Biostimulant composition

Abstract

Biostimulant compositions made by digestion of Ecklonia maxima feedstock, and methods of promoting plant growth by contacting a plant, plant seed or plant growth medium with such compositions.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 455,662, filed March 30, 2023, the entirety of which is incorporated by reference herein. BACKGROUND

[0002] The present disclosure relates generally to biostimulant compositions and methods of using such biostimulant compositions to promote plant growth.

[0003] For environmental and economic reasons, promoting efficient production of food crops and other crops is an important goal. Plant growth promoting products derived from organic materials can help promote crop growth, improve the efficacy of agricultural products such as fertilizers, and reduce the environmental impact of synthetic fertilizers and climate change. Thus, there is a need for plant growth promoting biostimulant compositions that utilize abundant and available organic feedstocks. SUMMARY

[0004] The present disclosure provides plant growth promoting biostimulant compositions made from giant kelp feedstocks and methods of using such biostimulant compositions.

[0005] Disclosed herein is a method of promoting plant growth, the method comprising contacting a plant, a plant seed, or a plant growth medium with a composition comprising a microbial digestion product of an organic feedstock, the organic feedstock comprising giant kelp seaweed (Ascophyllum nodosum). In some embodiments, the digestion is anaerobic digestion. In some embodiments, the digestion product is produced by endogenous microorganisms present within the giant kelp seaweed of the organic feedstock. In some embodiments, the digestion product comprises no more than 40 mol% of fucose relative to the concentration of all glycosyl residues in the composition. In some embodiments, the digestion product comprises no more than 15 mol% of xylose relative to the concentration of all glycosyl residues in the composition. In some embodiments, the digestion product comprises more than 7 mol% of mannose relative to the concentration of all glycosyl residues in the composition. In some embodiments, the digestion product comprises one or more of iso-butanol, pentadecanenitrile, pentadecanoic acid, 9-octadecenonitrile, hexadecanenitrile, or heneicosane. In some embodiments, the digestion product comprises one or more of the molecular species listed in Table 1. In some embodiments, the digestion product comprises one or more of the molecular species corresponding to one or more of peaks C, F, G, J, O, or P in the LC-MS chromatogram shown in FIG. 1. In some embodiments, the digestion product comprises one or more of the molecular species corresponding to one or more of peaks A, B, C, D, E, F, or G in the GC-MS chromatogram shown in FIG. 2. Ecklonia maxima kelp). In some embodiments, the digestion is anaerobic digestion. In some embodiments, the digestion product is produced by endogenous microorganisms present within the giant kelp seaweed of the organic feedstock. In some embodiments, the digestion product comprises no more than 40 mol% of fucose relative to the concentration of all glycosyl residues in the composition. In some embodiments, the digestion product comprises no more than 15 mol% of xylose relative to the concentration of all glycosyl residues in the composition. In some embodiments, the digestion product comprises more than 7 mol% of mannose relative to the concentration of all glycosyl residues in the composition. In some embodiments, the digestion product comprises one or more of iso-butanol, pentadecanenitrile, pentadecanoic acid, 9-octadecenonitrile, hexadecanenitrile, or heneicosane. In some embodiments, the digestion product comprises one or more of the molecular species listed in Table 1. In some embodiments, the digestion product comprises one or more of the molecular species corresponding to one or more of peaks C, F, G, J, O, or P in the LC-MS chromatogram shown in FIG. 1. In some embodiments, the digestion product comprises one or more of the molecular species corresponding to one or more of peaks A, B, C, D, E, F, or G in the GC-MS chromatogram shown in FIG. 2. Figure 39 kelp). In some embodiments, the digestion is anaerobic digestion. In some embodiments, the digestion product is produced by endogenous microorganisms present within the giant kelp seaweed of the organic feedstock. In some embodiments, the digestion product comprises no more than 40 mol% of fucose relative to the concentration of all glycosyl residues in the composition. In some embodiments, the digestion product comprises no more than 15 mol% of xylose relative to the concentration of all glycosyl residues in the composition. In some embodiments, the digestion product comprises more than 7 mol% of mannose relative to the concentration of all glycosyl residues in the composition. In some embodiments, the digestion product comprises one or more of iso-butanol, pentadecanenitrile, pentadecanoic acid, 9-octadecenonitrile, hexadecanenitrile, or heneicosane. In some embodiments, the digestion product comprises one or more of the molecular species listed in Table 1. In some embodiments, the digestion product comprises one or more of the molecular species corresponding to one or more of peaks C, F, G, J, O, or P in the LC-MS chromatogram shown in FIG. 1. In some embodiments, the digestion product comprises one or more of the molecular species corresponding to one or more of peaks A, B, C, D, E, F, or G in the GC-MS chromatogram shown in FIG. 2. Figure 3 kelp). In some embodiments, the digestion is anaerobic digestion. In some embodiments, the digestion product is produced by endogenous microorganisms present within the giant kelp seaweed of the organic feedstock. In some embodiments, the digestion product comprises no more than 40 mol% of fucose relative to the concentration of all glycosyl residues in the composition. In some embodiments, the digestion product comprises no more than 15 mol% of xylose relative to the concentration of all glycosyl residues in the composition. In some embodiments, the digestion product comprises more than 7 mol% of mannose relative to the concentration of all glycosyl residues in the composition. In some embodiments, the digestion product comprises one or more of iso-butanol, pentadecanenitrile, pentadecanoic acid, 9-octadecenonitrile, hexadecanenitrile, or heneicosane. In some embodiments, the digestion product comprises one or more of the molecular species listed in Table 1. In some embodiments, the digestion product comprises one or more of the molecular species corresponding to one or more of peaks C, F, G, J, O, or P in the LC-MS chromatogram shown in FIG. 1. In some embodiments, the digestion product comprises one or more of the molecular species corresponding to one or more of peaks A, B, C, D, E, F, or G in the GC-MS chromatogram shown in FIG. 2. Figure 4 kelp). In some embodiments, the digestion is anaerobic digestion. In some embodiments, the digestion product is produced by endogenous microorganisms present within the giant kelp seaweed of the organic feedstock. In some embodiments, the digestion product comprises no more than 40 mol% of fucose relative to the concentration of all glycosyl residues in the composition. In some embodiments, the digestion product comprises no more than 15 mol% of xylose relative to the concentration of all glycosyl residues in the composition. In some embodiments, the digestion product comprises more than 7 mol% of mannose relative to the concentration of all glycosyl residues in the composition. In some embodiments, the digestion product comprises one or more of iso-butanol, pentadecanenitrile, pentadecanoic acid, 9-octadecenonitrile, hexadecanenitrile, or heneicosane. In some embodiments, the digestion product comprises one or more of the molecular species listed in Table 1. In some embodiments, the digestion product comprises one or more of the molecular species corresponding to one or more of peaks C, F, G, J, O, or P in the LC-MS chromatogram shown in FIG. 1. In some embodiments, the digestion product comprises one or more of the molecular species corresponding to one or more of peaks A, B, C, D, E, F, or G in the GC-MS chromatogram shown in FIG. 2. Figure 5shown 1 The peaks A, B, C, D, or E in the H-NMR spectrum correspond to one or more molecular species. In some embodiments, the digestion product contains [a specific molecular species]. Figure 6 shown 13 One or more of peaks A or B in the C-NMR spectrum correspond to one or more molecular species.

[0006] In some embodiments, the composition further comprises endogenous microorganisms of *Lombyx mori* present in the organic raw material. In some embodiments, the microorganisms comprise sporulated microorganisms. In some embodiments, the dry weight percentage of microbial biomass in the composition is 0.071% to 0.714% relative to the total dry weight of the composition. In some embodiments, the microorganisms present in the composition comprise *Microbacterium amyloliquefaciens* (…). Microbacterium amylolyticum ), pyrolytic sugar heat anaerobic bacteria ( Thermoanaerobacterium thermosaccharolyticum ), Cellulosilyticum lentocellum Heat-resistant microbubbles ( Microbulbifer thermotolerans Collins strain ( Collinsella sp.), Acinetobacter species ( Acinetobacter spp.), Acinetobacter Thomsonii ( Acinetobacter towneri Lactobacillus bruneri ( Lentilactobacillus buchneri ), Lactobacillus barley ( Liquorilactobacillus hordei ) or secondary lactobacilli ( Secundilactobacillus paracollinoides One or more of the following. In some embodiments, the dry weight percentage of microbial biomass in the composition is less than 0.001 wt% relative to the total dry weight of the composition. In some embodiments, the composition does not contain microorganisms.

[0007] In some embodiments, promoting plant growth includes one or more of the following: enhancing seed germination, enhancing early plant development, improving root growth, increasing nutrient uptake, improving tolerance to abiotic stresses, mitigating transplant shock, improving plant reproduction, and improving soil microbial activity. In some embodiments, improving tolerance to abiotic stresses includes improving one or more of the following: salt tolerance, heat tolerance, cold tolerance, and drought tolerance. In some embodiments, the contact includes application in furrows, foliar spraying, application to the root zone, application to seeds, or mixing with a growth medium. In some embodiments, the growth medium is soil.

[0008] In some embodiments, the composition further comprises solid fertilizer granules. In some embodiments, the fertilizer granules are coated with digestion products. In some embodiments, the composition is a liquid. In some embodiments, the composition further comprises liquid fertilizer.

[0009] In some embodiments, the contact comprises applying the composition at a rate of 0.5 to 10 quarts per acre. In some embodiments, the contact comprises applying 0.14 to 6.7 g dry weight of the digested product per acre.

[0010] In some embodiments, the plants are experiencing drought conditions or at risk of experiencing drought conditions at the time of contact. In some embodiments, the growth medium is a high-salt soil. In some embodiments, the plants are experiencing freezing conditions or at risk of experiencing freezing conditions at the time of contact. In some embodiments, the plants are experiencing cold stress or at risk of experiencing cold stress at the time of contact. In some embodiments, the plants are experiencing heat stress or at risk of experiencing heat stress at the time of contact. In some embodiments, the plants have been transplanted. In some embodiments, the plants are maize, cotton, tomato, or sweet pepper. In some embodiments, the plants are cotton plants or maize plants, and the cotton plants or maize plants are under drought conditions at the time of contact.

[0011] This document also discloses a composition comprising a digestion product produced by microbial digestion of an organic feedstock (containing *Laminaria japonica*). In some embodiments, the microorganisms comprise endogenous *Laminaria japonica* microorganisms present in the organic feedstock. In some embodiments, the digestion product comprises fucose at a concentration not exceeding 40 mol% relative to the total concentration of all glycosyl residues in the composition. In some embodiments, the digestion product comprises xylose at a concentration exceeding 15 mol% relative to the total concentration of all glycosyl residues in the composition. In some embodiments, the digestion product comprises mannose at a concentration exceeding 7 mol% relative to the total concentration of all glycosyl residues in the composition. In some embodiments, the digestion product comprises one or more of isobutanol, pentadecanoic acid, pentadecanoic acid, 9-octadecenoic acid, hexadecanoic acid, or hexadecanoic acid. In some embodiments, the digestion product comprises... Figure 39 The molecular species listed herein. In some embodiments, the digestion products contain [elements related to...]. Figure 3 The peaks in the LC-MS chromatogram shown correspond to one or more molecular species from C, F, G, J, O, or P. In some embodiments, the digestion product contains [missing information - likely related to molecular species]. Figure 4 The peaks A, B, C, D, E, F, or G in the GC-MS chromatogram shown correspond to one or more molecular species. In some embodiments, the digestion product contains [missing information - likely related to molecular species]. Figure 5 shown 1 The peaks A, B, C, D, or E in the H-NMR spectrum correspond to one or more molecular species. In some embodiments, the digestion product contains [a specific molecular species]. Figure 6 shown 13 One or more of peaks A or B in the C-NMR spectrum correspond to one or more molecular species.

