9% or less, about 8% or less, about 7% or less, about 6%or less, about 5% or less, about 4% or less, about 3% or less, about 2% or less, or about 1% or less.

[0115] In some embodiments, biosolids are pulverized to at least 1 mesh size, at least 2 mesh size, at least 3 mesh size, at least 4 mesh size, at least 5 mesh size, at least 6 mesh size, at least 7 mesh size, at least 8 mesh size, at least 9 mesh size, at least 10 mesh size, at least 20 mesh size, at least 30 mesh size, at least 40 mesh size, at least 50 mesh size, at least 60 mesh size, at least 70 mesh size, at least 80 mesh size, at least 90 mesh size, at least 100 mesh size, at least 150 mesh size, at least 200 mesh size, at least 250 mesh size, and at least 300 mesh size.

[0116] In some embodiments, biosolids are pulverized to at least 10 mesh size and up to 300 mesh size to ensure even blending.

[0117] In some embodiments, biosolids are sourced from an Environmental Protection Agency (EP A) Class A exceptional quality rated facilities, with a pretreatment program in place to monitor any potential chemical inputs at concentrations that may be detrimental to soil and human health. To achieve exceptional quality status, the provider must not only meet, but exceed all standards for Class A biosolids including pathogen, metal, and vector attraction reduction parameters.

[0118] In further embodiments, biosolids are generated from pre and post-consumer solid food waste. The food wastes encompass (i) organic waste including, but not limited to food scraps, yard trimmings, paper, and cardboard, and (ii) industrial food waste including but not limited to leaves, peels, pomace, skins, rinds, cores, pits, pulp, stems, seeds, twigs, and spoiled fruits and vegetables produced as waste derived from industrial food processing and the creation of processed food. The food wastes may include, but are not limited to, fresh and processed fruits and vegetables, fluid dairy products, and meat (including poultry and fish), as well as grain products, caloric sweeteners, fats and oils, and may include a category of “other” foods (including eggs, peanuts, tree nuts, dry beans, peas, and lentils, and dairy products other than fluid milk). The food wastes may also include other forms of waste derived from the brewery sector as described above.

[0119] Similar to Class A EQ biosolids, food wastes can be anaerobically digested, dried down and sterilized through a source of heat, such as direct sunlight, a system that generates or cogenerates heat, or via a form of Pyrolysis. The processed food wastes may be blended in with or as an alternative to Class A EQ, Class A and Class B biosolids.

[0120] (ii) Grains

[0121] The present disclosure teaches a bio-nutrient composition comprises a grain, which include a dried distiller’s grain (DDG) or a dried brewer’s spent grain (DBSG). In some embodiments, the bio-nutrient composition comprises the DDG or DBSG at a percentage by weight of 0% to 100%, by weight of about 1% to about 99%, by weight of about 2% to about 95%, by weight of about 3% to about 90%, by weight of about 4% to about 85%, by weight of about 5% to about 80%, by weight of about 6% to about 75%, by weight of about 7% to about 70%, by weight of about 8% to about 65%, by weight of about 9% to about 60%, by weight of about 10% to about 55%, or by weight of about 10% to about 50%.

[0122] The present disclosure teaches that a bio-nutrient composition comprises a grain. In some embodiments, the bio-nutrient composition comprises the grain at a percentage by weight of 0%, by weight of about 1%, by weight of about 2%, by weight of about 3%, by weight of about 4%, by weight of about 5%, by weight of about 6%, by weight of about 7%, by weight of about 8%, by weight of about 9%, by weight of about 10%, by weight of about 11%, by weight of about 12%, by weight of about 13%, by weight of about 14%, by weight of about 15%, by weight of about 16%, by weight of about 17%, by weight of about 18%, by weight of about 19%, by weight of about 20%, by weight of about 21%, by weight of about 22%, by weight of about 23%, by weight of about 24%, by weight of about 25%, by weight of about 26%, by weight of about 27%, by weight of about 28%, by weight of about 29%, by weight of about 30%, by weight of about 31%, by weight of about 32%, by weight of about 33%, by weight of about 34%, by weight of about 35%, by weight of about 36%, by weight of about 37%, by weight of about 38%, by weight of about 39%, by weight of about 40%, by weight of about 41%, by weight of about 42%, by weight of about 43%, by weight of about 44%, by weight of about 45%, by weight of about 46%, by weight of about 47%, by weight of about 48%, by weight of about 49%, by weight of about 50%, by weight of about 51%, by weight of about 52%, by weight of about 53%, by weight of about 54%, by weight of about 55%, by weight of about 56%, by weight of about 57%, by weight of about 58%, by weight of about 59%, by weight of about 60%, by weight of about 61%, by weight of about 62%, by weight of about 63%, by weight of about 64%, by weight of about 65%, by weight of about 66%, by weight of about 67%, by weight of about 68%, by weight of about 69%, by weight of about 70%, by weight of about 71%, by weight of about 72%, by weight of about 73%, by weight of about 74%, by weight of about 75%, by weight of about 76%, by weight of about 77%, by weight of about 78%, by weight of about 79%, by weight of about 80%, by weight of about 81%, by weight of about 82%, by weight of about 83%, by weight of about 84%, by weight ofabout 85%, by weight of about 86%, by weight of about 87%, by weight of about 88%, by weight of about 89%, by weight of about 90%, by weight of about 91%, by weight of about 92%, by weight of about 93%, by weight of about 94%, by weight of about 95%, by weight of about 96%, by weight of about 97%, by weight of about 98%, by weight of about 99%, or by weight of 100%.

[0123] The term “grain” herewith refers to a type of grain that includes a brewers’ spent grain and a distiller’s grain, as well as a dried form thereof.

[0124] Distillers grains (DG) are a by-product of the distillation process that involves cereal grains. Brewer's spent grain (BSG) is produced as a by-product of brewing and is typically made from barley, while distillers grains (DG) are a mixture of com, rice, and other grains. This DG provides a better source of sugars from the com and other sweeter feedstock utilized in the process to make the bio-nutrient compositions taught herein. These grains can come from two primary sources: traditional brewers and ethanol biofuel plants. Distillers grains are made in distilleries by drying mash. For example, corn-based distillers grains are used as high protein feed for livestock, particularly ruminants.

[0125] There are two main types of distillers grains. Wet distillers grains (WDG) consist primarily of unfermented grain residues, such as protein, fiber, and fat, and have a moisture content of about 80%. Dried distillers grains (DDG) are WDG that has been dried with the concentrated thin stillage to about 30% or less, about 25% or less, about 20% or less, to about 15% or less, about 14% or less, about 13% or less, about 12% or less, about 11% or less, about 10% or less, about 9% or less, about 8% or less, about 7% or less, about 6% or less, about 5% or less, about 4% or less, about 3% or less, about 2% or less, or about 1% or less.

[0126] In some embodiments, the grain as a content of the bio-nutrient is the dried distillers grains (DDGs). In some embodiments, the grain as a content of the bio-nutrient is the dried brewer’s spent grains (DBSGs).

[0127] In some embodiments, the grain is dewatered to a moisture content of about 15% or less, or about 10% or less. In some embodiments, the biosolid is dried at a temperature between about 100°F. and about 300°F, between about 125° F. and about 275°F., between about 150°F and about 250°F, or between about 155° F. and about 250° F.

[0128] In some embodiments, pellets provide a storable, easily handled form of DDGs that are compressed into small nuggets that can be bagged or shipped in bulk. In some embodiments, DDGs substitute roughly 1-to-l for corn grain in feed rations.

[0129] In some embodiments, dried distillers grains (DDG) are prepared to maintain a moisture content is about 30% or less, about 25% or less, about 20% or less, to about 15% or less, about 14% or less, about 13% or less, about 12% or less, about 11% or less, about 10% or less, about 9% or less, about 8% or less, about 7% or less, about 6% or less, about 5% or less, about 4% or less, about 3% or less, about 2% or less, or about 1% or less.

[0130] In some embodiments, dried distillers grains are pulverized to at least 1 mesh size, at least 2 mesh size, at least 3 mesh size, at least 4 mesh size, at least 5 mesh size, at least 6 mesh size, at least 7 mesh size, at least 8 mesh size, at least 9 mesh size, at least 10 mesh size, at least 20 mesh size, at least 30 mesh size, at least 40 mesh size, at least 50 mesh size, at least 60 mesh size, at least 70 mesh size, at least 80 mesh size, at least 90 mesh size, at least 100 mesh size, at least 150 mesh size, at least 200 mesh size, at least 250 mesh size, and at least 300 mesh size. In some embodiments, the dried distillers grains are pulverized to at least 10 mesh size and up to 300 mesh size to ensure even blending.

[0131] Brewer's Spent Grain (BSG) is a by-product of the brewing industry. It is obtained as a mostly solid residue after wort production during the brewing process. Spent grain is commonly found wherever beer is consumed and often available at a low cost. Although it is initially wet and has a short shelf-life, it can be dried and processed in various ways to extend its preservation.

[0132] In some embodiments, BSG or DBSG is utilized as a cost-effective soil amendment for agricultural purposes. Its high protein content makes it an excellent source of nitrogen for many common crops. When used in combination with compost, BSG may increase the availability of organic matter in soil and improve the germination rate. The inclusion of BSG in soil may have a stronger positive effect on germination when combined with compost than compost alone.

[0133] The present disclosure teaches that any grains can be used in animal feed that comprising carbohydrate and protein, such as, for example, com, wheat, barley, rice husks. The present disclosure also teaches that other plant or processed food waste can be used as a substitute of grain waste, such as grape pumice, rinds, pulp, and so on. Fertilizer products made from dehydrated food waste are known in increasing soil health and crop productivity, as reported in O’Connor et al, 2022, Environmental Research volume 204, part A, 111927), which is incorporated by its entirety.

[0134] In some embodiments, brewers spent grains comprises malted barley, wheat, maize, com, rice, sorghum, yeast, and millet. The brewers spent grains are collected from one or moresources and dried to a moisture content of about 30% or less, about 25% or less, about 20% or less, to about 15% or less, about 14% or less, about 13% or less, about 12% or less, about 11% or less, about 10% or less, about 9% or less, about 8% or less, about 7% or less, about 6% or less, about 5% or less, about 4% or less, about 3% or less, about 2% or less, or about 1% or less. The dewatering process can be accomplished by one or more ways as known in the art, such as centrifugation, heat, and evaporation. Once the desired moisture content is reached, the spent brewers grains are transferred to a storage bin such as a Super Sack to be transferred to a Super Sack unloading machine, or a trailer to be transferred to a grain silo. Dried brewers grains are then conveyed via a screw auger into a pulverizer to render the dried grains into a fine powder, between 10-300 mesh size.

[0135] In some embodiments, grains used for the bio-nutrient composition is not excessively mouldy.

[0136] Brewers spent grains and wet distillers grains typically contain a moisture content near 80%. In some embodiments, brewers spent grains and wet distillers grains are dewatered using any combination of the following: a centrifuge, a rotary drum, a heat exchanger, roaster, dehydrator, solar drying pad, or other tools known in the art. Drying of brewers grains and wet distillers grains are done rapidly to avoid mold development and natural decomposition. In some embodiments, brewers spent grains are prepared to maintain a moisture content is about 30% or less, about 25% or less, about 20% or less, to about 15% or less, about 14% or less, about 13% or less, about 12% or less, about 11% or less, about 10% or less, about 9% or less, about 8% or less, about 7% or less, about 6% or less, about 5% or less, about 4% or less, about 3% or less, about 2% or less, or about 1% or less.

[0137] In some embodiments, dried brewers spent grains are pulverized to at least 1 mesh size, at least 2 mesh size, at least 3 mesh size, at least 4 mesh size, at least 5 mesh size, at least 6 mesh size, at least 7 mesh size, at least 8 mesh size, at least 9 mesh size, at least 10 mesh size, at least 20 mesh size, at least 30 mesh size, at least 40 mesh size, at least 50 mesh size, at least 60 mesh size, at least 70 mesh size, at least 80 mesh size, at least 90 mesh size, at least 100 mesh size, at least 150 mesh size, at least 200 mesh size, at least 250 mesh size, and at least 300 mesh size. In some embodiments, the dried brewers spent grains are pulverized to at least 10 mesh size and up to 300 mesh size to ensure even blending.

[0138] (Hi) Algae Biomass

[0139] The present disclosure teaches that algae and / or kelp are derived from fresh water sources, algae farms, salt or brackish water sources, or environmental cleanup sites and othercleanup and carbon capture technologies. The present disclosure teaches a bio-nutrient composition comprises an algae biomass or algae cell. In some embodiments, the algae cell comprises a dried algae cell. In some embodiments, the algae cell is present in algae process water. In some embodiments, the algae cell is present in a post-harvest post filtration undried algae solution.

[0140] In some embodiments, the algae process water contains algae cells. In some embodiments, the post-harvest post filtration undried algae solution contains algae cells.

[0141] In other embodiments, the algae process water and / or the post-harvest post filtration undried algae solution can be source of the algae cell, as one of components for the bio-nutrient composition of the present disclosure.

[0142] In some embodiments, the bio-nutrient composition comprises the algae cell at a percentage by weight of 0% to 100%, by weight of about 0.1% to about 90%, by weight of about 0.2% to about 80%, by weight of about 0.3% to about 70%, by weight of about 0.4% to about 60%, or by weight of about 0.5% to about 40%.

[0143] The present disclosure teaches that a bio-nutrient composition comprises an algae cell. In some embodiments, the bio-nutrient composition comprises the algae cell at a percentage by weight of 0%, by weight of about 1%, by weight of about 2%, by weight of about 3%, by weight of about 4%, by weight of about 5%, by weight of about 6%, by weight of about 7%, by weight of about 8%, by weight of about 9%, by weight of about 10%, by weight of about 11%, by weight of about 12%, by weight of about 13%, by weight of about 14%, by weight of about 15%, by weight of about 16%, by weight of about 17%, by weight of about 18%, by weight of about 19%, by weight of about 20%, by weight of about 21%, by weight of about 22%, by weight of about 23%, by weight of about 24%, by weight of about 25%, by weight of about 26%, by weight of about 27%, by weight of about 28%, by weight of about 29%, by weight of about 30%, by weight of about 31%, by weight of about 32%, by weight of about 33%, by weight of about 34%, by weight of about 35%, by weight of about 36%, by weight of about 37%, by weight of about 38%, by weight of about 39%, by weight of about 40%, by weight of about 41%, by weight of about 42%, by weight of about 43%, by weight of about 44%, by weight of about 45%, by weight of about 46%, by weight of about 47%, by weight of about 48%, by weight of about 49%, by weight of about 50%, by weight of about 51%, by weight of about 52%, by weight of about 53%, by weight of about 54%, by weight of about 55%, by weight of about 56%, by weight of about 57%, by weight of about 58%, by weight of about 59%, by weight of about 60%, by weight of about 61%, by weight of about 62%, byweight of about 63%, by weight of about 64%, by weight of about 65%, by weight of about 66%, by weight of about 67%, by weight of about 68%, by weight of about 69%, by weight of about 70%, by weight of about 71%, by weight of about 72%, by weight of about 73%, by weight of about 74%, by weight of about 75%, by weight of about 76%, by weight of about 77%, by weight of about 78%, by weight of about 79%, by weight of about 80%, by weight of about 81%, by weight of about 82%, by weight of about 83%, by weight of about 84%, by weight of about 85%, by weight of about 86%, by weight of about 87%, by weight of about 88%, by weight of about 89%, by weight of about 90%, by weight of about 91%, by weight of about 92%, by weight of about 93%, by weight of about 94%, by weight of about 95%, by weight of about 96%, by weight of about 97%, by weight of about 98%, by weight of about 99%, or by weight of 100%.

[0144] In some embodiments, the algae cell is pressed to isolate an algae cell wall. In some embodiments, the algae cells and / or the algae cell walls are dried to maintain a moisture content is about 30% or less, about 25% or less, about 20% or less, to about 15% or less, about 14% or less, about 13% or less, about 12% or less, about 11% or less, about 10% or less, about 9% or less, about 8% or less, about 7% or less, about 6% or less, about 5% or less, about 4% or less, about 3% or less, about 2% or less, or about 1% or less. In some embodiments, the algae cells and / or the algae cell walls are dried to maintain a moisture content is about 20% or less.

[0145] In some embodiments, dried algae cells and / or cell walls are pulverized to at least 1 mesh size, at least 2 mesh size, at least 3 mesh size, at least 4 mesh size, at least 5 mesh size, at least 6 mesh size, at least 7 mesh size, at least 8 mesh size, at least 9 mesh size, at least 10 mesh size, at least 20 mesh size, at least 30 mesh size, at least 40 mesh size, at least 50 mesh size, at least 60 mesh size, at least 70 mesh size, at least 80 mesh size, at least 90 mesh size, at least 100 mesh size, at least 150 mesh size, at least 200 mesh size, at least 250 mesh size, and at least 300 mesh size. In some embodiments, the dried distillers cells and / or cell walls are pulverized to at least 10 mesh size and up to 300 mesh size to ensure even blending.

[0146] Algae are ubiquitous organisms that can be found in various environments, and they possess unique features that can be utilized for several applications, including agricultural purposes such as bio-fertilizers and soil conditioning agents that enhance soil fertility and plant productivity (Chapman, 2013; Duarte et al., 2018). Soil algae are simple photosynthetic microorganisms that originate in the soil, and they thrive inches below the soil surface, making the soil a suitable habitat for algal growth and evolution (Duarte et al., 2018). Adding algae to different living organisms in different soil types and in various states can indicate the health ofthe soil environment. Algae can also contribute to the development of soil characteristics such as carbon content, texture, aeration, and nitrogen fixation. Additionally, the growth of algae in soil can help to reduce soil erosion by managing water flow. Algae also play a crucial role in soil reclamation, fertility enhancement, microbiological crust formation, biocontrol of agricultural pests, and treatment of agricultural wastewater.

[0147] The present disclosure teaches that the algae cell class is Cyanophyceae, Chlor ophyceae, Rhodophyceae , or Phaeophyceae.

[0148] In some embodiments, five species of algae are provided; cyanobacterium (also called blue-green algae) Arthrospira platensis (Spirulina), unicellular green algae Chlor ella sp., red seaweed Palmaria palmate, and brown seaweeds (Laminaria digitata and Ascophyllum nodosum). In some embodiments, Spirulina (A. platensis), Chlor ella sp., P. palmata, L. digitata, or A. nodosum can enhance or improve N, P, K, or C concentrations in the soil.

[0149] The present disclosure teaches that the genus of algae is Arthrospira genus, Chlorella genus, Palmaria genus, Lamanaria genus, or Ascophyllum genus.

[0150] In some embodiments, other green algae and cyanobacteria nc\u o_Aculodesmus dimorphus, Spirulina platensis, Chlorella vulgaris, Scenedesmus dimorphus, Anabaena azolla, Nostoc sp., Chlorella vulgaris, Sar gassum sp, Gracilaria verrucose, Scenedesmus obliquus, and Sargassum Polycystum.

[0151] The present disclosure further teaches that the genus of algae is Acutodesmus genus, Spirulina genus, Chlorella genus, Scenedesmus genus, Anabaena genus, Nostoc genus, Chlorella genus, Sargassum genus, Gracilaria genus, Scenedesmus genus, or Sargassum genus.

[0152] The disclosure described herein refers to a bio-nutrient compounds and the method of production thereof. In some embodiments, biosolids, dried brewers’ spent grains or dried distillers grains, and algae cell wall are collected, treated, and mixed to specific ratios to form bio-nutrient compounds, which are bio-fertilizers. The bio-nutrient compounds are added with a binder (e.g., lignosulfonate) to make a bio-nutrient formulation. Optionally, an additive and / or a biologic suppressants can be added to the bio-nutrient formulation.

[0153] The present disclosure teaches that the bio-nutrient compounds encompass slow release fertilizers, wherein a minimum of about 5%, about 10%, about 15%, or about 20% of total nitrogen, potassium, and phosphorous is biologically unavailable at the initial application.

[0154] The present disclosure teaches that each component of the bio-nutrient composition is collected from one or more sources. Components such as brewers spent grains or distillersgrains as well as algae are received having a high moisture content that can result in rampant mold growth or decomposition, thus spoiling the component. To dry, a number of methods can be used as described herein. Heat, if used, is kept below 250° F to prevent burning and to keep the total and available nutrients and compounds intact

[0155] In some embodiments, biosolid pellets are loaded into the super sack unloading machine and are then conveyed by screw auger into the pulverizer. The pulverized biosolids are conveyed to a separate super sack for storage prior to a blending stage

[0156] In some embodiments, manure and biosolids are used to meet the United States Environmental Protection Agencies (EP A) treatment standards for Class A Exceptional Quality. In further embodiments, manure is sourced from farms that allow animals free range and eating a natural diet. In further embodiments, biosolids are sourced from wastewater agencies that have about 80% or greater, about 85% or greater, about 90% or greater, about 91% or greater, about 92% or greater, about 93% or greater, about 94% or greater, or about 95% or greater residential customer base, and minimal toxic industrial and commercial customers.

[0157] Per US Environmental Protection Agency 40 CFR 503 regulations for Class A Exceptional Quality biosolids, sewage sludge is digested in an anaerobic (digester) fermenter, moved to a centrifuge for dewatering, and transferred through a heat source to ensure pathogen reduction and elimination, uniform size, and dewatering. Final moisture content is about 20% or less, about 15% or less, about 10% or less, 9% or less, 8% or less, 7% or less, 6% or less, or 5% or less.

[0158] In some embodiments, dried pellets are treated with a vegetable oil-based additive to prevent billowing of dust when they are transported and moved between containers and spaces. The dried pellets are then pulverized to at least 1 mesh size, at least 2 mesh size, at least 3 mesh size, at least 4 mesh size, at least 5 mesh size, at least 6 mesh size, at least 7 mesh size, at least 8 mesh size, at least 9 mesh size, at least 10 mesh size, at least 20 mesh size, at least 30 mesh size, at least 40 mesh size, at least 50 mesh size,, at least 60 mesh size, at least 70 mesh size, at least 80 mesh size, at least 90 mesh size, at least 100 mesh size, at least 150 mesh size, at least 200 mesh size, at least 250 mesh size, and at least 300 mesh size powder to be blended into the final bio-nutrient composition.

[0159] In some embodiments, a super sack containing dried algae cell wall is loaded into a super sack unloading machine and conveyed via a screw auger to a pulverizer. In someembodiments, dried algae are preprocessed to move through the screw auger without clogging. Pulverized algae are then conveyed into another super sack for storage prior to blending.

[0160] In some embodiments, the algae is harvested from a system that captures carbon dioxide, soot, heat, and other exhaust from coal fired power plants, or other sources of burned biomass, mined materials, timber, or natural resources. In some embodiments, the algae are pressed to extract lipids, leaving the algae cell wall post lipid extraction, thus isolating the algae cell wall. The retained algae cell wall is dehydrated using processes known in the art such that nutrient content is preserved. In some embodiments the algae are recovered from sludge from the ponds algae is grown in, recapturing the biomass shed in the growing process. The material is dehydrated using processes known in the art such that nutrient content is preserved. In some embodiments, the algae contains a bacteria count per cell too high for use as a veterinary, pharmaceutical or nutraceutical grade product and therefore has a reduced to non-viable market value in products created for human or animal consumption.

[0161] Each of the super sacks containing the dried brewers grain, pulverized biosolids, and pulverized algae cell wall are loaded into weighing and proportioning equipment as known in the art. Each component is mixed uniformly, which is more easily accomplished due to the drying and pulverizing pretreatment. Once fully blended, fertilizer composition is placed in a super sack storage container before allocation into marketable volumes.

[0162] In some embodiments, the final bio-nutrient composition in a formulation is comprised of biosolids at a range of 35% to 75% by weight, brewing grains at a range of 10% to 50% by weight, and dried algae cell wall at a range of 5% to 40% by weight.

[0163] As one example, the bio-nutrient composition or compound allows for faster metabolic breakdown and nutrient uptake by the crops. The specific blend, and mesh size of 10-300 allows for the composition to be utilized in a hydroponic system, wherein the composition is suspended in water and pumped through the hydroponic irrigation systemBio-nutrient formulation I

[0164] The present disclosure teaches a bio-nutrient formulation comprises (i) a bio-nutrient composition and (ii) a binder. In some embodiments, a binder is lignins, minerals, plant starches, or molasses.

[0165] In some embodiments, the bio-nutrient composition comprises: i) a biosolid at a final percentage-by-weight of about 35% to about 75% of the bio-nutrient composition; ii) a grain at a final percentage-by-weight of about 10% to about 50% of the bio-nutrient composition;and iii) an algae cell at a final percentage-by-weight of about 5% to about 40% of the bionutrient composition. In some embodiments, the bio-nutrient formulation comprises the bionutrient composition at a percentage at a percentage by weight of 0% to about 100%, by weight of about 0.1% to about 99%, by weight of about 0.2% to about 99%, by weight of about 0.3% to about 99%, by weight of about 0.4% to about 99%, by weight of about 0.5% to about 99%, by weight of about 1% to about 99%, by weight of about 5% to about 99%, by weight of about 10% to about 99%, by weight of about 20% to about 99%, by weight of about 30% to about 99%, by weight of about 40% to about 99%, or by weight of about 50% to about 99% of the formulation. In some embodiments, the bio-nutrient composition is passed through a heating source, thereby being dewatered.

[0166] In some embodiments, the biosolid is a sewage sludge, a manure or a food waste. In further embodiments, the biosolid is a Class A EQ biosolid, Class A biosolid, or a Class B biosolid. In some embodiments, the biosolid is digested in an anaerobic digester for at least 15 days. In some embodiments, the biosolid is dewatered to a moisture content of about 15% or less, or about 10% or less. In some embodiments, the biosolid is pulverized to a mesh size between about 10 and about 300. In some embodiments, the biosolid is dried at a temperature between about 155° F. and about 250° F. In some embodiments, the biosolid is treated with an oil based agent, wherein the oil based agent prevents billowing.

[0167] In some embodiments, the grain is dewatered to a moisture content of about 15% or less, or about 10% or less. In some embodiments, the grain is pulverized to a mesh size of about 10 to about 300. In some embodiments, the grain is dried at a temperature between about 155° F. and about 250° F.

[0168] In some embodiments, the algae cell is Cyanophyceae, Chlor ophyceae, Rhodophyceae, or Phaeophyceae . In some embodiments, the algae cell comprises a dried algae cell wall, algae process water, or a post-harvest post filtration undried algae solution. In some embodiments, the binder is at a percentage weight of about 1% to about 50% of said formulation. In some embodiments, the algae cell is pressed to isolate an algae cell wall. In some embodiments, the algae cell wall is pulverized to a mesh size of about 10 to about 300. In some embodiments, the algae cell wall is dried to a moisture content of about 20% or less.

[0169] In some embodiments, the bio-nutrient formulation is made by adding a binder to the bio-nutrient composition. In some embodiments, the binder is a lignin or a lignin derivative. In some embodiments, the lignin derivative is lignosulfonate. In some embodiments, the bio-nutrient formulation comprises the binder at a percentage at a percentage by weight of about 0.5% to about 50%, or by weight of about 1% to about 50% of the formulation.

