Biostimulant composition

Abstract

A biostimulant composition prepared by digesting Ecklonia maxima, and a method for promoting plant growth by contacting a plant, seed, or plant growth medium with such a composition.

Description

Technical Field

[0001] Cross-reference This application claims the benefit of U.S. Provisional Patent Application No. 63 / 455,662, filed Mar. 30, 2023, which is hereby incorporated by reference in its entirety.

Background Art

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

[0003] For environmental and economic reasons, promoting the efficient production of food crops and other crops is an important goal. Products for promoting plant growth supplied from organic materials can help enhance crop growth, improve the effectiveness of agricultural products such as fertilizers, and reduce the environmental impact of synthetic fertilizers and climate change. There is a need for biostimulant compositions that promote plant growth using abundant and available organic raw materials.

Summary of the Invention

[0004] The present disclosure provides a biostimulant composition for promoting plant growth made from Ecklonia Maxima raw materials and methods of using such biostimulant compositions.

[0005] A method for promoting plant growth is disclosed herein, which includes the step of contacting a plant, a plant seed, or a growth medium for a plant with a composition containing a microbial digestion product of an organic raw material containing Ecklonia maxima kelp. In some embodiments, the digestion is anaerobic digestion. In some embodiments, the digestion product is produced by microorganisms endogenous to Ecklonia maxima kelp present in the organic raw material. In some embodiments, the digestion product contains fucose at a concentration of 40 mol% or less relative to all glycosyl residues in the composition. In some embodiments, the digestion product contains xylose at a concentration of 15 mol% or less relative to all glycosyl residues in the composition. In some embodiments, the digestion product contains mannose at a concentration exceeding 7 mol% relative to all glycosyl residues in the composition. In some embodiments, the digestion product contains one or more of isobutanol, pentadecanenitrile, pentadecanoic acid, 9-octadecenenitrile, hexadecanenitrile, or heneicosane. In some embodiments, the digestion product contains the molecular species listed in Figure 39. In some embodiments, the digestion product contains 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 Figure 3. In some embodiments, the digestion product contains 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 Figure 4. In some embodiments, the digestion product contains the 1 one or more molecular species corresponding to one or more of peaks A, B, C, D, or E in the 1H-NMR spectrum. In some embodiments, the digestion product contains the 13 one or more molecular species corresponding to one or more of peaks A or B in the 13C-NMR spectrum.

[0006] In some embodiments, the composition further comprises microorganisms intrinsically present in Ecklonia maxima in the organic raw material. In some embodiments, the microorganisms include spore-forming microorganisms. In some embodiments, the dry weight percentage of microbial biomass in the composition is 0.071 to 0.714% of the total dry weight of the composition. In some embodiments, the microorganisms present in the composition include Microbacterium amylolyticum, Thermoanaerobacterium thermosaccharolyticum, Cellulosilyticum lentocellum, Microbulbifer thermotolerans, Collinsella sp., Acinetobacter spp., Acinetobacter towneri, Lentilactobacillus buchneri, Liquorilactobacillus hordei, or Secandelactobacillus paracolinoides It contains one or more paracollinoides. In some embodiments, the dry weight percentage of microbial biomass in the composition is less than 0.001 wt% of 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, promoting early plant development, improving root growth, increasing nutrient uptake, improving tolerance to abiotic stress, mitigating the effects of transplanting, improving plant reproduction, and improving soil microbial activity. In some embodiments, improving tolerance to abiotic stress includes improving one or more of the following: salt tolerance, heat tolerance, cold tolerance, and drought tolerance. In some embodiments, the contact step includes infallo application, foliar application, rhizosphere application, seed application, or mixing with a growth medium. In some embodiments, the growth medium is soil.

[0008] In some embodiments, the composition further comprises solid fertilizer particles. In some embodiments, the fertilizer particles are coated with digests. In some embodiments, the composition is liquid. In some embodiments, the composition further comprises liquid fertilizer.

[0009] In some embodiments, the contact step includes applying the composition at a rate of 0.5 to 10 quarts per acre. In some embodiments, the contact step includes applying 0.14 to 6.7 g of the digest by dry weight per acre.

[0010] In some embodiments, at the time of contact, the plants are experiencing or at risk of experiencing dehydration. In some embodiments, the growing medium is high-salinity soil. In some embodiments, at the time of contact, the plants are experiencing or at risk of experiencing freezing. In some embodiments, at the time of contact, the plants are experiencing or at risk of experiencing cold stress. In some embodiments, at the time of contact, the plants are experiencing or at risk of experiencing heat stress. In some embodiments, the plants are transplanted. In some embodiments, the plants are corn, cotton, tomato, or pepper. In some embodiments, the plants are cotton plants or maize plants, and the cotton plants or maize plants are dehydrated at the time of contact.

[0011] Also disclosed herein are compositions comprising digest products produced by the digestion of an organic raw material containing Ecklonia maxima kelp by microorganisms. In some embodiments, the microorganisms include those endemic to Ecklonia maxima kelp present in the organic raw material. In some embodiments, the digest product contains fucose at a concentration of 40 mol% or less relative to all glycosyl residues in the composition. In some embodiments, the digest product contains xylose at a concentration of more than 15 mol% relative to all glycosyl residues in the composition. In some embodiments, the digest product contains mannose at a concentration of more than 7 mol% relative to all glycosyl residues in the composition. In some embodiments, the digest product comprises one or more of isobutanol, pentadecanonitrile, pentadecanoic acid, 9-octadecennitrile, hexadecanenitrile, or heneicosane. In some embodiments, the digest product comprises molecular species listed in Figure 39. In some embodiments, the digested product includes one or more molecular species corresponding to one or more peaks C, F, G, J, O, or P in the LC-MS chromatogram shown in Figure 3. In some embodiments, the digested product includes one or more molecular species corresponding to one or more peaks A, B, C, D, E, F, or G in the GC-MS chromatogram shown in Figure 4. In some embodiments, the digested product is shown in Figure 5. 1 The digested product contains one or more molecular species corresponding to one or more peaks A, B, C, D, or E in the 1H-NMR spectrum. In some embodiments, the digested product is shown in Figure 6. 13 It contains one or more molecular species corresponding to one or more peaks A or B in the 1C-NMR spectrum.

[0012] In some embodiments, the composition further includes microorganisms endogenous to Ecklonia maxima present in the organic raw material. In some embodiments, the microorganisms include spore-forming microorganisms. In some embodiments, the dry weight percentage of microbial biomass in the composition is 0.071 to 0.714% of the total dry weight of the composition. In some embodiments, the microorganisms present in the composition include one or more of Microbacterium amiloristicum, Thermoanaerobacterium thermosaccharoristicum, Cellulosilicum lentocerum, Microbarbifer thermotolerance, Cholincera sp., Acinetobacter spp., Acinetobacter touneri, Lentilactobacillus buhineri, Lycorolactobacillus fordei, or Secandilactobacillus paracolinoides. In some embodiments, the microorganisms are removed from the composition. In some embodiments, the dry weight percentage of microbial biomass in the composition is less than 0.001% of the total dry weight of the composition. In some embodiments, the composition does not contain microorganisms.

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

[0014] In some embodiments, the composition further comprises microorganisms. In some embodiments, the microorganisms include spore-forming microorganisms. In some embodiments, the dry weight percentage of microbial biomass in the composition is 0.071 to 0.714% of the total dry weight of the composition. In some embodiments, the microorganisms present in the composition include one or more of the following: Microbacterium amiloristicum, Thermoanaerobacterium thermosaccharoristicum, Cellulosilicum lentocerum, Microbarbifer thermotolerance, Cholincera sp., Acinetobacter spp., Acinetobacter touneri, Lentilactobacillus buhineri, Lycorolactobacillus fordei, or Secandilactobacillus paracolinoides. In some embodiments, the dry weight percentage of microbial biomass in the composition is less than 0.001% of the total dry weight of the composition. In some embodiments, the composition does not contain microorganisms.

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

[0016] Plant treatment compositions comprising any of the above-described compositions and a fertilizer composition are also disclosed. In some embodiments, the fertilizer composition is liquid. In some embodiments, the fertilizer composition is solid. In some embodiments, the fertilizer composition is coated with any of the above-described biostimulant compositions.

[0017] Also disclosed are methods for promoting plant growth, comprising the step of contacting a plant, plant seeds, or a growing medium for plants with any of the above compositions. In some embodiments, promoting plant growth includes one or more of the following: promoting seed germination, promoting early plant development, improving root growth, increasing nutrient uptake, improving tolerance to abiotic stress, mitigating the effects of transplanting, improving plant reproduction, and improving soil microbial activity. In some embodiments, improving tolerance to abiotic stress includes improving one or more of the following: salt tolerance, heat tolerance, cold tolerance, and drought tolerance. In some embodiments, the contact step includes infallo application, foliar application, or application to the rhizosphere. In some embodiments, the contact step includes applying 0.14 to 6.7 g of the digest by dry weight per acre.

[0018] Reference All publications, patents, and patent applications referenced herein are incorporated herein by reference to the same extent as individual publications, patents, or patent applications are incorporated herein by reference specifically and individually. [Brief explanation of the drawing]

[0019] Novel features of this disclosure are described in particular in conjunction with the appended claims. A better understanding of the features and advantages of this disclosure can be obtained by referring to the following detailed description and accompanying drawings illustrating exemplary embodiments in which the principles of this disclosure are utilized.