[0012] In some embodiments, the composition further comprises endogenous microorganisms present in the organic feedstock. In some embodiments, the microorganisms comprise sporulating microorganisms. In some embodiments, the percentage of dry weight of the microorganism biomass in the composition is between 0.071% and 0.714% relative to the total dry weight of the composition. In some embodiments, the microorganisms present in the composition comprise one or more of Amyelois solitaria, Caloramator proteoclasticus, Clostridium thermosaccharolyticum, Moorella therma, Collinsella sp., Acinetobacter sp., Acinetobacter calcoaceticus, Lactobacillus buchneri, Lactobacillus farciminus, or Lactobacillus parafaricinium. In some embodiments, the microorganisms have been removed from the composition. In some embodiments, the percentage of dry weight of the microorganism biomass in the composition is less than 0.001% relative to the total dry weight of the composition. In some embodiments, the composition does not comprise microorganisms. Cellulosilyticum lentocellum In some embodiments, the composition further comprises endogenous microorganisms present in the organic feedstock. In some embodiments, the microorganisms comprise sporulating microorganisms. In some embodiments, the percentage of dry weight of the microorganism biomass in the composition is between 0.071% and 0.714% relative to the total dry weight of the composition. In some embodiments, the microorganisms present in the composition comprise one or more of Amyelois solitaria, Caloramator proteoclasticus, Clostridium thermosaccharolyticum, Moorella therma, Collinsella sp., Acinetobacter sp., Acinetobacter calcoaceticus, Lactobacillus buchneri, Lactobacillus farciminus, or Lactobacillus parafaricinium. In some embodiments, the microorganisms have been removed from the composition. In some embodiments, the percentage of dry weight of the microorganism biomass in the composition is less than 0.001% relative to the total dry weight of the composition. In some embodiments, the composition does not comprise microorganisms.

[0013] Also disclosed herein is a composition comprising one or more molecular species corresponding to one or more of peaks C, F, G, J, O, or P in the LC-MS chromatogram shown in FIG. 1. Figure 3 Also disclosed herein is a composition comprising one or more molecular species corresponding to one or more of peaks C, F, G, J, O, or P in the LC-MS chromatogram shown in FIG. 1. Figure 4 Also disclosed herein is a composition comprising one or more molecular species corresponding to one or more of peaks A, B, C, D, E, F, or G in the GC-MS chromatogram shown in FIG. 2. Figure 5 Also disclosed herein is a composition comprising one or more molecular species corresponding to one or more of peaks A, B, C, D, or E in the H-NMR spectrum shown in FIG. 3. 1 Also disclosed herein is a composition comprising one or more molecular species corresponding to one or more of peaks A, B, C, D, or E in the H-NMR spectrum shown in FIG. 3. Figure 6 Also disclosed herein is a composition comprising one or more molecular species corresponding to one or more of peaks A or B in the C-NMR spectrum shown in FIG. 4. 13 Also disclosed herein is a composition comprising one or more molecular species corresponding to one or more of peaks A or B in the C-NMR spectrum shown in FIG. 4. In some embodiments, the composition comprises xylose at a concentration of more than 15 mol% relative to all glycosyl residues in the composition. In some embodiments, the composition comprises mannose at a concentration of more than 7 mol% relative to all glycosyl residues in the composition. In some embodiments, the composition comprises one or more of isobutanol, pentadecanenitrile, pentadecanoic acid, 9-octadecenonitrile, hexadecanenitrile, or heneicosane. In some embodiments, the digestion product comprises one or more of Figure 39 Also disclosed herein is a composition comprising one or more molecular species corresponding to one or more of peaks A, B, C, D, or E in the H-NMR spectrum shown in FIG. 3.

[0014] In some embodiments, the composition further comprises microorganisms. In some embodiments, the microorganisms comprise sporulating microorganisms. In some embodiments, the percentage of dry weight of the microorganism biomass in the composition is between 0.071% and 0.714% relative to the total dry weight of the composition. In some embodiments, the microorganisms present in the composition comprise one or more of Amyelois solitaria, Caloramator proteoclasticus, Clostridium thermosaccharolyticum, Moorella therma, Collinsella sp., Acinetobacter sp., Acinetobacter calcoaceticus, Lactobacillus buchneri, Lactobacillus farciminus, or Lactobacillus parafaricinium. In some embodiments, the microorganisms have been removed from the composition. In some embodiments, the percentage of dry weight of the microorganism biomass in the composition is less than 0.001% relative to the total dry weight of the composition. In some embodiments, the composition does not comprise microorganisms.Cellulosilyticum lentocellum one or more of a heat-tolerant microbubble, a Collinsiella sp., an Acinetobacter sp., Acinetobacter thomasii, Lactobacillus buccalis, Lactobacillus hordeinum, or Lactobacillus paracasei. In some embodiments, the dry weight percentage of microbial biomass in the composition is less than 0.001% relative to the total dry weight of the composition. In some embodiments, the composition does not comprise a microbe.

[0015] In some embodiments, the composition is a liquid composition. In some embodiments, the digestate is present in the liquid composition at 0.06% to 0.08% by weight relative to the total weight of the liquid composition.

[0016] Also disclosed is a plant treatment composition comprising any of the above compositions and a fertilizer composition. In some embodiments, the fertilizer composition is a liquid. In some embodiments, the fertilizer composition is a solid. In some embodiments, the fertilizer composition is coated with any of the above bio-stimulant compositions.

[0017] Also disclosed is a method of promoting plant growth, the method comprising contacting a plant, a plant seed, or a plant growth medium with any of the above compositions. In some embodiments, promoting plant growth comprises one or more of: enhancing seed germination, enhancing early plant development, improving root growth, increasing nutrient uptake, improving tolerance to abiotic stress, reducing transplant shock, improving plant reproduction, and improving soil microbial activity. In some embodiments, improving tolerance to abiotic stress comprises improving one or more of: salt tolerance, heat tolerance, cold tolerance, and drought tolerance. In some embodiments, the contacting comprises in-furrow application, foliar spray application, or application to a root zone. In some embodiments, the contacting comprises applying 0.14 to 6.7 g dry weight of digestate per acre. INCORPORATION BY REFERENCE

[0018] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. BRIEF DESCRIPTION OF DRAWINGS

[0019] The novel features of the disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative

[0020] Figure 1 Ascophyllum nodosum (A.nodosum) Ascophyllum nodosum GC-MS chromatograms of Ascophyllum nodosum (A. nodosum) powder feedstock (top chromatogram) and E. cava powder feedstock (bottom chromatogram).

[0021] Figure 2 : Analysis of glycosyl residue content in MBT-A and MBT-E products.

[0022] Figure 3 : LC-MS chromatograms of MBT-A (top) and MBT-E (bottom) products.

[0023] Figure 4 : GC-MS chromatograms of MBT-E (top) and MBT-A (bottom) products. Arrows point to unique peaks in MBT-E that were identified by Agilent MassHunter Quantitative and Qualitative software based on matching coefficients > 70% and areas > 1 x 10 5 .

[0024] Figure 5 : H-NMR spectra of MBT-A (top) and MBT-E (bottom). 1 .

[0025] Figure 6 : C-NMR spectra of MBT-A (top) and MBT-E (bottom). 13 .

[0026] Figure 7 : Cotton leaf chlorophyll content under drought stress conditions (SPAD1 and SPAD2 recorded at 15 and 26 days after the start of the drought stress water regimen, respectively).

[0027] Figure 8 : Proline accumulation under drought stress conditions (Proline concentration recorded at 30 days after the start of the drought stress water regimen).

[0028] Figure 9 : Cotton plant height under drought stress conditions (Plant height recorded at 90 days after the start of the drought stress water regimen).

[0029] Figure 10 : Cotton boll yield under drought stress conditions (Cotton bolls counted at 80 days after the start of the drought stress water regimen). Cotton boll size was measured in centimeters (cm) and did not take into account cotton bolls with a size greater than or equal to 2 cm.

[0030] Figure 11 : Average cotton yield of MBT-E treated plants and untreated control plants.

[0031] Figure 12 : Relative water content of corn leaves under drought stress conditions measured at 26 days after the start of the drought stress water regimen.

[0032] Figure 13 Proline accumulation under drought stress conditions (Proline concentration was recorded 30 days after the beginning of the drought stress water regime).

[0033] Figure 14 Average ear length and weight measured just before harvest.

[0034] Figure 15 Corn yield resulting from the indicated treatments.

[0035] Figure 16 Shoot surface area resulting from the indicated treatments. Asterisks indicate statistical significance.

[0036] Figure 17 Root surface area resulting from the indicated treatments. Asterisks indicate statistical significance.

[0037] Figure 18 Root length resulting from the indicated treatments. Asterisks indicate statistical significance.

[0038] Figure 19 Shoot surface area resulting from the indicated treatments. Asterisks indicate statistical significance.

[0039] Figure 20A - C: (A) Growth rate under drought conditions resulting from the indicated treatments. (B) Growth rate after recovery resulting from the indicated treatments. (C) Growth rate resulting from the indicated treatments. Asterisks indicate statistical significance.

[0040] Figure 21 Leaf temperature resulting from the indicated treatments. UTC = untreated control.

[0041] Figure 22 Percent change in stomatal conductance over the indicated time resulting from the indicated treatments.

[0042] Figure 23 Percent change in stomatal conductance over the indicated time resulting from the indicated treatments.

[0043] Figure 24 Percent change in stomatal conductance over the indicated time resulting from the indicated treatments.

[0044] Figure 25 Percent change in stomatal conductance over the indicated time resulting from the indicated treatments.

[0045] Figure 26 Percent change in stomatal conductance over the indicated time resulting from the indicated treatments.

[0046] Figure 27 Cotton leaf chlorophyll content over the indicated time resulting from the indicated treatments.

[0047] Figure 28 : The tip surface area resulting from the treatment shown. An asterisk indicates statistical significance.

[0048] Figure 29 : Plant score resulting from the treatment shown. An asterisk indicates statistical significance.

[0049] Figure 30 : The weight of fresh shoots resulting from the treatment shown. An asterisk indicates statistical significance.

[0050] Figure 31 The treatments shown result in shoot surface area and growth rate. An asterisk indicates statistical significance.

[0051] Figure 32 : The tip surface area resulting from the treatment shown. An asterisk indicates statistical significance.

[0052] Figure 33 : The chlorophyll content (SPAD) resulting from the treatment. An asterisk indicates statistical significance.

[0053] Figure 34 The stem diameter resulting from the treatment shown is :.

[0054] Figure 35A -B: Harvest height resulting from treatment shown in (A). Total biomass resulting from treatment shown in (B).

[0055] Figure 36 The soil electrical conductivity resulting from the treatment shown is denoted as :.

[0056] Figure 37 The treatment shown represents the resulting bell pepper yield.

[0057] Figure 38 The nutrient content resulting from the treatment shown.

[0058] Figure 39 The unique peaks in MBT-E compared to the MBT-A GC-MS chromatogram.