[0170] Lignin functions as an essential polymer in plants, serving as the structural backbone of the plant body. Acting as a natural adhesive, it binds together the cellulosic fibers in the plant, providing rigidity, durability, strength and resilience. Lignin has great potential in the creation of biomaterials due to its abundance, non-toxicity, and ability to biodegrade.

[0171] As lignin has adhesive behavior and plays the part of a binder by holding cellulose fibers together, the present disclosure teaches a bio-nutrient formulation, which is made by the addition of a binder to a bio-nutrient composition.

[0172] In the present disclosure, lignin, such as lignosulfonate, play crucial roles in improving the physical properties and performance of bio-nutrient formulations. In one aspect, by acting as binders, they enhance granule formation, reduce dust, control nutrient release, and contribute to soil health. In another aspect, lignins and its derivatives can benefit and / or enhance plant growth, development, and stress defense by ensuring a steady supply of nutrients, improving soil structure and health, enhancing microbial activity, promoting better root development, and supporting osmotic regulation and antioxidant defense mechanisms. Thus, while the primary function of lignin and / or lignosulfonate is to enhance the physical characteristics of bio-nutrient compositions, lignin and lignosulfonate also play a significant role in creating favorable conditions for plant growth and resilience. While lignin and lignosulfonate themselves do not directly provide nutrients that plants use, their role in improving the delivery and availability of nutrients from bio-nutrient fertilizers indirectly supports plant growth and stress defense. By enhancing soil health and nutrient uptake, these binders contribute to overall plant vigor, making plants more capable of withstanding abiotic and biotic stresses.

[0173] The general response of plants under biotic and abiotic stress is the accumulation of reactive oxygen species (ROS), which mainly can damage cell walls. This phenomenon is accompanied by an increase in lignin accumulation (Moura et al., 2010). Lignin not only provides a physical barrier to insect and pathogen infestations, but it also stimulates hormone signal pathways to increase the plants’ resistance to them (Liu et al., 2018).

[0174] Lignosulfonates can promote rhizospheric and soil microbial populations, thus improving soil health and plant-environment interactions. A main component of lignosulfonates is sulfur (S), which is essential to the normal functioning of chlorophyll and plant proteins and will enhance photosynthesis in plants under salinity stress (Duncan et al., 2018). Calcium lignosulfonate (Ca-LGSN) has been shown to be quite effective at mitigatingsalinity stress in cereal crops such as barley, rice and com (Elsawy et al., 2022). The Ca in this compound is chemically active and can extract Na ions from soil particles in the rhizosphere. Additionally, it contains an active alkyl group that will bind to Na and convert it to an organic form which is not harmful to plants (Elsawy et al., 2022).

[0175] In some embodiments, lignin and / or lignosulfonate as a binder can help control the release rate of nutrients from the bio-nutrient fertilizer. This ensures a steady and prolonged supply of essential nutrients to the plants, supporting continuous growth and development. In other embodiments, lignosulfonate can chelate micronutrients (such as iron, zinc, and manganese), which are vital for the synthesis of antioxidants. These antioxidants protect plants from oxidative stress caused by environmental challenges like high light intensity, extreme temperatures, and pollution. This improves the efficiency of micronutrient use, which enhances various physiological functions and stress responses in plants.

[0176] In further embodiments, lignin and / or lignosulfonate can contribute to increase in organic matter to the soil as they decompose. Increased organic matter improves soil structure, water retention, and aeration, creating a better root environment and enhancing plant growth, as well as stimulate microbial activity in the soil.

[0177] The present disclosure provides that the bio-nutrient formulation comprises about 1% to about 50% of a binder, which is added to the bio-nutrient composition (about 50% to 99%) in order to form durable, separable form of the bio-nutrient formulation.

[0178] The present disclosure provides that the bio-nutrient formulation is used as a biostimulant, soil amendment, and / or source of nutrients.Bio-nutrient formulation II

[0179] The present disclosure provides a bio-nutrient formulation comprises (i) a bio-nutrient composition; (ii) a binder; and (c) optionally an additive. In some embodiments, the additive is present. In some embodiments, the additive is not present.

[0180] The bio-nutrient formulation has been associated with a strong, unpleasant smell of the product. The addition of odor reducing agents can address unpleasant odors and smells. In addition, various oils and other odor masking agents may provide potential enhancements to product efficacy and could possibly help with rodent and bug management.

[0181] In order to promote reduction in odor from the bio-nutrient formulation described above, an additive is added to or integrated into a powder form, a granular form, or a liquid form of the bio-nutrient formulation.

[0182] In some embodiments, the additive is an odor neutralizer as an absorbent; (i) an activated charcoal, which is known for its ability to absorb and neutralize odors and / or odorcausing molecules; (ii) zeolite, which is a natural mineral that can trap and neutralize odor and / or odor-causing molecules; and (iii) baking soda, which can absorb odors and neutralize acidic or basic odor molecules.

[0183] In some embodiments, the additive is an essential oil; (i) lavender oil, which is known for its pleasant, calming fragrance and antimicrobial properties that can help neutralize odorcausing bacteria and fungi; (ii) eucalyptus oil, which has a strong, fresh scent that can help mask unpleasant odors and possesses antibacterial and antifungal properties; (iii) peppermint oil, which offers a strong, refreshing, minty scent that can help neutralize strong smells with antibacterial properties; (iv) lemon oil, which provides a fresh, clean scent and has antibacterial properties; (v) tea tree oil, has strong antibacterial, antifungal, and antiviral properties, making it effective in eliminating the microbes responsible for bad odors; (vi) cinnamon oil, which is known for its warm and spicy scent with antimicrobial and antifungal properties that can help neutralize odors and eliminate odor-causing microbes; (vii) clove oil, which has a strong, spicy aroma and contains eugenol, which has antimicrobial properties that help in reducing odors; and (viii) orange oil, which has a sweet, citrus scent and contains limonene that is effective in breaking down and neutralizing odors. Essential oils are effective odor-removing agents due to their antimicrobial properties, strong fragrances, and ability to chemically neutralize odor molecules. In some embodiments, essential oils taught herein include, but are not limited to, lavender, tea tree, lemon, peppermint, eucalyptus, cinnamon, clove, and orange oils.

[0184] In some embodiments, the additive is an odor masking agent, including essential oils described above as well as botanical extracts; (i) vanilla extract, which can help mask odors with its sweet scent; and (ii) citrus / orange / lemon extracts, which can provide a pleasant fragrance.

[0185] In some embodiments, the additive is a microbial solution that uses beneficial microorganisms to neutralize odors by breaking down the organic matter that produces unpleasant smells: (i) bacterial solutions (such as Bacillus species o Pseudomonas species); (ii) fungal solutions (such as Trichoderma species); (iii) yeast solutions (such as Saccharomyces cerevisiae),' (iv) probiotic solutions (such as Lactobacillus species); and (v) combinations of (i)-(iv). Enzyme-Producing Bacteria (such as Pseudomonas species) are also utilized as a microbial solution. These Pseudomonas ox Bacillus bacteria produce enzymes thatbreak down proteins, fats, and carbohydrates, which are often the source of foul odors. The enzyme is a protease, a lipase, an amylase, a cellulase, a urease, an oxidoreductase, or a pectinase. By degrading these compounds, they help eliminate the odors.

[0186] In some embodiments, the additive is a chemical odor neutralizer: (i) odor neutralizing compounds containing cyclodextrins that trap and neutralize odor molecules; (ii) bio- enzymatic cleaners, which contain enzymes that break down the organic materials causing the odor.

[0187] In some embodiments, the additive is a pH adjuster; (i) acidulants lowering the pH with substances like citric acid or vinegar can help reduce odor by making the environment less favorable for odor-producing bacteria.

[0188] In some embodiments, the additive is an absorptive material; (i) clay (such as bentonite, zeolite or kaolin clay), having a high surface area and porous structure, which can absorb odors and trap odor molecules; and (ii) peat moss, which has a fibrous structure and high absorbency, making it effective at trapping odor molecules and absorbing moisture with antimicrobial properties that help reduce the growth of odor-causing microbes.

[0189] In some embodiments, the additive is an odor-reducing or odor-neutralizing agent. The additive is selected from the group consisting of: (i) an order neutralizer, which is an activated charcoal, a zeolite, a baking soda, a silica gel, or a compound comprising cyclodextrin; (ii) an odor-masking agent, which is a vanilla extract or a citrus extract; (iii) an essential oil, which is a lavender oil, a eucalyptus oil, a peppermint oil, a lemon oil, a tea tree oil, a clove oil, an orange oil, or a cinnamon oil, (iv) an odor absorbent is bentonite, kaolin clay, or peat moss; (v) an enzyme, which is a protease, a lipase, an amylase, a cellulase, a urease, an oxidoreductase, or a pectinase; and (vi) a odor-removing microbe, which is a species of genus Bacillus or Pseudomonas.

[0190] The present disclosure teaches that the bio-nutrient formulation is in a powder form, in a granular form, in a liquid form, or in a prill form. In some embodiments, the liquid form is a suspension or an extract.

[0191] In order to minimize offgassing in a liquid form of the bio-nutrient formulation, further additives and / or enhancers are added to or integrated into the liquid form of the bio-nutrient formulation.

[0192] In some embodiments, the additive or enhancer is a preservative; (i) sodium benzoate, which is commonly used in the food industry to inhibit microbial growth; (ii) potassiumsorbate, which is effective against molds, yeasts, and many bacteria; and (iii) citric acid, which helps to lower the pH, making the environment less favorable for microbial growth.

[0193] In some embodiments, the additive or enhancer is a stabilizer; (i) chelating agents, such as EDTA (ethylenediaminetetraacetic acid), can bind to metal ions that might catalyze unwanted reactions; and (ii) antioxidants, such as ascorbic acid or tocopherols, can prevent oxidation reactions that might cause offgassing.

[0194] In some embodiments, the additive or enhancer is a buffering agent; (i) phosphates, which can help maintain a stable pH, reducing the likelihood of reactions that produce gas; (ii) acetates, such as sodium acetate, which can also act as a buffering agent.

[0195] In some embodiments, the additive or enhancer is an essential oil; (i) thyme or oregano oil, which has antimicrobial properties and can be used in low concentrations; (ii) peppermint and other mints, which may also help manage rodent presence such as mice and bugs.

[0196] In some embodiments, the additive or enhancer is a silver nanoparticle, which is known for their antimicrobial properties but require careful handling and formulation.

[0197] In some embodiments, the additive or enhancer is a pH Adjuster; (i) hydrochloric acid, which can be used to lower the pH; (ii)sodium hydroxide, which can be used to raise the pH.

[0198] In some embodiments, the additive or enhancer is an anti-foaming agent; (i) silicone- based Antifoams, which can help to reduce the formation of foam which might trap gases.

[0199] In some embodiments, the additive or enhancer is a packaging solution, (i) barrier films, which use packaging materials with high gas barrier properties; and (ii) vented caps: These can allow gas to escape without letting contaminants in.)

[0200] In some embodiments, the additive or enhancer is a stabilizing agent, which is Xantham Gum.

[0201] The present disclosure provides that the liquid form of the bio-nutrient formulation further comprises an enhancer. In some embodiments, the enhancer is selected from the consisting of: a preservative, a stabilizer, a buffering agent, a microbial inhibitor, a pH adjuster, and an anti-foaming agent. In some embodiments, the preservative is sodium benzoate, potassium sorbate, or citric acid. In some embodiments, the stabilizer is a xanthan gum, a chelating agent or an antioxidant. In some embodiments, the chelating agent is ethylenediaminetetraacetic acid (EDTA). In some embodiments, the antioxidant is an ascorbic acid or a tocopherol. In some embodiments, the buffering agent is phosphate, phosphate derivative, acetate, or acetate derivative. In some embodiments, the microbial inhibitor is an essential oil or a silver nanoparticle. In some embodiments, the essential oil is a thyme oil, anoregano oil, a peppermint oil, or a mint oil. In some embodiments, the pH adjuster is hydrochloric acid or sodium hydroxide. In some embodiments, the anti-foaming agent is a silicone-based antifoam.Dry formulations

[0202] The present disclosure teaches that the bio-nutrient composition is in a powder form. For a better delivery mechanism of the bio-nutrient composition with product durability improved, the bio-nutrient composition is further processed to enhance the powder into a prill and / or granular form by adding a binder such as lignin. This binder provides a sticky substance that could melt, capture the powder, and agglomerate it into a granular “prill” form. In some embodiments, this prill form breaks down better than powder, avoids clumping associated with the powder form, and adds in another important source of nutrients for microbes by present of the sugars and nutrients from tree sap and timber industry waste. The term “prill” herein refers to a pellet or solid globule form of a bio-nutrient formulation as a final end product in the present disclosure. It may be formed using agglomeration methods such as a prill tower, a pan granulator or other granulation equipment using rotating methods for binding loose materials into a granular form.

[0203] The present disclosure teaches a bio-nutrient formulation, which is a prill or granular bio-nutrient formulation. The addition of a lignin binder to a bio-nutrient composition results in the binding of a powder form of the bio-nutrient composition into a final product in a prill or granular formulation.

[0204] Lignin is derived from the timber manufacturing process, specifically the sap, oil, and bark material waste not used in the production of finished timber. Lignin in this form when heated and combined with the powder form of the bio-nutrient composition is a means of combining, agglomerating and binding the powder into a fairly consistent, durable, and easy to apply material (i.e., prill). Utilizing lignin integrates natural sugars, carbohydrates and nutrients to supplement the carbohydrates found in the DBSG and DDG. Lignin can be bound to hemicelluloses or cellulose molecules to form lignin-carbohydrate complexes (LCC), which could be used in polymer composites.

[0205] The combination of lignin and dried grains is a key catalyst for enhancing microbes that are indigenous (naturally occurring and existing in a given plot of land), amended (added from soil inputs or other materials not native / indigenous to the soil), and engineered (genetically edited) to a given plot of soil.

[0206] In further embodiments, granulation technologies such as pan granulating can be utilized as a means of making a final product of the bio-nutrient formulation. This granulation step provides a similar size end product. Integrating a lignin or similar product (such as molasses) is an important step to integrate additional sugars and carbohydrates which function as feedstock for microbes and create the broad microbe responses.

[0207] The present disclosure teaches that the application of bio-nutrient formulations (comprising a bio-nutrient composition and a binder) enhances the biological activities and functions of microbes native / indigenous to soil, as well as microbes that are genetically edited and applied to plants. In some embodiments, the sugars and nutrients available in lignin can accelerate the microbe activity observed at the various intervals of the soil tests described in Examples and FIGs. 5-7. Also, the microbes demonstrate consistent increases when the bionutrient formulation (with lignin added) is applied, in comparison to the bio-nutrient composition (without lignin added). The presence of the lignin in the formulation provides additional sugars for microbes’ activities and functions in a positive manner.

[0208] Carbohydrates in the DDG and DBSG break down into glucose, glucose is what becomes the feedstock for microbes. Sugars already available in the Lignin from tree sap also become additional feedstock for microbes.

[0209] The enhancement of lignin has a positive impact in terms of efficacy with microbes, added nutrient value, ease of manufacturing, use and application, when compared to traditional sources of microbe food which would include but is not limited to sugar beet molasses, and other plant derived forms of molasses or other sugars.

[0210] The sugars and nutrients from the lignin as well as the product delivery form (i.e., prill) enhance the existing macronutrients, secondary and micronutrients, carbon, organic matter, and humic / fulvic acid complex that exists in the powder.

[0211] In further embodiments, the addition of lignin positively effects the root weight and density as measured against a synthetic fertilizer.Liquid formulations

[0212] The bi-nutrient formulation is also in a liquid form. The present disclosure teaches a method of making a liquid that involves (i) heating water, (ii) heating the powder form of the bio-nutrient composition into the water, (iii) removing the liquid and powder “tea” from heat, (iv) integrating a form of high-speed mixing / agitation into the production process to aid in the blending, and (v) steeping and disbursing the powder into the liquid. This is followed by (vi)straining the agitated “tea” through a substance fine enough to separate particles from liquid to result in a final “liquid” product.

[0213] In some embodiments, a cold water extract process is used to make a liquid formulation. Wet milling technologies are applied to reduce the particle size of the powder material, which may involve use of additional chemical or mechanical methods to extract nutrients and elements and / or suspend particulate matter in water at a size that will pass through irrigation system filters. The present disclosure teaches the liquid formulation prepared by suspending the powder form of the bio-nutrient composition and lignin into liquid and / or extracting nutrients and elements from the powder and lignin into liquid.

[0214] While the liquid form of the bio-nutrient composition is prepared, a lignin can be added to this liquid product in order to enhance the sugars / feedstock for microbes. In some embodiments, the adding of a small amount of the binder into the liquid formulation give positive benefits by increasing the sugars in the formulation, which can boost microbial activities or functions.

[0215] In some embodiments, lignin can be added to a liquid form of the bio-nutrient composition by about 1%, about 5%, about 10%, about 15%, or about 20% of the total volume of a liquid form of the bio-nutrient composition.

[0216] In other embodiments, the powder “tea” can be thickened to a point where it can be applied as a coating to existing granular products or seeds using suspension technologies.Application methods

[0217] The present disclosure teaches that a powder form of the bio-nutrient formulation is mixed into compost and topsoil as a pretreatment / soil charge prior to planting.

[0218] The present disclosure teaches that a granular form of the bio-nutrient formulation is applied by broadcast spreading, air seeder / drill, strip till, variable rate application (VRA), in furrow application, side dressing, hand spreader, belly grinder, hand broadcast, drop spreader, side dressing, banding, precision planter with fertilizer hoppers, attachment with transplanters, and / or fertilizer spikes. In some embodiments, granular formulation products can enhance the efficacy of existing products through co-formulation into existing fertilizer, chemical, biological or specialty chemical products such as herbicides, fungicides, pesticides, nematicides, adjuvants, or surfactants. This may be through various coformulation, mixing and blending technologies known in the art.

[0219] The present disclosure teaches that a granular form of the bio-nutrient formulation is applied by foliar spray, soil drench, seedling transplant / root soak, irrigation system integration / fertigation (including but not limited to drip irrigation, tape irrigation, microsprinkler irrigation), hose end sprayer, watering can, root feeder, and spray bottle, aerial application, boom sprayer, center pivot application, side dress application, variable rate technology (VRT), Y drop application, and in furrow application. In some embodiments, liquid formulation products can enhance efficacy of existing products through coformulation into existing fertilizer, chemical, biological or specialty chemical products such as but not limited to fungicides, herbicides, pesticides, nematicides, adjuvants, or surfactants. This may be through impregnation, coating, wet milling, and other mixing and blending technologies known in the art.Application rates

[0220] The present disclosure provides a granular product of the bio-nutrient formulation can be applied to soil, ground, or field at a rate of about 1 pounds / acre, about 10 pounds / acre, about 20 pounds / acre, about 30 pounds / acre, about 40 pounds / acre, about 50 pounds / acre, about 60 pounds / acre, about 70 pounds / acre, about 80 pounds / acre, about 90 pounds / acre, about 100 pounds / acre, about 150 pounds / acre, about 200 pounds / acre, about 250 pounds / acre, about 300 pounds / acre, about 350 pounds / acre, about 400 pounds / acre, about 450 pounds / acre, about 500 pounds / acre, about 550 pounds / acre, about 600 pounds / acre, about 650 pounds / acre, about 700 pounds / acre, about 750 pounds / acre, about 800 pounds / acre, about 850 pounds / acre, about 900 pounds / acre, about 950 pounds / acre, about 1,000 pounds / acre, about 1,000 pounds / acre, about 1,100 pounds / acre, about 1,200 pounds / acre, about 1,300 pounds / acre, about 1,400 pounds / acre, about 1,500 pounds / acre, about 1,600 pounds / acre, about 1,700 pounds / acre, about 1,800 pounds / acre, about 1,900 pounds / acre, about 2,000 pounds / acre, or about 2,500 pounds / acre.

[0221] In some embodiments, a granular product of the bio-nutrient formulation is applied to soil, ground, or field at a rate of about 50 pounds / acre to about 2,500 pounds / acre, about 100 pounds / acre to about 2,000 pounds / acre, or about 500 pounds / acre to about 1,5000 pounds / acre.

[0222] The present disclosure provides a liquid product of the bio-nutrient formulation can be applied to plants or crops at a rate of about 1 ounce / acre, about 2 ounces / acre, about 3 ounces / acre, about 4 ounces / acre, about 5 ounces / acre, about 6 ounces / acre, about 7ounces / acre, about 8 ounces / acre, about 9 ounces / acre, about 10 ounces / acre, about 11 ounces / acre, about 12 ounces / acre, about 13 ounces / acre, about 14 ounces / acre, about 15 ounces / acre, about 16 ounces / acre, about 17 ounces / acre, about 18 ounces / acre, about 19 ounces / acre, about 20 ounces / acre, about 21 ounces / acre, about 22 ounces / acre, about 23 ounces / acre, about 24 ounces / acre, about 25 ounces / acre, about 26 ounces / acre, about 27 ounces / acre, about 28 ounces / acre, about 29 ounces / acre, about 30 ounces / acre, about 31 ounces / acre, about 32 ounces / acre, about 33 ounces / acre, about 34 ounces / acre, about 35 ounces / acre, about 36 ounces / acre, about 37 ounces / acre, about 38 ounces / acre, about 39 ounces / acre, about 40 ounces / acre, about 41 ounces / acre, about 42 ounces / acre, about 43 ounces / acre, about 44 ounces / acre, about 45 ounces / acre, about 46 ounces / acre, about 47 ounces / acre, about 48 ounces / acre, about 49 ounces / acre, about 50 ounces / acre, about 51 ounces / acre, about 52 ounces / acre, about 53 ounces / acre, about 54 ounces / acre, about 55 ounces / acre, about 56 ounces / acre, about 57 ounces / acre, about 58 ounces / acre, about 59 ounces / acre, about 60 ounces / acre, about 61 ounces / acre, about 62 ounces / acre, about 63 ounces / acre, about 64 ounces / acre, about 65 ounces / acre, about 66 ounces / acre, about 67 ounces / acre, about 68 ounces / acre, about 69 ounces / acre, about 70 ounces / acre, about 71 ounces / acre, about 72 ounces / acre, about 73 ounces / acre, about 74 ounces / acre, about 75 ounces / acre, about 76 ounces / acre, about 77 ounces / acre, about 78 ounces / acre, about 79 ounces / acre, about 80 ounces / acre, about 81 ounces / acre, about 82 ounces / acre, about 83 ounces / acre, about 84 ounces / acre, about 85 ounces / acre, about 86 ounces / acre, about 87 ounces / acre, about 88 ounces / acre, about 89 ounces / acre, about 90 ounces / acre, about 91 ounces / acre, about 92 ounces / acre, about 93 ounces / acre, about 94 ounces / acre, about 95 ounces / acre, about 96 ounces / acre, about 97 ounces / acre, about 98 ounces / acre, about 99 ounces / acre, about 100 ounces / acre, about 101 ounces / acre, about 102 ounces / acre, about 103 ounces / acre, about 104 ounces / acre, about 105 ounces / acre, about 106 ounces / acre, about 107 ounces / acre, about 108 ounces / acre, about 109 ounces / acre, about 110 ounces / acre, about 111 ounces / acre, about 112 ounces / acre, about 113 ounces / acre, about 114 ounces / acre, about 115 ounces / acre, about 116 ounces / acre, about 117 ounces / acre, about 118 ounces / acre, about 119 ounces / acre, about 120 ounces / acre, about 121 ounces / acre, about 122 ounces / acre, about 123 ounces / acre, about 124 ounces / acre, about 125 ounces / acre, about 126 ounces / acre, about 127 ounces / acre, or about 128 ounces / acre.

[0223] The present disclosure provides a liquid product of the bio-nutrient formulation can be applied to plants or crops at a rate of about 1 gallon / acre, about 2 gallons / acre, about 3gallons / acre, about 4 gallons / acre, about 5 gallons / acre, about 6 gallons / acre, about 7 gallons / acre, about 8 gallons / acre, about 9 gallons / acre, about 10 gallons / acre, about 11 gallons / acre, about 12 gallons / acre, about 13 gallons / acre, about 14 gallons / acre, about 15 gallons / acre, about 16 gallons / acre, about 17 gallons / acre, about 18 gallons / acre, about 19 gallons / acre, about 20 gallons / acre, about 30 gallons / acre, about 40 gallons / acre, about 50 gallons / acre, about 60 gallons / acre, about 70 gallons / acre, about 80 gallons / acre, about 90 gallons / acre, or about 100 gallons / acre,

[0224] In some embodiments, a liquid product of the bio-nutrient formulation is applied to plants or crops at a rate of about 1 ounce / acre to about 100 gallons / acre, about 1 ounce / acre to about 90 gallons / acre, about 1 ounce / acre to about 80 gallons / acre, about 1 ounce / acre to about 70 gallons / acre, about 1 ounce / acre to about 60 gallons / acre, about 1 ounce / acre to about 50 gallons / acre, about 1 ounce / acre to about 40 gallons / acre, about 1 ounce / acre to about 30 gallons / acre, about 1 ounce / acre to about 20 gallons / acre, or about 1 ounce / acre to about 10 gallons / acre.

[0225] The present disclosure provides a liquid product of the bio-nutrient formulation can be applied to plants or crops at a rate of about 5ml / l, about 10ml / l, about 15ml / l, about 20ml / l, about 25ml / l, about 30ml / l, about 35ml / l, about 40ml / l, about 45ml / l, about 50ml / l, about55ml / l, about 60ml / l, about 65ml / l, about 70ml / l, about 75ml / l, about 80ml / l, about 85ml / l, about 90ml / l, about 95ml / l, about 100ml / l, about 105ml / l, about 110ml / l, about 115ml / l, about 120ml / l, about 125ml / l, about 130ml / l, about 135ml / l, about 140ml / l, about 145ml / l, about150ml / l, about 155ml / l, about 160ml / l, about 165ml / l, about 170ml / l, about 175ml / l, about180ml / l, about 185ml / l, about 190ml / l, about 195ml / l, about 200ml / l, about 205ml / l, about210ml / l, about 215ml / l, about 220ml / l, about 225ml / l, about 230ml / l, about 235ml / l, about240ml / l, about 245ml / l, about 250ml / l, about 255ml / l, about 260ml / l, about 265ml / l, about270ml / l, about 275ml / l, about 280ml / l, about 285ml / l, about 290ml / l, about 295ml / l, about300ml / l, about 305ml / l, about 310ml / l, about 315ml / l, about 320ml / l, about 325ml / l, about330ml / l, about 335ml / l, about 340ml / l, about 345ml / l, about 350ml / l, about 355ml / l, about360ml / l, about 365ml / l, about 370ml / l, about 375ml / l, about 380ml / l, about 385ml / l, about390ml / l, about 395ml / l, about 400ml / l, about 405ml / l, about 410ml / l, about 415ml / l, about420ml / l, about 425ml / l, about 430ml / l, about 435ml / l, about 440ml / l, about 445ml / l, about450ml / l, about 455ml / l, about 460ml / l, about 465ml / l, about 470ml / l, about 475ml / l, about 480ml / l, about 485ml / l, about 490ml / l, about 495ml / l, about 500ml / l, about 550ml / l, about600ml / l, about 650ml / l, about 700ml / l, about 750ml / l, about 800ml / l, about 850ml / l, about 900ml / l, about 950ml / l, or about l,000ml / l.