[0020] [Figure 1]These are GC-MS chromatograms of Ascophyllum nodosum powder (upper chromatogram) and Ecklonia maxima powder (lower chromatogram). [Figure 2] This shows the results of the analysis of glycosyl residue content in MBT-A and MBT-E products. [Figure 3] These are LC-MS chromatograms of the MBT-A (upper) product and the MBT-E (lower) product. [Figure 4] These are GC-MS chromatograms of MBT-E (upper) and MBT-A (lower) products. The arrows point to intrinsic peaks in MBT-E identified by Mass Hunter quantitative and qualitative software from Agilent Technologies (Palo Alto, California, USA), based on an agreement coefficient of over 70% and an area greater than 1 × 10⁵. [Figure 5] These are the 1H-NMR spectra of MBT-A (upper) and MBT-E (lower). [Figure 6] These are the 13C-NMR spectra of MBT-A (upper) and MBT-E (lower). [Figure 7] Chlorophyll content of cotton leaves under drought stress (SPAD1 and SPAD2 were recorded on day 15 and day 26, respectively, after the start of drought stress water management). [Figure 8] Proline accumulation under drought stress (proline concentration was recorded on day 30 after the start of drought stress water management). [Figure 9] Height of cotton plants under drought stress (plant height was recorded 90 days after the start of drought stress water management). [Figure 10] Cotton ball yield under drought stress (balls were counted 80 days after the start of drought stress moisture management). Cotton ball size was measured in centimeters (cm), and balls larger than 2 cm were not considered. [Figure 11] Average cotton production from MBT-E treated plants and untreated control plants. [Figure 12] Relative moisture content of corn leaves under drought stress, measured 26 days after the start of drought stress moisture management. [Figure 13] Proline accumulation under drought stress (proline concentration was recorded on day 30 after the start of drought stress water management). [Figure 14] Average length and weight of corn ears measured immediately before harvest. [Figure 15] The corn yield resulting from the indicated treatment. [Figure 16] The surface area of ​​the chute resulting from the indicated treatment. An asterisk indicates a statistically significant difference. [Figure 17] The root surface area resulting from the indicated treatment. An asterisk indicates a statistical difference. [Figure 18] Root length resulting from the indicated treatment. An asterisk indicates a statistical difference. [Figure 19] The surface area of ​​the chute resulting from the indicated treatment. An asterisk indicates a statistical difference. [Figure 20] (A) Growth rate in a dry state resulting from the indicated treatment. (B) Growth rate after recovery resulting from the indicated treatment. (C) Growth rate resulting from the indicated treatment. An asterisk indicates a statistical difference. [Figure 21] Leaf temperature resulting from the indicated treatment. UTC = untreated control. [Figure 22] The rate of change in stomatal conductance over the indicated time period, resulting from the indicated treatment. [Figure 23] The rate of change in stomatal conductance over the indicated time period, resulting from the indicated treatment. [Figure 24] The rate of change in stomatal conductance over the indicated time period, resulting from the indicated treatment. [Figure 25] The rate of change in stomatal conductance over the indicated time period, resulting from the indicated treatment. [Figure 26]The rate of change in stomatal conductance over the indicated time period, resulting from the indicated treatment. [Figure 27] The chlorophyll content of cotton leaves at the indicated time, resulting from the indicated treatment. [Figure 28] The surface area of ​​the chute resulting from the indicated treatment. An asterisk indicates a statistical difference. [Figure 29] The evaluation of plants resulting from the indicated treatment. Asterisks indicate statistical differences. [Figure 30] The weight of fresh shoots resulting from the indicated treatment. An asterisk indicates a statistical difference. [Figure 31] The surface area and growth rate of shoots resulting from the indicated treatment. Asterisks indicate statistical differences. [Figure 32] The surface area of ​​the chute resulting from the indicated treatment. An asterisk indicates a statistical difference. [Figure 33] Chlorophyll content (SPAD) resulting from the indicated treatment. Asterisks indicate statistically significant differences. [Figure 34] The diameter of the stem resulting from the indicated treatment. [Figure 35] (A) Height at harvest resulting from the indicated treatment. (B) Total biomass resulting from the indicated treatment. [Figure 36] The electrical conductivity of the soil resulting from the indicated treatment. [Figure 37] The yield of bell peppers resulting from the indicated treatment. [Figure 38] The nutrient content resulting from the indicated process. [Figure 39] Unique peaks in MBT-E compared to the GC-MS chromatogram of MBT-A. [Figure 40]Bacterial community analysis was performed on two seaweed samples: Ecklonia maxima (EMF) and Ascophyllum nodosum (ANF). DNA was extracted from the powdered seaweed samples, and the bacterial communities were characterized by amplicon sequencing of small ribosomal RNA genes (i.e., 16S rRNA genes). The bacterial community profiles were presented as UPGMA cluster analysis diagrams. [Figure 41] Bacterial community analysis of seaweed product solutions MBT-E and MBT-A. DNA was extracted from concentrated product solutions (4X), and bacterial communities were characterized by amplicon sequencing of small ribosomal RNA genes (i.e., 16S rRNA genes). Bacterial community profiles were presented as UPGMA cluster analysis diagrams. [Modes for carrying out the invention]

[0021] Embodiments described herein include biostimulant compositions and methods for promoting plant growth and increasing plant tolerance to abiotic stresses, such as drought, cold, heat, and salinity stress. The compositions include microbial digest products produced by the digestion of Ecklonia maxima kelp.

[0022] I. Digestive Process In some embodiments, the biostimulant composition is prepared by a digestion process of an organic raw material containing Ecklonia maxima kelp. In some embodiments, the organic raw material further comprises chitin and Saccharomyces cerevisiae yeast. The organic raw material may also be an aqueous slurry of powdered Ecklonia maxima kelp, chitin, and Saccharomyces yeast. In some embodiments, the digestion is anaerobic. While not bound by theory, it is believed that during the digestion process, microorganisms inherent in Ecklonia maxima kelp and chitin digest biomolecules and other nutrients present in the kelp, chitin, and yeast, producing digest products containing compounds that promote plant growth, abiotic stress tolerance, and soil health. Furthermore, the biostimulant may contain microorganisms that contribute to the plant-beneficial properties of the biostimulant product. The microorganisms in the biostimulant product may originate from the microbial population present in the kelp raw material.

[0023] The digestion process for producing a biostimulant may be carried out in a digestion system comprising a series of tanks through which raw materials flow continuously. The fluid flowing from the top of each tank may flow continuously into the next tank, and the rate of the outflowing product may match the rate of the incoming raw materials, providing a hydraulically balanced flow throughout the system. Each tank in the system may have its own stable microbial consortium with different physiological characteristics and digestive capabilities compared to the consortium in the other tanks in the system.

[0024] In some embodiments of the digestion process, powdered Ecklonia maxima kelp, chitin, and Saccharomyces cerevisiae yeast may be mixed with water to produce organic raw materials for an anaerobic digestion system. The anaerobic digestion system may include a mixing tank for mixing the organic raw materials to produce a homogeneous slurry. The slurry may then flow through four digestion tanks in a continuous and hydraulically balanced manner. More or fewer tanks may be used, and the hydraulic flow rate may be modified to obtain the desired result. In the first digestion tank, the slurry may be agitated at a rate that allows heavy or undigested solids to settle to the bottom. An outlet at the top of the first digestion tank may allow the fluid to flow into a second digestion tank. An outlet at the bottom of the first digestion tank may return the settled solids to the mixing tank. Each of the three digestion successor tanks, which may be called a packed-bed reactor, may have a submerged fixed medium substrate that provides a surface for biofilm growth. The flow rate of the digestion system is selected to allow sufficient residence time within each digestion tank so that a stable and unique microbial consortium can form within each tank. The microorganisms in the consortium may be derived from microorganisms originally present in the organic raw material. The microorganisms may digest Ecklonia maxima kelp, chitin, and yeast to produce digest products. As described in more detail below, the effluent from the top of the fourth digestion tank may be used as a biostimulant to promote plant growth or to improve soil quality.

[0025] In some embodiments, the biostimulant composition is prepared using Ecklonia maxima, chitin, and Saccharomyces cerevisiae yeast as raw materials by the process described in U.S. Patent Application Publication 2013 / 0324406, which is incorporated herein by reference in its entirety.

[0026] The biostimulant compositions produced by the digestion process described above may be used as is or may be further processed before use. For example, the effluent from the digestion system, referred herein as the "base product," may be concentrated, sterilized, filtered, pasteurized, or dehydrated before use, or any combination thereof. In some embodiments, the base product may be concentrated to 2×, 3×, 4×, 5×, 6×, 7×, 8×, 9×, 10×, or more. In some embodiments, the base product may be filtered and sterilized to remove any bacteria or other microorganisms in the composition.

[0027] Digestion system parameters, such as flow rate and solid content of organic raw materials, may be varied to achieve desired properties in the eluted biostimulant-based product.

[0028] II. Physical properties and compositions of biostimulants Embodiments described herein include biostimulant compositions comprising chemical species and / or microorganisms that promote plant growth, including by increasing the ability of plants to withstand abiotic stresses such as cold, heat, drought, and salinity. The biostimulant compositions described herein may include dead microorganisms, spore-forming microorganisms, fragments of dead microorganisms, viable microorganisms, microbial fermentation products, enzymes, biological plant growth regulators, organic acids, chelating agents, or combinations thereof.

[0029] The embodiments described herein also include biostimulant compositions comprising digestive products produced by the digestion of an organic raw material including Ecklonia maxima kelp. The biostimulant may include metabolites produced by microorganisms inherent in the organic raw material, and the microorganisms may be derived from the kelp raw material or other components of the organic raw material, such as chitin. Such metabolites may include, for example, sugars and fatty acids. The digestive products may also include dead microorganisms, fragments of dead microorganisms, microbial fermentation products, enzymes, biological plant growth regulators, organic acids, chelating agents, or combinations thereof.