[0059] Figure 40 Bacterial community analysis of two seaweed raw materials: *Laminaria japonica* (… Eklonia maxima Raw materials (EMF) and *Alternanthera philoxeroides* (Ascophylum nodosum) Raw material (ANF). DNA was extracted from powdered seaweed raw material, and the bacterial community was characterized by amplicon sequencing of the small ribosomal RNA gene (i.e., the 16S rRNA gene). The bacterial community map was presented in the form of a UPGMA cluster analysis tree.

[0060] Figure 41Bacterial community analysis of seaweed product solutions MBT-E and MBT-A. DNA was extracted from the concentrated product solutions (4-fold), and the bacterial community was characterized by amplicon sequencing of the small ribosomal RNA gene (i.e., the 16S rRNA gene). Bacterial community maps are presented in the form of UPGMA cluster analysis trees. Detailed Implementation

[0061] The embodiments described herein include biostimulant compositions and methods for promoting plant growth and enhancing plant tolerance to abiotic stresses, including, for example, drought, cold, heat, and salt stress. The compositions include microbial digestion products produced by digesting giant kelp.

[0062] I. Digestion Methods In some embodiments, the biostimulant composition is prepared by digesting an organic feedstock containing giant kelp. In some embodiments, the organic feedstock also contains chitin and brewer's yeast (Saccharomyces cerevisiae). Saccharomyces (Cerevisiae yeast). The organic feedstock can be an aqueous slurry of powdered kelp, chitin, and yeast. In some embodiments, the digestion is anaerobic. Not wishing to be bound by theory, in the digestion method, endogenous microorganisms of kelp and chitin are believed to digest the biomolecules and other nutrients present in the kelp, chitin, and yeast, producing digestion products that include compounds that promote plant growth, tolerance to abiotic stress, and soil health. The biostimulant may also contain microorganisms that contribute to the plant-beneficial properties of the biostimulant product. The microorganisms in the biostimulant product can be derived from the microbial population present in the kelp feedstock.

[0063] The digestion method for producing biostimulants can be carried out in a digestive system comprising a series of tanks through which the feedstock flows continuously. Liquid at the top of each tank can continuously flow into the next tank, and the rate of product outflow can be matched to the rate of feedstock inflow, thus providing a hydraulically balanced flow throughout the system. Each tank within the system can possess a unique and stable microbial community with different physiological characteristics and digestive capabilities compared to the communities in other tanks within the system.

[0064] In some embodiments of the digestion process, powdered giant kelp, chitin, and S. cerevisiae can be mixed with water to make an organic feedstock for an anaerobic digestion system. The anaerobic digestion system can include a mixing tank in which the organic feedstock is mixed to make a homogenous slurry. The slurry can then flow through a series of four digestion tanks in a continuous and hydraulically balanced manner. More or fewer digestion tanks can be used, and the hydraulic flow rate can be varied to achieve the desired results. In the first digestion tank, the slurry can be agitated at a rate that allows heavier or undigested solids to settle to the bottom. An outlet at the top of the first digestion tank can allow fluid to flow into the second digestion tank. An outlet at the bottom of the first digestion tank can deliver the settled solids back to the mixing tank. Each of the three subsequent digestion tanks (which can be referred to as packed bed reactors) can have an immersed stationary media matrix that provides a surface for biofilm growth. The flow rate of the digestion system can be selected to allow sufficient residence time in each digestion tank to allow a stable and unique microbial community to form in each digestion tank. The microorganisms in the community can originate from the microorganisms originally present in the organic feedstock. These microorganisms can digest the giant kelp, chitin, and yeast to produce digestion products. The effluent at the top of the fourth digestion tank can be used as a biostimulant to promote plant growth or to improve soil quality, as described in more detail below.

[0065] In some embodiments, the biostimulant composition is made by the method described in U.S. Application Publication No. 2013 / 0324406, which is hereby incorporated by reference in its entirety, using giant kelp, chitin, and S. cerevisiae as the feedstock.

[0066] The biostimulant composition produced by the digestion process described above can be used as is, or can be further processed prior to use. For example, the effluent of the digestion system (referred to herein as the "base product") can be concentrated, sterilized, filtered, pasteurized, or dehydrated, or any combination of these, prior to use. In some embodiments, the base product can be concentrated 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, or higher. In some embodiments, the base product can be filter sterilized to remove any bacteria or other microorganisms in the composition.

[0067] The parameters of the digestion system, such as the flow rate and the solids content of the organic feedstock, can be varied to achieve the desired properties of the effluent biostimulant base product.

[0068] II. Physical properties and composition of biostimulants Embodiments described herein include biostimulant compositions that include chemical substances and / or microorganisms that promote plant growth, including by increasing the ability of plants to tolerate abiotic stress, such as cold, heat, drought, and salt. The biostimulant compositions described herein can include dead microorganisms, sporulated microorganisms, dead microorganism fragments, live microorganisms, microbial fermentation products, enzymes, biological plant growth regulators, organic acids, chelating agents, or combinations thereof.

[0069] Embodiments described herein also include biostimulant compositions that include digestion products produced by digestion of organic feedstocks, including the giant kelp seaweed. The biostimulants can include metabolites produced by endogenous microorganisms within the organic feedstock, which can be derived from the seaweed feedstock or other components of the organic feedstock, such as, for example, chitin. Such metabolites can include, for example, sugars and fatty acids. The digestion products can also include dead microorganisms, dead microorganism fragments, microbial fermentation products, enzymes, biological plant growth regulators, organic acids, chelating agents, or combinations thereof.

[0070] The biostimulant compositions described herein can include one or more sugars. In some embodiments, the biostimulant compositions can be characterized by their sugar-based residue content. In some embodiments, the biostimulant compositions can include one or more of rhamnose, fucose, xylose, mannose, or glucose, or any combination thereof. In some embodiments, the biostimulant compositions do not include galactose, or include less than 1 mol% of galactose, as compared to other sugar-based residues present in the biostimulant composition. In some embodiments, the biostimulant compositions include less than about or about 40 mol%, 30 mol%, 20 mol%, or 15 mol% of fucose, as compared to all other sugar-based residues present in the biostimulant composition. In some embodiments, the biostimulant compositions include at least about or about 15 mol%, 20 mol%, 25 mol%, or 30 mol% of xylose, as compared to all other sugar-based residues present in the biostimulant composition. In some embodiments, the biostimulant compositions include at least about or about 6 mol%, 8 mol%, 10 mol%, 12 mol%, 14 mol%, 16 mol%, 18 mol%, or 20 mol% of mannose, as compared to other sugar-based residues present in the biostimulant composition.

[0071] In some embodiments, the biostimulant compositions described herein can be characterized by mass spectrometry or NMR spectroscopy. In some embodiments, the biostimulant compositions have an LC-MS chromatogram as shown in the bottom panel. In some embodiments, the biostimulant compositions include a peak at about 1 1 1 1 Da, as shown in the top panel. Figure 3 In some embodiments, the biostimulant compositions include a peak at about 1 1 1 1 Da, as shown in the top panel. Figure 3The LC-MS chromatogram shown on the bottom panel contains one or more peaks corresponding to one or more molecular species, or any combination of such molecular species. In some embodiments, the biostimulant composition comprises... Figure 3 The LC-MS chromatogram shown on the bottom panel contains molecular species corresponding to peaks labeled A, B, C, D, E, F, G, H, I, J, K, L, M, N, O, P, Q, R, S, T, U, or V, or any combination of such molecular species. In some embodiments, the biostimulant composition comprises molecules corresponding to... Figure 3 One or more peaks (which are not present in the LC-MS chromatogram shown in the bottom panel) Figure 3 (The LC-MS chromatogram shown in the top panel corresponds to one or more molecular species.) In some embodiments, the biostimulant composition contains molecular species corresponding to peaks labeled C, F, G, J, M, N, O, P, or Q, or any combination of such molecular species. In some embodiments, the biostimulant composition does not contain molecular species corresponding to peaks labeled C, F, G, J, M, N, O, P, or Q. Figure 3 The peaks present in the LC-MS chromatogram shown in the top panel (where) Figure 3 The molecular species that are not present in the LC-MS chromatogram shown in the bottom panel.

[0072] In some embodiments, the biostimulant composition has the following properties: Figure 4 The GC-MS chromatogram is shown in the top panel. In some embodiments, the biostimulant composition comprises one or more components such as... Figure 4 The molecular species corresponding to one or more peaks in the GC-MS chromatogram shown in the top panel, or any combination of such molecular species. In some embodiments, the biostimulant composition comprises... Figure 4 The peaks labeled A, B, C, D, E, F, or G in the GC-MS chromatogram shown in the top panel correspond to the molecular species, or any combination of such molecular species. In some embodiments, the biostimulant composition comprises one or more molecules that... Figure 4 One or more peaks in the GC-MS chromatogram shown in the top panel (which do not exist) Figure 4 The molecular species corresponding to the GC-MS chromatogram shown in the bottom panel. In some embodiments, the biostimulant composition does not contain the molecules corresponding to the GC-MS chromatogram shown in the bottom panel. Figure 4 The peaks present in the GC-MS chromatogram shown in the bottom panel (where) Figure 4 (The corresponding molecular species are not present in the GC-MS chromatogram shown in the top panel.) In some embodiments, the biostimulant composition contains... Figure 39 One or more of the molecular species listed, or any combination of such molecular species.

[0073] In some embodiments, the biostimulant composition has the following properties: Figure 5 The bottom spectrum shows 1 H-NMR spectroscopy. In some embodiments, the biostimulant composition comprises one or more [components / elements]. Figure 5 bottom 1 The molecular species corresponding to one or more peaks in the H-NMR spectrum, or any combination of such molecular species. In some embodiments, the biostimulant composition contains molecules corresponding to... Figure 5 bottom 1 The molecular species corresponding to peaks labeled A, B, C, D, E, F, G, H, I, J, or K in the H-NMR spectrum, or any combination of such molecular species. In some embodiments, the biostimulant composition comprises one or more molecules corresponding to... Figure 5 bottom 1 One or more peaks in the H-NMR spectrum (which do not exist in) Figure 5 top 1 The molecular species corresponding to (in H-NMR spectra). In some embodiments, the biostimulant composition contains molecules corresponding to (in H-NMR spectra). Figure 5 The molecular species corresponding to the peaks labeled A, C, D, or E, or any combination of such molecular species. In some embodiments, the biostimulant composition does not contain any molecules corresponding to the peaks labeled A, C, D, or E. Figure 5 top 1 The peaks present in the H-NMR spectrum (which are in) Figure 5 bottom 1 The corresponding molecular species are not present in the H-NMR spectrum.