[0226] In some embodiments, a liquid product of the bio-nutrient formulation is applied to plants or crops at a rate of about 5ml / l to about l,000ml / l, about 10ml / l to about 950ml / l, about 20ml / l to about 900ml / l, about 30ml / l to about 850ml / l, about 40ml / l to about 800ml / l, about 50ml / l to about 750ml / l, about 60ml / l to about 700ml / l, about 70ml / l to about 650ml / l, about 80ml / l to about 600ml / l, about 90ml / l to about 550ml / l, about 100ml / l to about 500ml / l, about 125ml / l to about 450ml / l, or about 150ml / l to about 400ml / l.Combinations of bio-nutrient formulations with other fertilizer products

[0227] The present disclosure further provides combinational use of bio-nutrient formulation as a final product with existing synthetic or organic soil inputs products that include, but not limited to, fertilizer blends, chemical, microbes, compost, and soil amendment products to enhance the effects of partner products.

[0228] In some embodiments, bio-nutrient formulations can be applied as a standalone product to a soil, a turfgrass, a plant, or an area around a grass, a plant, or a crop. In other embodiments, a dry form or a liquid form of bio-nutrient formulations can be applied into an existing soil, a turfgrass, or a plant where traditional organic and / or synthetic fertilizer and chemical products are treated for increasing soil health, positive microbial activities, and / or plant fertility.

[0229] In further embodiments, a final product of the bio-nutrient formulation is plantagnostic, so it can be utilized to any plant, crops or grasses to boost or enhance the underlying microbial functions that manage plant functions including but not limited to drought, salt, heat stress, disease pressure, promoting immune system mechanisms, macro, secondary and micronutrient uptake / regulation, carbon sequestration, metals solubilization, salt tolerance and mineralization / volatilization.

[0230] The present disclosure teaches that microbes affected by the bio-nutrient formulation taught herein are not specific to any given plant (crop or non-crop). That is, the bio-nutrient formulation can be effective on any plant (broadacre crops, specialty crops, trees, turf or ornamental crops, etc). As such, the bio-nutrient formulations can be plant agnostic - i.e. it is not meant for a specific crop or category of crops or plants. In some embodiments, the formulations can be used as universal fertilizer and / or biostimulant applied to any plants, crops, or grasses.

[0231] In other embodiments, the bio-nutrient formulation can be applied as a standalone product, in production agriculture to cover broadacre, or as part of a comprehensive and crop tailored fertilizer program.

[0232] In further embodiments, the bio-nutrient formation can be applied with another agricultural products sequentially or simultaneously, for example, in following combinations: (i) bio-nutrient formation + a traditional fertilizer source; (ii) bio-nutrient formation + chemical / pesticide, or (iii) bio-nutrient formation + a traditional fertilizer + chemical / pesticide, and so on. The agricultural products applied together with the bio-nutrient formulation includes, but are not limited to, a synthetic fertilizer, an organic fertilizer, a pesticide, an herbicide, a fungicide, a surfactant, a micronutrient, a biostimulant, a plurality of microbes (including indigenous or genetically engineered microbes), a microbial additive, compost, and topsoil.

[0233] The bio-nutrient formulation can be applied at intervals or specified windows in various amounts over the course of a growing season (from pre-plant up to or after harvest). It could be applied in a dry granular form, as a homogenous blend, as a heterogeneous blend, as a liquid application, or as a suspension technology / seed coating on other products.

[0234] This product and method may be included in a broader agronomic program with other chemical fertilizers, organic fertilizers, crop protection products, chemical or biological products and innovations.

[0235] In some embodiments, these inclusions could range from organic certified products (typically under a 10-10-10 NPK guaranteed analysis with some or no secondary or micronutrients, soil amendment qualities, and fungi or microbe additions) to synthetic products (varying NPK concentrations like polymer coated urea (such as 45-0-0 NPK), sulfate of potash (such as 0-0-50 NPK), muriate of potash (such as 0-0-60 NPK), super phosphate (such as 0- 42-0 NPK), and may include some or no secondary nutrients, micronutrients, herbicides, chemical products, or other inhibitors).

[0236] In some embodiments, the inclusions may also include crop protection products including but not limited to biostimulants, bionutrients, genetically edited microbes generated by Pivot Bio, Kula Bio, Sound Bio, Andes Bio, etc, or specific / colony / full spectrum microbial additives such as Holganix800, surfactants, adjuvants or other products designed to improve water absorption capacity or manage biotic and abiotic stress responses in plants.

[0237] In further embodiments, a final product of the bio-nutrient formulation is plantspecific, so it is utilized to boost or enhance fertility and reproduction of desired plants, crops, and / or grasses including turfgrasses.Methods of manufacturing bio-nutrient formulations

[0238] The present disclosure provides methods of making bio-nutrient formulations.

[0239] In step a), the biosolids, brewing grains, and dried algae cell wall demised to a powder form utilizing a variety of means of agitation. The powder form of the bio-nutrient composition is produced.

[0240] In step b), the resultant constituent powders are then blended utilizing a variety of blending technologies ranging from a ribbon blender to a rotating chamber. A binder is added at the granulation stage in order to form a durable and spreadable granule or prill of a bionutrient formulation.

[0241] As an alternative of the dry powder and granular forms, the blended constituent elements (biosolids, dried distillers or brewers spent grains, and dried algae cell) are mechanically and chemically treated in order to rehydrate to a liquid derivative and / or a suspension technology for application in various drip, backpack, irrigation, and sprayer applications, as well as coating on various granulated, prill or other soil input delivery technologies. In further embodiments, an enhanced soluble powder of the bio-nutrient composition is created that is condensed in a powder form, however, can be added to water in order to be delivered in a liquid form in various drip, backpack, irrigation and sprayer applications.

[0242] In some embodiments, this bio-nutrient formulation is manufactured through a low emissions process, helping to minimize greenhouse gas emissions including carbon dioxide, methane and nitrogen dioxide. This involves the reuse of organic waste and industrial food and ag waste, and the use of renewable fuel sources at all stages of the manufacturing process.

[0243] In other embodiments, this formulation effects a meaningful and measurable change in activity of soil microbiology, reducing the need for traditional fertilizer as well as improved growth to the point where many weeds are out-crowded. In further embodiments, this formulation improves nutritional status and growth of plants, which make a landscape (such as turf, lawn, or backyard) and a farm, garden or other operation where plants are grown and harvested supporting the production of food or processed food products more resilient to both biotic and abiotic challenges.

[0244] The present disclosure provides methods of making a granular formulation comprising the steps of: (a) dry milling a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell; (b) blending the dry milled bio-nutrient composition from step (a) in a powder form; (c) adding a binder to the blended bio-nutrient composition from step (b) to produce a formulation; (d) optionally, adding an additive; and (e) granulating the formulation from step (c) or (d). In some embodiments of the methods, the additive is added to the formulation from step (d). In other embodiments of the methods, the additive is not added to the formulation from step (d).

[0245] The present disclosure provides methods of making a liquid formulation comprising the steps of: (a) dry milling a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell; (b) agitating the dry milled bio-nutrient composition from step (a) in water with a speed mixer for wet milling; and (c) adding a liquid form of a binder to the wet milled bio-nutrient composition to produce a liquid formulation; (d) optionally adding an additive; and (e) filtering the formulation from step (c) or (d). In some embodiments of the methods, the additive is added to the formulation from step (d). In other embodiments of the methods, the additive is not added to the formulation from step (d).

[0246] In some embodiments of the methods, the bio-nutrient composition comprises: i) a biosolid at a final percentage-by-weight of about 35% to about 75% of the bio-nutrient composition; ii) a grain at a final percentage-by-weight of about 10% to about 50% of the bionutrient composition; and iii) an algae cell at a final percentage-by-weight of about 5% to about 40% of the bio-nutrient composition. In further embodiments of the methods, the binder is at a percentage weight of about 1% to about 50% of said formulation.Methods for improving soil health

[0247] The present disclosure provides methods for improving soil health, comprising: applying a bio-nutrient formulation to soil, thereby improving soil health. In some embodiments of the methods, the bio-nutrient formulation is (1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell. In other embodiments of the methods, the bio-nutrient formulation is (1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell; and (2) a binder. In further embodiments of the methods, the bionutrient formulation is (1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell; (2) a binder; and (3) an additive.

[0248] In some embodiments, the formulation improves a balanced ratio of essential nutrients selected from the group consisting of: nitrogen (N), Phosphorus (P), Potassium (K), Calcium (Ca), Magnesium (Mg), Sulfur (S), Carbon (C), Hydrogen (H), Oxygen (O), and micronutrient. In some embodiments, the micronutrient is selected from the group consisting of: Iron (Fe), Manganese (Mn), Zinc (Zn), Copper (Cu), Boron (B), Molybdenum (Mo), and Chlorine (Cl).

[0249] In some embodiments, the formulation increases activity and / or diversity of a plurality of beneficial microbes in soil. In some embodiments, the beneficial microbe is Pseudomonas sp, Achromobacter sp., Bacillus sp., Nitrobacter sp., Nitrosomonas sp., Nitrospira sp., Paenibacillus sp., Rhizobium sp., Streptomyces sp., Trichoderma sp., Alkalihalobacillus hwajinpoensis, Aureobasidium pullulans, Metarhizium anisopliae, Purpureocillium lilacinum, or Pythium oligandrum. In further embodiments, the beneficial microbe is an indigenous microbe species or a genetically engineered microbe. In some embodiments, the beneficial microbe enhances production of exopolysaccharides, salicylic acid, siderophores, or 1- aminocyclopropane-1 -carboxylate (ACC) deaminase, thereby improving soil health and promoting plant growth and stress tolerance.

[0250] In some embodiments, the formulation decreases activity and / or diversity of a plurality of non-beneficial microbes in soil. In some embodiments, the non-beneficial microbe is a soil pathogen selected from the group consisting of: Pythium sp., Colletotrichum sp., Fusarium sp., Curvularia sp., Sclerotinia sp., Sclerotinia homoeocarpa, Agrobacterium tumefaciens, Sclerotium rolfsii, or Leptosphaerulina americana.Methods for altering inicrobiome in soil

[0251] The present disclosure provides methods for altering microbiome in soil, comprising: applying a bio-nutrient formulation to soil, thereby altering microbiome in soil. In some embodiments of the methods, the bio-nutrient formulation is (1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell. In other embodiments of the methods, the bio-nutrient formulation is (1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell; and (2) a binder. In further embodiments of the methods, the bionutrient formulation is (1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell; (2) a binder; and (3) an additive.

[0252] In some embodiments, the formulation increases a plurality of beneficial microbes in soil. In some embodiments, the beneficial microbe is Pseudomonas sp, Achromobacter sp., Bacillus sp., Nitrobacter sp., Nitrosomonas sp., Nitrospira sp., Paenibacillus sp., Rhizobiumsp., Streptomyces sp., Trichoderma sp., Alkalihalobacillus hwajinpoensis, Aureobasidium pullulans, Metarhizium anisopliae, Purpureocillium lilacinum, or Pythium oligandrum. In some embodiments, the beneficial microbe is an indigenous microbe species or a genetically engineered microbe. In some embodiments, the beneficial microbe enhances production of exopolysaccharides, salicylic acid, siderophores, or 1 -aminocyclopropane- 1 -carboxylate (ACC) deaminase, thereby improving soil health and promoting plant growth and stress tolerance.

[0253] In some embodiments, the formulation decreases a plurality of non-beneficial microbes in soil. In some embodiments, the non-beneficial microbe is a soil pathogen selected from the group consisting of Pythium sp., Colletotrichum sp., Fusarium sp., Curvularia sp., or Sclerotinia sp., Sclerotinia homoeocarpa, Agrobacterium tumefaciens, Sclerotium r of sii, or Leptosphaerulina americana.Methods for inducing a stress-related defense mechanism in plants

[0254] The present disclosure provides methods for inducing a stress-related defense mechanism in a plant, comprising: applying a bio-nutrient formulation to a target, thereby improving plant defense response to biotic or abiotic stress. In some embodiments of the methods, the bio-nutrient formulation is (1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell. In other embodiments of the methods, the bio-nutrient formulation is (1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell; and (2) a binder. In further embodiments of the methods, the bio-nutrient formulation is (1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell; (2) a binder; and (3) an additive.

[0255] In some embodiments, the target is a plant, a plant part, or a plant cell thereof. In some embodiments, the plant is acacia, alfalfa, amaranth, apple, apricot, artichoke, ash tree, asparagus, avocado, banana, barley, beans, beet, birch, beech, blackberry, black raspberry, blueberry, broccoli, Brussel's sprouts, cabbage, cane berry, canola, cantaloupe, carrot, cassava, cauliflower, cedar, a cereal, celery, chestnut, cherry, Chinese cabbage, citrus, Clementine, clover, coffee, com, cotton, cowpea, cucumber, cypress, eggplant, elm, endive, eucalyptus, fennel, figs, fir, geranium, grape, grapefruit, groundnuts, ground cherry, gum hemlock, hickory, kale, kiwifruit, kohlrabi, larch, lettuce, leek, lemon, lime, locust, pine, maidenhair, maize, mango, maple, melon, millet, mushroom, mustard, nuts, oak, oats, oil palm, okra, onion, orange, an ornamental plant or flower or tree, papaya, palm, parsley, parsnip, pea, peach,peanut, pear, peat, pepper, persimmon, pigeon pea, peach, pine, pineapple, plantain, plum, pomegranate, potato, pumpkin, radicchio, radish, rapeseed, raspberry, rice, rye, sorghum, safflower, sallow, soybean, spinach, spruce, squash, strawberry, sugar beet, sugarcane, sunflower, sweet potato, sweet corn, tangerine, tea, tobacco, tomato, trees, triticale, turf grasses, turnips, vine, walnut, watercress, watermelon, wheat, wild strawberry, yams, yew, or zucchini.

[0256] In some embodiments, the plant is a grass. In some embodiments, the grass is a Poaceae family. In some embodiments, the grass is Agrostis genus, Poa genus, Lolium genus, Festuca genus, Cynodon genus, Stenotaphrum genus, or Zoysia genus. In some embodiments, the grass is a turfgrass, which is bentgrass, Kentucky bluegrass, ryegrass, tall fescue, Chewings fescue, Hard fescue, sheep fescue, creeping red fescue, bermudagrass, kikuyu grass, St. Augustine grass), buffalo grass, zoysia grass, zoyosa tenuifolia, or temple grass. In some embodiments, the target is a soil or an area where a plant is present.

[0257] In some embodiments, the induced stress-related defense mechanism is selected from the group consisting of: phenylpropanoid biosynthesis, glutathione metabolism, sesquiterpenoid and triterpenoid metabolism, plant-pathogen interaction, ascorbate and aldarate metabolism, carotenoid biosynthesis, plant hormone signal transduction, and linoleic acid metabolism. In some embodiments, the induced stress-related defense mechanism is associated with: (i) upregulation of the expression of a plurality of gene involved in phenylpropanoid biosynthesis, which is phenylalanine ammonia lyase, trans-cinnamate 4- monooxygenase, and / or caffeoylshikimate esterase, (ii) upregulation of the expression of a plurality of gene involved in glutathione metabolism, which is glutathione dehydroascorbate reductase 3, glutathione peroxidase, and / or gamma-glutamylcyclotransferase,' (iii) upregulation of the expression of a plurality of gene involved in sesquiterpenoid and triterpenoid metabolism, which is squalene synthase 1, squalene monooxygenase, and / or NAD+ dependent farnesal dehydrogenase, ' (iv) upregulation of the expression of a plurality of gene involved in plant-pathogen interaction, which is pathogenesis related 1, suppressor of G2 allele of SKP1, and / or respiratory burst oxidase,' (v) upregulation of the expression of a plurality of gene involved in ascorbate and aldarate metabolism, which is L-gulonolactone oxidase, GDP-L-galactose phosphorylase, and / or ascorbate peroxidase,' (vi) upregulation of the expression of a plurality of gene involved in carotenoid biosynthesis, which is ABA 8'- hydroxylase, lycopene beta-cyclase, and / or 9-cis-epoxycarotenoid dioxygenase,' (vii) upregulation of the expression of a plurality of gene involved in plant hormone signaltransduction, which is Not Responsive to PR1 protein (NPR1), jasmonic acid receptor, and / or abscisic acid receptor,' and / or (viii) upregulation of the expression of a plurality of gene involved in linoleic acid metabolism, which is linoleate 9S-lipoxygenase and / or linoleate 13S- Upoxygenase9.Methods for inducing a photosynthesis mechanism in plants

[0258] The present disclosure provides methods for inducing a photosynthesis mechanism in a plant, comprising: applying a bio-nutrient formulation to a target, thereby improving plant growth and development. In some embodiments of the methods, the bio-nutrient formulation is (1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell. In other embodiments of the methods, the bio-nutrient formulation is (1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell; and (2) a binder. In further embodiments of the methods, the bio-nutrient formulation is (1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell; (2) a binder; and (3) an additive.

[0259] In some embodiments, the target is a plant, a plant part, or a plant cell thereof. In some embodiments, the plant is acacia, alfalfa, amaranth, apple, apricot, artichoke, ash tree, asparagus, avocado, banana, barley, beans, beet, birch, beech, blackberry, black raspberry, blueberry, broccoli, Brussel's sprouts, cabbage, cane berry, canola, cantaloupe, carrot, cassava, cauliflower, cedar, a cereal, celery, chestnut, cherry, Chinese cabbage, citrus, Clementine, clover, coffee, com, cotton, cowpea, cucumber, cypress, eggplant, elm, endive, eucalyptus, fennel, figs, fir, geranium, grape, grapefruit, groundnuts, ground cherry, gum hemlock, hickory, kale, kiwifruit, kohlrabi, larch, lettuce, leek, lemon, lime, locust, pine, maidenhair, maize, mango, maple, melon, millet, mushroom, mustard, nuts, oak, oats, oil palm, okra, onion, orange, an ornamental plant or flower or tree, papaya, palm, parsley, parsnip, pea, peach, peanut, pear, peat, pepper, persimmon, pigeon pea, peach, pine, pineapple, plantain, plum, pomegranate, potato, pumpkin, radicchio, radish, rapeseed, raspberry, rice, rye, sorghum, safflower, sallow, soybean, spinach, spruce, squash, strawberry, sugar beet, sugarcane, sunflower, sweet potato, sweet corn, tangerine, tea, tobacco, tomato, trees, triticale, turf grasses, turnips, vine, walnut, watercress, watermelon, wheat, wild strawberry, yams, yew, or zucchini.

[0260] In some embodiments, the plant is a grass. In some embodiments, the grass is a Poaceae family. In some embodiments, the grass is Agrostis genus, Poa genus, Lolium genus,Festuca genus, Cynodon genus, Stenotaphrum genus, or Zoysia genus. In some embodiments, the grass is a turfgrass, which is bentgrass, Kentucky bluegrass, ryegrass, tall fescue, Chewings fescue, Hard fescue, sheep fescue, creeping red fescue, bermudagrass, kikuyu grass, St. Augustine grass, buffalo grass, zoysia grass, zoyosa tenuifolia, or temple grass. In some embodiments, the target is a soil or an area where a plant is present.

[0261] In some embodiments, the induced photosynthesis mechanism is associated with upregulation of the expression of a plurality of gene, which is photosystem II subunit R, photosystem I subunit IV, plastocyanin, light-harvesting complex I chlorophyll a / b binding protein 2, and / or light-harvesting complex II chlorophyll a / b binding protein 4.Methods for inducing nutrient uptake and assimilation in plants

[0262] The present disclosure provides methods for inducing nutrient uptake and assimilation in a plant, comprising: applying a bio-nutrient formulation to a target, thereby improving nitrogen assimilation or uptake of a plant. In some embodiments nutrient uptake and assimilation of the methods, the bio-nutrient formulation is (1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell. In other embodiments of the methods, the bio-nutrient formulation is (1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell; and (2) a binder. In further embodiments of the methods, the bionutrient formulation is (1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell; (2) a binder; and (3) an additive.

[0263] In some embodiments, the target is a plant, a plant part, or a plant cell thereof. In some embodiments, the plant is acacia, alfalfa, amaranth, apple, apricot, artichoke, ash tree, asparagus, avocado, banana, barley, beans, beet, birch, beech, blackberry, black raspberry, blueberry, broccoli, Brussel's sprouts, cabbage, cane berry, canola, cantaloupe, carrot, cassava, cauliflower, cedar, a cereal, celery, chestnut, cherry, Chinese cabbage, citrus, Clementine, clover, coffee, com, cotton, cowpea, cucumber, cypress, eggplant, elm, endive, eucalyptus, fennel, figs, fir, geranium, grape, grapefruit, groundnuts, ground cherry, gum hemlock, hickory, kale, kiwifruit, kohlrabi, larch, lettuce, leek, lemon, lime, locust, pine, maidenhair, maize, mango, maple, melon, millet, mushroom, mustard, nuts, oak, oats, oil palm, okra, onion, orange, an ornamental plant or flower or tree, papaya, palm, parsley, parsnip, pea, peach, peanut, pear, peat, pepper, persimmon, pigeon pea, peach, pine, pineapple, plantain, plum, pomegranate, potato, pumpkin, radicchio, radish, rapeseed, raspberry, rice, rye, sorghum, safflower, sallow, soybean, spinach, spruce, squash, strawberry, sugar beet, sugarcane,sunflower, sweet potato, sweet corn, tangerine, tea, tobacco, tomato, trees, triticale, turf grasses, turnips, vine, walnut, watercress, watermelon, wheat, wild strawberry, yams, yew, or zucchini.

[0264] In some embodiments, the plant is a grass. In some embodiments, the grass is a Poaceae family. In some embodiments, the grass is Agrostis genus, Poa genus, Lolium genus, Festuca genus, Cynodon genus, Stenotaphrum genus, or Zoysia genus. In some embodiments, the grass is a turfgrass, which is bentgrass, Kentucky bluegrass, ryegrass, tall fescue, Chewings fescue, Hard fescue, sheep fescue, creeping red fescue, bermudagrass, kikuyu grass, St. Augustine grass, buffalo grass, zoysia grass, zoyosa tenuifolia, or temple grass. In some embodiments, the target is a soil or an area where a plant is present.

[0265] In some embodiments, the induced nutrient uptake and assimilation is associated with upregulation of the expression of a plurality of gene, which is nitrate reductase, nitrite reductase, high-affinity nitrate transporter 2.1 (HAT2.1), high-affinity nitrate transporter 2.2 (HAT2.2), high-affinity nitrate transporter 2.3 (HAT2.3), glutamine synthetase, and / or mugineic acid-3 dioxygenase.

[0266] The disclosure has been described herein using specific embodiments for the purposes of illustration only. It will be readily apparent to one of ordinary skill in the art, however, that the principles of the disclosure can be embodied in other ways. Therefore, the disclosure should not be regarded as being limited in scope to the specific embodiments disclosed herein, but instead as being fully commensurate in scope with the following claims.EXAMPLES

[0267] The following examples are provided to illustrate further the various applications and are not intended to limit the disclosure beyond the limitations set forth in the appended claims.Example 1: Field Test Study of Granular Bio-Nutrient Formulation Products

[0268] A product used in the Examples is a dry granular bio-nutrient formulation of the present disclosure comprising a bio-nutrient composition and a binder (i.e., lignin).

[0269] In the first field test on turfgrass in Fresno, CA, the bio-nutrient formulation was compared to a slow-release synthetic polymer coated urea fertilizer (42-0-0; N-P-K) as the control as presented in FIG. 4. In the second field test on cotton in Auburn, AL, the bio-nutrient formulation was compared to the slow-release synthetic fertilizer as the control as presented in FIG. 4. Samples were collected before (TO) and after treatment applications (T1 = 14 daysafter; T2 = 30 days after; T3 = 60 days after; T4 = 90days after), with a total of 12 soil samples over one block. Soil samples were collected in triplicate, in order to ensure accuracy and account for the natural variability of the microbial communities across samples. Bulk soils were collected in sterile tubes according to BeCrop® Instructions, at a depth of 2-6 inches. Samples were collected as a composite of several cores in each block, in order to ensure representativity of the area.

[0270] The effect of the bio-nutrient formulation products on the soil microbial consortium was evaluated using Biome Makers’ platform and methodology. This approach includes a comparison between the status of the soil before and after an addition of the bio-nutrient formulation and then measures the developing analysis of the microbiome of treated and an untreated (control) blocks. The characterization of the soil microbial community was analyzed by amplicon-based metagenomics to define prokaryotes (16S amplicon) and fungi (ITS amplicon) populations. This comparison allowed us to avoid possible environmental changes in the microbial community and more specifically identify the changes caused by the product application.

[0271] After the granular bio-nutrient formulation products were treated to turfgrass field, significant impacts on turf, soil, and microbes were observed on 30, 60, and 90 days when compared to the synthetic fertilizer. As shown in shown in FIG. 5, turfgrass root weight and density were increased about 5% to 11% after treatments over the course of 90 days. This indicates that the bio-nutrient formulation products positively affect root size, which can boost plant growth and development.

[0272] The potential for nutrient cycling of bacterial communities associated with all samples across the one block were analysed using the abundance of each bacterium and the known metabolic enzyme compositions of their genomes. The results of major nutrients evolution were detected in carbon pathways associated with carbon fixation and methanogenesis (about 19% increase), macronutrient evolution (about 16% increase of calcium transport), micronutrient evolution (about 16% increase of zinc transport equilibrium; about 38% increase of manganese transport equilibrium), which improve soil structure, condition and health, as shown in FIG. 6.

[0273] The presence of beneficial organisms from both bacterial and fungal communities were analysed to identify those associated to the production of stress sensing and tolerance mechanisms in all samples across the three blocks. As presented in FIG. 6, increase in stress adaptation pathways are observed, such as exopolysaccharide (EPS) production (about 60% to160% increase), ACC deaminase (about 66% to 180% increase), heavy metal solubilization (about 70% to 144% increase), salicylic acid (SA) (about 198% to 380% increase), salt tolerance(about 12% to 15% increase), and siderophore production(up to about 390% increase). These adaptation pathways can be involved in alleviating water stress.

[0274] The presence of pathogens and biocontrol agents from both the bacterial and fungal communities were analyzed in all samples across the three blocks. Major changes to reduce disease pressure are observed in the bio-nutrient formulation treatments. For example, microbe populations that cause diseases such as Pythium blight (about 91% to 99% decrease), Pythium root rot (about 85% to 99% decrease), Anthracnose (about 47% to 98% decrease), Curvularia blight (about 55% to 99% decrease), leptosphaerulina leaf blight (about 55% to 85% decrease), dollar spot, Fusarium blight, bipolaris, Exserohilum leaf spot, and crown and root rot are significantly reduced with the bio-nutrient formulation applied, as presented in FIG. 6.

[0275] From the first turfgrass field test, the bio-nutrient formulation treatment of the present disclosure significantly increases Pseudomonas sp. Population (about 11,000% increase) along with increase in microbial phytohormones, auxin production (about 5% increase) and cytokinin production (about 12% increase), as shown in FIG. 6.

[0276] While no significant differences in nutrient pathways between the bio-nutrient formulation treatment and polymer coated urea treatments except for Zn mobilization at 90 days, the bio-nutrient formulation treatments significantly increase stress response pathways typically associated with tolerance or resistance to water stress. Also, the bio-nutrient formulation treatment significantly reduces disease pressure for Pythium, leaf blight, Fusarium, Curvularia, and Anthracnose .