[0030] The biostimulant compositions described herein may contain one or more sugars. In some embodiments, the biostimulant compositions may be characterized by their glycosyl residue content. In some embodiments, the biostimulant compositions may contain one or more of rhamnose, fucose, xylose, mannose, or glucose, or any combination thereof. In some embodiments, the biostimulant compositions may contain no galactose or less than 1 mol% of galactose compared to other glycosyl residues present in the biostimulant composition. In some embodiments, the biostimulant compositions may contain less than about 40, 30, 20, or 15 mol% of fucose, or about 40, 30, 20, or 15 mol% of fucose, compared to all other glycosyl residues present in the biostimulant composition. In some embodiments, the biostimulant compositions may contain at least about 15, 20, 25, or 30 mol% of xylose, or about 15, 20, 25, or 30 mol% of xylose, compared to all other glycosyl residues present in the biostimulant composition. In some embodiments, the biostimulant composition contains at least about 6, 8, 10, 12, 14, 16, 18, or 20 mol% of mannose, or about 6, 8, 10, 12, 14, 16, 18, or 20 mol% of mannose, compared to other glycosyl residues present in the biostimulant composition.

[0031] In some embodiments, the biostimulant composition may be characterized by mass spectrometry or NMR spectroscopy. In some embodiments, the biostimulant composition has an LC-MS chromatogram as shown in the lower panel of Figure 3. In some embodiments, the biostimulant composition includes one or more molecular species, or any combination of such molecular species, corresponding to one or more peaks in the LC-MS chromatogram as shown in the lower panel of Figure 3. In some embodiments, the biostimulant composition includes molecular species, or any combination of such 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 in the LC-MS chromatogram shown in the lower panel of Figure 3. In some embodiments, the biostimulant composition includes one or more molecular species, corresponding to one or more peaks in the LC-MS chromatogram shown in the lower panel of Figure 3 that are not present in the LC-MS chromatogram shown in the upper panel of Figure 3. In some embodiments, the biostimulant composition includes 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 include molecular species corresponding to peaks present in the LC-MS chromatogram shown in the upper panel of Figure 3 but not present in the LC-MS chromatogram shown in the lower panel of Figure 3.

[0032] In some embodiments, the biostimulant composition has a GC-MS chromatogram as shown in the upper panel of FIG. 4. In some embodiments, the biostimulant composition includes one or more molecular species corresponding to any one of the one or more peaks in the GC-MS chromatogram as shown in the upper panel of FIG. 4, or any combination of such molecular species. In some embodiments, the biostimulant composition includes molecular species corresponding to the peaks labeled A, B, C, D, E, F, or G in the GC-MS chromatogram shown in the upper panel of FIG. 4, or any combination of such molecular species. In some embodiments, the biostimulant composition includes one or more molecular species corresponding to one or more peaks in the GC-MS chromatogram shown in the upper panel of FIG. 4 that are not present in the GC-MS chromatogram shown in the lower panel of FIG. 4. In some embodiments, the biostimulant composition does not include molecular species corresponding to peaks that are present in the GC-MS chromatogram shown in the lower panel of FIG. 4 but not present in the GC-MS chromatogram shown in the upper panel of FIG. 4. In some embodiments, the biostimulant composition includes one or more molecular species listed in FIG. 39, or any combination of such molecular species.

[0033] In some embodiments, the biostimulant composition has an 1 H-NMR spectrum as shown in the lower spectrum of FIG. 5. In some embodiments, the biostimulant composition includes one or more molecular species corresponding to any one of the one or more peaks in the 1 H-NMR spectrum of the lower spectrum of FIG. 5, or any combination of such molecular species. In some embodiments, the biostimulant composition includes molecular species corresponding to the peaks labeled A, B, C, D, E, F, G, H, I, J, or K in the 1 H-NMR spectrum of the lower spectrum of FIG. 5, or any combination of such molecular species. In some embodiments, the biostimulant composition includes 1 in the H-NMR spectrum of the lower spectrum of FIG. 5, the upper 1The biostimulant composition contains one or more molecular species corresponding to one or more peaks that are not present in the H-NMR spectrum. In some embodiments, the biostimulant composition contains molecular species, or any combination of such molecular species, corresponding to the peaks labeled A, C, D, or E in Figure 5. In some embodiments, the biostimulant composition contains the top of Figure 5 1 It is present in H-NMR, at the bottom of Figure 5. 1 The H-NMR spectrum does not contain molecular species corresponding to peaks that are not present.

[0034] In some embodiments, the biostimulant composition is as shown in the lower spectrum of Figure 6. 13 It has a 1C-NMR spectrum. In some embodiments, the biostimulant composition is shown in the lower part of Figure 6. 13 The biostimulant composition includes one or more molecular species, or any combination thereof, corresponding to one or more peaks in the 1C-NMR spectrum. In some embodiments, the biostimulant composition is shown at the bottom of Figure 6. 13 The composition includes molecular species corresponding to peaks labeled A, B, C, D, E, or F in the 13C-NMR spectrum, or any combination of such molecular species. In some embodiments, the biostimulant composition is shown in the lower part of Figure 6. 13 In the 1C NMR spectrum, the upper part of Figure 6 13 The biostimulant composition contains one or more molecular species corresponding to one or more peaks that are not present in the 1C-NMR spectrum. In some embodiments, the biostimulant composition contains molecular species corresponding to the peaks labeled A or B in Figure 6, or any combination of such molecular species. In some embodiments, the biostimulant composition contains the top of Figure 6 13 It is present in C-NMR, at the bottom of Figure 6. 13 The C-NMR spectrum does not contain molecular species corresponding to peaks that are not present.

[0035] In some embodiments, the biostimulant composition contains viable microorganisms. In some embodiments, the microorganisms include bacteria derived from the bacterial population present in the Ecklonia maxima kelp raw material. The bacteria may 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 the microbial digests of other kelp species, such as Ascophyllum nodulum. In some embodiments, the biostimulant includes Microbacterium amiloristicum, Thermoanaerobacterium thermosaccharoristicum, Cellulosilicum lentocerum, Microbarbifer thermotolerance, Cholincera sp., Acinetobacter spp., Acinetobacter touneri, Lentilactobacillus buhineri, Lycorolactobacillus fordei, or Secandilactobacillus paracholinoides, or any combination thereof. In some embodiments, any one of these bacterial species comprises 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.

[0036] In some embodiments, the biostimulant is filtered and sterilized to be free of viable microorganisms. In some embodiments, the dry weight of the microbial biomass is less than 0.0001% of the total dry weight of the composition.

[0037] In some embodiments, the biostimulant contains 0.05–0.8% by dry weight of microbial biomass relative to the total dry weight of the biostimulant composition. In some embodiments, the dry weight percentage is at least about 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, or 0.8%, and at most about 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, or 0.8%, 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.

[0038] In some embodiments, the biostimulant is 100-5 × 10 5 Contains bacteria at CFU / ml. In some embodiments, the biostimulant is at least about 100, 500, 1 × 10 3 , 5×10 3 , 1 x 10 4 , 5×10 4 , or 1 x 10 5 CFU / ml, or up to approximately 100, 500, or 1 x 10⁶. 3 , 5×10 3 , 1 x 10 4 , 5×10 4 , or 1 x 10 5 CFU / ml, or approximately 100, 500, 1 x 10 3 , 5×10 3 , 1 x 10 4 , 5×10 4 , or 1 x 10 5 Contains bacteria in a range of CFU / ml, or between any two of these values.

[0039] In some embodiments, the biostimulant has a pH of 7.5–8.5. In some embodiments, the electrical conductivity of the biostimulant is about 900, 950, 1000, 1050, or 1100 μS / cm. In some embodiments, the density of the biostimulant is about 0.997–0.999 g / cm³. 3 Or approximately 0.998 g / cm³ 3In some embodiments, the biostimulant has a solids content of 0.01–2%. In some embodiments, the solids content is in the range of approximately 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 any two of these values. In some embodiments, the chemical oxygen demand (COD) of the biostimulant is 10–200 mg / L. In some embodiments, the COD is 10, 20, 30, 40, 50, 100, 125, 150, 175, or 200 mg / L, or any two of these values. Conductivity and COD values ​​vary with the concentration of the biostimulant, increasing as the concentration increases.

[0040] III.How to use Embodiments of the biostimulant composition may be used in methods to promote the growth of various different plants and plants in different conditions. In some embodiments, contact of plants, seeds, or growing media with the biostimulant promotes plant growth by increasing, for example, growth rate, yield at harvest, productivity, stem diameter, fruit quantity and / or size, grain yield, leaf surface area, root surface area, root length, root depth, shoot thickness, or total mass compared to untreated plants. In some embodiments, promoting plant growth includes one or more of the following: promoting seed germination, promoting early plant development, increasing nutrient uptake, mitigating the effects of transplanting, improving plant reproduction, and improving soil microbial activity. In some embodiments, one or more of these plant qualities are increased by at least 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50% or more compared to untreated plants. In some embodiments, contacting plants, seeds, or growing media with a biostimulant promotes plant growth, for example, by increasing the plant's tolerance to abiotic stress. Such abiotic stresses may include drought stress, heat stress, cold stress, or stress due to high salinity. In some embodiments, increasing tolerance to one or more abiotic stresses increases growth rate, harvest yield, productivity, stem diameter, fruit quantity and / or size, grain yield, leaf surface area, root surface area, root length, root depth, shoot thickness, or total mass compared to the same plant under similar stress conditions but untreated. In some embodiments, treatment with a biostimulant promotes plant growth by increasing the plant's ability to recover from abiotic stress more quickly than it would in the untreated state.