[0074] In some embodiments, the biostimulant composition has the following properties: Figure 6 The bottom spectrum shows 13 C-NMR spectroscopy. In some embodiments, the biostimulant composition comprises one or more compounds related to... Figure 6 bottom 13 The molecular species corresponding to any one or more peaks in the C-NMR spectrum, or any combination of such molecular species. In some embodiments, the biostimulant composition comprises a molecular species corresponding to any one or more peaks in the C-NMR spectrum. Figure 6 bottom 13 The molecular species corresponding to peaks labeled A, B, C, D, E, or F in the C-NMR spectrum, or any combination of such molecular species. In some embodiments, the biostimulant composition comprises one or more molecules corresponding to... Figure 6 bottom 13 One or more peaks in the C-NMR spectrum (which do not exist in) Figure 6 top 13 (In C-NMR spectra) corresponding molecular species. In some embodiments, the biostimulant composition contains molecules corresponding to... Figure 6molecular species corresponding to peaks labeled A or B, or any combination of such molecular species. In some embodiments, the biostimulant composition does not include a molecular species corresponding to a peak Figure 6 at the top of a C-NMR spectrum 13 at the bottom of a C-NMR spectrum Figure 6 at the bottom of a C-NMR spectrum 13 at the bottom of a C-NMR spectrum

[0075] In some embodiments, the biostimulant composition includes live microorganisms. In some embodiments, the microorganisms include bacteria derived from a population of bacteria present in the laminaria hyperborea seaweed feedstock. These bacteria can include one or more of the bacteria listed in Table 1. In some embodiments, the biostimulant includes one or more bacterial species not found in products derived from microbial digestion products of other seaweed species, such as Ascophyllum nodosum. In some embodiments, the biostimulant includes Cellulosilyticum lentocellum Olsenella, Collinsella, Acinetobacter, Acinetobacter thomasii, Lactobacillus buchneri, Lactobacillus farciminus, or Paralactobacillus parafarciminus, or any combination thereof. In some embodiments, any of these bacterial species is included in at least 0.00001, 0.00005, 0.0001, 0.0005, 0.001, 0.0015, or 0.002% of the bacterial species present in the biostimulant, as determined by metagenomic sequencing.

[0076] In some embodiments, the biostimulant is filter-sterilized and does not include live microorganisms. In some embodiments, the dry weight of the microbial biomass is less than 0.0001% of the total dry weight of the composition.

[0077] In some embodiments, the biostimulant includes between 0.05 and 0.8% dry weight of microbial biomass relative to the total dry weight of the biostimulant composition. In some embodiments, the percentage by dry weight is at least about, at most about, or about 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, or 0.8%, or a range between any two of these values.

[0078] In some embodiments, the biostimulant includes between 100 and 5x10 5 CFU / ml of bacteria. In some embodiments, the biostimulant includes at least about, at most about, or about 100, 500, 1x10 3 , 5x10 3 , 1x10 4 , 5x10 4 , or 1x10 5 CFU / ml of bacteria, or a range between any two of these values.

[0079] In some embodiments, the biostimulant has a pH of 7.5 to 8.5. In some embodiments, the biostimulant has a conductivity of about 900, 950, 1000, 1050, or 1100 μ8 / αη. In some embodiments, the biostimulant has a density of about 0.997 to 0.999 g / cm3, or about 0.998 g / cm3. 3 3 In some embodiments, the biostimulant has a solids content of 0.01 to 2%. In some embodiments, the solids content is about 0.01, 0.05, 0.1, 0.15, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, or 2.0, or a range between any two of these values. In some embodiments, the biostimulant has a chemical oxygen demand (COD) of 10 to 200 mg / L. In some embodiments, the COD is 10, 20, 30, 40, 50, 100, 125, 150, 175, or 200 mg / L, or a range between any two of these values. Conductivity and COD values vary with the concentration of the biostimulant and increase with increasing concentration.

[0080] III. Methods of use ​Embodiments of the biostimulant compositions can be used in methods for promoting plant growth under a variety of different plants and conditions. In some embodiments, contacting a plant, seed, or growth medium with a biostimulant promotes plant growth by, for example, increasing growth rate, yield at harvest, yield, stem girth, fruit abundance and / or size, grain yield, leaf surface area, root surface area, root length, root depth, shoot girth, or total mass, as compared to a plant that did not receive the treatment. In some embodiments, promoting plant growth comprises one or more of: enhancing seed germination, promoting early plant development, increasing nutrient uptake, reducing transplant shock, improving plant reproduction, and improving soil microbial activity. In some embodiments, any one or more of these plant qualities can be increased by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% or more, as compared to a plant of the same species that did not receive the treatment. In some embodiments, contacting a plant, seed, or growth medium with a biostimulant promotes plant growth by, for example, increasing the plant's tolerance to abiotic stress. Such abiotic stress can include drought stress, heat stress, cold stress, or stress caused by high salt concentrations. In some embodiments, increasing tolerance to one or more abiotic stresses can result in an increase in growth rate, yield at harvest, yield, stem girth, fruit abundance and / or size, grain yield, leaf surface area, root surface area, root length, root depth, shoot girth, or total mass, as compared to a plant of the same species that did not receive the treatment and was under similar stress conditions. In some embodiments, treatment with a biostimulant promotes plant growth by increasing the plant's ability to recover from abiotic stress, such that it recovers faster than it would have without the treatment.

[0081] In some embodiments, a plant or growth medium is contacted with a biostimulant composition prior to, during, or after abiotic stress. For example, in some embodiments, treatment with a biostimulant prior to abiotic stress can enable a plant to better tolerate the abiotic stress than a similar plant that did not receive the treatment. In some embodiments, a plant or growth medium is contacted with a biostimulant when the plant is at risk of experiencing abiotic stress, but before the abiotic stress occurs. A plant can be determined to be at risk of abiotic stress based on, for example, weather patterns or forecasts for the location where the plant is growing. In some embodiments, depending on the geographic location of the plant, a treatment to help alleviate cold stress can be applied during a period of the year when frost is more likely to occur, such as early spring or late fall. In some embodiments, a treatment to help alleviate heat stress or drought stress can be applied at the end of summer when the plant is at risk of experiencing relatively high temperatures.

[0082] One of skill in the art will be able to determine the relative risk of a plant to certain abiotic stresses based on the type of plant, the geographic location of the plant, and the local weather patterns and forecasts for that location.

[0083] In some embodiments, the biostimulant is administered to a plant when it is experiencing an abiotic stress. One skilled in the art can determine whether a plant is experiencing an abiotic stress based on the type of plant and the particular environment in which it is growing. For example, drought stress can be determined based on observing the soil water content and the condition of the plant. Because some plant species and varieties are naturally more drought tolerant than others, soil and air humidity conditions that cause stress to one species or variety can not stress another species or variety. The same applies to other potential stresses, such as heat, cold, and salt stress.

[0084] In some embodiments, the biostimulant is administered when the plant has experienced, is experiencing, or is predicted to experience a temperature of about 15°C, 10°C, 5°C, or 0°C or below. In some embodiments, the biostimulant is administered when the plant has experienced, is experiencing, or is predicted to experience a temperature of about 20°C, 25°C, 30°C, 35°C, or 40°C or above. In some embodiments, the biostimulant is administered when the plant has experienced, is experiencing, or is predicted to experience a soil water content of less than about 30%, 25%, 20%, 15%, 10%, 5%, or 1% for a duration of at least about 6, 12, 24, or 48 hours or 3, 4, 5, 6, 7, 8, 9, or 10 days.

[0085] In some embodiments, the biostimulant is administered within 12, 24, 36, or 48 hours, or 3, 4, 5, 6, 7, 8, 9, or 10 days of the plant experiencing or being predicted to experience an abiotic stress. In some embodiments, the biostimulant is administered when it is determined that there is at least about 30%, 40%, 50%, 60%, 70%, 80%, or 90% probability that the plant will experience an abiotic stress within 12, 24, 36, or 48 hours, or 3, 4, 5, 6, 7, 8, 9, or 10 days after administration.

[0086] In some embodiments, the biostimulant is administered when the plant is not experiencing or is not predicted to experience an abiotic stress. In addition to increasing tolerance to abiotic stress, embodiments of the biostimulant compositions disclosed herein can also promote plant growth in the absence of abiotic stress.

[0087] In some embodiments, the biostimulant is administered to the plant or growth medium prior to transplanting the plant. In some embodiments, the biostimulant is administered to the plant or growth medium after transplanting the plant. In some embodiments, the biostimulant is administered to the plant or growth medium at the time of transplanting the plant. In some embodiments, the biostimulant is administered to the growth medium (e.g., soil) into which the plant is to be transplanted.

[0088] In some embodiments, the plants treated with the biostimulant composition can be, for example, a crop, a vegetable, a flower, a foliage plant, a turfgrass, a tree, a shrub, and the like. Non-limiting examples of crops include corn, rice, wheat, barley, rye, oat, sorghum, cotton, soybean, peanut, buckwheat, sugar beet, rapeseed, sunflower, sugarcane, hemp, and tobacco. Non-limiting examples of vegetables include Solanaceae vegetables (eggplant, tomato, sweet pepper, hot pepper, potato, and the like), Cucurbitaceae vegetables (cucumber, pumpkin, zucchini, watermelon, melon, squash, and the like), Brassicaceae vegetables (Japanese radish, white radish, horseradish, kohlrabi, Chinese cabbage, cabbage, mustard, broccoli, cauliflower, and the like), Asteraceae vegetables (burdock, chervil, artichoke, lettuce, and the like), Liliaceae vegetables (onion, garlic, and asparagus), ammiaceous vegetables (carrot, parsley, celery, parsnip, and the like), Chenopodiaceae vegetables (spinach, Swiss chard, and the like), Lamiaceae vegetables (perilla, mint, basil, and the like), strawberry, sweet potato, yam, and taro. Non-limiting examples of fruits include pome fruits (apple, pear, Japanese pear, papaya, Japanese pear, and the like), stone fruits (peach, plum, nectarine, plum, cherry, apricot, prune, and the like), citrus fruits (Satsuma mandarin, orange, lemon, lime, grapefruit, and the like), nuts (chestnut, walnut, hazelnut, almond, pistachio, cashew, macadamia, and the like), berries (blueberry, cranberry, blackberry, raspberry, and the like), grape, persimmon, olive, Japanese plum, banana, coffee, date palm, and coconut. Non-limiting examples of trees include fruit trees, tea trees, mulberry trees, flowering plants, and street trees (ash tree, birch tree, dogwood tree, eucalyptus tree, ginkgo tree, lilac tree, maple tree, oak tree, poplar tree, Judas tree, sweetgum tree, sycamore tree, Japanese arborvitae, fir tree, hemlock tree, juniper tree, pine tree, spruce tree, and Taxus cuspidate). The term "plant" refers to native plants and genetically engineered plants. In some embodiments, the biostimulant is applied to seeds of any of the above plants.

[0089] In certain embodiments, the biostimulant compositions described herein can be applied to soil, applied to fertilizers that are applied to plants, applied directly to plants, or applied to both soil and plants. The compositions can also be applied directly to plant seeds. In addition to soil, the biostimulant compositions can also be applied to other plant growth media, such as hydroponic growth media. The compositions can be used for in-furrow applications, foliar applications, or both. In some embodiments, the biostimulant compositions are applied alone. When applied alone, in some embodiments, the compositions are applied before or after the application of conventional fertilizers and / or pesticides. When applied before or after the application of conventional fertilizers and / or pesticides, the compositions are applied close enough in time to the conventional fertilizer and / or pesticide application so that the formulation can have its intended effect of enhancing the effect of the conventional fertilizer and / or pesticide. In some embodiments, the compositions are applied in conjunction with conventional fertilizers and / or pesticides. The compositions can be mixed with conventional fertilizers and / or pesticides, or applied simultaneously with conventional fertilizers and / or pesticides.