[0277] FIG. 6 summarizes the test results on turfgrass field treated with the bio-nutrient formulation in comparison to the synthetic fertilizer, including about 11% increase in root size, about 38% increase in soil improvement, about 390% increase in stress tolerance (less water, tolerating weather extremes, reducing salts and minerals), about 45% to about 99% reduction in diseases (eliminating diseases before becoming diseases, more effective than actual herbicides and fungicides), and about 12% increase in plant hormones (building up the plant’s immune system and natural antibiotics) as well as about 11,000% increase for Pseudomonas production.

[0278] The granular bio-nutrient formulation treatment significantly increases Pseudomonas population in soil and stress pathways associated with alleviating water stress, as well as reduces disease pressure such as Pythium, leaf blight, Fusarium, Curvularia, and Anthracnose .

[0279] The second field tests were conducted using the granular bio-nutrient formulation product in cotton (Auburn, AL). The results were strikingly similar to the first field test in turfgrass. The turf trial was sampled at 30, 60 and 90 days after treatment and demonstrated a "major change" increase in microbes that contribute to stress tolerance via the production of exopolysaccharides, ACC deaminase, salicylic acid and siderophore production. In addition, levels of key soil pathogens were reduced (Pythium sp., Colletotrichum sp., Fusarium sp., and Sclerotinia sp.).

[0280] The cotton trial was sampled at approximately 30 days after treatment. This trial also demonstrated a "major increase" in microbes that produce ACC Deaminase (for ethylene production), salicylic acid, siderophore production and microbes that generally contribute to stress tolerance. The cotton trial results showed a consistent increase in hormones that contribute to stress tolerance (cytokinin and auxins). This trial also showed a decrease in similar key pathogens, Sclerotinia sp., Fusarium sp. and Pythium sp. The increase in microbes (including Pseudomonas sp.) that produce auxins would contribute to a more robust root system which would improve the ability of the crop to uptake nutrients and water, as well as enhance drought tolerance.

[0281] The same trial cotton trial referenced above conducted in Auburn, AL took leaf sap samples at approximately 68 days after treatment. There was an improvement in levels of the N, P and K in the sap of young leaves in the bio-nutrient formulation treated compared to the standard which was treated with microalgae. In addition, there were increases in sap levels of Mg, Zn and Cu in the sap of young leaves compared to the standard.

[0282] Taken together, these first and second trials indicate the bio-nutrient formulation product of the present disclosure has a significant impact on the soil microbiome (as presented in Table 1) of increasing levels of microbes that help crops tolerate abiotic stress as well as reducing levels of key fungal pathogens.

[0283] Table 1. Top candidate microbes effecting biological responses

[0284] Additional tests were further conducted at sites by comparing the bio-nutrient formulation product with Hydretain®, NatureSafe® (organic fertilizer), a surfactant as a control, and compost. The most meaningful results are the impact on bacteria colonies, fungal colonies, and micronutrients (Sulfur, Chloride, Magnesium, and Calcium in particular. Zinc had meaningful results in another test).

[0285] Another third field test set from these properties indicate a consistent reduction in salt levels up to 38% in 90 days after one treatment.

[0286] This dry bio-nutrient formulation product has demonstrated in field tests to influence, signal, and proliferate microbes associated with drought, salt, disease pressure, heat, macro, secondary and micronutrient nutrient mobilization and availability into the plant, plant immune responses, heavy metals solubilization, and the like.Example 2: Plant / Crop Study of Liquid Bio-Nutrient Formulation Products

[0287] A product used in the Examples is a liquid bio-nutrient formulation of the present disclosure comprising a bio-nutrient composition and a binder (i.e., lignin).

[0288] Example 1 suggested indirect effects on plant defense response by promoting the colonization of salicylic acid-producing microbes and ethylene precursor-producing microbes (e.g., Pseudomonas sp. and Bacillus sp.) in the soils of treated plants. The objectives of this Example were to investigate the effects and identify candidate modes of action of a liquid bionutrient formulation production on treated com plants through transcriptomic analysis. This type of analysis is used to identify the effects of treatments or challenges on the relative gene expression levels of protein-coding genes in a species in comparison to untreated controls. Another objective was to validate findings from Example 1 to identify and characterize defense response elicited in corn plants following foliar treatment with the liquid bio-nutrient formulation product.

[0289] Corn plants (n=4) were grown in pots in a controlled greenhouse environment and foliar sprayed to complete coverage with two rates of liquid bio-nutrient formulation product when the plants reached the V4 growth stage. The “high rate” (H) plants received a 35% dilution of liquid bio-nutrient formulation product in Deionized (DI) water, the “low rate” (LR” plants received a 17.5% dilution of liquid bio-nutrient formulation product in DI water, and the Untreated Control (UTC) plants received a treatment of only DI water. The uppermost leaves of treated plants were sampled for RNA sequencing at 24 hours and 72 hours following the treatment. 14 inch leaf discs were collected. Total RNA was extracted, and libraries for sequencing were prepared with Poly-A identification for mRNA selection. 150 bp, paired-end sequencing (20 million reads depth) was performed using Novoseq Plus™ platform. Quantitative bioinformatics analysis was performed to identify differentially expressed genes and enriched pathways among treatment groups.

[0290] There are significant changes in the effects over time, with distinct differences between LR and HR, and UTC at 24 hours and all three groups at 72 hours. Time affected UTC samples as well, though UTC samples at both time points are distinctly different from both treatment groups at both respective time points.

[0291] Both groups elicit similar abiotic and biotic stress responses, including jasmonic acid, ABA, salicylic acid, ROS production, phenylpropanoid, and antioxidant responses. These stress responses last longer in the HR group than the LR group, and the HR more strongly induces stress responses.

[0292] Both HR and LR groups upregulate nitrogen metabolism (by upregulating expression of nitrate reductase, nitrite reductase, and / or high-affinity nitrate transporter 2.2 (HAT2.2) genes) and the effects continue through 72 hours. The HR induces this response more strongly and continues to express more strongly at 72 hours versus the LR.

[0293] Both groups upregulate Mugineic acid metabolism (a key component of Fe and Zn uptake, assimilation, and exudates). At 24 hours, parts of this pathway are more highly expressed in the LR group, but the effects last longer in the HR. Nitrogen and Mugineic acid related expression is still strongly upregulated at 72 hours.

[0294] HR group shows more robust and stronger reactions - such as reactive oxygen species (ROS) signalling, ABA biosynthesis and signalling, and biosynthesis and signalling of jasmonic acid and salicylic acid.

[0295] Some downstream growth hormone related signalling - auxin, cytokinin, and Gibberellin related gene expression at 24 hours. Generally, applications at the higher rate resulted in longer lasting expression of noted genes (72 hours after treatment).

[0296] At 72 hours, the HR group is still in “stress” defense mode, while the LR group is quicker to recover and has increased photosynthesis and growth-related gene expression versus the UTC and the HR.

[0297] KEGG (Kyoto Encyclopedia of Genes and Genomes) Enrichment analysis showed statistically significant enrichment of(1) several key stress and defense-related pathways (Phenylpropanoid Biosynthesis (zma00940), Glutathione Metabolism (zma00480), Sesquiterpenoid and Triterpenoid metabolism (zma00909), Plant-pathogen Interaction (zma04626), Ascorbate and Aldarate metabolism (zma00053), Carotenoid Biosynthesis (zma00906), Plant Hormone Signal Transduction (zma04075), Linoleic acid metabolism (zma00591)), as presented in FIG. 8;(2) growth and development related pathways (Photosynthesis (zma00195), Photosynthesis - antennae proteins (zma00196)) , as presented in FIG. 9; and(3) nutrient metabolism-related pathways (Nitrogen Metabolism (zma00910), Biosynthesis of various plant secondary metabolites (zma00999)), as presented in FIG. 10.

[0298] These findings suggest that both rates of liquid bio-nutrient formulation product elicit multiple stresses and defense response, promote growth through increased expression of photosynthesis-related genes, and promote improved nutrient uptake and assimilation through increased expression of nitrogen and metal ion uptake and assimilation genes. These effects could improve plant resistance and recovery to abiotic and biotic stresses, increase plant growththrough improved photosynthetic efficiency, and increase plant absorption of available nutrients in the soil for increased nutrient uptake and use efficiencies. Further research should investigate the longer-term effects of these responses, as well as the variability of responses that may occur in different crop species or when applied at different crop growth stages.

[0299] Table 2. Genes related to KEGG pathway identified in FIGs. 8-10Example 3: Plant Field Study of Bio-Nutrient Formulation Product

[0300] A field study using the bio-nutrient formulation production of Example 2 was conducted on lettuce that has the irrigation water amended with high salt levels in Somis, CA. Salty soils reduce soil microbial activity.

[0301] There were two controls; (i) untreated control - No salt added and (ii) salt (NaCl) added control. Then, (iii) Salt (NaCl) + Low rate of the bio-nutrient formulation treated lettuce grown in salty conditions by mixing 175 ml formulation in 1 liter of deionized water and applied as a drench at transplanting and again 2 weeks later, (iv) Salt (NaCl) + High rate of bio-nutrient formulation treated lettuce grown in salty conditions by mixing 350 ml bio-nutrient formulation in 1 liter of deionized water and applied as a drench at transplanting and again 2 weeks later.

[0302] The bio-nutrient formulation treatments were applied as a drench at transplanting (2 weeks prior to salt initiation) and again foliar 2 weeks post-transplant. Treatments were made assuming 30 GPA mix size. Salt (NaCl) was added to treatments 2-4 beginning through the drip irrigation 2 weeks post-transplant. Trial limited irrigation to create water stress that should exacerbate salt toxicity symptoms.

[0303] The addition of salt successfully created differences between salted treatments and the unsalted control. The two bio-nutrient formulation treatments resulted in the highest levels of salt in the soil. This indicates that the bio-nutrient formulation treated plants took up fewer salts than did the salted control plants. The bio-nutrient formulation product enhanced the plant’s ability to “exclude” salts.

[0304] Both bio-nutrient formulation treatments had less sodium in the sap compared to the salted control with the low rate having less sodium than both the salted and unsalted controls. This indicates that sodium exclusion may be a part of the mode of action with a low rate better than high rate for this function.

[0305] Salting the soil resulted in less nitrate compared to the unsalted control. The bionutrient formulation treatments resulted in lower nitrate levels in the soil compared to both the unsalted and salted controls.

[0306] Salty soils reduce nitrate levels in the soil, but treatment with bio-nutrient formulation at either rate resulted in higher levels of nitrate in the leaf sap. This could be related to the upregulation of nitrogen transporter genes (HAT2.3 and HAT2.1).

[0307] Treatment with bio-nutrient formulation at the low rate greatly increased levels of ammonium in the leaf sap. This would be due to the upregulation of Ferredoxin-ni trite reductase gene which reduces nitrite to ammonia in the chloroplast.

[0308] Treatment with bio-nutrient formulation at the high rate increased Fe levels in the leaf sap. This may be due to the upregulation of mugineic acid-3 dioxygenase which is involved in final steps of the pathway to convert methionine to mugineic acid. This is a key component of Fe and Zn uptake, assimilation, and exudates. The formulation increases Fe and / or Zn update and assimilation in plants.

[0309] This trial was conducted to see if upregulation of genes that were discovered in RNAseq analysis resulted in salinity tolerance or improved growth in live plants exposed to salty soil. The results show the improved salt tolerance as well as improved nitrogen, iron or zinc uptake or transport, despite the salt stress, when the liquid bio-nutrient formulation product was applied to lettuce plants and soil.

[0310] Table 3. Genes related to desirable traits identified in Example 3Example 4: Combinational Use of Bio-Nutrient Formulation Product

[0311] As described in Examples 1 and 2, significant changes were observed in soil microbes treated with dry and / or liquid bio-nutrient formulation products of the present disclosure. Based on these results, these microbes are not specific to any given plant (crop or non-crop). Thus, the bio-nutrient formulation may be effective on any plants, crops, or grasses.

[0312] The bio-nutrient formulation can be not only applied as a standalone product, but also applied in any combination with following products such as traditional fertilizers, bulk minerals and nutrients, pesticides, herbicides, organic fertilizers, crop protection products (such asincluding but not limited to biostimulants and bionutrients), genetically edited microbes, specific / colony / full spectrum microbial additives (such as Holganix800), surfactants, adjuvants or other products designed to improve water absorption capacity, or manage biotic and abiotic stress responses in plants.INCORPORATION BY REFERENCE

[0313] All references, articles, publications, patents, patent publications, and patent applications cited herein are incorporated by reference in their entireties for all purposes. However, mention of any reference, article, publication, patent, patent publication, and patent application cited herein is not, and should not be taken as, an acknowledgment or any form of suggestion that they constitute valid prior art or form part of the common general knowledge in any country in the world.REFERENCESUS10,351,483R.L. Chapman, Algae: The world’s most important “plants”-an introductionMitig Adapt Strateg. Glob. Chang, 18 (2013), pp. 5-12I. Duarte, S. Hernandez, A. Ibanez, A. Canto, Macroalgae as Soil Conditioners or Growth Promoters of Pisum sativum (L), Annu. Res. Rev. Biol., 27 (2018), pp. 1-8N. Abdel-Raouf, A. Al-Homaidan, I. Ibraheem, Agricultural importance of algae, African J. Biotechnol., 11 (2016), pp. 11648-11658James O'Connor, Son A. Hoang, Lauren Bradney, Jorg Rinklebe, M.B. Kirkham, Nanthi S. Bolan, Value of dehydrated food waste fertiliser products in increasing soil health and crop productivity, Environmental Research, Volume 204, Part A, 2022, 111927Liu Q, Luo L, Zheng L. Lignins: Biosynthesis and Biological Functions in Plants. Int J Mol Sci. 2018 Jan 24; 19(2):335.Moura JC, Bonine CA, de Oliveira Fernandes Viana J, Domelas MC, Mazzafera P. Abiotic and biotic stresses and changes in the lignin content and composition in plants. J Integr Plant Biol. 2010 Apr;52(4):360-76. Duncan, E., Deng, Y., Bao, J., Yuan, F., Liang, X., & Feng, Z. (2018). Exogenous hydrogen sulfide alleviates salt stress in wheat seedlings by decreasing Na+ content. Plant Growth Regulation, 79, 391-399Elsawy, H.I.A.; Alharbi, K.; Mohamed, A.M.M.; Ueda, A.; AlKahtani, M.; AlHusnain, L.;Attia, K.A.; Abdelaal, K.; Shahein, A.M.E.A. Calcium Lignosulfonate Can Mitigate theImpact of Salt Stress on Growth, Physiological, and Yield Characteristics of Two Barley Cultivars (Hordeum vulgare L.). Agriculture 2022, 12, 1459.NUMBERED EMBODIMENTS OF THE DISCLOSURE