[0041] In some embodiments, plants or growing media are exposed to a biostimulant before, during, or after abiotic stress. For example, treatment with a biostimulant before abiotic stress may, in some embodiments, enable plants to tolerate abiotic stress better than similar plants that have not been treated. In some embodiments, plants or growing media are exposed to a biostimulant before the abiotic stress occurs, even though the plants are at risk of abiotic stress. Plants may be determined to be at risk of abiotic stress based, for example, on weather patterns or forecasts in the location where the plants are growing. In some embodiments, treatments to help mitigate cold stress may be applied during frost-prone periods of the year, such as early spring or late autumn, depending on the geographical location of the plants. In some embodiments, treatments to help mitigate heat or drought stress may be applied in late summer when plants are at risk of relatively high temperatures.

[0042] Those skilled in the art will be able to determine the relative risk of a plant to a particular abiotic stress based on the plant species, its geographical location, and the local weather patterns and forecasts for that location.

[0043] In some embodiments, a biostimulant is applied to a plant while it is experiencing abiotic stress. Whether a plant is experiencing abiotic stress may be determined by those skilled in the art, based on the type of plant and the specific conditions under which it is growing. For example, drought stress may be determined based on observations of soil moisture content and the plant's condition. Some plant species and varieties are inherently more drought-tolerant than others, so soil and air humidity conditions that stress one species or variety may not stress another. The same applies to other potential stresses such as high temperature, low temperature, and salinity stress.

[0044] In some embodiments, a biostimulant is applied when a plant is expected to have been, received, or received a temperature of approximately 15, 10, 5, or 0°C, or less than approximately 15, 10, 5, or 0°C. In some embodiments, a biostimulant is applied when a plant is expected to have been, received, or received a temperature of approximately 20, 25, 30, 35, or 40°C, or more than approximately 20, 25, 30, 35, or 40°C. In some embodiments, a biostimulant is applied when a plant is expected to have been, received, or received a soil water content of approximately 30, 25, 20, 15, 10, 5, or 1% or less for at least approximately 6, 12, 24, or 48 hours or 3, 4, 5, 6, 7, 8, 9, or 10 days.

[0045] In some embodiments, the biostimulant is applied within 12, 24, 36, or 48 hours or 3, 4, 5, 6, 7, 8, 9, or 10 days from when the plant has experienced or is expected to experience abiotic stress. In some embodiments, the biostimulant is applied when it is determined that there is at least about 30, 40, 50, 60, 70, 80, or 90% of the probability that the plant will experience abiotic stress within 12, 24, 36, or 48 hours or 3, 4, 5, 6, 7, 8, 9, or 10 days after treatment.

[0046] In some embodiments, the biostimulant is applied when the plant is not experiencing or is not expected to experience abiotic stress. In addition to increasing tolerance to abiotic stress, embodiments of the biostimulant compositions disclosed herein can promote plant growth in the absence of abiotic stress.

[0047] In some embodiments, the biostimulant is applied to the plant or growing medium before transplanting the plant. In some embodiments, the biostimulant is applied to the plant or growing medium after transplanting the plant. In some embodiments, the biostimulant is applied to the plant or growing medium while the plant is being transplanted. In some embodiments, the biostimulant is applied to the growing medium (e.g., soil) to which the plant is transplanted.

[0048] In some embodiments, the plants treated with the biostimulant composition may be, for example, crops, vegetables, flowers, ornamental plants, turfgrass, trees, shrubs, etc. Non-limiting examples of crops include corn, rice, wheat, barley, rye, oats, sorghum, cotton, soybeans, peanuts, buckwheat, beets, rapeseed, sunflowers, sugarcane, marijuana, and tobacco. Non-exclusive examples of vegetables include nightshade vegetables (eggplant, tomato, pimento, pepper, potato, etc.), cucurbitaceous vegetables (cucumber, pumpkin, zucchini, watermelon, melon, squash, etc.), cruciferous vegetables (radish, white turnip, horseradish, kohlrabi, Chinese cabbage, cabbage, mustard greens, broccoli, cauliflower, etc.), daisy family vegetables (burdock, garland chrysanthemum, artichoke, lettuce, etc.), lily family vegetables (leeks, onions, garlic, and asparagus), parsley family vegetables (carrots, parsley, celery, parsnips, etc.), amaranth family vegetables (spinach, Swiss chard, etc.), mint family vegetables (perilla, mint, basil, etc.), strawberries, sweet potatoes, yams, and taro. Non-exclusive examples of fruits include apples (apples, pears, Japanese pears, quince, European quince, etc.), fleshy drupes (peaches, plums, nectarines, Japanese apricots, cherries, apricots, prunes, etc.), citrus fruits (Satsuma oranges, oranges, lemons, limes, grapefruit, etc.), nuts (chestnuts, walnuts, hazelnuts, almonds, pistachios, cashews, macadamia nuts, etc.), berries (blueberries, cranberries, blackberries, raspberries, etc.), grapes, persimmons, olives, plums, bananas, coffee, dates, and coconuts. Non-limiting examples of trees include fruit trees, tea, mulberry, flowering plants, and street trees (ash, birch, dogwood, eucalyptus, ginkgo, lilac, maple, oak, poplar, redbud, sweetgum, plane tree, zelkova, cypress, fir, hemlock, juniper, pine, spruce, and yew). The term "plant" or "plants" refers to both naturally occurring plants and genetically modified plants. In some embodiments, the biostimulant is applied to the seeds of any of the above plants.

[0049] In certain embodiments, the biostimulant compositions described herein may be applied to soil, to fertilizers used to fertilize plants, to plants directly, or to both soil and plants. The compositions may be applied directly to plant seeds. In addition to soil, the biostimulant compositions may be applied to other plant growing media, such as hydroponic growing media. The compositions may be used in infall application, foliar application, 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 in sufficiently close temporal proximity to the conventional fertilizers and / or pesticides so that the formulation may have its desired effect, such as enhancing the effect of the conventional fertilizers and / or pesticides. In some embodiments, the compositions are applied in combination with conventional fertilizers and / or pesticides. The compositions may be mixed with conventional fertilizers and / or pesticides, or applied simultaneously with them.

[0050] In some embodiments, the biostimulant compositions described herein are mixed with conventional fertilizers or pesticides in a ratio of biostimulant to conventional fertilizers or pesticides of about 3:1 to about 1:100. In some embodiments, the biostimulant compositions are mixed with conventional fertilizers or pesticides in a ratio of biostimulant to conventional fertilizers or pesticides of about 1:20. The biostimulant compositions described herein may be coated onto particles of conventional fertilizers or pesticides. The fertilizer or pesticide particles may be coated, for example, by spray-drying the biostimulant onto the surface of the fertilizer, or by mixing the biostimulant in a dehydrated powder form with or without a binder or carrier with the particles.

[0051] In certain embodiments, the conventional fertilizer is a starter fertilizer. In some embodiments, the conventional fertilizer includes at least one of ammonia, urea, ammonium nitrate, ammonium sulfate, ammonium thiosulfate, monoammonium phosphate (MAP), diammonium phosphate (DAP), potassium chloride (MOP), potassium sulfate (SOP), and potassium nitrate (NOP). In some embodiments, the starter fertilizer is a 10-34-0 starter fertilizer.

[0052] 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 applied biostimulant composition. The dry weight of a given volume of liquid biostimulant composition is the weight of all substances in the volume of biostimulant other than water. In some embodiments, an amount of biostimulant applied that provides 0.10 to 10 g of digested product per acre is used. In some embodiments, the amount of biostimulant applied provides at least about 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 g per acre as dry weight of digested product, and up to about 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 g, or about 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 g, or a range between any two of these values. In some embodiments, the amount of biostimulant applied provides a range of at least about 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 g per acre as the dry weight of the biostimulant component applied, up to a maximum of about 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 g, or about 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 g, or any two of these values. The amount of the biostimulant composition applied may also be characterized in relation to the number of bacterial colonizing units applied. In some embodiments, the amount of biostimulant applied is at least about 5 × 10 as the CFU of bacteria applied per acre. 3 , 1 x 10 4 , 5×10 4 , 1 x 10 5 , 5×10 5 , 1 x 10 6 , or 5 x 10 6 , up to approximately 5 x 10 3, 1 x 10 4 , 5×10 4 , 1 x 10 5 , 5×10 5 , 1 x 10 6 , or ×10 6 , or approximately 5 x 10 3 , 1 x 10 4 , 5×10 4 , 1 x 10 5 , 5×10 5 , 1 x 10 6 , or 5 x 10 6 , or a range between any two of these values ​​is provided.

[0053] In some embodiments, the biostimulant compositions described herein may be applied in a dry form. The biostimulant base product may be dehydrated to produce a powder product that can be applied to a growth medium (e.g., soil), to plants, or to seeds.

[0054] In some embodiments, the amount of biostimulant applied is an effective amount to achieve the desired plant growth-promoting effect. For example, an effective amount of a biostimulant-based product, such as the MBT-E product described in the following examples, to increase the height of cotton plants compared to untreated plants is 0.5 or 1 quart (qt. / A) per acre. In some embodiments, the biostimulant composition is applied in an effective amount to increase the plant's tolerance to drought, salinity, high temperature, or low temperature stress, or to achieve any other desired result described herein that the biostimulant composition can achieve.