[0090] In some embodiments, the biostimulant compositions described herein are mixed with conventional fertilizers or pesticides in a ratio of about 3: 1 to about 1 : 100 (biostimulant: conventional fertilizer or pesticide). In some embodiments, the biostimulant compositions are mixed with conventional fertilizers or pesticides in a ratio of about 1 :20 (biostimulant: conventional fertilizer or pesticide). The biostimulant compositions described herein can also be coated on the granules of conventional fertilizers or pesticides. The granules of the fertilizer or pesticide can be coated by, for example, spray drying the biostimulant onto the surface of the fertilizer, or mixing the biostimulant in dehydrated powder form with the granules (with or without a binder or carrier).

[0091] In certain embodiments, the conventional fertilizer is a starter fertilizer. In some embodiments, the conventional fertilizer includes at least one of the following: ammonia, urea, ammonium nitrate, ammonium sulfate, ammonium thiosulfate, monoammonium phosphate (MAP), diammonium phosphate (DAP), muriate of potash (MOP), sulfate of potash (SOP), nitrate of potash (NOP). In some embodiments, the starter fertilizer is a 10-34-0 starter fertilizer.

[0092] In certain embodiments, the biostimulant compositions described herein are applied to soil or plants in an amount of about 0.5 to about 10 quarts per acre. In some embodiments, the formulation is applied in an amount of about 4 quarts per acre. In some embodiments, the biostimulant composition is applied in an amount of about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 quarts per acre. In some embodiments, the amount of biostimulant composition applied is characterized by the dry weight of the substances present in the biostimulant composition applied. The dry weight of a given volume of a liquid biostimulant composition is the weight of all substances in that volume of biostimulant other than water. In some embodiments, the amount of biostimulant applied provides 0.10 to 10 g dry weight of digestion products per acre applied. In some embodiments, the amount of biostimulant applied provides at least about, at most about, or about 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 g dry weight of digestion products per acre applied, or a range between any two of these values. In some embodiments, the amount of biostimulant applied provides at least about, at most about, or about 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 g dry weight of biostimulant components per acre applied, or a range between any two of these values. The amount of biostimulant composition applied can also be characterized by the number of colony forming units of bacteria applied. In some embodiments, the amount of biostimulant applied provides at least about, at most about, or about 5 x 10 3 , 1 x 10 4 , 5 x 10 4 , 1 x 10 5 , 5 x 10 5 , 1 x 10 6 , or 5 x 10 6 CFU of bacteria per acre applied, or a range between any two of these values.

[0093] In some embodiments, the biostimulant compositions described herein can be applied in a dry form. The biostimulant base product can be dehydrated to make a powdered product, which is applied to a growth medium (e.g., soil), a plant, or a seed.

[0094] In some embodiments, the amount of biostimulant applied is an effective amount to achieve a desired plant growth promoting effect. For example, an effective amount of a biostimulant base product (such as the MBT-E product described in the Examples below) to increase cotton plant height compared to untreated plants is 0.5 or 1 quart per acre (qt. / A). In some embodiments, the biostimulant composition is applied in an amount effective to increase plant tolerance to drought, salt, heat, or cold stress, or to achieve any other desired result that the biostimulant composition described herein is capable of achieving.

[0095] Compositions comprising a biostimulant composition and other ingredients described herein (e.g., a fertilizer) can be formed by mixing the ingredients in a tank (i.e., tank mix). After mixing, the formulation can be bottled or otherwise packaged (e.g., tanked), applied to a field or crop, or mixed with other ingredients. After bottling or otherwise packaging, the end user can mix the formulation with other ingredients prior to application. The biostimulant composition can be mixed with conventional fertilizers by tank mix (including splash mix, with minimal further mixing), or can be incorporated into conventional fertilizers.

[0096] In some embodiments, the biostimulant is applied only once. In some embodiments, a single application is sufficient to promote plant growth as described herein. In some embodiments, the biostimulant composition is applied 1, 2, 3, 4, or 5 times in a growing season. In some embodiments, the applications are spaced 1, 2, 3, 4, 5, or 6 weeks apart.

[0097] IV. Certain Definitions In the foregoing description, certain specific details are set forth in order to provide a thorough understanding of various embodiments. However, one skilled in the relevant art will recognize that the embodiments provided can be practiced without one or more of the specific details, or with other methods, components, materials, etc. In other instances, well-known structures have not been described in detail in order to avoid obscuring aspects of the application. Unless otherwise specified, the word "comprise," and variations such as "comprises" or "comprising," will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps. As used herein, the term "or" is generally employed in its sense of "and / or" unless the content clearly dictates otherwise. In addition, the headings provided herein are for convenience only and are not to be construed as limiting the scope of the claimed embodiments.

[0098] The term "about" means within an acceptable range of error for the particular value as determined by one of ordinary skill in the art to which the claim pertains, which will depend in part on how the value is measured or determined, e.g., the limitations of the measurement system. For example, "about" can mean within 1 standard deviation, or more than 1 standard deviation, as is applied to a given value in practice. When particular values from a claim are described, unless otherwise indicated, the term "about" shall be presumed to indicate an acceptable range of error around that particular value.

[0099] Examples The following examples are provided to further illustrate some embodiments of the present disclosure, but are not intended to limit the scope of the present disclosure; it is understood that other procedures, methods, or techniques known to those skilled in the art can alternatively be used by their exemplary nature.

[0100] Example 1: Microbial digestion of giant kelp and characterization of digestion products Powdered giant kelp, chitin, and S. cerevisiae were mixed with water to make an organic feedstock for an anaerobic digestion system. The anaerobic digestion system included a first tank in which the organic feedstock was mixed to make a homogenous slurry. The slurry then flowed through a series of four digestion tanks in a continuous and hydraulically balanced manner. In the first digestion tank, the slurry was agitated at a rate that allowed heavier or undigested solids to settle to the bottom. An outlet at the top of the first digestion tank flowed fluid into the second digestion tank. An outlet at the bottom of the first digestion tank flowed the settled solids back into the first digestion tank. Each of the three subsequent tanks, referred to as packed bed reactors, had an immersed fixed media substrate that provided a surface for biofilm growth. The flow rate of the digestion system allowed for sufficient residence time within each digestion tank, thereby allowing for the formation of stable and unique microbial communities within each digestion tank. The microbes in the communities originated from the microbes originally present in the organic feedstock. These microbes digested the giant kelp, chitin, and yeast to produce digestion products. The effluent at the top of the fourth digestion tank, referred to herein as MBT-E base product (BP), was a clear liquid with a light tan color.

[0101] The giant kelp powder used as feedstock was evaluated by GC-MS and compared to Ascophyllum powder used as feedstock for Loveland AgriProducts commercial product sold as Maritime TM Bio-Products (also referred to herein as MBT-A). The GC-MS chromatograms are shown in Figure 1 FIG. 1, with the top chromatogram from Ascophyllum and the bottom chromatogram from giant kelp. Each chromatogram has unique peaks, as indicated by the arrows. The chromatograms also indicate that the relative abundance of the chemicals common to both seaweed feeds is different.

[0102] The chemical composition of Maritime TM Bio-Products (MBT-A) and MBT-E BP was also analyzed and compared. As previously by Santander et al. (2013) Microbiology159:1471 The sugar residues present in the base products were analyzed by GC-MS analysis of the per-oxytrimethylsilyl (TMS) derivatives of the monosaccharide methyl glycosides generated from the HCI methanolysis of the samples. Myo-inositol was added to each sample as an internal standard. After lyophilization and derivatization, the samples were extracted with hexane and GC-MS analysis of the TMS methyl glycosides was performed using an Agilent 7890A GC connected to a 5975C MSD equipped with a Supelco Equity - 1 fused silica capillary column (30 m x 0.25 mm ID). The results are shown in FIG. 1472. Figure 2 The LC-MS chromatograms of the two base products are shown in FIG. 1473. Figure 3 where MBT-A is on top and MBT-E is on the bottom.

[0103] The dichloromethane extracts of MBT-A and MBT-E were analyzed by GC-MS following the procedure: 0.25 L of each sample was extracted with CH2Cl2(0.25 L x 2 times) and a mixture of CH2Cl2:MeOH (2:1) (0.25 L x 2 times). The solvent extracts were filtered and dried under vacuum to obtain the dry material. To compare the chemical profile of different batches, GC-MS analysis was performed. The samples were derivatized with N, O-bis(trimethylsilyl) trifluoroacetamide (BSTFA) + 1% TMCS and analyzed using a palmitic acid- 13 C14 as an internal standard. GC-MS analysis was performed using an Agilent (Palo Alto, CA, USA) 8890 series GC system equipped with a CTC-Pal autosampler, a 5977B network mass selective detector, and a DB-1MS column (J&W, Palo Alto, CA, USA) (60 m x 0.25 mm id, 0.25 um film thickness). Data collection and analysis were performed using Agilent Mass Hunter Quantitative and Qualitative software by Agilent Technologies (Palo Alto, CA, USA). The GC-MS chromatograms of MBT-A and MBT-E are shown in FIG. 1474. Figure 4 where unique peaks are identified with arrows. Figure 39 The molecular species found only in MBT-E are listed. These were identified by the Mass Hunter Quantitative and Qualitative software by Agilent Technologies (Palo Alto, CA, USA) based on a matching factor > 70% and area > 1 x 10 5 .

[0104] The methanol extracts of the base products were analyzed by 1 H-NMR and 13 C-NMR, the spectra of which are shown in FIG. 1475.Figure 5 and Figure 6 Arrows indicate selected unique peaks.

[0105] MBT-E base product was treated with reverse osmosis to generate 4-fold and 8-fold concentrated versions of MBT-E.

[0106] Bacteria present in the giant kelp raw material and MBT-E base product were identified by 16S rRNA sequencing. Table 1 below shows bacteria present in both the raw material and MBT-E product.

[0107] Table 1: Bacteria identified in the giant kelp raw material and MBT-E product

[0108] Metagenomic sequencing was performed to identify spore-forming bacteria in the MBT-E product. The spore-forming bacteria content by metagenomic sequencing constituted approximately 0.7% of the total population (1 x 10 2 - 3 x 10 3 CFU / ml based on total bacterial count). Operational taxonomic units of spore-forming bacteria included Bacillus spp., Thermoanaerobacterium thermosulfurigenes, Paenibacillus Bacillus bacteriophores, T. peptonophilum, and T. indicum), psychrophilic Bacillus sp., and Paenibacillus. Aneurinibacillus thermoaerophilus Virgibacillus V. phasianinus V. dokdonensis Psychrobacillus Paenibacillus sphorae

[0109] Bacteria identified in the MBT-E product by metagenomic sequencing were compared to those present in MBT-A / Maritime™. The following list includes members of the MBT-E community that were present in statistically significantly higher numbers than in the MBT-A product. The percentages listed represent the percentage of the total population present in MBT-E, and the “x” numbers listed represent how many times higher the microbial population was in MBT-E compared to MBT-A.

[0110] • Sugar-degrading and cellulose community: Amycolatopsis sp. (0.0018%), Thermoanaerobacterium thermosaccharolyticum (0.0002%), Caldicellulosiruptor Cellulosilyticum lentocellum (0.00005%), Thermoanaerobacterium thermohydrosulfuricum (.0015%, 1.38x), an genus that can also degrade complex carbohydrates such as cellulose, alginate, and chitin.

[0111] • Collinsella sp. (.00003%, 2.19x): a genus that can produce NADPH-dependent 7 ​​​​​​- the genus of hydroxysteroid dehydrogenases that degrade bile acids (e.g., cholic acid) to secondary bile acids.