[0314] Subject matter contemplated by the present disclosure is set out in the following numbered embodiments:1. A formulation comprising:(a) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell;(b) a binder; and(c) optionally an additive.2. The formulation of embodiment 1, wherein the additive is present.3. The formulation of embodiment 1, wherein the additive is not present.4. The formulation of embodiment 1, wherein the bio-nutrient composition comprises: i) a biosolid at a final percentage-by-weight of about 35% to about 75% of the bio-nutrient composition; ii) a grain at a final percentage-by-weight of about 10% to about 50% of the bio-nutrient composition; and iii) an algae cell at a final percentage-by-weight of about 5% to about 40% of the bionutrient composition.5. The formulation of embodiment 1 or 4, wherein the biosolid is a sewage sludge, a manure or a food waste.6. The formulation of any one of embodiments 1 and 4-5, wherein the biosolid is a Class A EQ biosolid, Class A biosolid, or a Class B biosolid.7. The formulation of embodiment 1 or 4, wherein the grain is a brewers’ spent grain (BSG) or a distillers grain (DG).8. The formulation of any one of embodiments 1, 4, and 7, wherein the grain is a dried brewers’ spent grain (DBSG) or a dried distillers grain (DDG).9. The formulation of embodiment 1 or 4, wherein the algae cell is CyanophyceaexChlor ophyceae. Rhodophyceae , or Phaeophyceae .10. The formulation of any one of embodiments 1, 4, and 9, wherein the algae cell comprises a dried algae cell wall, algae process water, or a post-harvest post filtration undried algae solution.11. The formulation of embodiment 1, wherein the binder is at a percentage weight of about 1% to about 50% of said formulation.The formulation of embodiment 1 or 11, wherein the binder is a lignin, a mineral, a plant starch, or a molasses. The formulation of any one of embodiments 1 and 11-12, wherein the binder is a lignin or a lignin derivative. The formulation of embodiment 13, wherein the lignin derivative is lignosulfonate. The formulation of any one of embodiments 1-3, wherein the additive is an odor-reducing or odor-neutralizing agent. The formulation of any one of embodiments 1-3 and 15, wherein the additive is selected from the group consisting of: an order neutralizer, an odor-masking agent, an essential oil, an odor absorbent, an enzyme, or a microbe. The formulation of embodiment 16, wherein the odor neutralizer is an activated charcoal, a zeolite, a baking soda, a silica gel, or a compound comprising cyclodextrin. The formulation of embodiment 16, wherein the odor-masking agent is a vanilla extract or a citrus extract. The formulation of embodiment 16, wherein the essential oil is a lavender oil, a eucalyptus oil, a peppermint oil, a lemon oil, a tea tree oil, a clove oil, an orange oil, or a cinnamon oil. The formulation of embodiment 16, wherein the odor absorbent is bentonite, kaolin clay, or peat moss. The formulation of embodiment 16, wherein the enzyme is a protease, a lipase, an amylase, a cellulase, a urease, an oxidoreductase, or a pectinase. The formulation of embodiment 16, wherein the microbe is a species of genus Bacillus or Pseudomonas. The formulation of any one of embodiments 1-22, wherein the formulation is in a powder form. The formulation of any one of embodiments 1-22, wherein the formulation is in a granular form. The formulation of any one of embodiments 1-22, wherein the formulation is in a liquid form. The formulation of embodiment 24, wherein the granular form is a prill form. The formulation of embodiment 25, wherein the liquid form of said formation is a suspension or an extract.The formulation of embodiment 25, wherein the liquid form of the formulation further comprises an enhancer. The formulation of embodiment 28, wherein the enhancer is selected from the consisting of: a preservative, a stabilizer, a buffering agent, a microbial inhibitor, a pH adjuster, and an anti -foaming agent. The formulation of embodiment 29, wherein the preservative is sodium benzoate, potassium sorbate, or citric acid. The formulation of embodiment 29, wherein the stabilizer is a xanthan gum, a chelating agent or an antioxidant. The formulation of embodiment 31, wherein the chelating agent is ethylenediaminetetraacetic acid (EDTA). The formulation of embodiment 31, wherein the antioxidant is an ascorbic acid or a tocopherol. The formulation of embodiment 29, wherein the buffering agent is phosphate, phosphate derivative, acetate, or acetate derivative. The formulation of embodiment 29, wherein the microbial inhibitor is an essential oil or a silver nanoparticle. The formulation of embodiment 35, wherein the essential oil is a thyme oil, an oregano oil, a peppermint oil, or a mint oil. The formulation of embodiment 29, wherein the pH adjuster is hydrochloric acid or sodium hydroxide. The formulation of embodiment 29, wherein the anti-foaming agent is a silicone-based antifoam. The formulation of any one of embodiments 1-10, wherein the biosolid is digested in an anaerobic digester for at least 15 days. The formulation of any one of embodiments 1-10, wherein the biosolid is dewatered to a moisture content of about 10% or less. The formulation of any one of embodiments 1-10, wherein the biosolid is pulverized to a mesh size between about 10 and about 300. The formulation of any one of embodiments 1-10, wherein the biosolid is treated with an oil based agent, wherein the oil based agent prevents billowing.The formulation of any one of embodiments 1-10, wherein the biosolid and / or the grain are dried at a temperature between about 155° F. and about 250° F. The formulation of any one of embodiments 1-10, wherein the grain is dewatered to a moisture content of about 10% or less. The formulation of any one of embodiments 1-10, wherein the grain is pulverized to a mesh size of about 10 to about 300. The formulation of any one of embodiments 1-10, wherein the algae cell is pressed to isolate an algae cell wall. The formulation of any one of embodiments 1-10, wherein the algae cell wall is dried to a moisture content of about 20% or less. The formulation of any one of embodiments 1-10, wherein the algae cell wall is pulverized to a mesh size of about 10 to about 300. The formulation of embodiment 1, wherein the bio-nutrient composition is passed through a heating source, thereby being dewatered. The formulation of any one of embodiments 1-49, wherein the formulation enhances activity and / or diversity of a plurality of beneficial microbes in soil to which the formulation is applied, when comparing to the soil untreated with the formulation. The formulation of embodiment 50, wherein the beneficial microbe is Pseudomonas sp, Achromobacter sp., Bacillus sp., Nitrobacter sp., Nitrosomonas sp., Nitrospira sp., Paenibacillus sp., Rhizobium sp., Streptomyces sp., Trichoderma sp., Alkalihalobacillus hwajinpoensis, Aureobasidium pullulans, Metarhizium anisopliae, Purpureocillium lilacinum, or Pythium oligandrum. The formation of embodiment 50 or 51, wherein the beneficial microbe is an indigenous microbe species or a genetically engineered microbe. The formation of embodiment 50 or 51, wherein the beneficial microbe enhances production of exopolysaccharides, salicylic acid, siderophores, or 1 -aminocyclopropane- 1- carboxylate (ACC) deaminase, thereby improving soil health and promoting plant growth and stress tolerance. The formation of embodiment 50 or 51, wherein the formulation inhibits a plurality of soil pathogens. The formation of embodiment 54, wherein the soil pathogen is Pythium sp., Col etotrichum sp., Fusarium sp., Curvularia sp., Sclerotinia sp., Sclerotinia homoeocarpa, Agrobacterium tumefaciens, Sclerotium rolfsii, or Leptosphaerulina americana.The formulation of any one of embodiments 1-49, wherein the formulation induces expression of a plurality of genes involved in a stress-related defense mechanism in plants to which the formulation is applied, when comparing to plants untreated with the formulation. The formulation of embodiment 56, wherein the induced stress-related defense mechanism is selected from the group consisting of: phenylpropanoid biosynthesis, glutathione metabolism, sesquiterpenoid and triterpenoid metabolism, plant-pathogen interaction, ascorbate and aldarate metabolism, carotenoid biosynthesis, plant hormone signal transduction, and linoleic acid metabolism. The formulation of embodiment 56 or 57, wherein the induced stress-related defense mechanism is associated with:(i) upregulation of the expression of a plurality of gene involved in phenylpropanoid biosynthesis, which is phenylalanine ammonia lyase, trans-cinnamate 4-monooxygenase , and / or caffeoylshikimate esterase,(ii) upregulation of the expression of a plurality of gene involved in glutathione metabolism, which is glutathione dehydroascorbate reductase3, glutathione peroxidase, and / or gamma-glutamylcyclotransferase, '(iii) upregulation of the expression of a plurality of gene involved in sesquiterpenoid and triterpenoid metabolism, which is squalene synthase 1, squalene monooxygenase, and / or NAD+ dependent fame sal dehydrogenase,'(iv) upregulation of the expression of a plurality of gene involved in plant-pathogen interaction, which is pathogenesis related 1, suppressor of G2 allele of SKP1, and / or respiratory burst oxidase,'(v) upregulation of the expression of a plurality of gene involved in ascorbate and aldarate metabolism, which is L-gulonolactone oxidase, GDP-L-galactose phosphorylase, and / or ascorbate peroxidase, '(vi) upregulation of the expression of a plurality of gene involved in carotenoid biosynthesis, which is ABA 8' -hydroxylase, lycopene beta-cyclase, and / or 9-cis- epoxy carotenoid dioxygenase, '(vii) upregulation of the expression of a plurality of gene involved in plant hormone signal transduction, which is Not Responsive to PR1 protein (NPR1), jasmonic acid receptor, and / or abscisic acid receptor, ' and / or(viii) upregulation of the expression of a plurality of gene involved in linoleic acid metabolism, which is linoleate 9S-lipoxygenase and / or linoleate 13S-lipoxygenase9. The formulation of any one of embodiments 1-49, wherein the formulation induces expression of a plurality of genes involved in photosynthesis mechanism for increasing plant growth and development in plants to which the formulation is applied, when comparing to plants untreated with the formulation. The formulation of embodiment 59, wherein the induced photosynthesis mechanism is associated with upregulation of the expression of a plurality of gene, which is photosystem II subunit R, photosystem I subunit IV, plastocyanin, light-harvesting complex I chlorophyll a / b binding protein 2, and / or light-harvesting complex II chlorophyll a / b binding protein 4. The formulation of any one of embodiments 1-49, wherein the formulation induces expression of a plurality of genes involved in nutrient uptake and assimilation in plants to which the formulation is applied, when comparing to plants untreated with the formulation. The formulation of embodiment 61, wherein the induced nutrient uptake and assimilation is associated with upregulation of the expression of a plurality of gene, which is nitrate reductase, nitrite reductase, high-affinity nitrate transporter 2.1 (HAT2.1), high-affinity nitrate transporter 2.2 (HAT2.2), high-affinity nitrate transporter 2.3 (HAT2.3), glutamine synthetase, and / or mugineic acid- 3 dioxygenase. The formulation of any one of embodiments 1-62, wherein the formulation is applied with an agricultural product sequentially or simultaneously. The formation of embodiment 63, wherein the agricultural product is a mineral based fertilizer, synthetic fertilizer, an organic fertilizer, a micronutrient, a biostimulant, a plurality of microbes, a microbial additive, or an agricultural or landscaping chemical selected from the group consisting of a pesticide, an herbicide, a fungicide, and a surfactant. A method of making a granular formulation comprising the steps of:(a) dry milling a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell;(b) blending the dry milled bio-nutrient composition from step (a) in a powder form;(c) adding a binder to the blended bio-nutrient composition from step (b) to produce a formulation;(d) optionally, adding an additive; and(e) granulating the formulation from step (c) or (d).The method of embodiment 65, wherein the additive is added to the formulation from step (d). The method of embodiment 65, wherein the additive is not added to the formulation from step (d). The method of embodiment 65, wherein the bio-nutrient composition comprises: i) a biosolid at a final percentage-by-weight of about 35% to about 75% of the bio-nutrient composition; ii) a grain at a final percentage-by-weight of about 10% to about 50% of the bio-nutrient composition; and iii) an algae cell at a final percentage-by-weight of about 5% to about 40% of the bionutrient composition. The method of embodiment 65 or 68, wherein the biosolid is a sewage sludge, a manure or a food waste. The method of any one of embodiments 65 and 68-69, wherein the biosolid is a Class A EQ biosolid, Class A biosolid, or a Class B biosolid. The method of embodiment 65 or 68, wherein the grain is a brewers’ spent grain (BSG) or a distillers grain (DG). The method of any one of embodiments 65, 68, and 71, wherein the grain is a dried brewers’ spent grain (DBSG) or a dried distillers grain (DDG). The method of embodiment 65 or 68, wherein the algae cell is Cyanophyceae, Chlor ophyceae. Rhodophyceae , or Phaeophyceae . The method of any one of embodiments 65, 68, and 73, wherein the algae cell comprises a dried algae cell wall. The method of embodiment 65, wherein the binder is at a percentage weight of about 1% to about 50% of said formulation. The method of embodiment 65 or 75, wherein the binder is a lignin, a mineral, a plant starch, or a molasses. The method of any one of embodiments 65 and 75-76, wherein the binder is a lignin or a lignin derivative. The method of embodiment 77, wherein the lignin derivative is lignosulfonate. The method of any one of embodiments 65-67, wherein the additive is an odor-reducing or odor-neutralizing agent.The method of any one of embodiments 65-67 and 79, wherein the additive is selected from the group consisting of: an order neutralizer, an odor-masking agent, an essential oil, an odor absorbent, an enzyme, or a microbe. The method of embodiment 80, wherein the odor neutralizer is an activated charcoal, a zeolite, a baking soda, a silica gel, or a compound comprising cyclodextrin. The method of embodiment 80, wherein the odor-masking agent is a vanilla extract or a citrus extract. The method of embodiment 80, wherein the essential oil is a lavender oil, a eucalyptus oil, a peppermint oil, a lemon oil, a tea tree oil, a clove oil, an orange oil, or a cinnamon oil. The method of embodiment 80, wherein the odor absorbent is bentonite, kaolin clay, or peat moss. The method of embodiment 80, wherein the enzyme is a protease, a lipase, an amylase, a cellulase, a urease, an oxidoreductase, or a pectinase. The method of embodiment 80, wherein the microbe is a species of genus Bacillus or Pseudomonas. The method of any one of embodiments 65-74, wherein the biosolid is digested in an anaerobic digester for at least 15 days. The method of any one of embodiments 65-74, wherein the biosolid is dewatered to a moisture content of about 10% or less. The method of any one of embodiments 65-74, wherein the biosolid is pulverized to a mesh size between about 10 and about 300. The method of any one of embodiments 65-74, wherein the biosolid is treated with an oil based agent, wherein the oil based agent prevents billowing. The method of any one of embodiments 65-74, wherein the biosolid and / or the grain are dried at a temperature between about 155° F. and about 250° F. The method of any one of embodiments 65-74, wherein the grain is dewatered to a moisture content of about 10% or less. The method of any one of embodiments 65-74, wherein the grain is pulverized to a mesh size of about 10 to about 300. The method of any one of embodiments 65-74, wherein the algae cell is pressed to isolate an algae cell wall.The method of any one of embodiments 65-74, wherein the algae cell wall is dried to a moisture content of about 20% or less. The method of any one of embodiments 65-74, wherein the algae cell wall is pulverized to a mesh size of about 10 to about 300. The method of embodiment 65, wherein the bio-nutrient composition is passed through a heating source, thereby being dewatered. A method of making a liquid formulation comprising the steps of:(a) dry milling a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell;(b) agitating the dry milled bio-nutrient composition from step (a) in water with a speed mixer for wet milling; and(c) adding a liquid form of a binder to the wet milled bio-nutrient composition to produce a liquid formulation;(d) optionally adding an additive; and(e) filtering the formulation from step (c) or (d). The method of embodiment 98, wherein the additive is added to the formulation from step (d). . The method of embodiment 98, wherein the additive is not added to the formulation from step (d). . The method of embodiment 98, wherein the bio-nutrient composition comprises: i) a biosolid at a final percentage-by-weight of about 35% to about 75% of the bio-nutrient composition; ii) a grain at a final percentage-by-weight of about 10% to about 50% of the bio-nutrient composition; and iii) an algae cell at a final percentage-by-weight of about 5% to about 40% of the bionutrient composition. . The method of embodiment 98 or 101, wherein the biosolid is a sewage sludge, a manure or a food waste. . The method of any one of embodiments 98 and 101-102, wherein the biosolid is a Class A EQ biosolid, Class A biosolid, or a Class B biosolid. . The method of embodiment 98 or 101, wherein the grain is a brewers’ spent grain (BSG) or a distillers grain (DG).. The method of any one of embodiments 98, 101, and 104, wherein the grain is a dried brewers’ spent grain (DBSG) or a dried distillers grain (DDG). . The method of embodiment 98 or 101, wherein the algae cell is Cyanophyceae, Chlor ophyceae. Rhodophyceae , or Phaeophyceae . . The method of any one of embodiments 98, 101, and 106, wherein the algae cell comprises a dried algae cell wall or a post-harvest post filtration undried algae solution.. The method of embodiment 98, wherein the binder is at a percentage weight of about 1% to about 50% of said formulation. . The method of embodiment 98 or 108, wherein the binder is a lignin, a mineral, a plant starch, or a molasses. . The method of any one of embodiments 98 and 108-109, wherein the binder is a lignin or a lignin derivative. . The method of embodiment 110, wherein the lignin derivative is lignosulfonate. . The method of embodiment 98, wherein the additive is an odor-reducing or odorneutralizing agent. . The method of any one of embodiments 98-100 and 112, wherein the additive is selected from the group consisting of: an order neutralizer, an odor-masking agent, an essential oil, an odor absorbent, an enzyme, or a microbe. . The method of embodiment 113, wherein the odor neutralizer is an activated charcoal, a zeolite, a baking soda, a silica gel, or a compound comprising cyclodextrin. . The method of embodiment 113, wherein the odor-masking agent is a vanilla extract or a citrus extract. . The method of embodiment 113, wherein the essential oil is a lavender oil, a eucalyptus oil, a peppermint oil, a lemon oil, a tea tree oil, a clove oil, an orange oil, or a cinnamon oil. . The method of embodiment 113, wherein the odor absorbent is bentonite, kaolin clay, or peat moss. . The method of embodiment 113, wherein the enzyme is a protease, a lipase, an amylase, a cellulase, a urease, an oxidoreductase, or a pectinase. . The method of embodiment 113, wherein the microbe is a species of genus Bacillus orPseudomonas .. The method of any one of embodiments 98-107, wherein the biosolid is digested in an anaerobic digester for at least 15 days. . The method of any one of embodiments 98-107, wherein the biosolid is dewatered to a moisture content of about 10% or less. . The method of any one of embodiments 98-107, wherein the biosolid is pulverized to a mesh size between about 10 and about 300. . The method of any one of embodiments 98-107, wherein the biosolid is treated with an oil based agent, wherein the oil based agent prevents billowing. . The method of any one of embodiments 98-107, wherein the biosolid and / or the grain are dried at a temperature between about 155° F. and about 250° F. . The method of any one of embodiments 98-107, wherein the grain is dewatered to a moisture content of about 10% or less. . The method of any one of embodiments 98-107, wherein the grain is pulverized to a mesh size of about 10 to about 300. . The method of any one of embodiments 98-107, wherein the algae cell is pressed to isolate an algae cell wall. . The method of any one of embodiments 98-107, wherein the algae cell wall is dried to a moisture content of about 20% or less. . The method of any one of embodiments 98-107, wherein the algae cell wall is pulverized to a mesh size of about 10 to about 300. . The method of embodiment 98, wherein the bio-nutrient composition is passed through a heating source, thereby being dewatered. . The method of embodiment 98, wherein the liquid formulation further comprises an enhancer. . The method of embodiment 131, wherein the enhancer is selected from the consisting of: a preservative, a stabilizer, a buffering agent, a microbial inhibitor, a pH adjuster, and an anti -foaming agent. . The method of embodiment 132, wherein the preservative is sodium benzoate, potassium sorbate, or citric acid. . The method of embodiment 132, wherein the stabilizer is a xanthan gum, a chelating agent or an antioxidant. . The method of embodiment 134, wherein the chelating agent is ethylenediaminetetraacetic acid (EDTA).. The method of embodiment 134, wherein the antioxidant is an ascorbic acid or a tocopherol. . The method of embodiment 132 wherein the buffering agent is phosphate, phosphate derivative, acetate, or acetate derivative. . The method of embodiment 132, wherein the microbial inhibitor is an essential oil or a silver nanoparticle. . The method of embodiment 138, wherein the essential oil is a thyme oil, an oregano oil, a peppermint oil, or a mint oil. . The method of embodiment 132, wherein the pH adjuster is hydrochloric acid or sodium hydroxide. . The method of embodiment 132, wherein the anti-foaming agent is a silicone-based antifoam. . A method for improving soil health, comprising: applying the formulation of any one of embodiments 1-49 to soil, thereby improving soil health. . The method of embodiment 142, wherein the formulation improves a balanced ratio of essential nutrients selected from the group consisting of: nitrogen (N), Phosphorus (P), Potassium (K), Calcium (Ca), Magnesium (Mg), Sulfur (S), Carbon (C), Hydrogen (H), Oxygen (O), and micronutrient. . The method of embodiment 143, wherein the micronutrient is selected from the group consisting of: Iron (Fe), Manganese (Mn), Zinc (Zn), Copper (Cu), Boron (B), Molybdenum (Mo), and Chlorine (Cl). . The method of embodiment 142, wherein the formulation increases activity and / or diversity of a plurality of beneficial microbes in soil. . The method of embodiment 145, wherein the beneficial microbe is Pseudomonas sp, Achromobacter sp., Bacillus sp., Nitrobacter sp., Nitrosomonas sp., Nitrospira sp., Paenibacillus sp., Rhizobium sp., Streptomyces sp., Trichoderma sp., Alkalihalobacillus hwajinpoensis, Aureobasidium pullulans, Metarhizium anisopliae, Purpureocillium lilacinum, or Pythium oligandrum. . The method of embodiment 145 or 146, wherein the beneficial microbe is an indigenous microbe species or a genetically engineered microbe. . The method of embodiment 145 or 146, wherein the beneficial microbe enhances production of exopolysaccharides, salicylic acid, siderophores, or 1 -aminocyclopropane- 1-carboxylate (ACC) deaminase, thereby improving soil health and promoting plant growth and stress tolerance. . The method of embodiment 142, wherein the formulation decreases activity and / or diversity of a plurality of non-beneficial microbes in soil. . The method of embodiment 142, wherein the non-beneficial microbe is a soil pathogen selected from the group consisting of: Pythium sp., Colletotrichum sp., Fusarium sp., Curvularia sp., or Sclerotinia sp., Sclerotinia homoeocarpa, Agrobacterium tumefaciens, Sclerotium rolfsii, or Leptosphaerulina americana. . A method for altering microbiome in soil, comprising: applying the formulation of any one of embodiments 1-49 to soil, thereby altering microbiome in soil. . The method of embodiment 151, wherein the formulation increases a plurality of beneficial microbes in soil. . The method of embodiment 152, wherein the beneficial microbe Pseudomonas sp, Achromobacter sp., Bacillus sp., Nitrobacter sp., Nitrosomonas sp., Nitrospira sp., Paenibacillus sp., Rhizobium sp., Streptomyces sp., Trichoderma sp., Alkalihalobacillus hwajinpoensis, Aureobasidium pullulans, Metarhizium anisopliae, Purpureocillium lilacinum, or Pythium oligandrum. . The method of embodiment 152 or 153, wherein the beneficial microbe is an indigenous microbe species or a genetically engineered microbe. . The method of embodiment 152 or 153, wherein the beneficial microbe enhances production of exopolysaccharides, salicylic acid, siderophores, or 1 -aminocyclopropane- 1- carboxylate (ACC) deaminase, thereby improving soil health and promoting plant growth and stress tolerance. . The method of embodiment 151, wherein the formulation decreases a plurality of non- beneficial microbes in soil. . The method of embodiment 156, wherein the non-beneficial microbe is a soil pathogen selected from the group consisting of: Pythium sp., Colletotrichum sp., Fusarium sp., Curvularia sp., or Sclerotinia sp., Sclerotinia homoeocarpa, Agrobacterium tumefaciens, Sclerotium rolfsii, or Leptosphaerulina americana. . A method for inducing a stress-related defense mechanism in a plant, comprising: applying the formulation of any one of embodiments 1-49 to a target, thereby improving plant defense response to biotic or abiotic stress.. The method of embodiment 158, wherein the target is a plant, a plant part, or a plant cell thereof. . The method of embodiment 159, wherein the plant is acacia, alfalfa, amaranth, apple, apricot, artichoke, ash tree, asparagus, avocado, banana, barley, beans, beet, birch, beech, blackberry, black raspberry, blueberry, broccoli, Brussel's sprouts, cabbage, cane berry, canola, cantaloupe, carrot, cassava, cauliflower, cedar, a cereal, celery, chestnut, cherry, Chinese cabbage, citrus, Clementine, clover, coffee, corn, cotton, cowpea, cucumber, cypress, eggplant, elm, endive, eucalyptus, fennel, figs, fir, geranium, grape, grapefruit, groundnuts, ground cherry, gum hemlock, hickory, kale, kiwifruit, kohlrabi, larch, lettuce, leek, lemon, lime, locust, pine, maidenhair, maize, mango, maple, melon, millet, mushroom, mustard, nuts, oak, oats, oil palm, okra, onion, orange, an ornamental plant or flower or tree, papaya, palm, parsley, parsnip, pea, peach, peanut, pear, peat, pepper, persimmon, pigeon pea, peach, pine, pineapple, plantain, plum, pomegranate, potato, pumpkin, radicchio, radish, rapeseed, raspberry, rice, rye, sorghum, safflower, sallow, soybean, spinach, spruce, squash, strawberry, sugar beet, sugarcane, sunflower, sweet potato, sweet com, tangerine, tea, tobacco, tomato, trees, triticale, turf grasses, turnips, vine, walnut, watercress, watermelon, wheat, wild strawberry, yams, yew, or zucchini. . The method of embodiment 159, wherein the plant is a grass. . The method of embodiment 161, wherein the grass is Poaceae family. . The method of embodiment 161 or 162, wherein the grass is Agrostis genus, Poa genus, Lolium genus, Festuca genus, Cynodon genus, Stenotaphrum genus, or Zoysia genus.. The method of any one of embodiments 161-163, wherein the grass is a turfgrass, which is bentgrass, Kentucky bluegrass, ryegrass, tall fescue, Chewings fescue, Hard fescue, sheep fescue, creeping red fescue, bermudagrass, kikuyu grass, St. Augustine grass), buffalo grass, zoysia grass, zoyosa tenuifolia, or temple grass. . The method of embodiment 158, wherein the target is a soil or an area where a plant is present. . The method of embodiment 158, wherein the induced stress-related defense mechanism is selected from the group consisting of: phenylpropanoid biosynthesis, glutathione metabolism, sesquiterpenoid and triterpenoid metabolism, plant-pathogen interaction, ascorbate and aldarate metabolism, carotenoid biosynthesis, plant hormone signal transduction, and linoleic acid metabolism.. The method of embodiment 158 or 166, wherein the induced stress-related defense mechanism is associated with:(i) upregulation of the expression of a plurality of gene involved in phenylpropanoid biosynthesis, which is phenylalanine ammonia lyase, trans-cinnamate 4-monooxygenase , and / or caffeoylshikimate esterase,(ii) upregulation of the expression of a plurality of gene involved in glutathione metabolism, which is glutathione dehydroascorbate reductase3, glutathione peroxidase, and / or gamma-glutamylcyclotransferase, '(iii) upregulation of the expression of a plurality of gene involved in sesquiterpenoid and triterpenoid metabolism, which is squalene synthase 1, squalene monooxygenase, and / or NAD+ dependent fame sal dehydrogenase,'(iv) upregulation of the expression of a plurality of gene involved in plant-pathogen interaction, which is pathogenesis related 1, suppressor of G2 allele of SKP1, and / or respiratory burst oxidase,'(v) upregulation of the expression of a plurality of gene involved in ascorbate and aldarate metabolism, which is L-gulonolactone oxidase, GDP-L-galactose phosphorylase, and / or ascorbate peroxidase, '(vi) upregulation of the expression of a plurality of gene involved in carotenoid biosynthesis, which is ABA 8'-hydroxylase, lycopene beta-cyclase, and / or 9-cis- epoxy carotenoid dioxygenase, '(vii) upregulation of the expression of a plurality of gene involved in plant hormone signal transduction, which is Not Responsive to PR1 protein (NPR1), jasmonic acid receptor, and / or abscisic acid receptor, ' and / or(viii) upregulation of the expression of a plurality of gene involved in linoleic acid metabolism, which is linoleate 9S-lipoxygenase and / or linoleate 13S-lipoxygenase9. . A method for inducing a photosynthesis mechanism in a plant, comprising: applying the formulation of any one of embodiments 1-49 to a target, thereby improving plant growth and development. . The method of embodiment 168, wherein the target is a plant, a plant part, or a plant cell thereof. . The method of embodiment 169, wherein the plant is acacia, alfalfa, amaranth, apple, apricot, artichoke, ash tree, asparagus, avocado, banana, barley, beans, beet, birch, beech, blackberry, black raspberry, blueberry, broccoli, Brussel's sprouts, cabbage, cane berry,canola, cantaloupe, carrot, cassava, cauliflower, cedar, a cereal, celery, chestnut, cherry, Chinese cabbage, citrus, Clementine, clover, coffee, corn, cotton, cowpea, cucumber, cypress, eggplant, elm, endive, eucalyptus, fennel, figs, fir, geranium, grape, grapefruit, groundnuts, ground cherry, gum hemlock, hickory, kale, kiwifruit, kohlrabi, larch, lettuce, leek, lemon, lime, locust, pine, maidenhair, maize, mango, maple, melon, millet, mushroom, mustard, nuts, oak, oats, oil palm, okra, onion, orange, an ornamental plant or flower or tree, papaya, palm, parsley, parsnip, pea, peach, peanut, pear, peat, pepper, persimmon, pigeon pea, peach, pine, pineapple, plantain, plum, pomegranate, potato, pumpkin, radicchio, radish, rapeseed, raspberry, rice, rye, sorghum, safflower, sallow, soybean, spinach, spruce, squash, strawberry, sugar beet, sugarcane, sunflower, sweet potato, sweet com, tangerine, tea, tobacco, tomato, trees, triticale, turf grasses, turnips, vine, walnut, watercress, watermelon, wheat, wild strawberry, yams, yew, or zucchini. . The method of embodiment 169, wherein the plant is a grass. . The method of embodiment 171, wherein the grass is Poaceae family. . The method of embodiment 171 or 172, wherein the grass is Agrostis genus, Poa genus, Lolium genus, Festuca genus, Cynodon genus, Stenotaphrum genus, or Zoysia genus.. The method of any one of embodiments 171-173, wherein the grass is a turfgrass, which is bentgrass, Kentucky bluegrass, ryegrass, tall fescue, Chewings fescue, Hard fescue, sheep fescue, creeping red fescue, bermudagrass, kikuyu grass, St. Augustine grass, buffalo grass, zoysia grass, zoyosa tenuifolia, or temple grass. . The method of embodiment 168, wherein the target is a soil or an area where a plant is present. . The method of embodiment 168, wherein the induced photosynthesis mechanism is associated with upregulation of the expression of a plurality of gene, which is photosystem II subunit R, photosystem I subunit IV, plastocyanin, light-harvesting complex I chlorophyll a / b binding protein 2, and / or light-harvesting complex II chlorophyll a / b binding protein 4. . A method for inducing nutrient uptake and assimilation in a plant, comprising: applying the formulation of any one of embodiments 1-49 to a target, thereby improving nitrogen assimilation or uptake of a plant. . The method of embodiment 177, wherein the target is a plant, a plant part, or a plant cell thereof.179. The method of embodiment 178, wherein the plant is acacia, alfalfa, amaranth, apple, apricot, artichoke, ash tree, asparagus, avocado, banana, barley, beans, beet, birch, beech, blackberry, black raspberry, blueberry, broccoli, Brussel's sprouts, cabbage, cane berry, canola, cantaloupe, carrot, cassava, cauliflower, cedar, a cereal, celery, chestnut, cherry, Chinese cabbage, citrus, Clementine, clover, coffee, corn, cotton, cowpea, cucumber, cypress, eggplant, elm, endive, eucalyptus, fennel, figs, fir, geranium, grape, grapefruit, groundnuts, ground cherry, gum hemlock, hickory, kale, kiwifruit, kohlrabi, larch, lettuce, leek, lemon, lime, locust, pine, maidenhair, maize, mango, maple, melon, millet, mushroom, mustard, nuts, oak, oats, oil palm, okra, onion, orange, an ornamental plant or flower or tree, papaya, palm, parsley, parsnip, pea, peach, peanut, pear, peat, pepper, persimmon, pigeon pea, peach, pine, pineapple, plantain, plum, pomegranate, potato, pumpkin, radicchio, radish, rapeseed, raspberry, rice, rye, sorghum, safflower, sallow, soybean, spinach, spruce, squash, strawberry, sugar beet, sugarcane, sunflower, sweet potato, sweet com, tangerine, tea, tobacco, tomato, trees, triticale, turf grasses, turnips, vine, walnut, watercress, watermelon, wheat, wild strawberry, yams, yew, or zucchini.180. The method of embodiment 178, wherein the plant is a grass.181. The method of embodiment 180, wherein the grass is a Poaceae family.182. The method of embodiment 180 or 181 , wherein the grass is Agrostis genus, Poa genus, Lolium genus, Festuca genus, Cynodon genus, Stenotaphrum genus, or Zoysia genus.183. The method of any one of embodiments 180-182, wherein the grass is a turfgrass, which is bentgrass, Kentucky bluegrass, ryegrass, tall fescue, Chewings fescue, Hard fescue, sheep fescue, creeping red fescue, bermudagrass, kikuyu grass, St. Augustine grass, buffalo grass, zoysia grass, zoyosa tenuifolia, or temple grass.184. The method of embodiment 177, wherein the target is a soil or an area where a plant is present.185. The method of embodiment 177, wherein the induced nutrient uptake and assimilation is associated with upregulation of the expression of a plurality of gene, which is nitrate reductase, nitrite reductase, high-affinity nitrate transporter 2.1 (HAT2.1), high-affinity nitrate transporter 2.2 (HAT2.2), high-affinity nitrate transporter 2.3 (HAT2.3), glutamine synthetase, and / or mugineic acid- 3 dioxygenase.Use of Bio-nutrient compositionA method for improving soil health, comprising: applying to soil a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell, thereby improving soil health. The method of embodiment 186, wherein the bio-nutrient composition improves a balanced ratio of essential nutrients selected from the group consisting of: nitrogen (N), Phosphorus (P), Potassium (K), Calcium (Ca), Magnesium (Mg), Sulfur (S), Carbon (C), Hydrogen (H), Oxygen (O), and micronutrient. The method of embodiment 187, wherein the micronutrient is selected from the group consisting of: Iron (Fe), Manganese (Mn), Zinc (Zn), Copper (Cu), Boron (B), Molybdenum (Mo), and Chlorine (Cl). The method of embodiment 1, wherein the bio-nutrient composition increases activity and / or diversity of a plurality of beneficial microbes in soil. The method of embodiment 4, wherein the beneficial microbe is Pseudomonas sp, Achromobacter sp., Bacillus sp., Nitrobacter sp., Nitrosomonas sp., Nitrospira sp., Paenibacillus sp., Rhizobium sp., Streptomyces sp., Trichoderma sp., Alkalihalobacillus hwajinpoensis, Aureobasidium pullulans, Metarhizium anisopliae, Purpureocillium lilacinum, or Pythium oligandrum. The method of embodiment 4 or 5, wherein the beneficial microbe is an indigenous microbe species or a genetically engineered microbe. The method of embodiment 4 or 5, wherein the beneficial microbe enhances production of exopolysaccharides, salicylic acid, siderophores, or 1 -aminocyclopropane- 1 -carboxylate (ACC) deaminase, thereby improving soil health and promoting plant growth and stress tolerance. The method of embodiment 1, wherein the bio-nutrient composition decreases activity and / or diversity of a plurality of non-beneficial microbes in soil. The method of embodiment 1, wherein the non-beneficial microbe is a soil pathogen selected from the group consisting of: Pythium sp., Colletotrichum sp., Fusarium sp., Curvularia sp., or Sclerotinia sp., Sclerotinia homoeocarpa, Agrobacterium tumefaciens, Sclerotium rolfsii, or Leptosphaerulina americana. The method of embodiment 1, wherein the bio-nutrient composition comprises: i) a biosolid at a final percentage-by-weight of about 35% to about 75% of the bio-nutrient composition; ii) a grain at a final percentage-by-weight of about 10% to about 50% of the bio-nutrient composition; andiii) an algae cell at a final percentage-by-weight of about 5% to about 40% of the bionutrient composition. The method of embodiment 1 or 10, wherein the biosolid is a sewage sludge, a manure or a food waste. The method of any one of embodiments 1 and 10-11, wherein the biosolid is a Class A EQ biosolid, Class A biosolid, or a Class B biosolid. The method of embodiment 1 or 10, wherein the grain is a brewers’ spent grain (BSG) or a distillers grain (DG). The method of any one of embodiments 1, 10, and 13, wherein the grain is a dried brewers’ spent grain (DBSG) or a dried distillers grain (DDG). The method of embodiment 1 or 10, wherein the algae cell is Cyanophyceae^ Chlor ophyceae. Rhodophyceae , or Phaeophyceae . The method of any one of embodiments 1, 10, and 15, wherein the algae cell comprises a dried algae cell wall, algae process water, or a post-harvest post filtration undried algae solution. The method of any one of embodiments 1-16, wherein the biosolid is digested in an anaerobic digester for at least 15 days. The method of any one of embodiments 1-16, wherein the biosolid is dewatered to a moisture content of about 10% or less. The method of any one of embodiments 1-16, wherein the biosolid is pulverized to a mesh size between about 10 and about 300. The method of any one of embodiments 1-16, wherein the biosolid is treated with an oil based agent, wherein the oil based agent prevents billowing. The method of any one of embodiments 1-16, wherein the biosolid and / or the grain are dried at a temperature between about 155° F. and about 250° F. The method of any one of embodiments 1-16, wherein the grain is dewatered to a moisture content of about 10% or less. The method of any one of embodiments 1-16, wherein the grain is pulverized to a mesh size of about 10 to about 300. The method of any one of embodiments 1-16, wherein the algae cell is pressed to isolate an algae cell wall. The method of any one of embodiments 1-16, wherein the algae cell wall is dried to a moisture content of about 20% or less.The method of any one of embodiments 1-16, wherein the algae cell wall is pulverized to a mesh size of about 10 to about 300. The method of embodiment 1, wherein the bio-nutrient composition is passed through a heating source, thereby being dewatered. A method for altering microbiome in soil, comprising: applying to soil a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell, thereby altering microbiome in soil. The method of embodiment 28, wherein the bio-nutrient composition increases a plurality of beneficial microbes in soil. The method of embodiment 29, wherein the beneficial microbe Pseudomonas sp, Achromobacter sp., Bacillus sp., Nitrobacter sp., Nitrosomonas sp., Nitrospira sp., Paenibacillus sp., Rhizobium sp., Streptomyces sp., Trichoderma sp., Alkalihalobacillus hwajinpoensis, Aureobasidium pullulans, Metarhizium anisopliae, Purpureocillium lilacinum, or Pythium oligandrum. The method of embodiment 29 or 30, wherein the beneficial microbe is an indigenous microbe species or a genetically engineered microbe. The method of embodiment 29 or 30, wherein the beneficial microbe enhances production of exopolysaccharides, salicylic acid, siderophores, or 1 -aminocyclopropane- 1 -carboxylate (ACC) deaminase, thereby improving soil health and promoting plant growth and stress tolerance. The method of embodiment 28, wherein the bio-nutrient composition decreases a plurality of non-beneficial microbes in soil. The method of embodiment 33, wherein the non-beneficial microbe is a soil pathogen selected from the group consisting of: Pythium sp., Colletotrichum sp., Fusarium sp., Curvularia sp., or Sclerotinia sp., Sclerotinia homoeocarpa, Agrobacterium tumefaciens, Sclerotium rolfsii, or Leptosphaerulina americana. The method of embodiment 28, wherein the bio-nutrient composition comprises: i) a biosolid at a final percentage-by-weight of about 35% to about 75% of the bio-nutrient composition; ii) a grain at a final percentage-by-weight of about 10% to about 50% of the bio-nutrient composition; and iii) an algae cell at a final percentage-by-weight of about 5% to about 40% of the bionutrient composition.The method of embodiment 28 or 35, wherein the biosolid is a sewage sludge, a manure or a food waste. The method of any one of embodiments 28 and 35-36, wherein the biosolid is a Class A EQ biosolid, Class A biosolid, or a Class B biosolid. The method of embodiment 28 or 35, wherein the grain is a brewers’ spent grain (BSG) or a distillers grain (DG). The method of any one of embodiments 28, 35, and 38, wherein the grain is a dried brewers’ spent grain (DBSG) or a dried distillers grain (DDG). The method of embodiment 28 or 35, wherein the algae cell is Cyanophyceae^ Chlor ophyceae. Rhodophyceae , or Phaeophyceae . The method of any one of embodiments 28, 35, and 40, wherein the algae cell comprises a dried algae cell wall, algae process water, or a post-harvest post filtration undried algae solution. The method of any one of embodiments 28-41, wherein the biosolid is digested in an anaerobic digester for at least 15 days. The method of any one of embodiments 28-41, wherein the biosolid is dewatered to a moisture content of about 10% or less. The method of any one of embodiments 28-41, wherein the biosolid is pulverized to a mesh size between about 10 and about 300. The method of any one of embodiments 28-41, wherein the biosolid is treated with an oil based agent, wherein the oil based agent prevents billowing. The method of any one of embodiments 28-41, wherein the biosolid and / or the grain are dried at a temperature between about 155° F. and about 250° F. The method of any one of embodiments 28-41, wherein the grain is dewatered to a moisture content of about 10% or less. The method of any one of embodiments 28-41, wherein the grain is pulverized to a mesh size of about 10 to about 300. The method of any one of embodiments 28-41, wherein the algae cell is pressed to isolate an algae cell wall. The method of any one of embodiments 28-41, wherein the algae cell wall is dried to a moisture content of about 20% or less. The method of any one of embodiments 28-41, wherein the algae cell wall is pulverized to a mesh size of about 10 to about 300.The method of embodiment 28, wherein the bio-nutrient composition is passed through a heating source, thereby being dewatered. A method for inducing a stress-related defense mechanism in a plant, comprising: applying to a target a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell, thereby improving plant defense response to biotic or abiotic stress. The method of embodiment 53, wherein the target is a plant, a plant part, or a plant cell thereof. The method of embodiment 54, wherein the plant is acacia, alfalfa, amaranth, apple, apricot, artichoke, ash tree, asparagus, avocado, banana, barley, beans, beet, birch, beech, blackberry, black raspberry, blueberry, broccoli, Brussel's sprouts, cabbage, cane berry, canola, cantaloupe, carrot, cassava, cauliflower, cedar, a cereal, celery, chestnut, cherry, Chinese cabbage, citrus, Clementine, clover, coffee, corn, cotton, cowpea, cucumber, cypress, eggplant, elm, endive, eucalyptus, fennel, figs, fir, geranium, grape, grapefruit, groundnuts, ground