[0055] Compositions comprising biostimulant compositions and other components described herein (e.g., fertilizers) can be formed by mixing the components in a tank (i.e., tank mix). After mixing, the formulation can be bottled or packaged in another manner (e.g., in a drum) and applied to a field or crop, or mixed with other components. If bottled or packaged in another manner, the end user can mix the formulation with other components before application. Biostimulant compositions can be mixed with conventional fertilizers by tank mix, including splash mix with minimal further mixing, or blended with conventional fertilizers.

[0056] 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 one, two, three, four, or five times during the growing season. In some embodiments, the applications are spaced one, two, three, four, five, or six weeks apart.

[0057] IV. Specific Definitions The above description includes certain details to fully understand the various embodiments. However, those skilled in the art will understand that the embodiments provided can be carried out even without these details. Throughout the following specification and claims, unless the context specifically requires otherwise, the word “comprise” and its variations (e.g., “comprises” or “comprising”) shall be interpreted in an open, comprehensive sense, i.e., “including, but not limited to.” As used in this specification and the appended claims, the singular “one” (“a”, “an”, and “the”) includes plural nouns unless otherwise explicitly stated. It should also be noted that the term “or” is usually used to mean “and / or” unless its content explicitly indicates something else. Furthermore, the headings provided herein are for convenience only and do not imply any scope or meaning of the embodiments of the subject matter.

[0058] The terms “about” or “approximately” mean within an acceptable margin of error for a particular value, as determined by those skilled in the art, and this depends in part on how that value is measured or determined, for example, on the limitations of the measuring system. For example, “about” could mean a standard deviation of 1 or more than 1 for a given value. Where a particular value is described in this application and claims, unless otherwise specified, the term “about” should be assumed to mean within an acceptable margin of error for that particular value. [Examples]

[0059] The following embodiments are provided to further illustrate some embodiments of the present disclosure, but are not intended to limit the scope of the present disclosure. It will be understood that, by their exemplary nature, other procedures, methods, or techniques known to those skilled in the art may be used instead.

[0060] Example 1: Microbial digestion of Ecklonia maxima kelp and characterization of digested products Powdered Ecklonia maxima kelp, chitin, and Saccharomyces cerevisiae yeast were mixed with water to prepare organic raw materials for an anaerobic digestion system. The anaerobic digestion system included a first tank containing the mixed organic raw materials to produce a homogeneous slurry. The slurry was then flowed through four digestion tanks in a continuous and hydraulically balanced manner. In the first digestion tank, the slurry was agitated at a rate that allowed heavy or undigested solids to settle to the bottom. The fluid flowed into the second digestion tank through an outlet at the top of the first digestion tank. The settled solids were returned to the first tank through an outlet at the bottom of the first digestion tank. Each of the three subsequent tanks, called packed-bed reactors, had a submerged, fixed culture medium substrate providing a surface for biofilm growth. The flow rate of the digestion system allowed for sufficient residence time in each digestion tank to allow a stable and unique microbial consortium to form within each tank. The microorganisms in the consortium originated from microorganisms naturally present in the organic raw materials. The microorganisms digested Ecklonia maxima kelp, chitin, and yeast to produce digest products. The effluent from the top of the fourth digestion tank, referred herein as MBT-E base product (BP), was a clear, light brownish liquid.

[0061] Ecklonia maxima kelp used as a raw material was evaluated by GC-MS and compared to Ascophyllum nodosum powder used as a raw material in a commercially available product sold by Loveland Agri Products as Maritime® (also known herein as MBT-A). GC-MS chromatograms are shown in Figure 1, with the upper chromatogram for Ascophyllum nodosum and the lower chromatogram for Ecklonia maxima. Each chromatogram has its own distinct peaks, as indicated by the arrows. The chromatograms also show that the two kelp feedstocks possess chemical species they have in common in different relative abundances.

[0062] The chemical compositions of Maritime™ BP (MBT-A) and MBT-E BP were also analyzed and compared. Sugar residues present in the base products were analyzed by GC-MS of per-O-trimethylsilyl (TMS) derivatives of monosaccharide methyl glycosides generated from the samples by HCl methanolysis, as previously described by Santander et al. (2013) Microbiology 159: 1471. Inositol was added to each sample as an internal standard. After lyophilization and derivatization, the samples were extracted with hexane, and the TMS methyl glycoside was analyzed by GC-MS using an Agilent 7890A GC connected to a 5975C MSD equipped with a Supelco Equity-1 fused silica capillary column (30 m × 0.25 mm ID). The results are shown in Figure 2. LC-MS chromatograms of the two base products are shown in Figure 3, with MBT-A at the top and MBT-E at the bottom.

[0063] Dichloromethane extracts of MBT-A and MBT-E were analyzed by GC-MS according to the following 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 dried material. GC-MS analysis was performed to compare the chemical profiles of different batches. The samples were derivatized using N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) + 1% TMCS, and palmitate- 13 C 14A standard was used as the internal standard. GC-MS analysis was performed using an Agilent (Palo Alto, California, USA) 8890 series GC system equipped with a CTC-Pal injector, a 5977B Network Mass Selective Detector, and a DB-1MS column (J&W, Palo Alto, California, USA) (60m inner diameter × 0.25mm, 0.25um film thickness). Data acquisition and analysis were performed using Agilent Mass Hunter quantitative and qualitative software from Agilent Technologies (Palo Alto, California, USA). GC-MS chromatograms of MBT-A and MBT-E are shown in Figure 4, with characteristic peaks identified by arrows. Figure 39 lists the molecular species found only in MBT-E. These have an agreement coefficient of over 70% and 1x10⁻⁶. 5 Based on the area of ​​the superstructure, it was identified using Mass Hunter quantitative and qualitative software from Agilent Technologies (Palo Alto, California, USA).

[0064] methanol extract of the base product 1 H-NMR and 13 The spectra were analyzed by 13C-NMR and are shown in Figures 5 and 6, respectively. The arrows indicate selected intrinsic peaks.

[0065] MBT-E-based products were treated by reverse osmosis to produce 4× and 8× enriched versions of MBT-E.

[0066] Bacteria present in the Ecklonia maxima raw material and MBT-E base product were identified by 16S rRNA sequencing. Table 1 below shows the bacteria present in both the raw material and the MBT-E product.

[0067] [Table 1]

[0068] Metagenomic sequencing was performed to identify spore-forming bacteria in MBT-E products. The spore-forming bacteria, as determined by metagenomic sequencing, constituted approximately 0.7% of the total population (based on the percentage of the total bacterial population, 1 × 10⁻⁶). 2 -3 × 10 3 (CFU / ml). The operational taxonomic units of spore-forming bacteria include Bacillus spp., Aneurinibacillus thermoaerophilus, Bulgibacillus spp. (including V. phasianinus and V. dokdonensis), Cyclobacillus sp., and Paenibacillus sphorae.

[0069] Bacteria identified in MBT-E products by metagenomic sequencing were compared to those present in MBT-A / Maritime™. The following list includes members of the MBT-E community that are present in statistically significantly higher amounts than those in MBT-A products. The listed percentages represent the percentage of the total population present in MBT-E, and the number of "×" symbols indicates how many times higher the microbial population is in MBT-E compared to MBT-A. • Carbohydrate-degrading and cellulosic community: Microbacterium amiloristicum (0.0018%), Thermoanaerobacterium thermosaccharoticum (0.0002%), Cellulosilicum lentocerum (0.00005%), Microbarbifer thermotolerance (0.0015%, 1.38×), a genus that can also degrade complex carbohydrates such as cellulose, alginates, and chitin. • Cholinethera sp. (0.00003%, 2.19×): A genus capable of breaking down bile acids (e.g., cholic acid) into secondary bile acids through the production of NADPH-dependent 7β-hydroxysteroid dehydrogenase. Acinetobacter spp., including A. toneri (0.00043%, 2.6×), is an aromatic compound degrader that enables the solubilization of iron and zinc and the release of nutrients through siderophore production. It also contains fungal inhibitory genes and contributes to pathogen-suppressing soil. Several non-spore-forming lactic acid bacteria (LAB) communities include Lentilactobacillus buhineri (0.00005%, 2.41×), Lycorolactobacillus fordei (0.000023%, 4.01%), and Secandilactobacillus paracolinoides (0.00013%, 1.83×).

[0070] 4×concentrated MBT-E also had the following characteristics: a pale yellow liquid with a pH range of 7.5 to 8.5, electrical conductivity of 900 to 1100 (μS / cm), density of 0.998 (g / cm3), solids content of 0.07%, viscosity of 1.29 (cP), COD of 20 to 150 (mg / L), and 5.0×10 4 ~5.0×10 5 Total bacterial count and spore-forming organism count in the range of CFU / ml (2 × 10⁻⁶) 2 ~3×10 3 CFU / ml.

[0071] Example 2: Properties of MBT-E that promote plant growth Two experiments were conducted using Denton urban soil (compost biosolids) and Whitesboro soil to mitigate drought stress in maize and cotton in rain shelters. Maize and cotton plants were thinned based on their uniform growth after germination.

[0072] material and method: Plant physiological traits: After initiating drought stress water management and drenching pots to a soil moisture capacity of 20-30%, plant physiological traits such as stomatal conductance (gsw), transpiration rate (E), chlorophyll, 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, Nebraska, USA). Ambient leaf temperature was measured as follows: Ambient leaf temperature (T) amb )=T leaf -T ref On days 15 and 26 after the start of drought stress water management, the physiological parameters of the plants were measured between 11:00 and 15:00 for fully swollen cotton leaves of each plant. On days 7, 17, 26, and 32 after the start of drought stress water management, the physiological parameters of the plants were measured between 11:00 and 15:00 for fully swollen corn leaves of each plant.