[0112] • Acinetobacter sp. (0.00043%, 2.6x), which includes A. towneri is an aromatic compound degrader, solubilizes iron and zinc through siderophore production and releases nutrients, contains a fungal inhibiting gene, and helps form a soil that suppresses pathogens.

[0113] • several non-spore forming lactic acid bacteria (LAB) communities including Lactobacillus buchneri (0.00005%, 2.41x), Lactobacillus bavaricus (0.000023%, 4.01x), and Lactobacillus paralimentarius (0.00013%, 1.83x).

[0114] 4x concentrated MBT-E also has the following properties: a light yellow liquid with a pH range of 7.5-8.5, a conductivity of 900-1100 (uS / cm), a density of 0.998 (g / cm 3 ), a solids content of 0.07%, a viscosity of 1.29 (cP), a COD of 20-150 (mg / L), a total bacterial count in the range of 5.0x10 4 to 5.0x10 5 CFU / ml, and a spore former count in the range of 2x10 2 to 3x10 3 CFU / ml.

[0115] Example 2: Plant growth promotion properties of MBT-E Two experiments were conducted in a rain shelter using drought stress mitigation of corn and cotton using Denton City soil (composted biosolids) and Whitestown soil. Corn and cotton plants were thinned after germination based on their uniform growth.

[0116] Materials and Methods: Plant physiological properties: After the start of the drought stress water regime and pot irrigation to 20-30% soil water capacity, plant physiological properties such as stomatal conductance (gsw), transpiration rate (E), chlorophyll a fluorescence / quantum yield (QY), electron transport rate (ETR), and leaf temperature (T) were measured using a LICOR 600 porometer / fluorometer portable photosynthesis system (Li-Cor, Inc. Lincoln, NE, USA). Ambient leaf temperature was measured as follows: Ambient leaf temperature (T amb ) = T 叶片 – T 参考Plant physiology parameters were measured on fully expanded cotton leaf blades of each plant between 11 :00 and 15:00 on days 15 and 26 after the start of the drought stress water regime. Plant physiology parameters were measured on fully expanded corn leaf blades of each plant between 11 :00 and 15:00 on days 7, 17, 26, and 32 after the start of the drought stress water regime.

[0117] Leaf chlorophyll content: The chlorophyll content was measured from fully expanded leaves using a chlorophyll meter SPAD (Soil Plant Analysis Development-502, Konica Minolta, Tokyo, Japan). The middle leaf position of the leaf was selected for measuring the leaf chlorophyll content to prevent variation.

[0118] Proline assay: Extraction and determination of the proline assay was performed using the method described by Carillo and Yves, PROTOCOL: Extraction and determination of proline Proline assay: Extraction and determination of the proline assay was performed using the method described by Carillo and Yves,

[0119] Relative water content (RWC): To determine the relative water content of the leaves, fresh fully expanded leaves were collected and measured using the following equation described by Teulat et al., QTL for relative water content in field-grown barley and their stability across Mediterranean environments. Theor. Appl. Genet. 2003, 108: 181-188 (2003). RWC (%) = (fresh weight - dry weight) / (fully turgid weight - dry weight) X 100 Statistical Analysis and Experimental Design: In the rain shelter experiment, a randomized complete block design (RCBD) was used to arrange the flowerpots. For the maize experiment, there were six treatments and 20 replicates, and for the cotton experiment, there were eight treatments and 20 replicates. Each replicate consisted of a single plant in a single flowerpot. JMP16 software (SAS Institute, Cary, NC, USA) was used to analyze the data on plant physiological parameters, leaf chlorophyll content, proline determination, biomass, and meta-analysis at a significance level of p < 0.1.

[0120] result Cotton drought stress relief experiment: In a rain shelter experiment, plants treated with MBT-E BP at ratios of 0.5 and 1 qt. / A showed better drought stress relief than the untreated control. During drought stress, MBT-E treatment improved all plant physiological parameters, such as stomatal conductance, transpiration rate, quantum yield, electron transport rate, leaf chlorophyll content, and proline. The effect on cotton leaf temperature was also discussed. Figure 21 and Figure 22 As shown. The effect on matrix conductivity is as follows. Figure 23 As shown. The effect on transpiration rate is as follows. Figure 24 As shown. The effect on quantum yield (the percentage of captured light energy used for photosynthesis and not lost as heat) is as follows. Figure 25 As shown. The effect on electron transport rate is as follows. Figure 26 As shown. The effect on leaf chlorophyll content is as follows. Figure 27 As shown. SPAD readings (leaf chlorophyll content) ranged from 44 to 51, and it was found that the leaves of plants treated with MBT-E had the highest chlorophyll content under drought stress. The SPAD readings for MBT-E recorded 15 days after the start of the drought stress water program were 50.46 (at a 0.5 qt. / A ratio) and 51.25 (at a 1 qt. / A ratio). Figure 7 (Table 2). Compared with the untreated control, the leaf temperature of MBT-E-treated plants was lower. The ambient leaf temperature of MBT-E-treated plants was 2.13 °C (at a ratio of 0.5 qt. / A) and 2.31 °C (at a ratio of 1 qt. / A). The proline concentration of MBT-E-treated plants was higher than that of the untreated control. At 30 days after the start of the drought stress water program, the proline concentration of MBT-E-treated plants was 28.66 (at a ratio of 0.5 qt. / A) and 27.75 (at a ratio of 1 qt. / A). Figure 8). Cotton boll production in plants treated with MBT-E at 1 qt. / A rate increased more than the untreated control. The average cotton plant height in MBT-E treated plants was 61.15 cm (at 0.5 qt. / A rate) and 62.35 cm (at 1 qt. / A rate) (Table 1). Figure 9 ). The average cotton bolls produced were 45 per plant (MBT-E at 1 qt. / A rate) and 44 per plant (MBT-E at 0.5 qt. / A rate) (Table 2). Figure 10 ). Cotton yield was significantly increased in MBT-E treated plants at 0.5 and 1 qt. / A (Table 3). Figure 11

[0121] Corn drought stress mitigation trial: In corn, the same trend was observed in terms of drought stress mitigation by applying MBT-E at the vegetative growth 6 (V6) stage and the vegetative tassel (Vt) stage. The stomatal conductance in MBT-E treated plants was higher than the untreated control plants at the V6 growth stage. The transpiration rate in MBT-E treated plants increased more than the untreated control plants at the V6+Vt growth stage. The quantum yield and electron transport rate in MBT-A and MBT-E treated plants were higher than the untreated control. The leaf temperature in all MBT-E treated plants was reduced during drought stress. The leaf temperature in MBT-E treated plants was lower than the untreated control at the Vt growth stage. The ambient leaf temperature for MBT-E was 0.38, 0.37, 0.11, and 0.15 °C at the Vt growth stage (Table 3). The SPAD readings (leaf chlorophyll content) ranged from 18 to 51, and the leaf chlorophyll content was the highest in plants treated with MBT-E at the V6+Vt growth stage under drought stress conditions (Table 4). The leaf relative water content (%) in MBT-E treated plants was higher than the untreated control at the V6 and V6+Vt growth stages (Table 5). Figure 12 ). The proline concentration in MBT-E was 21.50 (at the Vt growth stage) and 22.44 (at the V6+Vt growth stage) 30 days after the start of the drought stress water regime (Table 6). Figure 13 ). MBT-E treatment increased the ear length and dry weight in corn (Table 7). Figure 14 ). The highest average corn grain yield was observed in plants treated with MBT-E at the V6+Vt growth stage (Table 8). Figure 15

[0122] ​​In another experiment, the effect of MBT-E BP, 4x and 8x treatments on the growth rate of corn under drought stress conditions was tested by the following experiment: Corn (variety WS095 2021) was germinated in Berger general grow mix in a growth chamber at 22°C for 14 days. They were kept at 100% moisture capacity. After 14 days, they were fertilized with Jacks fertilizer. Treatments were applied as foliar treatments, with a total of 10 ml per plant for each treatment. The treated plants were placed in a lighted growth chamber at 22°C for 14 days. Each plant was kept at 30% moisture capacity to create moisture stress. All treatment solutions were filter sterilized and filter sterilized water was used for application at a dilution of 0.8%. The treatment solutions were: 1) water (as a negative control), 2) Accomplish LM TM (as a positive control), MBT-E base product (BP), MBT-E BP concentrated 4x (4xCP), and MBT-E BP concentrated 8x (8xCP). Each plant was a replicate, and each treatment was run 7 replicates. Plants were analyzed for indicators after 14 days. The results are shown in Table 2. Figure 20A

[0123] Example 3: Testing plant growth promoting properties of MBT-E in Arabidopsis The MS medium used in these examples was prepared by adding 4.43 g MS basal salts (Murashige and Skoog basal medium, Sigma Aldrich, M5519), 0.2 g myo-inositol, 1 g MES, and 20 g sucrose to 2 L of water. The pH was then adjusted to 5.7. 500 ml of the resulting solution was poured into each of 4 glass bottles that were filled with 1.77 g of Phytagel. The bottles were autoclaved, and the medium was poured into Petri dishes and allowed to set. If the goal was to collect data from the roots, the treated seedlings were grown in 0.01% MS medium with the addition of 0.66 g / l CaCl2 and 3.4 g / l Phytagel, adjusted to 5.7 pH before autoclaving. The medium was then poured into Petri dishes for setting. If the goal was to collect shoot data, the MS medium used in the treatment preparation was liquid MS medium consisting of MS basal salts plus 2 g / l MES.

[0124] ​The effect of MBT-E BP, 4x and 8x treatments on Arabidopsis shoot surface area was tested by the following experiment: Surface sterilized Arabidopsis seeds were placed on plates containing MS medium solidified with Phytagar and germinated at 20°C for 7 days. The seeds were then placed on rockwool cubes which were wetted with 40 ml of each treatment solution. All treatment solutions were filter sterilized and applied at a dilution of 0.8% with filter-sterilized water. The treatment solutions were: 1) water (as a negative control), 2) Accomplish LM TM (as a positive control), MBT-E base product (BP), 4x concentrated MBT-E BP (4xCP), and 8x concentrated MBT-E BP (8xCP), all diluted to 0.8% in liquid MS medium. Four rockwool cubes, each containing one seedling, constituted a replicate. Four replicates were performed for each treatment. The treated plants were placed on an LED growth cart in a completely randomized block design and grown at approximately 20°C for 14 days. The cubes were kept moist by the addition of 8 ml of water every 2 days. After 14 days, the leaf area of each plant was measured using ImageJ software by taking photographic images. The results are shown in Figure 16 All MBT-E treatments resulted in a significant increase in shoot surface area.

[0125] The effect of MBT-E BP, 4x and 8x treatments on Arabidopsis root length was tested by the following experiment: Surface sterilized Arabidopsis seeds were placed on plates containing MS medium solidified with Phytagar and germinated at 20°C for 6 days. The seeds were treated by immersing the roots of each seedling in its respective treatment solution. The treatment solutions were: 1) water (as a negative control), 2) Accomplish LM TM (as a positive control), MBT-E base product (BP), 4x concentrated MBT-E BP (4xCP), and 8x concentrated MBT-E BP (8xCP), all diluted to 0.8% in liquid MS medium. The treated seedlings were then placed on water agar plates containing 0.01% v / v bromocresol purple (3 seedlings per plate constituted a replicate). Four replicates were performed for each treatment. The treatments and replicates were placed on an LED growth cart in a completely randomized block design and grown at approximately 20°C for 12 days. After 7 days, the root area of each plant was measured by scanning and using WinRhizo software. The results are shown in Figure 17 and Figure 18 Statistical significance is indicated by an asterisk.