cherry, gum hemlock, hickory, kale, kiwifruit, kohlrabi, larch, lettuce, leek, lemon, lime, locust, pine, maidenhair, maize, mango, maple, melon, millet, mushroom, mustard, nuts, oak, oats, oil palm, okra, onion, orange, an ornamental plant or flower or tree, papaya, palm, parsley, parsnip, pea, peach, peanut, pear, peat, pepper, persimmon, pigeon pea, peach, pine, pineapple, plantain, plum, pomegranate, potato, pumpkin, radicchio, radish, rapeseed, raspberry, rice, rye, sorghum, safflower, sallow, soybean, spinach, spruce, squash, strawberry, sugar beet, sugarcane, sunflower, sweet potato, sweet com, tangerine, tea, tobacco, tomato, trees, triticale, turf grasses, turnips, vine, walnut, watercress, watermelon, wheat, wild strawberry, yams, yew, or zucchini. The method of embodiment 54, wherein the plant is a grass. The method of embodiment 56, wherein the grass is Poaceae family. The method of embodiment 56 or 57, wherein the grass is Agrostis genus, Poa genus, Lolium genus, Festuca genus, Cynodon genus, Stenotaphrum genus, or Zoysia genus. The method of any one of embodiments 56-58, wherein the grass is a turfgrass, which is bentgrass, Kentucky bluegrass, ryegrass, tall fescue, Chewings fescue, Hard fescue, sheep fescue, creeping red fescue, bermudagrass, kikuyu grass, St. Augustine grass), buffalo grass, zoysia grass, zoyosa tenuifolia, or temple grass. The method of embodiment 53, wherein the target is a soil or an area where a plant is present.The method of embodiment 53, wherein the induced stress-related defense mechanism is selected from the group consisting of: phenylpropanoid biosynthesis, glutathione metabolism, sesquiterpenoid and triterpenoid metabolism, plant-pathogen interaction, ascorbate and aldarate metabolism, carotenoid biosynthesis, plant hormone signal transduction, and linoleic acid metabolism. The method of embodiment 53 or 62, wherein the induced stress-related defense mechanism is associated with:(i) upregulation of the expression of a plurality of gene involved in phenylpropanoid biosynthesis, which is phenylalanine ammonia lyase, trans-cinnamate 4-monooxygenase , and / or caffeoylshikimate esterase,(ii) upregulation of the expression of a plurality of gene involved in glutathione metabolism, which is glutathione dehydroascorbate reductase3, glutathione peroxidase, and / or gamma-glutamylcyclotransferase, '(iii) upregulation of the expression of a plurality of gene involved in sesquiterpenoid and triterpenoid metabolism, which is squalene synthase 1, squalene monooxygenase, and / or NAD+ dependent fame sal dehydrogenase,'(iv) upregulation of the expression of a plurality of gene involved in plant-pathogen interaction, which is pathogenesis related 1, suppressor of G2 allele of SKP1, and / or respiratory burst oxidase,'(v) upregulation of the expression of a plurality of gene involved in ascorbate and aldarate metabolism, which is L-gulonolactone oxidase, GDP-L-galactose phosphorylase, and / or ascorbate peroxidase, '(vi) upregulation of the expression of a plurality of gene involved in carotenoid biosynthesis, which is ABA 8'-hydroxylase, lycopene beta-cyclase, and / or 9-cis- epoxy carotenoid dioxygenase, '(vii) upregulation of the expression of a plurality of gene involved in plant hormone signal transduction, which is Not Responsive to PR1 protein (NPR1), jasmonic acid receptor, and / or abscisic acid receptor, ' and / or(viii) upregulation of the expression of a plurality of gene involved in linoleic acid metabolism, which is linoleate 9S-lipoxygenase and / or linoleate 13S-lipoxygenase9. The method of embodiment 53, wherein the bio-nutrient composition comprises: i) a biosolid at a final percentage-by-weight of about 35% to about 75% of the bio-nutrient composition;ii) a grain at a final percentage-by-weight of about 10% to about 50% of the bio-nutrient composition; and iii) an algae cell at a final percentage-by-weight of about 5% to about 40% of the bionutrient composition. The method of embodiment 53 or 63, wherein the biosolid is a sewage sludge, a manure or a food waste. The method of any one of embodiments 53 and 63-64, wherein the biosolid is a Class A EQ biosolid, Class A biosolid, or a Class B biosolid. The method of embodiment 53 or 63, wherein the grain is a brewers’ spent grain (BSG) or a distillers grain (DG). The method of any one of embodiments 53, 63, and 66, wherein the grain is a dried brewers’ spent grain (DBSG) or a dried distillers grain (DDG). The method of embodiment 53 or 63, wherein the algae cell is Cyanophyceae^ Chlor ophyceae. Rhodophyceae , or Phaeophyceae . The method of any one of embodiments 53, 63, and 68, wherein the algae cell comprises a dried algae cell wall, algae process water, or a post-harvest post filtration undried algae solution. The method of any one of embodiments 53-69, wherein the biosolid is digested in an anaerobic digester for at least 15 days. The method of any one of embodiments 53-69, wherein the biosolid is dewatered to a moisture content of about 10% or less. The method of any one of embodiments 53-69, wherein the biosolid is pulverized to a mesh size between about 10 and about 300. The method of any one of embodiments 53-69, wherein the biosolid is treated with an oil based agent, wherein the oil based agent prevents billowing. The method of any one of embodiments 53-69, wherein the biosolid and / or the grain are dried at a temperature between about 155° F. and about 250° F. The method of any one of embodiments 53-69, wherein the grain is dewatered to a moisture content of about 10% or less. The method of any one of embodiments 53-69, wherein the grain is pulverized to a mesh size of about 10 to about 300. The method of any one of embodiments 53-69, wherein the algae cell is pressed to isolate an algae cell wall.The method of any one of embodiments 53-69, wherein the algae cell wall is dried to a moisture content of about 20% or less. The method of any one of embodiments 53-69, wherein the algae cell wall is pulverized to a mesh size of about 10 to about 300. The method of embodiment 53, wherein the bio-nutrient composition is passed through a heating source, thereby being dewatered. A method for inducing a photosynthesis mechanism in a plant, comprising: applying to a target a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell, thereby improving plant growth and development. The method of embodiment 81, wherein the target is a plant, a plant part, or a plant cell thereof. The method of embodiment 82, wherein the plant is acacia, alfalfa, amaranth, apple, apricot, artichoke, ash tree, asparagus, avocado, banana, barley, beans, beet, birch, beech, blackberry, black raspberry, blueberry, broccoli, Brussel's sprouts, cabbage, cane berry, canola, cantaloupe, carrot, cassava, cauliflower, cedar, a cereal, celery, chestnut, cherry, Chinese cabbage, citrus, Clementine, clover, coffee, corn, cotton, cowpea, cucumber, cypress, eggplant, elm, endive, eucalyptus, fennel, figs, fir, geranium, grape, grapefruit, groundnuts, ground cherry, gum hemlock, hickory, kale, kiwifruit, kohlrabi, larch, lettuce, leek, lemon, lime, locust, pine, maidenhair, maize, mango, maple, melon, millet, mushroom, mustard, nuts, oak, oats, oil palm, okra, onion, orange, an ornamental plant or flower or tree, papaya, palm, parsley, parsnip, pea, peach, peanut, pear, peat, pepper, persimmon, pigeon pea, peach, pine, pineapple, plantain, plum, pomegranate, potato, pumpkin, radicchio, radish, rapeseed, raspberry, rice, rye, sorghum, safflower, sallow, soybean, spinach, spruce, squash, strawberry, sugar beet, sugarcane, sunflower, sweet potato, sweet com, tangerine, tea, tobacco, tomato, trees, triticale, turf grasses, turnips, vine, walnut, watercress, watermelon, wheat, wild strawberry, yams, yew, or zucchini. The method of embodiment 82, wherein the plant is a grass. The method of embodiment 84, wherein the grass is Poaceae family. The method of embodiment 84 or 85, wherein the grass is Agrostis genus, Poa genus, Lolium genus, Festuca genus, Cynodon genus, Stenotaphrum genus, or Zoysia genus. The method of any one of embodiments 84-86, wherein the grass is a turfgrass, which is bentgrass, Kentucky bluegrass, ryegrass, tall fescue, Chewings fescue, Hard fescue, sheepfescue, creeping red fescue, bermudagrass, kikuyu grass, St. Augustine grass, buffalo grass, zoysia grass, zoyosa tenuifolia, or temple grass. The method of embodiment 81, wherein the target is a soil or an area where a plant is present. The method of embodiment 81, wherein the induced photosynthesis mechanism is associated with upregulation of the expression of a plurality of gene, which is photosystem II subunit R, photosystem I subunit IV, plastocyanin, light-harvesting complex I chlorophyll a / b binding protein 2, and / or light-harvesting complex II chlorophyll a / b binding protein 4. The method of embodiment 81, wherein the bio-nutrient composition comprises: i) a biosolid at a final percentage-by-weight of about 35% to about 75% of the bio-nutrient composition; ii) a grain at a final percentage-by-weight of about 10% to about 50% of the bio-nutrient composition; and iii) an algae cell at a final percentage-by-weight of about 5% to about 40% of the bionutrient composition. The method of embodiment 81 or 90, wherein the biosolid is a sewage sludge, a manure or a food waste. The method of any one of embodiments 81 and 90-91, wherein the biosolid is a Class A EQ biosolid, Class A biosolid, or a Class B biosolid. The method of embodiment 81 or 90, wherein the grain is a brewers’ spent grain (BSG) or a distillers grain (DG). The method of any one of embodiments 81, 90, and 83, wherein the grain is a dried brewers’ spent grain (DBSG) or a dried distillers grain (DDG). The method of embodiment 81 or 90, wherein the algae cell is Cyanophyceae^ Chlor ophyceae, Rhodophyceae, or Phaeophyceae . The method of any one of embodiments 81, 90, and 95, wherein the algae cell comprises a dried algae cell wall, algae process water, or a post-harvest post filtration undried algae solution. The method of any one of embodiments 81-96, wherein the biosolid is digested in an anaerobic digester for at least 15 days. The method of any one of embodiments 81-96, wherein the biosolid is dewatered to a moisture content of about 10% or less.The method of any one of embodiments 81-96, wherein the biosolid is pulverized to a mesh size between about 10 and about 300. . The method of any one of embodiments 81-96, wherein the biosolid is treated with an oil based agent, wherein the oil based agent prevents billowing. . The method of any one of embodiments 81-96, wherein the biosolid and / or the grain are dried at a temperature between about 155° F. and about 250° F. . The method of any one of embodiments 81-96, wherein the grain is dewatered to a moisture content of about 10% or less. . The method of any one of embodiments 81-96, wherein the grain is pulverized to a mesh size of about 10 to about 300. . The method of any one of embodiments 81-96, wherein the algae cell is pressed to isolate an algae cell wall. . The method of any one of embodiments 81-96, wherein the algae cell wall is dried to a moisture content of about 20% or less. . The method of any one of embodiments 81-96, wherein the algae cell wall is pulverized to a mesh size of about 10 to about 300. . The method of embodiment 81, wherein the bio-nutrient composition is passed through a heating source, thereby being dewatered. . A method for inducing nutrient uptake and assimilation in a plant, comprising: applying to a target a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell, thereby improving nitrogen assimilation or uptake of a plant. . The method of embodiment 108, wherein the target is a plant, a plant part, or a plant cell thereof. . The method of embodiment 109, wherein the plant is acacia, alfalfa, amaranth, apple, apricot, artichoke, ash tree, asparagus, avocado, banana, barley, beans, beet, birch, beech, blackberry, black raspberry, blueberry, broccoli, Brussel's sprouts, cabbage, cane berry, canola, cantaloupe, carrot, cassava, cauliflower, cedar, a cereal, celery, chestnut, cherry, Chinese cabbage, citrus, Clementine, clover, coffee, corn, cotton, cowpea, cucumber, cypress, eggplant, elm, endive, eucalyptus, fennel, figs, fir, geranium, grape, grapefruit, groundnuts, ground cherry, gum hemlock, hickory, kale, kiwifruit, kohlrabi, larch, lettuce, leek, lemon, lime, locust, pine, maidenhair, maize, mango, maple, melon, millet, mushroom, mustard, nuts, oak, oats, oil palm, okra, onion, orange, an ornamental plant or flower or tree, papaya, palm, parsley, parsnip, pea, peach, peanut, pear, peat, pepper,persimmon, pigeon pea, peach, pine, pineapple, plantain, plum, pomegranate, potato, pumpkin, radicchio, radish, rapeseed, raspberry, rice, rye, sorghum, safflower, sallow, soybean, spinach, spruce, squash, strawberry, sugar beet, sugarcane, sunflower, sweet potato, sweet com, tangerine, tea, tobacco, tomato, trees, triticale, turf grasses, turnips, vine, walnut, watercress, watermelon, wheat, wild strawberry, yams, yew, or zucchini. . The method of embodiment 109, wherein the plant is a grass. . The method of embodiment 111, wherein the grass is a Poaceae family. . The method of embodiment 111 orl 12, wherein the grass is Agrostis genus, Poa genus, Lolium genus, Festuca genus, Cynodon genus, Stenotaphrum genus, or Zoysia genus.. The method of any one of embodiments 111-113, wherein the grass is a turfgrass, which is bentgrass, Kentucky bluegrass, ryegrass, tall fescue, Chewings fescue, Hard fescue, sheep fescue, creeping red fescue, bermudagrass, kikuyu grass, St. Augustine grass, buffalo grass, zoysia grass, zoyosa tenuifolia, or temple grass. . The method of embodiment 108, wherein the target is a soil or an area where a plant is present. . The method of embodiment 108, wherein the induced nutrient uptake and assimilation is associated with upregulation of the expression of a plurality of gene, which is nitrate reductase, nitrite reductase, high-affinity nitrate transporter 2.1 (HAT2.1), high-affinity nitrate transporter 2.2 (HAT2.2), high-affinity nitrate transporter 2.3 (HAT2.3), glutamine synthetase, and / or mugineic acid- 3 dioxygenase. . The method of embodiment 108, wherein the bio-nutrient composition comprises: i) a biosolid at a final percentage-by-weight of about 35% to about 75% of the bio-nutrient composition; ii) a grain at a final percentage-by-weight of about 10% to about 50% of the bio-nutrient composition; and iii) an algae cell at a final percentage-by-weight of about 5% to about 40% of the bionutrient composition. . The method of embodiment 108 or 117, wherein the biosolid is a sewage sludge, a manure or a food waste. . The method of any one of embodiments 108 and 117-118, wherein the biosolid is a Class A EQ biosolid, Class A biosolid, or a Class B biosolid. . The method of embodiment 108 or 117, wherein the grain is a brewers’ spent grain(BSG) or a distillers grain (DG).121. The method of any one of embodiments 108, 117, and 120, wherein the grain is a dried brewers’ spent grain (DBSG) or a dried distillers grain (DDG).122. The method of embodiment 108 or 117, wherein the algae cell is Cyanophyceae^ Chlor ophyceae. Rhodophyceae , or Phaeophyceae .123. The method of any one of embodiments 108, 117, and 122, wherein the algae cell comprises a dried algae cell wall, algae process water, or a post-harvest post filtration undried algae solution.124. The method of any one of embodiments 108-123, wherein the biosolid is digested in an anaerobic digester for at least 15 days.125. The method of any one of embodiments 108-123, wherein the biosolid is dewatered to a moisture content of about 10% or less.126. The method of any one of embodiments 108-123, wherein the biosolid is pulverized to a mesh size between about 10 and about 300.127. The method of any one of embodiments 108-123, wherein the biosolid is treated with an oil based agent, wherein the oil based agent prevents billowing.128. The method of any one of embodiments 108-123, wherein the biosolid and / or the grain are dried at a temperature between about 155° F. and about 250° F.129. The method of any one of embodiments 108-123, wherein the grain is dewatered to a moisture content of about 10% or less.130. The method of any one of embodiments 108-123, wherein the grain is pulverized to a mesh size of about 10 to about 300.131. The method of any one of embodiments 108-123, wherein the algae cell is pressed to isolate an algae cell wall.132. The method of any one of embodiments 108-123, wherein the algae cell wall is dried to a moisture content of about 20% or less.133. The method of any one of embodiments 108-123, wherein the algae cell wall is pulverized to a mesh size of about 10 to about 300.134. The method of embodiment 108, wherein the bio-nutrient composition is passed through a heating source, thereby being dewatered.Use of formulation comprising a bio-nutrient composition and a binderA method for improving soil health, comprising: applying to soil a formulation comprising(1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell and(2) a binder, thereby improving soil health. The method of embodiment 1, wherein the formulation improves a balanced ratio of essential nutrients selected from the group consisting of: nitrogen (N), Phosphorus (P), Potassium (K), Calcium (Ca), Magnesium (Mg), Sulfur (S), Carbon (C), Hydrogen (H), Oxygen (O), and micronutrient. The method of embodiment 2, wherein the micronutrient is selected from the group consisting of: Iron (Fe), Manganese (Mn), Zinc (Zn), Copper (Cu), Boron (B), Molybdenum (Mo), and Chlorine (Cl). The method of embodiment 1, wherein the formulation increases activity and / or diversity of a plurality of beneficial microbes in soil. The method of embodiment 4, wherein the beneficial microbe is Pseudomonas sp, Achromobacter sp., Bacillus sp., Nitrobacter sp., Nitrosomonas sp., Nitrospira sp., Paenibacillus sp., Rhizobium sp., Streptomyces sp., Trichoderma sp., Alkalihalobacillus hwajinpoensis, Aureobasidium pullulans, Metarhizium anisopliae, Purpureocillium lilacinum, or Pythium oligandrum. The method of embodiment 4 or 5, wherein the beneficial microbe is an indigenous microbe species or a genetically engineered microbe. The method of embodiment 4 or 5, wherein the beneficial microbe enhances production of exopolysaccharides, salicylic acid, siderophores, or 1 -aminocyclopropane- 1 -carboxylate (ACC) deaminase, thereby improving soil health and promoting plant growth and stress tolerance. The method of embodiment 1, wherein the formulation decreases activity and / or diversity of a plurality of non-beneficial microbes in soil. The method of embodiment 8, wherein the non-beneficial microbe is a soil pathogen selected from the group consisting of: Pythium sp., Colletotrichum sp., Fusarium sp., Curvularia sp., or Sclerotinia sp., Sclerotinia homoeocarpa, Agrobacterium tumefaciens, Sclerotium rolfsii, or Leptosphaerulina americana. The method of embodiment 1, wherein the bio-nutrient composition comprises: i) a biosolid at a final percentage-by-weight of about 35% to about 75% of the bio-nutrient composition;ii) a grain at a final percentage-by-weight of about 10% to about 50% of the bio-nutrient composition; and iii) an algae cell at a final percentage-by-weight of about 5% to about 40% of the bionutrient composition. The method of embodiment 1 or 10, wherein the biosolid is a sewage sludge, a manure or a food waste. The method of any one of embodiments 1 and 10-11, wherein the biosolid is a Class A EQ biosolid, Class A biosolid, or a Class B biosolid. The method of embodiment 1 or 10, wherein the grain is a brewers’ spent grain (BSG) or a distillers grain (DG). The method of any one of embodiments 1, 10, and 13, wherein the grain is a dried brewers’ spent grain (DBSG) or a dried distillers grain (DDG). The method of embodiment 1 or 10, wherein the algae cell is Cyanophyceae^ Chlor ophyceae. Rhodophyceae , or Phaeophyceae . The method of any one of embodiments 1, 10, and 15, wherein the algae cell comprises a dried algae cell wall, algae process water, or a post-harvest post filtration undried algae solution. The method of any one of embodiments 1-16, wherein the biosolid is digested in an anaerobic digester for at least 15 days. The method of any one of embodiments 1-16, wherein the biosolid is dewatered to a moisture content of about 10% or less. The method of any one of embodiments 1-16, wherein the biosolid is pulverized to a mesh size between about 10 and about 300. The method of any one of embodiments 1-16, wherein the biosolid is treated with an oil based agent, wherein the oil based agent prevents billowing. The method of any one of embodiments 1-16, wherein the biosolid and / or the grain are dried at a temperature between about 155° F. and about 250° F. The method of any one of embodiments 1-16, wherein the grain is dewatered to a moisture content of about 10% or less. The method of any one of embodiments 1-16, wherein the grain is pulverized to a mesh size of about 10 to about 300. The method of any one of embodiments 1-16, wherein the algae cell is pressed to isolate an algae cell wall.The method of any one of embodiments 1-16, wherein the algae cell wall is dried to a moisture content of about 20% or less. The method of any one of embodiments 1-16, wherein the algae cell wall is pulverized to a mesh size of about 10 to about 300. The method of embodiment 1, wherein the bio-nutrient composition is passed through a heating source, thereby being dewatered. The method of claim 1, wherein the binder is at a percentage weight of about 1% to about 50% of said formulation. The method of claim 1 or 28, wherein the binder is a lignin, a mineral, a plant starch, or a molasses. The method of any one of claims 1 and 28-29, wherein the binder is a lignin or a lignin derivative. The method of claim 30, wherein the lignin derivative is lignosulfonate. A method for altering microbiome in soil, comprising: applying to soil a formulation comprising (1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell and (2) a binder, thereby altering microbiome in soil. The method of embodiment 32, wherein the formulation composition increases a plurality of beneficial microbes in soil. The method of embodiment 33, wherein the beneficial microbe Pseudomonas sp, Achromobacter sp., Bacillus sp., Nitrobacter sp., Nitrosomonas sp., Nitrospira sp., Paenibacillus sp., Rhizobium sp., Streptomyces sp., Trichoderma sp., Alkalihalobacillus hwajinpoensis, Aureobasidium pullulans, Metarhizium anisopliae, Purpureocillium lilacinum, or Pythium oligandrum. The method of embodiment 33 or 34, wherein the beneficial microbe is an indigenous microbe species or a genetically engineered microbe. The method of embodiment 33 or 34, wherein the beneficial microbe enhances production of exopolysaccharides, salicylic acid, siderophores, or 1 -aminocyclopropane- 1 -carboxylate (ACC) deaminase, thereby improving soil health and promoting plant growth and stress tolerance. The method of embodiment 32, wherein the formulation decreases a plurality of non- beneficial microbes in soil. The method of embodiment 37, wherein the non-beneficial microbe is a soil pathogen selected from the group consisting of: Pythium sp., Colletotrichum sp., Fusarium sp.,Curvularia sp., or Sclerotinia sp., Sclerotinia homoeocarpa, Agrobacterium tumefaciens, Sclerotium rolfsii, or Leptosphaerulina americana. The method of embodiment 32, wherein the bio-nutrient composition comprises: i) a biosolid at a final percentage-by-weight of about 35% to about 75% of the bio-nutrient composition; ii) a grain at a final percentage-by-weight of about 10% to about 50% of the bio-nutrient composition; and iii) an algae cell at a final percentage-by-weight of about 5% to about 40% of the bionutrient composition. The method of embodiment 32 or 39, wherein the biosolid is a sewage sludge, a manure or a food waste. The method of any one of embodiments 32 and 39-40, wherein the biosolid is a Class A EQ biosolid, Class A biosolid, or a Class B biosolid. The method of embodiment 32 or 39, wherein the grain is a brewers’ spent grain (BSG) or a distillers grain (DG). The method of any one of embodiments 32, 39, and 42, wherein the grain is a dried brewers’ spent grain (DBSG) or a dried distillers grain (DDG). The method of embodiment 32 or 39, wherein the algae cell is Cyanophyceae^ Chlor ophyceae, Rhodophyceae , or Phaeophyceae . The method of any one of embodiments 32, 39, and 44, wherein the algae cell comprises a dried algae cell wall, algae process water, or a post-harvest post filtration undried algae solution. The method of any one of embodiments 32-45, wherein the biosolid is digested in an anaerobic digester for at least 15 days. The method of any one of embodiments 32-45, wherein the biosolid is dewatered to a moisture content of about 10% or less. The method of any one of embodiments 32-45, wherein the biosolid is pulverized to a mesh size between about 10 and about 300. The method of any one of embodiments 32-45, wherein the biosolid is treated with an oil based agent, wherein the oil based agent prevents billowing. The method of any one of embodiments 32-45, wherein the biosolid and / or the grain are dried at a temperature between about 155° F. and about 250° F.The method of any one of embodiments 32-45, wherein the grain is dewatered to a moisture content of about 10% or less. The method of any one of embodiments 32-45, wherein the grain is pulverized to a mesh size of about 10 to about 300. The method of any one of embodiments 32-45, wherein the algae cell is pressed to isolate an algae cell wall. The method of any one of embodiments 32-45, wherein the algae cell wall is dried to a moisture content of about 20% or less. The method of any one of embodiments 32-45, wherein the algae cell wall is pulverized to a mesh size of about 10 to about 300. The method of embodiment 32, wherein the bio-nutrient composition is passed through a heating source, thereby being dewatered. The method of claim 32, wherein the binder is at a percentage weight of about 1% to about 50% of said formulation. The method of claim 32 or 57, wherein the binder is a lignin, a mineral, a plant starch, or a molasses. The method of any one of claims 32 and 57-58, wherein the binder is a lignin or a lignin derivative. The method of claim 59, wherein the lignin derivative is lignosulfonate. A method for inducing a stress-related defense mechanism in a plant, comprising: applying to a target a formulation comprising (1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell and (2) a binder, thereby improving plant defense response to biotic or abiotic stress. The method of embodiment 61, wherein the target is a plant, a plant part, or a plant cell thereof. The method of embodiment 62, wherein the plant is acacia, alfalfa, amaranth, apple, apricot, artichoke, ash tree, asparagus, avocado, banana, barley, beans, beet, birch, beech, blackberry, black raspberry, blueberry, broccoli, Brussel's sprouts, cabbage, cane berry, canola, cantaloupe, carrot, cassava, cauliflower, cedar, a cereal, celery, chestnut, cherry, Chinese cabbage, citrus, Clementine, clover, coffee, corn, cotton, cowpea, cucumber, cypress, eggplant, elm, endive, eucalyptus, fennel, figs, fir, geranium, grape, grapefruit, groundnuts, ground cherry, gum hemlock, hickory, kale, kiwifruit, kohlrabi, larch, lettuce, leek, lemon, lime, locust, pine, maidenhair, maize, mango, maple, melon, millet,mushroom, mustard, nuts, oak, oats, oil palm, okra, onion, orange, an ornamental plant or flower or tree, papaya, palm, parsley, parsnip, pea, peach, peanut, pear, peat, pepper, persimmon, pigeon pea, peach, pine, pineapple, plantain, plum, pomegranate, potato, pumpkin, radicchio, radish, rapeseed, raspberry, rice, rye, sorghum, safflower, sallow, soybean, spinach, spruce, squash, strawberry, sugar beet, sugarcane, sunflower, sweet potato, sweet com, tangerine, tea, tobacco, tomato, trees, triticale, turf grasses, turnips, vine, walnut, watercress, watermelon, wheat, wild strawberry, yams, yew, or zucchini. The method of embodiment 62, wherein the plant is a grass. The method of embodiment 64, wherein the grass is Poaceae family. The method of embodiment 64 or 65, wherein the grass is Agrostis genus, Poa genus, Lolium genus, Festuca genus, Cynodon genus, Stenotaphrum genus, or Zoysia genus. The method of any one of embodiments 64-66, wherein the grass is a turfgrass, which is bentgrass, Kentucky bluegrass, ryegrass, tall fescue, Chewings fescue, Hard fescue, sheep fescue, creeping red fescue, bermudagrass, kikuyu grass, St. Augustine grass), buffalo grass, zoysia grass, zoyosa tenuifolia, or temple grass. The method of embodiment 61, wherein the target is a soil or an area where a plant is present. The method of embodiment 61, wherein the induced stress-related defense mechanism is selected from the group consisting of: phenylpropanoid biosynthesis, glutathione metabolism, sesquiterpenoid and triterpenoid metabolism, plant-pathogen interaction, ascorbate and aldarate metabolism, carotenoid biosynthesis, plant hormone signal transduction, and linoleic acid metabolism. The method of embodiment 61 or 69, wherein the induced stress-related defense mechanism is associated with:(i) upregulation of the expression of a plurality of gene involved in phenylpropanoid biosynthesis, which is phenylalanine ammonia lyase, trans-cinnamate 4-monooxygenase, and / or caffeoylshikimate esterase,(ii) upregulation of the expression of a plurality of gene involved in glutathione metabolism, which is glutathione dehydroascorbate reductase3, glutathione peroxidase, and / or gamma-glutamylcyclotransferase, '(iii) upregulation of the expression of a plurality of gene involved in sesquiterpenoid and triterpenoid metabolism, which is squalene synthase 1, squalene monooxygenase, and / or(iv) upregulation of the expression of a plurality of gene involved in plant-pathogen interaction, which is pathogenesis related 1, suppressor of G2 allele of SKP1, and / or respiratory burst oxidase,'(v) upregulation of the expression of a plurality of gene involved in ascorbate and aldarate metabolism, which is L-gulonolactone oxidase, GDP-L-galactose phosphorylase, and / or ascorbate peroxidase,(vi) upregulation of the expression of a plurality of gene involved in carotenoid biosynthesis, which is ABA 8'-hydroxylase, lycopene beta-cyclase, and / or 9-cis- epoxy carotenoid dioxygenase, '(vii) upregulation of the expression of a plurality of gene involved in plant hormone signal transduction, which is Not Responsive to PR1 protein (NPR1), jasmonic acid receptor, and / or abscisic acid receptor, ' and / or(viii) upregulation of the expression of a plurality of gene involved in linoleic acid metabolism, which is linoleate 9S-lipoxygenase and / or linoleate 13S-lipoxygenase9. The method of embodiment 61, wherein the bio-nutrient composition comprises: i) a biosolid at a final percentage-by-weight of about 35% to about 75% of the bio-nutrient composition; ii) a grain at a final percentage-by-weight of about 10% to about 50% of the bio-nutrient composition; and iii) an algae cell at a final percentage-by-weight of about 5% to about 40% of the bionutrient composition. The method of embodiment 61 or 71, wherein the biosolid is a sewage sludge, a manure or a food waste. The method of any one of embodiments 61 and 71-72, wherein the biosolid is a Class A EQ biosolid, Class A biosolid, or a Class B biosolid. The method of embodiment 61 or 71, wherein the grain is a brewers’ spent grain (BSG) or a distillers grain (DG). The method of any one of embodiments 61, 71, and 74, wherein the grain is a dried brewers’ spent grain (DBSG) or a dried distillers grain (DDG). The method of embodiment 61 or 71, wherein the algae cell is Cyanophyceae^ Chlor ophyceae, Rhodophyceae , or Phaeophyceae .The method of any one of embodiments 61, 71, and 76, wherein the algae cell comprises a dried algae cell wall, algae process water, or a post-harvest post filtration undried algae solution. The method of any one of embodiments 61-77, wherein the biosolid is digested in an anaerobic digester for at least 15 days. The method of any one of embodiments 61-77, wherein the biosolid is dewatered to a moisture content of about 10% or less. The method of any one of embodiments 61-77, wherein the biosolid is pulverized to a mesh size between about 10 and about 300. The method of any one of embodiments 61-77, wherein the biosolid is treated with an oil based agent, wherein the oil based agent prevents billowing. The method of any one of embodiments 61-77, wherein the biosolid and / or the grain are dried at a temperature between about 155° F. and about 250° F. The method of any one of embodiments 61-77, wherein the grain is dewatered to a moisture content of about 10% or less. The method of any one of embodiments 61-77, wherein the grain is pulverized to a mesh size of about 10 to about 300. The method of any one of embodiments 61-77, wherein the algae cell is pressed to isolate an algae cell wall. The method of any one of embodiments 61-77, wherein the algae cell wall is dried to a moisture content of about 20% or less. The method of any one of embodiments 61-77, wherein the algae cell wall is pulverized to a mesh size of about 10 to about 300. The method of embodiment 61, wherein the bio-nutrient composition is passed through a heating source, thereby being dewatered. The method of claim 61, wherein the binder is at a percentage weight of about 1% to about 50% of said formulation. The method of claim 61 or 89, wherein the binder is a lignin, a mineral, a plant starch, or a molasses. The method of any one of claims 61 and 89-90, wherein the binder is a lignin or a lignin derivative. The method of claim 91, wherein the lignin derivative is lignosulfonate.A method for inducing a photosynthesis mechanism in a plant, comprising: applying to a target a formulation comprising (1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell and (2) a binder, thereby improving plant growth and development. The method of embodiment 93, wherein the target is a plant, a plant part, or a plant cell thereof. The method of embodiment 94, wherein the plant is acacia, alfalfa, amaranth, apple, apricot, artichoke, ash tree, asparagus, avocado, banana, barley, beans, beet, birch, beech, blackberry, black raspberry, blueberry, broccoli, Brussel's sprouts, cabbage, cane berry, canola, cantaloupe, carrot, cassava, cauliflower, cedar, a cereal, celery, chestnut, cherry, Chinese cabbage, citrus, Clementine, clover, coffee, corn, cotton, cowpea, cucumber, cypress, eggplant, elm, endive, eucalyptus, fennel, figs, fir, geranium, grape, grapefruit, groundnuts, ground cherry, gum hemlock, hickory, kale, kiwifruit, kohlrabi, larch, lettuce, leek, lemon, lime, locust, pine, maidenhair, maize, mango, maple, melon, millet, mushroom, mustard, nuts, oak, oats, oil palm, okra, onion, orange, an ornamental plant or flower or tree, papaya, palm, parsley, parsnip, pea, peach, peanut, pear, peat, pepper, persimmon, pigeon pea, peach, pine, pineapple, plantain, plum, pomegranate, potato, pumpkin, radicchio, radish, rapeseed, raspberry, rice, rye, sorghum, safflower, sallow, soybean, spinach, spruce, squash, strawberry, sugar beet, sugarcane, sunflower, sweet potato, sweet com, tangerine, tea, tobacco, tomato, trees, triticale, turf grasses, turnips, vine, walnut, watercress, watermelon, wheat, wild strawberry, yams, yew, or zucchini. The method of embodiment 94, wherein the plant is a grass. The method of embodiment 96, wherein the grass is Poaceae family. The method of embodiment 96 or 97, wherein the grass is Agrostis genus, Poa genus, Lolium genus, Festuca genus, Cynodon genus, Stenotaphrum genus, or Zoysia genus. The method of any one of embodiments 96-98, wherein the grass is a turfgrass, which is bentgrass, Kentucky bluegrass, ryegrass, tall fescue, Chewings fescue, Hard fescue, sheep fescue, creeping red fescue, bermudagrass, kikuyu grass, St. Augustine grass, buffalo grass, zoysia grass, zoyosa tenuifolia, or temple grass. . The method of embodiment 93, wherein the target is a soil or an area where a plant is present. . The method of embodiment 93, wherein the induced photosynthesis mechanism is associated with upregulation of the expression of a plurality of gene, which is photosystemII subunit R, photosystem I subunit IV, plastocyanin, light-harvesting complex I chlorophyll a / b binding protein 2, and / or light-harvesting complex II chlorophyll a / b binding protein 4. . The method of embodiment 93, wherein the bio-nutrient composition comprises: i) a biosolid at a final percentage-by-weight of about 35% to about 75% of the bio-nutrient composition; ii) a grain at a final percentage-by-weight of about 10% to about 50% of the bio-nutrient composition; and iii) an algae cell at a final percentage-by-weight of about 5% to about 40% of the bionutrient composition. . The method of embodiment 93 or 102, wherein the biosolid is a sewage sludge, a manure or a food waste. . The method of any one of embodiments 93 and 102-103, wherein the biosolid is a Class A EQ biosolid, Class A biosolid, or a Class B biosolid. . The method of embodiment 93 or 102, wherein the grain is a brewers’ spent grain (BSG) or a distillers grain (DG). . The method of any one of embodiments 93, 102, and 105, wherein the grain is a dried brewers’ spent grain (DBSG) or a dried distillers grain (DDG). . The method of embodiment 93 or 102, wherein the algae cell is Cyanophyceae^ Chlor ophyceae, Rhodophyceae , or Phaeophyceae . . The method of any one of embodiments 93, 102, and 107, wherein the algae cell comprises a dried algae cell wall, algae process water, or a post-harvest post filtration undried algae solution. . The method of any one of embodiments 93-108, wherein the biosolid is digested in an anaerobic digester for at least 15 days. . The method of any one of embodiments 93-108, wherein the biosolid is dewatered to a moisture content of about 10% or less. . The method of any one of embodiments 93-108, wherein the biosolid is pulverized to a mesh size between about 10 and about 300. . The method of any one of embodiments 93-108, wherein the biosolid is treated with an oil based agent, wherein the oil based agent prevents billowing. . The method of any one of embodiments 93-108, wherein the biosolid and / or the grain are dried at a temperature between about 155° F. and about 250° F.. The method of any one of embodiments 93-108, wherein the grain is dewatered to a moisture content of about 10% or less. . The method of any one of embodiments 93-108, wherein the grain is pulverized to a mesh size of about 10 to about 300. . The method of any one of embodiments 93-108, wherein the algae cell is pressed to isolate an algae cell wall. . The method of any one of embodiments 93-108, wherein the algae cell wall is dried to a moisture content of about 20% or less. . The method of any one of embodiments 93-108, wherein the algae cell wall is pulverized to a mesh size of about 10 to about 300. . The method of embodiment 93, wherein the bio-nutrient composition is passed through a heating source, thereby being dewatered. . The method of claim 93, wherein the binder is at a percentage weight of about 1% to about 50% of said formulation. . The method of claim 93 or 120, wherein the binder is a lignin, a mineral, a plant starch, or a molasses. . The method of any one of claims 93 and 120-121, wherein the binder is a lignin or a lignin derivative. . The method of claim 122, wherein the lignin derivative is lignosulfonate. . A method for inducing nutrient uptake and assimilation in a plant, comprising: applying to a target a formulation comprising (1) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell and (2) a binder, thereby improving nitrogen assimilation or uptake of a plant. . The method of embodiment 124, wherein the target is a plant, a plant part, or a plant cell thereof. . The method of embodiment 125, wherein the plant is acacia, alfalfa, amaranth, apple, apricot, artichoke, ash tree, asparagus, avocado, banana, barley, beans, beet, birch, beech, blackberry, black raspberry, blueberry, broccoli, Brussel's sprouts, cabbage, cane berry, canola, cantaloupe, carrot, cassava, cauliflower, cedar, a cereal, celery, chestnut, cherry, Chinese cabbage, citrus, Clementine, clover, coffee, corn, cotton, cowpea, cucumber, cypress, eggplant, elm, endive, eucalyptus, fennel, figs, fir, geranium, grape, grapefruit, groundnuts, ground cherry, gum hemlock, hickory, kale, kiwifruit, kohlrabi, larch, lettuce, leek, lemon, lime, locust, pine, maidenhair, maize, mango, maple, melon, millet,mushroom, mustard, nuts, oak, oats, oil palm, okra, onion, orange, an ornamental plant or flower or tree, papaya, palm, parsley, parsnip, pea, peach, peanut, pear, peat, pepper, persimmon, pigeon pea, peach, pine, pineapple, plantain, plum, pomegranate, potato, pumpkin, radicchio, radish, rapeseed, raspberry, rice, rye, sorghum, safflower, sallow, soybean, spinach, spruce, squash, strawberry, sugar beet, sugarcane, sunflower, sweet potato, sweet com, tangerine, tea, tobacco, tomato, trees, triticale, turf grasses, turnips, vine, walnut, watercress, watermelon, wheat, wild strawberry, yams, yew, or zucchini. . The method of embodiment 125, wherein the plant is a grass. . The method of embodiment 127, wherein the grass is Poaceae family. . The method of embodiment 127 or 128, wherein the grass is Agrostis genus, Poa genus, Lolium genus, Festuca genus, Cynodon genus, Stenotaphrum genus, or Zoysia genus.. The method of any one of embodiments 127-129, wherein the grass is a turfgrass, which is bentgrass, Kentucky bluegrass, ryegrass, tall fescue, Chewings fescue, Hard fescue, sheep fescue, creeping red fescue, bermudagrass, kikuyu grass, St. Augustine grass, buffalo grass, zoysia grass, zoyosa tenuifolia, or temple grass. . The method of embodiment 124, wherein the target is a soil or an area where a plant is present. . The method of embodiment 124, wherein the induced nutrient uptake and assimilation is associated with upregulation of the expression of a plurality of gene, which is nitrate reductase, nitrite reductase, high-affinity nitrate transporter 2.1 (HAT2.1), high-affinity nitrate transporter 2.2 (HAT2.2), high-affinity nitrate transporter 2.3 (HAT2.3), glutamine synthetase, and / or mugineic acid- 3 dioxygenase. . The method of embodiment 124, wherein the bio-nutrient composition comprises: i) a biosolid at a final percentage-by-weight of about 35% to about 75% of the bio-nutrient composition; ii) a grain at a final percentage-by-weight of about 10% to about 50% of the bio-nutrient composition; and iii) an algae cell at a final percentage-by-weight of about 5% to about 40% of the bionutrient composition. . The method of embodiment 124 or 133, wherein the biosolid is a sewage sludge, a manure or a food waste. . The method of any one of embodiments 124 and 133-134, wherein the biosolid is a Class A EQ biosolid, Class A biosolid, or a Class B biosolid.I l l. The method of embodiment 124 or 133, wherein the grain is a brewers’ spent grain(BSG) or a distillers grain (DG). . The method of any one of embodiments 124, 133, and 136, wherein the grain is a dried brewers’ spent grain (DBSG) or a dried distillers grain (DDG). . The method of embodiment 124 or 133, wherein the algae cell is Cyanophyceae^ Chlor ophyceae. Rhodophyceae , or Phaeophyceae . . The method of any one of embodiments 124, 133, and 138, wherein the algae cell comprises a dried algae cell wall, algae process water, or a post-harvest post filtration undried algae solution. . The method of any one of embodiments 124-139, wherein the biosolid is digested in an anaerobic digester for at least 15 days. . The method of any one of embodiments 124-139, wherein the biosolid is dewatered to a moisture content of about 10% or less. . The method of any one of embodiments 124-139, wherein the biosolid is pulverized to a mesh size between about 10 and about 300. . The method of any one of embodiments 124-139, wherein the biosolid is treated with an oil based agent, wherein the oil based agent prevents billowing. . The method of any one of embodiments 124-139, wherein the biosolid and / or the grain are dried at a temperature between about 155° F. and about 250° F. . The method of any one of embodiments 124-139, wherein the grain is dewatered to a moisture content of about 10% or less. . The method of any one of embodiments 124-139, wherein the grain is pulverized to a mesh size of about 10 to about 300. . The method of any one of embodiments 124-139, wherein the algae cell is pressed to isolate an algae cell wall. . The method of any one of embodiments 124-139, wherein the algae cell wall is dried to a moisture content of about 20% or less. . The method of any one of embodiments 124-139, wherein the algae cell wall is pulverized to a mesh size of about 10 to about 300. . The method of embodiment 124, wherein the bio-nutrient composition is passed through a heating source, thereby being dewatered. . The method of claim 124, wherein the binder is at a percentage weight of about 1% to about 50% of said formulation.. The method of claim 124 or 151, wherein the binder is a lignin, a mineral, a plant starch, or a molasses. . The method of any one of claims 124 and 151-152, wherein the binder is a lignin or a lignin derivative. . The method of claim 153, wherein the lignin derivative is lignosulfonate.