[0073] Leaf chlorophyll content: Leaf chlorophyll content was measured from fully swollen leaves using a chlorophyll meter, SPAD (Soil Plant Analysis Development-502, Konica Minolta, Tokyo, Japan). To prevent variability, the central position of the leaf was selected for measuring leaf chlorophyll content.

[0074] Proline assay: Proline assay extraction and determination were performed using the method described by Carillo and Yves, PROTOCOL: Extraction and determination of proline (2011), to determine the amount of proline produced in leaves during drought stress. The proline assay was performed 30 days after the start of drought stress water management. Leaf discs were collected, measured, and homogenized as extracts in 100% ethanol (e.g., 0.015 g / 0.5 mL). Standards known as proline solutions were prepared in the range of 0.01–0.1 mM in 100% ethanol. Reaction mixtures were prepared using 60% (v / v) acetic acid and 1% (w / v) ninhydrin in 20% ethanol. The combined mixtures were transferred to a 96-well plate, heated in a water bath at 95°C for 20 minutes, and cooled to room temperature. The 96-well plate was read at 520 nm using a plate reader.

[0075] Relative Water Content (RWC): To determine the relative water content of a leaf, fresh, fully swollen leaves were collected and measured using the following equation, as 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 swollen weight - dry weight) × 100

[0076] Statistical Analysis and Experimental Design: In the rain shelter trials, pots were placed in a randomized complete block design (RCBD) with 6 treatments and 20 replicates in the corn experiment and 8 treatments and 20 replicates in the cotton experiment, with each replicate containing a single plant in a single pot. Data from plant physiological parameters, leaf chlorophyll content, proline assays, biomass, and meta-analysis were analyzed using JMP16 software (SAS Institute, Cary, North Carolina, USA) with a Fit model at a significance level of p<0.1.

[0077] result Drought Stress Mitigation in Cotton Experiments: MBT-E BP-treated plants performed better than untreated controls in drought stress mitigation in cotton in rain shelter tests at rates of 0.5 and 1 qt. / A. Physiological parameters of the plants, including stomatal conductance, transpiration rate, quantum yield, electron transport rate, leaf chlorophyll content, and proline, were all increased by treating the plants with MBT-E during periods of drought stress. The effects on cotton leaf temperature are shown in Figures 21 and 22. The effects on stomatal conductance are shown in Figure 23. The effects on transpiration rate are shown in Figure 24. The effects on quantum yield (percentage of blocked light energy used in photosynthesis and not lost as heat) are shown in Figure 25. The effects on electron transport rate are shown in Figure 26. The effects on leaf chlorophyll content are shown in Figure 27. SPAD readings (leaf chlorophyll content) ranged from 44 to 51, with the highest leaf chlorophyll content found in plants treated with MBT-E under drought stress. SPAD readings for MBT-E were 50.46 at a 0.5 qt. / A rate and 51.25 at a 1 qt. / A rate, recorded 15 days after the start of drought stress water management (Figure 7; Table 2). Leaf temperature was lower in plants treated with MBT-E compared to the untreated control. Ambient leaf temperature was 2.13°C at a 0.5 qt. / A rate and 2.31°C at a 1 qt. / A rate for MBT-E treated plants. MBT-E treated plants had higher proline concentrations than the untreated control. Proline concentrations in MBT-E treated plants were 28.66 at a 0.5 qt. / A rate and 27.75 at a 1 qt. / A rate 30 days after the start of drought stress water management (Figure 8). Cotton ball yield increased in plants treated with MBT-E at a 1 qt. / A rate compared to the untreated control. The average height of cotton plants was 61.15 cm at a rate of 0.5 qt. / A and 62.35 cm at a rate of 1 qt. / A for MBT-E treated plants (Figure 9). For MBT-E, the average number of cotton balls produced per plant was 45 (Figure 10). MBT-E treated plants significantly increased cotton production at rates of 0.5 and 1 qt. / A (Figure 11).

[0078] Mitigation of drought stress in maize experiments: The same trend was observed in mitigating drought stress in maize by applying MBT-E during the vegetative growth 6 (V6) and vegetative tasseling (Vt) stages. Stomatal conductance in MBT-E treated plants was higher than that of untreated control plants during the V6 growth stage. Transpiration rates in MBT-E treated plants increased compared to untreated control plants during the V6+Vt growth stages. Quantum yield and electron transport rates increased in MBT-A and MBT-E treated plants compared to untreated control plants. Leaf temperature decreased during drought stress in all MBT-E treated plants. MBT-E treated plants had lower leaf temperatures during the Vt growth stage than untreated control plants. Ambient leaf temperatures were 0.38, 0.37, 0.11, and 0.15°C during the MBT-E Vt growth stage (Table 3). SPAD readings (leaf chlorophyll content) ranged from 18 to 51, with the highest leaf chlorophyll content found in plants treated with MBT-E under drought stress conditions during the V6+Vt growth stage (Table 4). Relative leaf moisture content (%) in MBT-E treated plants was higher than in untreated controls during the V6 and V6+Vt growth stages (Figure 12). Proline concentration in MBT-E was 21.50 in the Vt growth stage and 22.44 in the V6+Vt growth stage 30 days after the start of drought stress water management (Figure 13). MBT-E treatment increased maize ear length and dry weight (Figure 14). The highest average maize kernel yield was observed in plants treated with MBT-E during the V6+Vt growth stage (Figure 15).

[0079] In a separate experiment, the effects of MBT-E BP, 4×, and 8× treatments on corn growth rate under drought stress were tested by the following experiment: Corn variety WS095 2021 was germinated in Berger general-purpose potting soil in a growth chamber at 22°C for 14 days. They were maintained at 100% water volume. On day 14, they were fertilized with Jacks fertilizer. The treatment was applied as a foliar treatment, with a total of 10 ml per plant per treatment. The treated plants were placed in a lit growth chamber at 22°C for 14 days. Each plant was maintained at 30% water volume to induce water stress. All treatments were filter-sterilized and applied after dilution to 0.8% with filter-sterilized water. The treatments were as follows: 1) water as a negative control, 2) AccomplishLM®, MBT-E base product (BP), MBT-E BP 4x concentrate (4×CP), and MBT-E BP 8x concentrate (8×CP) as positive controls. Each individual plant was used as an replicate, with seven replicates per treatment. Plant metrics were analyzed after 14 days. The results are shown in Figures 20A-C.

[0080] Example 3: Plant growth-promoting properties of MBT-E tested in Arabidopsis thaliana. The MS medium used in these examples was prepared by adding 4.43 g of MS basal salt (Murashige and Skoog basal medium, Sigma Aldrich, M5519), 0.2 g of myo-inositol, 1 g of MES, and 20 g of sucrose to 2 L of water. The pH was then adjusted to 5.7. 500 ml of the resulting solution was poured into four glass bottles, each containing 1.77 g of Phytagel. The bottles were autoclaved, and the medium was poured into Petri dishes for solidification. When the objective was to collect data from the roots, treated seedlings were grown on 0.01% MS medium supplemented with 0.66 g / l of CaCl2 and 3.4 g / l of Phytagel, adjusted to pH 5.7 before autoclaving. The medium was then poured into Petri dishes for solidification. When the objective was to collect data from the shoots, the MS medium used in the processed preparation was liquid MS medium containing MS base salts + 2 g / l MES.

[0081] The effects of MBT-E BP, 4×, and 8× treatments on shoot surface area in Arabidopsis thaliana were tested by the following experiment: Surface-sterilized Arabidopsis thaliana seeds were germinated for 7 days at 20°C on plates containing MS medium solidified with phytagel. The seeds were then placed on rockwool cubes moistened with 40 ml of each treatment. All treatments were filter-sterilized and applied after dilution to 0.8% with filter-sterilized water. The treatments were as follows: 1) water as a negative control, 2) a commercially available biostimulant, AccomplishLM® as a positive control, MBT-E base product (BP), MBT-E BP 4x concentrate (4×CP), MBT-E BP 8x concentrate (8×CP), diluted to 0.8% in liquid MS medium. Four rockwool cubes, each containing one seedling, constituted one replicate. Each treatment was repeated four times. The treated plants were placed on LED grow carts with a completely randomized block design and grown at approximately 20°C for 14 days. The cubes were kept moist by adding 8 mL of water every two days. After 14 days, the leaf area of ​​each plant was measured using ImageJ software on shoot photographs. The results are shown in Figure 16. All MBT-E treatments resulted in a significant increase in shoot surface area.

[0082] The effects of MBT-E BP, 4×, and 8× treatments on root length in Arabidopsis thaliana were tested by the following experiment: Surface-sterilized Arabidopsis thaliana seeds were germinated at 20°C for 6 days on plates containing MS medium solidified with phytagel. Seeds were treated by immersing the roots of each seedling in their respective treatments. The treatments were as follows: 1) water as a negative control, 2) AccomplishLM®, MBT-E base product (BP), MBT-E BP 4x concentrate (4×CP), and MBT-E BP 8x concentrate (8×CP) as positive controls, 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 as replicates). There were 4 replicates per treatment. The treatments and replicates were placed on LED cultivation carts in a fully randomized block design and grown at approximately 20°C for 12 days. Seven days later, the root area of ​​each plant was measured using WinRhizo software via scanning. The results are shown in Figures 17 and 18. Asterisks indicate statistically significant differences.