[0126] The effects of MBT-E BP, 4x and 8x treatments on Arabidopsis under drought stress conditions were tested in the following experiment: Surface-sterilized Arabidopsis seeds were placed on plates with MS medium solidified with Phytagar and germinated for 7 days at 20°C. Each treated seed was then placed in a separate petri dish with sphagnum moss, which was wetted with 30 ml of treatment solution. All treatment solutions were filter-sterilized and applied at a dilution of 0.8% with filter-sterilized water. The treatment solutions were: 1) water (as a negative control), 2) Accomplish LM TM (as a positive control), MBT-E base product (BP), 4x concentrated MBT-E BP (4xCP), and 8x concentrated MBT-E BP (8xCP). Four petri dishes, each with one seedling, constituted one replicate. Each treatment was run in four replicates. The treated plants were placed in a 22°C light growth chamber (Percival model 136LL) for 14 days. Water stress was created by maintaining the seedlings at 20% water capacity. The LED growth cart was arranged in a completely randomized block design and grown for 14 days at approximately 20°C. After 12 days, leaf area of each plant was measured using ImageJ software to take photographic images. The results are shown in Figure 19 . Asterisks indicate statistical significance.

[0127] Example 4: Effects of MBT-E under cold stress conditions The effects of MBT-E on Arabidopsis under cold stress conditions were tested in the following experiment: Surface-sterilized Arabidopsis seeds were placed on plates with MS medium solidified with Phytagar and germinated for 7 days at 20°C. Each treated seed was then placed in a separate petri dish with sphagnum moss, which was wetted with 30 ml of treatment solution. All treatment solutions were filter-sterilized and applied at a dilution of 0.8% with filter-sterilized water. The treatment solutions were: 1) water (as a negative control), 2) Accomplish LM TM (as a positive control), MBT-E base product (BP), 4x concentrated MBT-E BP (4xCP), and 8x concentrated MBT-E BP (8xCP). Four petri dishes, each with one seedling, constituted one replicate. Each treatment was run in four replicates. The treated plants were placed in a 12°C light growth chamber (Percival model LT41VL) for 21 days using a completely randomized block design. Observations were made at 14 and 21 days. Leaf area of each plant was measured using ImageJ software to take photographic images. The results of shoot surface area after cold treatment are shown in Figure 28 . Asterisks indicate statistical significance.

[0128] The effect of MBT-E on tomato under cold stress conditions was tested by the following experiment: Rutgers variety of tomato was germinated in Berger's General Mix and grown for 14 days in a growth chamber (Percival model 136LL) at 22°C. They were kept at 100% moisture capacity. Fourteen days after planting, they were fertilized with Jacks fertilizer. Treatments were provided as foliar applications, with each plant receiving a total of 10 ml of treatment solution. The treatment solutions were: 1) water (as a negative control), 2) MBT-E base product (BP), 3) MBT-E BP concentrated 4x (4xCP), 4) MBT-E BP concentrated 8x (8xCP). The treated plants were placed in a lighted growth chamber (Percival model LT41VL) programmed to provide cold stress by first providing 16°C for 1 hour, 8°C for 1 hour, 4°C for 2 hours, and -4°C for 2 hours. After this cold regime, the plants were evaluated using a cold stress rating scale of 0-5, where 0 indicates no observed shoot stress, and 5 indicates complete shoot death. The results of the cold stress rating are shown in Figure 29 The results of the fresh weight recovery are shown in Figure 30

[0129] Further cold stress effects in Arabidopsis were tested by the following experiment: Surface-sterilized Arabidopsis seeds were placed on plates with MS medium solidified with Phytagel and germinated for 7 days at 20°C. Each treated seed was then placed individually into a medicine cup with peat moss, which was moistened with 30 ml of treatment solution. All treatment solutions were filter-sterilized and applied at a dilution of 0.8% with filter-sterilized water. The treatment solutions were: water (as a negative control), MBT-E base product (BP), and MBT-E BP concentrated 4x (4xCP). Four medicine cups, each with one seedling, constituted one replicate. Each treatment was conducted in four replicates. The treated plants were placed in a lighted growth chamber at 12°C for 21 days, using a completely randomized block design. Observations were made at 14, 21, and 27 days after treatment (DAT). Leaf area of each plant was measured using ImageJ software, which takes photographic images. The results of shoot surface area are shown in Figure 31

[0130] Example 5: Effect of MBT-E on salt tolerance ​​The effect of MBT-E on salt tolerance was tested by the following experiment: Surface sterilized Arabidopsis seeds were placed on plates with MS medium solidified with Phytagar and germinated for 7 days at 20°C. The seeds were then placed on rock wool blocks, which were wetted with 40 ml of each treatment. All treatments were filter sterilized and applied at a dilution of 0.2% with filter sterilized water. The treatments were: water (as a negative control), MBT-E base product (BP), and 4x concentrated MBT-E BP (4xCP). All treatments also contained 75 mM NaCl. Four rock wool blocks, each containing one seedling, constituted one replicate. Four replicates were performed for each treatment. The treated plants were placed on an LED grow cart in a completely randomized block design and grown for 14 days at approximately 20°C. The blocks were kept moist by adding 8 ml of water every 2 days. After 14 days, leaf area of each plant was measured using ImageJ by taking photographic images. The results are shown in Figure 32

[0131] The effect of MBT-E on salt tolerance in corn was tested by the following experiment: Dynagro corn seeds were planted in 4: 1 MVP turface / Sungro black peat medium. The corn was thinned 12 days after planting to maintain uniformity, fertilized with Jack’s 20-20-20 at 25 pounds N / A, and salt stressed with 75 millimoles of NaCl. MBT-E was applied foliarly at 1 qt / A and 2 qt / A 21 days after planting. SPAD, imaging, and LiCor measurements were taken for biomass at 3 dates prior to corn harvest. The results for leaf chlorophyll content (SPAD) are shown in Figure 33

[0132] The effect of MBT-E on salt tolerance in zinnias was tested by the following experiment: Dwarf zinnia seeds were planted in 3: 1 Isolite / Sunshine Mix LC1 peat medium, thinned to maintain uniformity, and fertilized with Jack’s 20-20-20 at 50 pounds / A. Soil electrical conductivity (EC) measurements were taken prior to planting and throughout the experiment. Sodium chloride was applied in three applications for a total of 100 millimoles. MBT-E was applied foliarly at 1 qt / A and 2 qt / A rates 34 days after planting. Indicators for this experiment included stem diameter, height, total biomass, and final EC reading of the soil at harvest. The results for stem diameter at harvest are shown in Figure 34 Figure 35A Figure 36

[0133] Example 6: Field trial of MBT-E to promote growth of sweet pepper plants​​​​​ Pepper seedlings were transplanted into raised beds in Yuma, Arizona. Prior to planting, the beds were provided with MAP (monoammonium phosphate) fertilizer at a rate of 300 pounds per acre. The control treatment (no MBT-E added) was planted in two beds in four replicates, each replicate 75 feet long. The MBT-E treatment was planted in four beds in eight replicates, each replicate 75 feet long. Three weeks after planting, the plants were fertilized with UAN 32 (urea ammonium nitrate) fertilizer using an underground drip irrigation system, each treatment (no MBT-E added or 2 qt per acre added MBT-E). The no MBT-E and MBT-E treatments were provided only at the first fertilization. Thereafter, the plants were provided with two additional fertilizations during the growing season. The plants were harvested on June 17, 2022. After harvesting, yield ( Figure 37 ) and tissue nutrient data ( Figure 38 ) were collected. Leaf tissue from plants treated with MBT-E showed increased levels of nitrogen, phosphorus, and potassium relative to leaf tissue from control plants.

[0134] Example 7: Microbial population analysis of seaweed feedstock and biostimulant products Two batches of different lots of seaweed feedstock powder (Ecklonia maxima for MBT-E and Ascophyllum nodosum for MBT-A) were sampled, with five or three technical replicates per batch, respectively. DNA was extracted from 0.025 g of powder using the bead beating extraction method and phenol-chloroform purification. Two or three different lots of solution were sampled for MBT-A 4X and MBT-E, respectively, with four or three technical replicates. 100 ml of concentrated product solution was filtered, and bacterial cells were collected from the filter and then DNA was extracted using the MP Biomedicals DNA Soil Pro kit.

[0135] Amplification-based DNA sequencing for samples was completed by Molecular Research Corporation (MRDNA, Shallowater, TX) using their standard methods for bacterial analysis, with 16S-515F primers, with 20,000 reads per sample on an Illumina NovaSeq 6000 system. Additionally, MRDNA performed QA / QC, chimera checking, and OTU (operational taxonomic unit) binning. MRDNA’s output was analyzed in the statistical analysis platform R using the vegan package for displaying community analysis profiles as UPGMA-based cluster analysis trees.

[0136] Cluster analysis trees for Ascophyllum nodosum and Ecklonia maxima feedstocks are shown in FIGS. 6A and 6B, respectively. Figure 40The EMF and AMF raw material powder bacterial communities are clearly different. All EMF community samples cluster together and originate from the same branch and show some slight separation and differences between the two batches of EMF analyzed (1 and 2). However, these EMF samples are very similar to each other as they branch off from each other. For all AMF community samples, they cluster together and originate from the same branch. The “height” scale on the left is analogous to the percent difference between samples. The longer the branch, the greater the difference in samples. Since the EMF and AMF raw material powder communities do not overlap in the same series of branches, it can be concluded that the EMF and AMF raw material powders have different microbial communities.

[0137] The cluster analysis tree for MBT-A and MBT-E is shown in FIG. 4. Figure 41 The MBT-E 4X and MBT-A 4X bacterial communities are clearly different. The MBT-E and MBT-A communities are clearly separated and do not overlap in a series of branches. The three batches of MBT-E are very similar to each other and the MBT-A-1 and 2 communities are more similar to each other than MBT-A-3. These MBT-E batch communities do overlap in a series of branches.

[0138] Table 2. Plant physiological parameters, plant height, and boll yield recorded under cotton drought stress conditions (1 and 2 indicate two different data recording dates: 14 and 25 days after treatment application) (stomatal conductance - gsw, transpiration rate - E, quantum yield - QY, electron transport rate - ETR, leaf chlorophyll content - SPAD, relative water content - RWC, and temperature - T)

[0139] Table 3. Plant physiological parameters recorded under corn drought stress conditions (1, 2, 3, and 4 indicate four different data recording dates: 10, 16, 30, and 37 days after treatment application)

[0140] Table 4. Plant physiological parameters, leaf relative water content, and proline accumulation recorded under corn drought stress conditions (1, 2, 3, and 4 indicate four different data recording dates in 2021: 10, 16, 30, and 37 days after treatment application)

[0141] While preferred embodiments of the present disclosure have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the disclosure. It should be understood that in the practice of the present disclosure, various alternatives to the embodiments of the disclosure can be employed. It is intended that the following claims define the scope of the disclosure and that methods and structures within the scope of these claims and their equivalents be covered thereby.

Claims

1. A method of promoting plant growth, the method comprising contacting a plant, a seed of the plant, or a growth medium of the plant with a composition comprising a microbial digest product of an organic feedstock, the organic feedstock comprising E. maxima kelp.