Claims

CLAIMS:

1. A formulation comprising:(a) a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell;(b) a binder; and(c) optionally an additive.

2. The formulation of claim 1, wherein the additive is present.

3. The formulation of claim 1, wherein the additive is not present.

4. The formulation of claim 1, wherein the bio-nutrient composition comprises: i) a biosolid at a final percentage-by-weight of about 35% to about 75% of the bio-nutrient composition; ii) a grain at a final percentage-by-weight of about 10% to about 50% of the bio-nutrient composition; and iii) an algae cell at a final percentage-by-weight of about 5% to about 40% of the bionutrient composition.

5. The formulation of claim 1 or 4, wherein the biosolid is a sewage sludge, a manure or a food waste.

6. The formulation of any one of claims 1 and 4-5, wherein the biosolid is a Class A EQ biosolid, Class A biosolid, or a Class B biosolid.

7. The formulation of claim 1 or 4, wherein the grain is a brewers’ spent grain (BSG) or a distillers grain (DG).

8. The formulation of any one of claims 1, 4, and 7, wherein the grain is a dried brewers’ spent grain (DBSG) or a dried distillers grain (DDG).

9. The formulation of claim 1 or 4, wherein the algae cell is Cyanophyceae^ Chlor ophyceae. Rhodophyceae, or Phaeophyceae .

10. The formulation of any one of claims 1, 4, and 9, wherein the algae cell comprises a dried algae cell wall, algae process water, or a post-harvest post filtration undried algae solution.

11. The formulation of claim 1, wherein the binder is at a percentage weight of about 1% to about 50% of said formulation.

12. The formulation of claim 1 or 11, wherein the binder is a lignin, a mineral, a plant starch, or a molasses.

13. The formulation of any one of claims 1 and 11-12, wherein the binder is a lignin or a lignin derivative.

14. The formulation of claim 13, wherein the lignin derivative is lignosulfonate.

15. The formulation of any one of claims 1-3, wherein the additive is an odor-reducing or odorneutralizing agent.

16. The formulation of claim any one of claims 1-3 and 15, wherein the additive is selected from the group consisting of: an order neutralizer, an odor-masking agent, an essential oil, an odor absorbent, an enzyme, or a microbe.

17. The formulation of claim 16, wherein the odor neutralizer is an activated charcoal, a zeolite, a baking soda, a silica gel, or a compound comprising cyclodextrin.

18. The formulation of claim 16, wherein the odor-masking agent is a vanilla extract or a citrus extract.

19. The formulation of claim 16, wherein the essential oil is a lavender oil, a eucalyptus oil, a peppermint oil, a lemon oil, a tea tree oil, a clove oil, an orange oil, or a cinnamon oil.

20. The formulation of claim 16, wherein the odor absorbent is bentonite, kaolin clay, or peat moss.

21. The formulation of claim 16, wherein the enzyme is a protease, a lipase, an amylase, a cellulase, a urease, an oxidoreductase, or a pectinase.

22. The formulation of claim 16, wherein the microbe is a species of genus Bacillus or Pseudomonas.

23. The formulation of any one of claims 1-22, wherein the formulation is in a powder form.

24. The formulation of any one of claims 1-22, wherein the formulation is in a granular form.

25. The formulation of any one of claims 1-22, wherein the formulation is in a liquid form.

26. The formulation of claim 24, wherein the granular form is a prill form.

27. The formulation of claim 25, wherein the liquid form of said formation is a suspension or an extract.

28. The formulation of claim 25, wherein the liquid form of the formulation further comprises an enhancer.

29. The formulation of claim 28, wherein the enhancer is selected from the consisting of: a preservative, a stabilizer, a buffering agent, a microbial inhibitor, a pH adjuster, and an antifoaming agent.

30. The formulation of claim 29, wherein the preservative is sodium benzoate, potassium sorbate, or citric acid.

31. The formulation of claim 29, wherein the stabilizer is a xanthan gum, a chelating agent or an antioxidant.

32. The formulation of claim 31, wherein the chelating agent is ethylenediaminetetraacetic acid (EDTA).

33. The formulation of claim 31, wherein the antioxidant is an ascorbic acid or a tocopherol.

34. The formulation of claim 29, wherein the buffering agent is phosphate, phosphate derivative, acetate, or acetate derivative.

35. The formulation of claim 29, wherein the microbial inhibitor is an essential oil or a silver nanoparticle.

36. The formulation of claim 35, wherein the essential oil is a thyme oil, an oregano oil, a peppermint oil, or a mint oil.

37. The formulation of claim 29, wherein the pH adjuster is hydrochloric acid or sodium hydroxide.

38. The formulation of claim 29, wherein the anti-foaming agent is a silicone-based antifoam.

39. The formulation of any one of claims 1-10, wherein the biosolid is digested in an anaerobic digester for at least 15 days.

40. The formulation of any one of claims 1-10, wherein the biosolid is dewatered to a moisture content of about 10% or less.

41. The formulation of any one of claims 1-10, wherein the biosolid is pulverized to a mesh size between about 10 and about 300.

42. The formulation of any one of claims 1-10, wherein the biosolid is treated with an oil based agent, wherein the oil based agent prevents billowing.

43. The formulation of any one of claims 1-10, wherein the biosolid and / or the grain are dried at a temperature between about 155° F. and about 250° F.

44. The formulation of any one of claims 1-10, wherein the grain is dewatered to a moisture content of about 10% or less.

45. The formulation of any one of claims 1-10, wherein the grain is pulverized to a mesh size of about 10 to about 300.

46. The formulation of any one of claims 1-10, wherein the algae cell is pressed to isolate an algae cell wall.

47. The formulation of any one of claims 1-10, wherein the algae cell wall is dried to a moisture content of about 20% or less.

48. The formulation of any one of claims 1-10, wherein the algae cell wall is pulverized to a mesh size of about 10 to about 300.

49. The formulation of claim 1, wherein the bio-nutrient composition is passed through a heating source, thereby being dewatered.

50. The formulation of any one of claims 1-49, wherein the formulation enhances activity and / or diversity of a plurality of beneficial microbes in soil to which the formulation is applied, when comparing to the soil untreated with the formulation.

51. The formulation of claim 50, wherein the beneficial microbe is Pseudomonas sp, Achromobacter sp., Bacillus sp., Nitrobacter sp., Nitrosomonas sp., Nitrospira sp., Paenibacillus sp., Rhizobium sp., Streptomyces sp., Trichoderma sp., Alkalihalobacillus hwajinpoensis, Aureobasidium pullulans, Metarhizium anisopliae, Purpureocillium lilacinum, or Pythium oligandrum.

52. The formation of claim 50 or 51, wherein the beneficial microbe is an indigenous microbe species or a genetically engineered microbe.

53. The formation of claim 50 or 51, wherein the beneficial microbe enhances production of exopolysaccharides, salicylic acid, siderophores, or 1 -aminocyclopropane- 1 -carboxylate (ACC) deaminase, thereby improving soil health and promoting plant growth and stress tolerance.

54. The formation of claim 50 or 51, wherein the formulation inhibits a plurality of soil pathogens.

55. The formation of claim 54, wherein the soil pathogen is Pythium sp., Colle totrichum sp., Fusarium sp., Curvularia sp., Sclerotinia sp., Sclerotinia homoeocarpa, Agrobacterium tumefaciens, Sclerotium rolfsii, or Leptosphaerulina americana.

56. The formulation of any one of claims 1-49, wherein the formulation induces expression of a plurality of genes involved in a stress-related defense mechanism in plants to which the formulation is applied, when comparing to plants untreated with the formulation.