[0083] The effects of MBT-E BP, 4×, and 8× treatments on Arabidopsis thaliana under drought stress were tested by the following experiment: Surface-sterilized Arabidopsis thaliana seeds were germinated for 7 days at 20°C on plates containing MS medium solidified with phytagel. Then, one treated seed was placed in a medicine cup containing 30 ml of peat moss moistened with the treatment. All treatments were filter-sterilized and applied after being diluted 0.8% with filter-sterilized water. The treatments were as follows: 1) water as a negative control, 2) AccomplishLM®, MBT-E base product (BP), MBT-E BP 4x concentrate (4×CP), and MBT-E BP 8x concentrate (8×CP) as positive controls. Four medicine cups, each containing one seedling, formed one replicate. Each treatment was repeated four times. Treated plants were placed in an illuminated growth chamber (Percival Model 136LL) at 22°C for 14 days. Water stress was induced by maintaining seedlings at 20% water capacity. They were grown for 14 days at approximately 20°C in LED grow carts with a fully randomized block design. After 12 days, the leaf area of ​​each plant was measured using ImageJ software on shoot photographs. The results are shown in Figure 19. Asterisks indicate statistically significant differences.

[0084] Example 4: Effects of MBT-E under low-temperature stress conditions The effect of MBT-E on Arabidopsis thaliana under cold stress was tested by the following experiment: Surface-sterilized Arabidopsis thaliana seeds were germinated at 20°C for 7 days on plates containing MS medium solidified with phytagel. Then, one treated seed was placed in a medicine cup containing 30 ml of peat moss moistened with the treatment. All treatments were filter-sterilized and applied after being diluted 0.8% with filter-sterilized water. The treatments were as follows: 1) water as a negative control, 2) AccomplishLM®, MBT-E base product (BP), MBT-E BP 4x concentrate (4×CP), and MBT-E BP 8x concentrate (8×CP) as positive controls. Four medicine cups, each containing one seedling, formed one replicate. Each treatment was repeated four times. Treated plants were placed in an illuminated growth chamber (Percival Model LT41VL) at 12°C for 21 days using a fully randomized block design. Observations were made on days 14 and 21. The leaf area of ​​each plant was measured using ImageJ software on shoot photographs. The results for shoot surface area after low-temperature treatment are shown in Figure 28. Asterisks indicate statistically significant differences.

[0085] The effect of MBT-E on tomatoes under cold stress was tested by the following experiment: Tomato variety Rutgers was germinated in Berger general-purpose potting soil and grown in a growth chamber (Percival Model 136LL) at 22°C for 14 days. They were maintained at 100% water capacity. Fourteen days after planting, they were fertilized with Jacks fertilizer. The treatment was provided as a foliar application, and each plant received a total of 10 ml of the treatment solution. The treatments were as follows: 1) water as a negative control, 2) MBT-E base product (BP), 3) MBT-E BP 4x concentrate (4×CP), and 4) MBT-E BP 8x concentrate (8×CP). Treated plants were placed in a fully randomized block design in a lit growth chamber (Percival Model LT41VL) programmed to provide cold stress by first providing 1 hour at 16°C, 1 hour at 8°C, 2 hours at 4°C, and 2 hours at -4°C. After this low-temperature management, the plants were evaluated using a low-temperature stress rating scale from 0 to 5, where 0 indicated no shoot stress and 5 indicated complete shoot death. The results of the low-temperature stress evaluation are shown in Figure 29. The results of the recovered fresh weight are shown in Figure 30.

[0086] Further cold stress effects in Arabidopsis thaliana were tested by the following experiment: Surface-sterilized Arabidopsis thaliana seeds were germinated at 20°C for 7 days on plates containing MS medium solidified with phytagel. Then, one treated seed was placed in a medicine cup containing 30 ml of peat moss moistened with the treatment. All treatments were filter-sterilized and applied after being diluted 0.8% with filter-sterilized water. The treatments were as follows: water, MBT-E base product (BP), and MBT-E BP 4x concentrate (4×CP) as negative controls. Four medicine cups, each containing one seedling, formed one replicate. Each treatment was repeated four times. Treated plants were placed in a lit grow chamber at 12°C for 21 days using a fully randomized block design. Observations were made 14, 21, and 27 days after treatment (DAT). The leaf area of ​​each plant was measured using ImageJ software on shoot photographic images. The results for shoot surface area are shown in Figure 31.

[0087] 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 thaliana seeds were germinated for 7 days at 20°C on plates containing MS medium solidified with phytagel. The seeds were then placed on rockwool cubes moistened with 40 ml of each treatment. All treatments were filter-sterilized and applied after being diluted 0.2% with filter-sterilized water. The treatments were as follows: water, MBT-E base product (BP), and MBT-E BP 4-fold concentrate (4×CP) as negative controls. All treatments also contained 75 mM NaCl. Four rockwool cubes, each containing one seedling, formed one replicate. Each treatment was repeated four times. Treated plants were placed on LED grow carts in a fully randomized block design and grown at approximately 20°C for 14 days. The cubes were kept moist by adding 8 ml of water every two days. After 14 days, the leaf area of ​​each plant was measured using ImageJ software on shoot photographic images. The results are shown in Figure 32.

[0088] The effect of MBT-E on salt tolerance in maize was tested by the following experiment: Dynagro maize seeds were planted in a 4:1 MVP turf / Sungro Blackgold peat medium. Twelve days after planting, the maize was thinned for uniformity, fertilized with Jack's 20-20-20 at 25 lbs N / A, and subjected to salt stress with 75 mmol NaCl. MBT-E was applied to the leaves at 1 qt / A and 2 qt / A 21 days after planting. SPAD, imaging, and LiCor measurements were performed three days before harvesting the maize for biomass. The results for leaf chlorophyll content (SPAD) are shown in Figure 33.

[0089] The effect of MBT-E on salt tolerance in zinnia was tested by the following experiment: Dwarf zinnia (Zinnia elegans) seeds were planted in a 3:1 Isolite / Sunshine Mix LC1 peat medium, thinned for uniformity, and fertilized with Jack's 20-20-20 at 50 lbs / A. Soil electrical conductivity (EC) measurements were performed multiple times before seed planting and throughout the experiment. NaCl was applied three times at a total of 100 mmol. MBT-E was applied as a foliar treatment at a rate of 1 qt / A and 2 qt / A 34 days after sowing. Metrics for this experiment included stem diameter, height, total biomass, and final soil EC readings at harvest. The results for stem diameter at harvest are shown in Figure 34. The results for height and total dry biomass at harvest are shown in Figures 35A-B. The results for soil electrical conductivity after MBT-E treatment are shown in Figure 36.

[0090] Example 6: MBT-E field test for promoting bell pepper plant growth Pepper seedlings were transplanted into raised beds in Yuma, Arizona. Prior to planting, the beds were fertilized with MAP (monoammonium phosphate) at a rate of 300 lbs / acre. The control treatment (no MBT-E added) was planted in two beds as four replicates, each 75 feet long. The MBT-E treatment was planted in four beds as eight replicates, each 75 feet long. Three weeks after planting, the plants were grown in a subterranean drip system containing UAN32 (ammonium urea nitrate) fertilizer for each treatment (no MBT-E added, or with MBT-E added at 2 qts per acre). The no-MBT-E and MBT-E treatments were provided only during this initial fertilization period. The plants were then fertilized two more times during the growing season. The plants were harvested on June 17, 2022. Post-harvest 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 compared to leaf tissue from control plants.

[0091] Example 7: Microbial community analysis of kelp raw materials and biostimulant products Two different batches of seaweed raw material powder, each with 5 or 3 technical replicates (Ecklonia maxima for MBT-E and Ascophyllum nodosum for MBT-A), were sampled. DNA was extracted from 0.025 g of powder using bead beet extraction and phenol chloroform cleanup. For MBT-A 4X and MBT-E, two or three different batches of solution were sampled with 4 or 3 technical replicates, respectively. 100 ml of concentrated product solution was filtered, bacterial cells were recovered from the filter, and DNA was extracted using the MP Biomedicals DNA Soil Pro Kit.

[0092] Amplicon-based DNA sequencing of the samples was completed by Molecular Research (MRDNA, Shallowater, TX) using their standard method for bacterial analysis on an Illumina NovaSeq 6000 system with 16S-515F primers and 20,000 reads per sample. Furthermore, MRDNA underwent QA / QC, chimera checks, and OTU (Surgical Taxonomy Unit) binning. Outputs from MRDNA were analyzed in the statistical analysis platform R using the vegan package to display the community analysis profile as a UPGMA-based cluster analysis diagram.

[0093] Cluster analysis diagrams of Ascophyllum and Ecklonia raw materials are shown in Figure 40. The bacterial communities of the EMF and AMF raw material powders are clearly different. All EMF community samples are grouped together and originate from the same branch, showing 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 because they branch from one another. All AMF community samples are grouped together and originate from the same one. The "height" scale on the left is similar to the percentage between samples. The longer the branch, the more different samples there are. 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.

[0094] The cluster analysis diagrams for MBT-A and MBT-E are shown in 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 MBT-A-2 communities are more similar to each other than MBT-A-3. These MBT-E batch communities overlap in a series of branches.

[0095] [Table 2]

[0096] [Table 3]

[0097] [Table 4]

[0098] 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 only as examples. Those skilled in the art will be able to conceive of numerous variations, modifications, and substitutions without departing from the present disclosure. It should be understood that various alternatives to the embodiments of the present disclosure may be used in the practice of the present disclosure. The following claims define the scope of the present disclosure, and the methods and structures within these claims, as well as their equivalents, are intended to be encompassed thereby.