2. The method of claim 1, wherein the digest product is produced by endogenous microorganisms of the E. maxima kelp present in the organic feedstock.

3. The method of claim 1, wherein the digest product comprises no more than 40 mol% of fucose relative to the concentration of all glycosyl residues in the composition.

4. The method of claim 1, wherein the digest product comprises more than 15 mol% of xylose relative to the concentration of all glycosyl residues in the composition.

5. The method of claim 1, wherein the digest product comprises more than 7 mol% of mannose relative to the concentration of all glycosyl residues in the composition.

6. The method of claim 1, wherein the digest product comprises one or more of iso-butanol, pentadecanenitrile, pentadecanoic acid, 9-octadecenonitrile, hexadecanenitrile, or heneicosane.

7. The method of claim 6, wherein the digest product comprises the molecular species listed in FIG.

39.

8. The method of claim 1, wherein the digest product comprises one or more molecular species corresponding to one or more of peaks C, F, G, J, O, or P in the LC-MS chromatogram shown in FIG.

3.

9. The method of claim 1, wherein the digest product comprises one or more molecular species corresponding to one or more of peaks A, B, C, D, E, F, or G in the GC-MS chromatogram shown in FIG.

4.

10. The method of claim 1, wherein the digestion product comprises one or more molecular species corresponding to one or more of peaks A, B, C, D, or E in the H-NMR spectrum shown in Figure 5. 1 one or more of peaks A, B, C, D, or E in the H-NMR spectrum.

11. The method of claim 1, wherein the digestion product comprises one or more molecular species corresponding to one or more of peaks A or B in the C-NMR spectrum shown in Figure 6. 13 C-NMR spectrum.

12. The method of claim 1, wherein the composition further comprises the endogenous microorganisms of the E. maxima kelp present in the organic feedstock.

13. The method of claim 12, wherein the microorganisms comprise sporulating microorganisms.

14. The method of claim 12, wherein the percentage of dry weight of microbial biomass in the composition is from 0.071% to 0.714% relative to the total dry weight of the composition.

15. The method of claim 12, wherein the microorganisms present in the composition comprise *Microbacterium amyloliquefaciens*, *Anaerobic bacillus pyrolyticus*, and... Cellulosilyticumlentocellum One or more of the following: heat-resistant microbubbles, Collins bacteria, Acinetobacter species, Acinetobacter tangs, Lactobacillus brunelli, Lactobacillus barleyii, or secondary Lactobacillus parasiticus.

16. The method of claim 1, wherein the percentage of dry weight of microbial biomass in the composition is less than 0.001 wt% relative to the total dry weight of the composition.

17. The method of claim 1, wherein the composition does not comprise microorganisms.

18. The method of claim 1, wherein promoting plant growth comprises one or more of: enhancing seed germination, enhancing early plant development, improving root growth, increasing nutrient uptake, improving tolerance to abiotic stress, reducing transplant shock, improving plant reproduction, and improving soil microbial activity.

19. The method of claim 18, wherein improving tolerance to abiotic stress comprises improving one or more of: salt tolerance, heat tolerance, cold tolerance, and drought tolerance.

20. The method of claim 1, wherein said contacting comprises in-ditch application, foliar spray application, application to a root zone, application to a seed, or mixing with a growth medium.

21. The method of claim 1, wherein said growth medium is soil.

22. The method of claim 1, wherein said composition further comprises solid fertilizer particles.

23. The method of claim 22, wherein said fertilizer particles are coated with said digestion product.

24. The method of claim 1, wherein said composition is a liquid.

25. The method of claim 24, wherein said composition further comprises a liquid fertilizer.

26. The method of claim 24, wherein said contacting comprises applying said composition at a rate of 0.5 to 10 quarts per acre.

27. The method of claim 1, wherein said contacting comprises applying 0.14 to 6.7 g dry weight of said digestion product per acre.

28. The method of claim 1, wherein said plant is experiencing or at risk of experiencing drought conditions at the time said contacting is performed.

29. The method of claim 1, wherein said growth medium is a high-salt soil.

30. The method of claim 1, wherein said plant is experiencing or at risk of experiencing freezing conditions at the time said contacting is performed.

31. The method of claim 1, wherein said plant is experiencing or at risk of experiencing cold stress at the time said contacting is performed.

32. The method of claim 1, wherein said plant is experiencing or at risk of experiencing heat stress at the time said contacting is performed.

33. The method of claim 1, wherein said plant has been transplanted.

34. The method of claim 1, wherein said plant is corn, cotton, tomato, or pepper.

35. The method of claim 1, wherein said plant is a cotton plant or a corn plant, and wherein said cotton plant or corn plant is under drought conditions at the time said contacting is performed.

36. A composition comprising a digestion product produced from digestion of an organic feedstock by a microorganism, said organic feedstock comprising the giant kelp seaweed.

37. The composition of claim 36, wherein said microorganism comprises endogenous microorganisms of said giant kelp seaweed present in said organic feedstock.

38. The composition of claim 36, wherein said digestion product comprises no more than 40 mol% of fucose relative to the concentration of all glycosyl residues in said composition.

39. The composition of claim 36, wherein said digestion product comprises more than 15 mol% of xylose relative to the concentration of all glycosyl residues in said composition.

40. The composition of claim 36, wherein said digestion product comprises more than 7 mol% of mannose relative to the concentration of all glycosyl residues in said composition.

41. The composition of claim 36, wherein the digestion products comprise one or more of isobutanol, pentadecanenitrile, pentadecanoic acid, 9-octadecenonitrile, hexadecanenitrile, or heneicosane.

42. The composition of claim 40, wherein the digestion products comprise the molecular species listed in FIG.

39.

43. The composition of claim 36, wherein the digestion products comprise one or more molecular species corresponding to one or more of peaks C, F, G, J, O, or P in the LC-MS chromatogram shown in FIG.

3.

44. The composition of claim 36, wherein the digestion products comprise one or more molecular species corresponding to one or more of peaks A, B, C, D, E, F, or G in the GC-MS chromatogram shown in FIG.

4.

45. The composition of claim 36, wherein the digestion product comprises one or more of the molecular species corresponding to one or more of peaks A, B, C, D, or E shown in FIG.

5. 1 one or more of peaks A, B, C, D, or E in the H-NMR spectrum.

46. The composition of claim 36, wherein the digestion product comprises one or more molecular species corresponding to one or more of peaks A or B in the C-NMR spectrum shown in Figure 6. 13 one or more of peaks A or B in the C-NMR spectrum.

47. The composition of claim 36, further comprising the E. maxima endogenous microorganism present in the organic feedstock.

48. The composition of claim 47, wherein the microorganism comprises a sporulating microorganism.

49. The composition of claim 47, wherein the percent dry weight of the microbial biomass in the composition is between 0.071% and 0.714% relative to the total dry weight of the composition.

50. The composition of claim 47, wherein the microorganisms present in the composition comprise *Microbacterium amyloliquefaciens*, *Anaerobic bacillus pyrolyticus*, and... Cellulosilyticumlentocellum One or more of the following: heat-resistant microbubbles, Collins bacteria, Acinetobacter species, Acinetobacter tangs, Lactobacillus brunelli, Lactobacillus barleyii, or secondary Lactobacillus parasiticus.

51. The composition of claim 36, wherein the microorganism has been removed from the composition.

52. The composition of claim 36, wherein the percent dry weight of the microbial biomass in the composition is less than 0.001% relative to the total dry weight of the composition.

53. The composition of claim 36, wherein the composition does not comprise a microorganism.

54. A composition comprising one or more molecular species corresponding to one or more of peaks C, F, G, J, O, or P in the LC-MS chromatogram shown in FIG.

3.

55. The composition of claim 54, wherein the composition comprises one or more molecular species corresponding to one or more of peaks A, B, C, D, E, F, or G in the GC-MS chromatogram shown in FIG.

4.

56. The composition of claim 54, wherein the composition comprises one or more molecular species corresponding to one or more of peaks A, B, C, D, or E shown in FIG.

5. 1 one or more of peaks A, B, C, D, or E in the H-NMR spectrum.

57. The composition of claim 54, wherein the composition comprises one or more molecular species corresponding to one or more of peaks A or B in the C-NMR spectrum shown in Figure 6. 13 one or more of the one or more molecular species corresponding to one or more of peaks A or B in the C-NMR spectrum.

58. The composition of claim 54, wherein the composition comprises xylose at a concentration of more than 15 mol% relative to all glycosyl residues in the composition.

59. The composition of claim 54, wherein the composition comprises mannose at a concentration of more than 7 mol% relative to all glycosyl residues in the composition.

60. The composition of claim 54, wherein the digestion products comprise one or more of isobutanol, pentadecanenitrile, pentadecanoic acid, 9-octadecenonitrile, hexadecanenitrile, or heneicosane.

61. The composition of claim 60, wherein the digestion products comprise the molecular species listed in FIG.

39.

62. The composition of claim 54, further comprising a microorganism.

63. The composition of claim 62, wherein the microorganism comprises a sporulating microorganism.

64. The composition of claim 62, wherein the percentage of dry weight of the microbial biomass in the composition is from 0.071% to 0.714% relative to the total dry weight of the composition.

65. The composition of claim 62, wherein the microorganisms present in the composition comprise one or more of Tyzzerella praeacuta, Caldicellulosiruptor obscurus, Thermoanaerobacterium sp., Collinsella sp., Acinetobacter sp., Acinetobacter tonkinensis, Lactobacillus buccae, Lactobacillus farciminis, Lactobacillus britannicus, or Lactobacillus paracasei. Cellulosilyticumlentocellum , Thermoanaerobacterium sp., Collinsella sp., Acinetobacter sp., Acinetobacter tonkinensis, Lactobacillus buccae, Lactobacillus farciminis, Lactobacillus britannicus, or Lactobacillus paracasei.

66. The composition of claim 54, wherein the percentage of dry weight of the microbial biomass in the composition is less than 0.001% relative to the total dry weight of the composition.

67. The composition of claim 54, wherein the composition does not comprise microorganisms.

68. The composition of claim 54, wherein the composition is a liquid composition.

69. The composition of claim 68, wherein the digestion product is present in the liquid composition at from 0.06% to 0.08% by weight relative to the total weight of the liquid composition.

70. A plant treatment composition comprising the composition of claim 36 and a fertilizer composition.

71. The plant treatment composition of claim 70, wherein the fertilizer composition is a liquid.

72. The plant treatment composition of claim 70, wherein the fertilizer composition is a solid.

73. The plant treatment composition of claim 72, wherein the fertilizer composition is coated with the composition of claim 36.

74. A method of promoting plant growth, the method comprising contacting a plant, a seed of the plant, or a growth medium of the plant with the composition of claim 36.

75. The method of claim 74, wherein promoting plant growth comprises one or more of: enhancing seed germination, enhancing early plant development, improving root growth, increasing nutrient uptake, improving tolerance to abiotic stress, reducing transplant shock, improving plant reproduction, and improving soil microbial activity.

76. The method of claim 75, wherein improving tolerance to abiotic stress comprises improving one or more of: salt tolerance, heat tolerance, cold tolerance, and drought tolerance.

77. The method of claim 74, wherein the contacting comprises in-furrow application, foliar spray application, or application to a root zone.

78. The method of claim 70, wherein the contacting comprises applying from 0.14 to 6.7 g of dry weight of the digestion product per acre.

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