57. The formulation of claim 56, wherein the induced stress-related defense mechanism is selected from the group consisting of: phenylpropanoid biosynthesis, glutathione metabolism, sesquiterpenoid and triterpenoid metabolism, plant-pathogen interaction, ascorbate and aldarate metabolism, carotenoid biosynthesis, plant hormone signal transduction, and linoleic acid metabolism.

58. The formulation of claim 56 or 57, wherein the induced stress-related defense mechanism is associated with:(i) upregulation of the expression of a plurality of gene involved in phenylpropanoid biosynthesis, which is phenylalanine ammonia lyase, trans-cinnamate 4-monooxygenase , and / or caffeoylshikimate esterase,(ii) upregulation of the expression of a plurality of gene involved in glutathione metabolism, which is glutathione dehydroascorbate reductase3, glutathione peroxidase, and / or gamma-glutamylcyclotransferase, '(iii) upregulation of the expression of a plurality of gene involved in sesquiterpenoid and triterpenoid metabolism, which is squalene synthase 1, squalene monooxygenase, and / or NAD+ dependent fame sal dehydrogenase,'(iv) upregulation of the expression of a plurality of gene involved in plant-pathogen interaction, which is pathogenesis related 1, suppressor of G2 allele of SKP1, and / or respiratory burst oxidase,'(v) upregulation of the expression of a plurality of gene involved in ascorbate and aldarate metabolism, which is L-gulonolactone oxidase, GDP-L-galactose phosphorylase, and / or ascorbate peroxidase, '(vi) upregulation of the expression of a plurality of gene involved in carotenoid biosynthesis, which is ABA 8'-hydroxylase, lycopene beta-cyclase, and / or 9-cis- epoxy carotenoid dioxygenase, '(vii) upregulation of the expression of a plurality of gene involved in plant hormone signal transduction, which is Not Responsive to PR1 protein (NPR1), jasmonic acid receptor, and / or abscisic acid receptor, ' and / or(viii) upregulation of the expression of a plurality of gene involved in linoleic acid metabolism, which is linoleate 9S-lipoxygenase and / or linoleate 13S-lipoxygenase9.

59. The formulation of any one of claims 1-49, wherein the formulation induces expression of a plurality of genes involved in photosynthesis mechanism for increasing plant growth and development in plants to which the formulation is applied, when comparing to plants untreated with the formulation.

60. The formulation of claim 59, wherein the induced photosynthesis mechanism is associated with upregulation of the expression of a plurality of gene, which is photosystem II subunit R, photosystem I subunit IV, plastocyanin, light-harvesting complex I chlorophyll a / b binding protein 2, and / or light-harvesting complex II chlorophyll a / b binding protein 4.

61. The formulation of any one of claims 1-49, wherein the formulation induces expression of a plurality of genes involved in nutrient uptake and assimilation in plants to which the formulation is applied, when comparing to plants untreated with the formulation.

62. The formulation of claim 61, wherein the induced nutrient uptake and assimilation is associated with upregulation of the expression of a plurality of gene, which is nitrate reductase, nitrite reductase, high-affinity nitrate transporter 2.1 (HAT2.1), high-affinity nitrate transporter 2.2 (HAT2.2), high-affinity nitrate transporter 2.3 (HAT2.3), glutamine synthetase, and / or mugineic acid- 3 dioxygenase.

63. The formulation of any one of claims 1-62, wherein the formulation is applied with an agricultural product sequentially or simultaneously.

64. The formation of claim 63, wherein the agricultural product is a mineral based fertilizer, synthetic fertilizer, an organic fertilizer, a micronutrient, a biostimulant, a plurality of microbes, a microbial additive, or an agricultural or landscaping chemical selected from the group consisting of a pesticide, an herbicide, a fungicide, and a surfactant.

65. A method of making a granular formulation comprising the steps of:(a) dry milling a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell;(b) blending the dry milled bio-nutrient composition from step (a) in a powder form;(c) adding a binder to the blended bio-nutrient composition from step (b) to produce a formulation;(d) optionally, adding an additive; and(e) granulating the formulation from step (c) or (d).

66. The method of claim 65, wherein the additive is added to the formulation from step (d).

67. The method of claim 65, wherein the additive is not added to the formulation from step (d).

68. The method of claim 65, wherein the bio-nutrient composition comprises: i) a biosolid at a final percentage-by-weight of about 35% to about 75% of the bio-nutrient composition; ii) a grain at a final percentage-by-weight of about 10% to about 50% of the bio-nutrient composition; and iii) an algae cell at a final percentage-by-weight of about 5% to about 40% of the bionutrient composition.

69. The method of claim 65 or 68, wherein the biosolid is a sewage sludge, a manure or a food waste.

70. The method of any one of claims 65 and 68-69, wherein the biosolid is a Class A EQ biosolid, Class A biosolid, or a Class B biosolid.

71. The method of claim 65 or 68, wherein the grain is a brewers’ spent grain (BSG) or a distillers grain (DG).

72. The method of any one of claims 65, 68, and 71, wherein the grain is a dried brewers’ spent grain (DBSG) or a dried distillers grain (DDG).

73. The method of claim 65 or 68, wherein the algae cell is Cyanophyceae, Chlor ophyceae. Rhodophyceae, or Phaeophyceae .

74. The method of any one of claims 65, 68, and 73, wherein the algae cell comprises a dried algae cell wall.

75. The method of claim 65, wherein the binder is at a percentage weight of about 1% to about 50% of said formulation.

76. The method of claim 65 or 75, wherein the binder is a lignin, a mineral, a plant starch, or a molasses.

77. The method of any one of claims 65 and 75-76, wherein the binder is a lignin or a lignin derivative.

78. The method of claim 77, wherein the lignin derivative is lignosulfonate.

79. The method of any one of claims 65-67, wherein the additive is an odor-reducing or odorneutralizing agent.

80. The method of any one of claims 65-67 and 79, wherein the additive is selected from the group consisting of: an order neutralizer, an odor-masking agent, an essential oil, an odor absorbent, an enzyme, or a microbe.

81. The method of claim 80, wherein the odor neutralizer is an activated charcoal, a zeolite, a baking soda, a silica gel, or a compound comprising cyclodextrin.

82. The method of claim 80, wherein the odor-masking agent is a vanilla extract or a citrus extract.

83. The method of claim 80, wherein the essential oil is a lavender oil, a eucalyptus oil, a peppermint oil, a lemon oil, a tea tree oil, a clove oil, an orange oil, or a cinnamon oil.

84. The method of claim 80, wherein the odor absorbent is bentonite, kaolin clay, or peat moss.

85. The method of claim 80, wherein the enzyme is a protease, a lipase, an amylase, a cellulase, a urease, an oxidoreductase, or a pectinase.

86. The method of claim 80, wherein the microbe is a species of genus Bacillus orPseudomonas .

87. The method of any one of claims 65-74, wherein the biosolid is digested in an anaerobic digester for at least 15 days.

88. The method of any one of claims 65-74, wherein the biosolid is dewatered to a moisture content of about 10% or less.

89. The method of any one of claims 65-74, wherein the biosolid is pulverized to a mesh size between about 10 and about 300.

90. The method of any one of claims 65-74, wherein the biosolid is treated with an oil based agent, wherein the oil based agent prevents billowing.

91. The method of any one of claims 65-74, wherein the biosolid and / or the grain are dried at a temperature between about 155° F. and about 250° F.

92. The method of any one of claims 65-74, wherein the grain is dewatered to a moisture content of about 10% or less.

93. The method of any one of claims 65-74, wherein the grain is pulverized to a mesh size of about 10 to about 300.

94. The method of any one of claims 65-74, wherein the algae cell is pressed to isolate an algae cell wall.

95. The method of any one of claims 65-74, wherein the algae cell wall is dried to a moisture content of about 20% or less.

96. The method of any one of claims 65-74, wherein the algae cell wall is pulverized to a mesh size of about 10 to about 300.

97. The method of claim 65, wherein the bio-nutrient composition is passed through a heating source, thereby being dewatered.

98. A method of making a liquid formulation comprising the steps of:(a) dry milling a bio-nutrient composition comprising i) a biosolid, ii) a grain, and iii) an algae cell;(b) agitating the dry milled bio-nutrient composition from step (a) in water with a speed mixer for wet milling; and(c) adding a liquid form of a binder to the wet milled bio-nutrient composition to produce a liquid formulation;(d) optionally adding an additive; and(e) filtering the formulation from step (c) or (d).

99. The method of claim 98, wherein the additive is added to the formulation from step (d).

100. The method of claim 98, wherein the additive is not added to the formulation from step(d).

101. The method of claim 98, wherein the bio-nutrient composition comprises: i) a biosolid at a final percentage-by-weight of about 35% to about 75% of the bio-nutrient composition; ii) a grain at a final percentage-by-weight of about 10% to about 50% of the bio-nutrient composition; and iii) an algae cell at a final percentage-by-weight of about 5% to about 40% of the bionutrient composition.

102. The method of claim 98 or 101, wherein the biosolid is a sewage sludge, a manure or a food waste.

103. The method of any one of claims 98 and 101-102, wherein the biosolid is a Class A EQ biosolid, Class A biosolid, or a Class B biosolid.

104. The method of claim 98 or 101, wherein the grain is a brewers’ spent grain (BSG) or a distillers grain (DG).

105. The method of any one of claims 98, 101, and 104, wherein the grain is a dried brewers’ spent grain (DBSG) or a dried distillers grain (DDG).

106. The method of claim 98 or 101, wherein the algae cell is Cyanophyceae, Chlor ophyceae. Rhodophyceae , or Phaeophyceae .

107. The method of any one of claims 98, 101, and 106, wherein the algae cell comprises a dried algae cell wall or a post-harvest post filtration undried algae solution.

108. The method of claim 98, wherein the binder is at a percentage weight of about 1% to about 50% of said formulation.

109. The method of claim 98 or 108, wherein the binder is a lignin, a mineral, a plant starch, or a molasses.

110. The method of any one of claims 98 and 108-109, wherein the binder is a lignin or a lignin derivative.

111. The method of claim 110, wherein the lignin derivative is lignosulfonate.

112. The method of claim 98, wherein the additive is an odor-reducing or odor-neutralizing agent.

113. The method of any one of claims 98-100 and 112, wherein the additive is selected from the group consisting of: an order neutralizer, an odor-masking agent, an essential oil, an odor absorbent, an enzyme, or a microbe.

114. The method of claim 113, wherein the odor neutralizer is an activated charcoal, a zeolite, a baking soda, a silica gel, or a compound comprising cyclodextrin.

115. The method of claim 113, wherein the odor-masking agent is a vanilla extract or a citrus extract.

116. The method of claim 113, wherein the essential oil is a lavender oil, a eucalyptus oil, a peppermint oil, a lemon oil, a tea tree oil, a clove oil, an orange oil, or a cinnamon oil.

117. The method of claim 113, wherein the odor absorbent is bentonite, kaolin clay, or peat moss.

118. The method of claim 113, wherein the enzyme is a protease, a lipase, an amylase, a cellulase, a urease, an oxidoreductase, or a pectinase.

119. The method of claim 113, wherein the microbe is a species of genus Bacillus or Pseudomonas.

120. The method of any one of claims 98-107, wherein the biosolid is digested in an anaerobic digester for at least 15 days.

121. The method of any one of claims 98-107, wherein the biosolid is dewatered to a moisture content of about 10% or less.

122. The method of any one of claims 98-107, wherein the biosolid is pulverized to a mesh size between about 10 and about 300.

123. The method of any one of claims 98-107, wherein the biosolid is treated with an oil based agent, wherein the oil based agent prevents billowing.

124. The method of any one of claims 98-107, wherein the biosolid and / or the grain are dried at a temperature between about 155° F. and about 250° F.

125. The method of any one of claims 98-107, wherein the grain is dewatered to a moisture content of about 10% or less.

126. The method of any one of claims 98-107, wherein the grain is pulverized to a mesh size of about 10 to about 300.

127. The method of any one of claims 98-107, wherein the algae cell is pressed to isolate an algae cell wall.

128. The method of any one of claims 98-107, wherein the algae cell wall is dried to a moisture content of about 20% or less.

129. The method of any one of claims 98-107, wherein the algae cell wall is pulverized to a mesh size of about 10 to about 300.

130. The method of claim 98, wherein the bio-nutrient composition is passed through a heating source, thereby being dewatered.

131. The method of claim 98, wherein the liquid formulation further comprises an enhancer.

132. The method of claim 131, wherein the enhancer is selected from the consisting of: a preservative, a stabilizer, a buffering agent, a microbial inhibitor, a pH adjuster, and an antifoaming agent.

133. The method of claim 132, wherein the preservative is sodium benzoate, potassium sorbate, or citric acid.

134. The method of claim 132, wherein the stabilizer is a xanthan gum, a chelating agent or an antioxidant.

135. The method of claim 134, wherein the chelating agent is ethylenediaminetetraacetic acid (EDTA).

136. The method of claim 134, wherein the antioxidant is an ascorbic acid or a tocopherol.

137. The method of claim 132 wherein the buffering agent is phosphate, phosphate derivative, acetate, or acetate derivative.

138. The method of claim 132, wherein the microbial inhibitor is an essential oil or a silver nanoparticle.

139. The method of claim 138, wherein the essential oil is a thyme oil, an oregano oil, a peppermint oil, or a mint oil.

140. The method of claim 132, wherein the pH adjuster is hydrochloric acid or sodium hydroxide.

141. The method of claim 132, wherein the anti-foaming agent is a silicone-based antifoam.

142. A method for improving soil health, comprising: applying the formulation of any one of claims 1-49 to soil, thereby improving soil health.

143. The method of claim 142, wherein the formulation improves a balanced ratio of essential nutrients selected from the group consisting of: nitrogen (N), Phosphorus (P), Potassium (K), Calcium (Ca), Magnesium (Mg), Sulfur (S), Carbon (C), Hydrogen (H), Oxygen (O), and micronutrient.

144. The method of claim 143, wherein the micronutrient is selected from the group consisting of: Iron (Fe), Manganese (Mn), Zinc (Zn), Copper (Cu), Boron (B), Molybdenum (Mo), and Chlorine (Cl).

145. The method of claim 142, wherein the formulation increases activity and / or diversity of a plurality of beneficial microbes in soil.

146. The method of claim 145, wherein the beneficial microbe is Pseudomonas sp, Achromobacter sp., Bacillus sp., Nitrobacter sp., Nitrosomonas sp., Nitrospira sp., Paenibacillus sp., Rhizobium sp., Streptomyces sp., Trichoderma sp., Alkalihalobacillus hwajinpoensis, Aureobasidium pullulans, Metarhizium anisopliae, Purpureocillium lilacinum, or Pythium oligandrum.

147. The method of claim 145 or 146, wherein the beneficial microbe is an indigenous microbe species or a genetically engineered microbe.

148. The method of claim 145 or 146, wherein the beneficial microbe enhances production of exopolysaccharides, salicylic acid, siderophores, or 1 -aminocyclopropane- 1 -carboxylate (ACC) deaminase, thereby improving soil health and promoting plant growth and stress tolerance.

149. The method of claim 142, wherein the formulation decreases activity and / or diversity of a plurality of non-beneficial microbes in soil.

150. The method of claim 142, wherein the non-beneficial microbe is a soil pathogen selected from the group consisting of: Pythium sp., Colletotrichum sp., Fusarium sp., Curvularia sp., Sclerotinia sp., Sclerotinia homoeocarpa, Agrobacterium tumefaciens, Sclerotium rolfsii, or Leptosphaerulina americana.

151. A method for altering microbiome in soil, comprising: applying the formulation of any one of claims 1-49 to soil, thereby altering microbiome in soil.

152. The method of claim 151, wherein the formulation increases a plurality of beneficial microbes in soil.

153. The method of claim 152, wherein the beneficial microbe is Pseudomonas sp, Achromobacter sp., Bacillus sp., Nitrobacter sp., Nitrosomonas sp., Nitrospira sp., Paenibacillus sp., Rhizobium sp., Streptomyces sp., Trichoderma sp., Alkalihalobacillus hwajinpoensis, Aureobasidium pullulans, Metarhizium anisopliae, Purpureocillium lilacinum, or Pythium oligandrum.

154. The method of claim 152 or 153, wherein the beneficial microbe is an indigenous microbe species or a genetically engineered microbe.

155. The method of claim 152 or 153, wherein the beneficial microbe enhances production of exopolysaccharides, salicylic acid, siderophores, or 1 -aminocyclopropane- 1 -carboxylate (ACC) deaminase, thereby improving soil health and promoting plant growth and stress tolerance.

156. The method of claim 151, wherein the formulation decreases a plurality of non- beneficial microbes in soil.

157. The method of claim 156, wherein the non-beneficial microbe is a soil pathogen selected from the group consisting of: Pythium sp., Colletotrichum sp., Fusarium sp., Curvularia sp., or Sclerotinia sp., Sclerotinia homoeocarpa, Agrobacterium tumefaciens, Sclerotium rolfsii, or Leptosphaerulina americana.

158. A method for inducing a stress-related defense mechanism in a plant, comprising: applying the formulation of any one of claims 1-49 to a target, thereby improving plant defense response to biotic or abiotic stress.

159. The method of claim 158, wherein the target is a plant, a plant part, or a plant cell thereof.

160. The method of claim 159, wherein the plant is acacia, alfalfa, amaranth, apple, apricot, artichoke, ash tree, asparagus, avocado, banana, barley, beans, beet, birch, beech, blackberry, black raspberry, blueberry, broccoli, Brussel's sprouts, cabbage, cane berry, canola, cantaloupe, carrot, cassava, cauliflower, cedar, a cereal, celery, chestnut, cherry, Chinese cabbage, citrus, Clementine, clover, coffee, corn, cotton, cowpea, cucumber, cypress, eggplant, elm, endive, eucalyptus, fennel, figs, fir, geranium, grape, grapefruit, groundnuts, ground cherry, gum hemlock, hickory, kale, kiwifruit, kohlrabi, larch, lettuce, leek, lemon, lime, locust, pine, maidenhair, maize, mango, maple, melon, millet, mushroom, mustard, nuts, oak, oats, oil palm, okra, onion, orange, an ornamental plant or flower or tree, papaya, palm, parsley, parsnip, pea, peach, peanut, pear, peat, pepper, persimmon, pigeon pea, peach, pine, pineapple, plantain, plum, pomegranate, potato, pumpkin, radicchio, radish, rapeseed, raspberry, rice, rye, sorghum, safflower, sallow, soybean, spinach, spruce, squash, strawberry, sugar beet, sugarcane, sunflower, sweet potato, sweet com, tangerine, tea, tobacco, tomato, trees, triticale, turf grasses, turnips, vine, walnut, watercress, watermelon, wheat, wild strawberry, yams, yew, or zucchini.

161. The method of claim 159, wherein the plant is a grass.

162. The method of claim 161, wherein the grass is a Poaceae family.

163. The method of claim 161 or 162, wherein the grass is Agrostis genus, Poa genus, Lolium genus, Festuca genus, Cynodon genus, Stenotaphrum genus, or Zoysia genus.

164. The method of any one of claims 161-163, wherein the grass is a turfgrass, which is bentgrass, Kentucky bluegrass, ryegrass, tall fescue, Chewings fescue, Hard fescue, sheepfescue, creeping red fescue, bermudagrass, kikuyu grass, St. Augustine grass), buffalo grass, zoysia grass, zoyosa tenuifolia, or temple grass.

165. The method of claim 158, wherein the target is a soil or an area where a plant is present.

166. The method of claim 158, wherein the induced stress-related defense mechanism is selected from the group consisting of: phenylpropanoid biosynthesis, glutathione metabolism, sesquiterpenoid and triterpenoid metabolism, plant-pathogen interaction, ascorbate and aldarate metabolism, carotenoid biosynthesis, plant hormone signal transduction, and linoleic acid metabolism.

167. The method of claim 158 or 166, wherein the induced stress-related defense mechanism is associated with:(i) upregulation of the expression of a plurality of gene involved in phenylpropanoid biosynthesis, which is phenylalanine ammonia lyase, trans-cinnamate 4-monooxygenase , and / or caffeoylshikimate esterase,(ii) upregulation of the expression of a plurality of gene involved in glutathione metabolism, which is glutathione dehydroascorbate reductase3, glutathione peroxidase, and / or gamma-glutamylcyclotransferase, '(iii) upregulation of the expression of a plurality of gene involved in sesquiterpenoid and triterpenoid metabolism, which is squalene synthase 1, squalene monooxygenase, and / or NAD+ dependent fame sal dehydrogenase,'(iv) upregulation of the expression of a plurality of gene involved in plant-pathogen interaction, which is pathogenesis related 1, suppressor of G2 allele of SKP1, and / or respiratory burst oxidase,'(v) upregulation of the expression of a plurality of gene involved in ascorbate and aldarate metabolism, which is L-gulonolactone oxidase, GDP-L-galactose phosphorylase, and / or ascorbate peroxidase, '(vi) upregulation of the expression of a plurality of gene involved in carotenoid biosynthesis, which is ABA 8'-hydroxylase, lycopene beta-cyclase, and / or 9-cis- epoxy carotenoid dioxygenase, '(vii) upregulation of the expression of a plurality of gene involved in plant hormone signal transduction, which is Not Responsive to PR1 protein (NPR1), jasmonic acid receptor, and / or abscisic acid receptor, ' and / or(viii) upregulation of the expression of a plurality of gene involved in linoleic acid metabolism, which is linoleate 9S-lipoxygenase and / or linoleate 13S-lipoxygenase9.

168. A method for inducing a photosynthesis mechanism in a plant, comprising: applying the formulation of any one of claims 1-49 to a target, thereby improving plant growth and development.

169. The method of claim 168, wherein the target is a plant, a plant part, or a plant cell thereof.

170. The method of claim 169, wherein the plant is acacia, alfalfa, amaranth, apple, apricot, artichoke, ash tree, asparagus, avocado, banana, barley, beans, beet, birch, beech, blackberry, black raspberry, blueberry, broccoli, Brussel's sprouts, cabbage, cane berry, canola, cantaloupe, carrot, cassava, cauliflower, cedar, a cereal, celery, chestnut, cherry, Chinese cabbage, citrus, Clementine, clover, coffee, corn, cotton, cowpea, cucumber, cypress, eggplant, elm, endive, eucalyptus, fennel, figs, fir, geranium, grape, grapefruit, groundnuts, ground cherry, gum hemlock, hickory, kale, kiwifruit, kohlrabi, larch, lettuce, leek, lemon, lime, locust, pine, maidenhair, maize, mango, maple, melon, millet, mushroom, mustard, nuts, oak, oats, oil palm, okra, onion, orange, an ornamental plant or flower or tree, papaya, palm, parsley, parsnip, pea, peach, peanut, pear, peat, pepper, persimmon, pigeon pea, peach, pine, pineapple, plantain, plum, pomegranate, potato, pumpkin, radicchio, radish, rapeseed, raspberry, rice, rye, sorghum, safflower, sallow, soybean, spinach, spruce, squash, strawberry, sugar beet, sugarcane, sunflower, sweet potato, sweet com, tangerine, tea, tobacco, tomato, trees, triticale, turf grasses, turnips, vine, walnut, watercress, watermelon, wheat, wild strawberry, yams, yew, or zucchini.

171. The method of claim 169, wherein the plant is a grass.

172. The method of claim 171, wherein the grass is a Poaceae family.

173. The method of claim 171 or 172, wherein the grass is Agrostis genus, Poa genus, Lolium genus, Festuca genus, Cynodon genus, Stenotaphrum genus, or Zoysia genus.

174. The method of any one of claims 171-173, wherein the grass is a turfgrass, which is bentgrass, Kentucky bluegrass, ryegrass, tall fescue, Chewings fescue, Hard fescue, sheep fescue, creeping red fescue, bermudagrass, kikuyu grass, St. Augustine grass, buffalo grass, zoysia grass, zoyosa tenuifolia, or temple grass.

175. The method of claim 168, wherein the target is a soil or an area where a plant is present.

176. The method of claim 168, wherein the induced photosynthesis mechanism is associated with upregulation of the expression of a plurality of gene, which is photosystem II subunit R, photosystem I subunit IV, plastocyanin, light-harvesting complex I chlorophyll a / b binding protein 2, and / or light-harvesting complex II chlorophyll a / b binding protein 4.

177. A method for inducing nutrient uptake and assimilation in a plant, comprising: applying the formulation of any one of claims 1-49 to a target, thereby improving nutrient uptake and assimilation of a plant.

178. The method of claim 177, wherein the target is a plant, a plant part, or a plant cell thereof.

179. The method of claim 178, wherein the plant is acacia, alfalfa, amaranth, apple, apricot, artichoke, ash tree, asparagus, avocado, banana, barley, beans, beet, birch, beech, blackberry, black raspberry, blueberry, broccoli, Brussel's sprouts, cabbage, cane berry, canola, cantaloupe, carrot, cassava, cauliflower, cedar, a cereal, celery, chestnut, cherry, Chinese cabbage, citrus, Clementine, clover, coffee, corn, cotton, cowpea, cucumber, cypress, eggplant, elm, endive, eucalyptus, fennel, figs, fir, geranium, grape, grapefruit, groundnuts, ground cherry, gum hemlock, hickory, kale, kiwifruit, kohlrabi, larch, lettuce, leek, lemon, lime, locust, pine, maidenhair, maize, mango, maple, melon, millet, mushroom, mustard, nuts, oak, oats, oil palm, okra, onion, orange, an ornamental plant or flower or tree, papaya, palm, parsley, parsnip, pea, peach, peanut, pear, peat, pepper, persimmon, pigeon pea, peach, pine, pineapple, plantain, plum, pomegranate, potato, pumpkin, radicchio, radish, rapeseed, raspberry, rice, rye, sorghum, safflower, sallow, soybean, spinach, spruce, squash, strawberry, sugar beet, sugarcane, sunflower, sweet potato, sweet com, tangerine, tea, tobacco, tomato, trees, triticale, turf grasses, turnips, vine, walnut, watercress, watermelon, wheat, wild strawberry, yams, yew, or zucchini.

180. The method of claim 178, wherein the plant is a grass.

181. The method of claim 180, wherein the grass is a Poaceae family.

182. The method of claim 180 or 181 , wherein the grass is Agrostis genus, Poa genus, Lolium genus, Festuca genus, Cynodon genus, Stenotaphrum genus, or Zoysia genus.

183. The method of any one of claims 180-182, wherein the grass is a turfgrass, which is bentgrass, Kentucky bluegrass, ryegrass, tall fescue, Chewings fescue, Hard fescue, sheep fescue, creeping red fescue, bermudagrass, kikuyu grass, St. Augustine grass, buffalo grass, zoysia grass, zoyosa tenuifolia, or temple grass.

184. The method of claim 177, wherein the target is a soil or an area where a plant is present.

185. The method of claim 177, wherein the induced nutrient uptake and assimilation is associated with upregulation of the expression of a plurality of gene, which is nitrate reductase, nitrite reductase, high-affinity nitrate transporter 2.1 (HAT2.1), high-affinitynitrate transporter 2.2 (HAT2.2), high-affinity nitrate transporter 2.3 (HAT2.3), glutamine synthetase, and / or mugineic acid- 3 dioxygenase.

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