Claims

1. A method for promoting plant growth, comprising the step of contacting a plant, the seeds of the plant, or a growth medium for the plant with a composition comprising a microbial digest product of an organic raw material including Ecklonia maxima kelp.

2. The method according to claim 1, wherein the digested product is produced by microorganisms inherent in the Ecklonia maxima kelp present in the organic raw material.

3. The method according to claim 1, wherein the digested product contains fucose at a concentration of 40 mol% or less relative to all glycosyl residues in the composition.

4. The method according to claim 1, wherein the digested product contains xylose at a concentration of more than 15 mol% relative to all glycosyl residues in the composition.

5. The method according to claim 1, wherein the digested product contains mannose at a concentration of more than 7 mol% relative to all glycosyl residues in the composition.

6. The method according to claim 1, wherein the digested product comprises one or more of isobutanol, pentadecanonitrile, pentadecanoic acid, 9-octadecennitrile, hexadecanenitrile, or heneikosan.

7. The method according to claim 6, wherein the digested product includes molecular species listed in Figure 39.

8. The method according to claim 1, wherein the digested product comprises one or more molecular species corresponding to one or more of the peaks C, F, G, J, O, or P in the LC-MS chromatogram shown in Figure 3.

9. The method according to claim 1, wherein the digested product comprises one or more molecular species corresponding to one or more of the peaks A, B, C, D, E, F, or G in the GC-MS chromatogram shown in Figure 4.

10. The digested products are shown in Figure 5. １ The method according to claim 1, comprising one or more molecular species corresponding to one or more peaks A, B, C, D, or E in the 1H-NMR spectrum.

11. The digested products are shown in Figure 6. １３ The method according to claim 1, comprising one or more molecular species corresponding to one or more peaks A or B in the 13C-NMR spectrum.

12. The method according to claim 1, wherein the composition further comprises microorganisms inherent in Ecklonia maxima present in the organic raw material.

13. The method according to claim 12, wherein the microorganism includes a spore-forming microorganism.

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

15. The microorganisms present in the composition include Microbacterium amyloricum, Thermoanaerobacterium thermosaccharolicum, Cellulosililycum lentocellum, Microbarbifer thermotolerans, Collinsella sp., and Acinetobacter sp. The method according to claim 12, comprising one or more of the following: Acinetobacter spp., Acinetobacter towneri, Lentilactobacillus buchneri, Liquoricactobacillus hordei, or Secundilactobacillus paracolinoides.

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

17. The method according to claim 1, wherein the composition does not contain microorganisms.

18. The method according to claim 1, wherein promoting plant growth includes one or more of the following: increasing seed germination, promoting early plant development, improving root growth, increasing nutrient intake, improving tolerance to abiotic stress, mitigating the effects of transplanting, improving plant reproduction, and improving soil microbial activity.

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

20. The method according to claim 1, wherein the contact step includes infall application, foliar application, application to the rhizosphere, application to seeds, or mixing with a growth medium.

21. The method according to claim 1, wherein the growth medium is soil.

22. The method according to claim 1, wherein the composition further comprises solid fertilizer particles.

23. The method according to claim 22, wherein the fertilizer particles are coated with digested products.

24. The method according to claim 1, wherein the composition is a liquid.

25. The method according to claim 24, wherein the composition further comprises a liquid fertilizer.

26. The method according to claim 24, wherein the contact step includes applying the composition at a rate of 0.5 to 10 quarts per acre.

27. The method according to claim 1, wherein the contact step includes applying a dry weight of digested product at a rate of 0.14 to 6.7 g per acre.

28. The method according to claim 1, wherein at the time of the contact step, the plant is subjected to or at risk of being subjected to a drying state.

29. The method according to claim 1, wherein the growth medium is high-salinity soil.

30. The method according to claim 1, wherein at the time of the contact step, the plant is subjected to or at risk of being subjected to a freezing condition.

31. The method according to claim 1, wherein at the time of the contact step, the plant is under cold stress or is at risk of being subjected to cold stress.

32. The method according to claim 1, wherein at the time of the contact step, the plant is under heat stress or is at risk of being subjected to heat stress.

33. The method according to claim 1, wherein the plant is transplanted.

34. The method according to claim 1, wherein the plant is corn, cotton, tomato, or bell pepper.

35. The method according to claim 1, wherein the plant is a cotton plant or a maize plant, and the cotton plant or maize plant is in a dry state at the time of contact.

36. A composition comprising digested products produced by the digestion of organic raw materials, including Ecklonia maxima kelp, by microorganisms.

37. The composition according to claim 36, wherein the microorganisms include microorganisms inherent in the Ecklonia maxima kelp present in the organic raw material.

38. The composition according to claim 36, wherein the digested product contains fucose at a concentration of 40 mol% or less relative to all glycosyl residues in the composition.

39. The composition according to claim 36, wherein the digested product contains xylose at a concentration of more than 15 mol% relative to all glycosyl residues in the composition.

40. The composition according to claim 36, wherein the digested product contains mannose at a concentration of more than 7 mol% relative to all glycosyl residues in the composition.

41. The composition according to claim 36, wherein the digested product comprises one or more of isobutanol, pentadecanonitrile, pentadecanoic acid, 9-octadecenenitrile, hexadecanenitrile, or heneikosan.

42. The composition according to claim 40, wherein the digested product comprises the molecular species listed in Figure 39.

43. The composition according to claim 36, wherein the digested product comprises one or more molecular species corresponding to one or more of the peaks C, F, G, J, O, or P in the LC-MS chromatogram shown in Figure 3.

44. The composition according to claim 36, wherein the digested product comprises one or more molecular species corresponding to one or more of the peaks A, B, C, D, E, F, or G in the GC-MS chromatogram shown in Figure 4.

45. The digested product is shown in Figure 5. １ The composition according to claim 36, comprising one or more molecular species corresponding to one or more peaks A, B, C, D, or E in the 1H-NMR spectrum.

46. The digested product is shown in Figure 6. １３ The composition according to claim 36, comprising one or more molecular species corresponding to one or more peaks A or B in the 13C-NMR spectrum.

47. The composition according to claim 36, further comprising microorganisms inherent in the Ecklonia maxima kelp present in the organic raw material.

48. The composition according to claim 47, wherein the microorganism includes a spore-forming microorganism.

49. The composition according to claim 47, wherein the dry weight percentage of microbial biomass in the composition is 0.071 to 0.714% of the total dry weight of the composition.

50. The microorganisms present in the composition include Microbacterium amyloricum, Thermoanaerobacterium thermosaccharolicum, Cellulosililycum lentocellum, Microbarbifer thermotolerans, Collinsella sp., and Acinetobacter sp. The composition according to claim 47, comprising one or more of the following: Acinetobacter spp., Acinetobacter towneri, Lentilactobacillus buchneri, Liquoricactobacillus hordei, or Seculindilactobacillus paracolinoides.

51. The composition according to claim 36, wherein the microorganisms are removed from the composition.

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

53. The composition according to claim 36, which does not contain microorganisms.

54. A composition comprising one or more molecular species corresponding to one or more peaks C, F, G, J, O, or P in the LC-MS chromatogram shown in Figure 3.

55. The composition according to claim 54, comprising one or more molecular species corresponding to one or more of the peaks A, B, C, D, E, F, or G in the GC-MS chromatogram shown in Figure 4.

56. Figure 5 １ The composition according to claim 54, comprising one or more molecular species corresponding to one or more peaks A, B, C, D, or E in the 1H-NMR spectrum.

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

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

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

60. The composition according to claim 54, comprising one or more of isobutanol, pentadecanonitrile, pentadecanoic acid, 9-octadecennitrile, hexadecanenitrile, or heneikosan.

61. The composition according to claim 60, comprising the molecular species listed in Figure 39.

62. The composition according to claim 54, further comprising microorganisms.

63. The composition according to claim 62, wherein the microorganisms include spore-forming microorganisms.

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

65. The microorganisms present in the composition include Microbacterium amyloricum, Thermoanaerobacterium thermosaccharolicum, Cellulosililycum lentocellum, Microbarbifer thermotolerans, Collinsella sp., and Acinetobacter sp. The composition according to claim 62, comprising one or more of the following: Acinetobacter spp., Acinetobacter towneri, Lentilactobacillus buchneri, Liquoricactobacillus hordei, or Seculindilactobacillus paracolinoides.

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

67. The composition according to claim 54, which does not contain microorganisms.

68. The composition according to claim 54, which is a liquid composition.

69. The composition according to claim 68, wherein the digested product is present in the liquid composition in an amount of 0.06% to 0.08% by weight relative to the total weight of the liquid composition.

70. A plant treatment composition comprising the composition and fertilizer composition described in claim 36.

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

72. The plant treatment composition according to claim 70, wherein the fertilizer composition is solid.

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

74. A method for promoting plant growth, comprising the step of bringing a plant, the seeds of the plant, or a growth medium for the plant into contact with the composition described in claim 36.

75. The method according to claim 74, wherein promoting plant growth includes one or more of the following: increasing seed germination, promoting early plant development, improving root growth, increasing nutrient intake, improving tolerance to abiotic stress, mitigating the effects of transplanting, improving plant reproduction, and improving soil microbial activity.

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

77. The method according to claim 74, wherein the contact step includes infarlow application, foliar application, or application to the rhizosphere.

78. The method according to claim 70, wherein the contact step includes applying a dry weight of digested product at a rate of 0.14 to 6.7 g per acre.

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