WO2024054500A1 - Compositions for enhancing plant growth based on microbe-free broth - Google Patents

Abstract

The presently disclosed methods, compositions, and systems are useful in enhancing plant growth and/or a plant characteristic in plants treated with the composition relative to untreated plants. Enhanced plant growth and/or plant characteristics include, a decrease in the concentration of sodium, an increase in fresh weight, and an increase in the growth of roots. In many embodiments, the disclosed compositions may comprise, filtered, fermented broth and may include one or both of a supplement and a material, which may be in dry or liquid form.

Description

COMPOSITIONS FOR ENHANCING PLANT GROWTH BASED ON MICROBE-FREE BROTH

[0001] This application claims benefit of priority pursuant to 35 U.S.C. § 1 19(e) of U.S. provisional patent application No. 63/404,881 , filed September 8, 2022, which is incorporated herein by reference in its entirety.

FIELD

[0002] The disclosed fermented filtrate compositions, processes, methods, and systems are directed to enhancing at least one characteristic of a plant.

SUMMARY

[0003] Disclosed herein are compositions comprising a fermented filtrate from a bacterial culture comprising at least one gram negative (-) bacterial species and a feedstock, wherein the fermented filtrate may comprise a supplement and is substantially sterile and free of microbes.

[0004] In some embodiments, the gram negative (-) bacterial culture comprises a species selected from Citrobacter sp., Comamonas sp., Enterobacter sp., Ensifer sp., Pseudomonas sp., and Rhizobium sp. In some embodiments, the bacterial culture comprises one or more of Citrobacter sp., Comamonas sp., Enterobacter sp., Ensifer sp., Pseudomonas sp., and Rhizobium sp., e.g., two or more of Citrobacter freundii, Enterobacter cloacae, Comamonas testosteroni, Pseudomonas putida, Pseudomonas fluorescens, Rhizobium tibeticum, and Ensifer adherens.

[0005] In some embodiments the composition further comprises one or more materials selected from the group consisting of a fertilizer, worm casting tea, rooting hormone, growth hormone, triglycerides, fatty acids, lipids, carbohydrates, simple sugars, amino acids, proteins, yeast, organic materials, fulvic acids, humates, and a vitamin, for example worm casting tea or a fertilizer.

[0006] In some embodiments the supplement is citric acid at a concentration of from 0.5% to 2.0%, or from 0.5M to 1.5. In some embodiments, the composition further comprises betaine at a concentration of from 0.5M to 1 .5M. In some embodiments the composition further comprises arginine or another supplement at a concentration of about from 0.5M to 1.5M.

[0007] The feedstock may be a plant-based extract derived from alfalfa, soybean hulls, wheat bran, beet pulp, spent hops, rice bran, grape skin, grass clippings or oat bran.

[0008] The composition may be effective to, (a) decrease the concentration of sodium in plants treated with the composition and/or (b) increase the fresh weight in plants treated with the composition relative to untreated plants, and/or (c) increase the growth of roots in plants treated with the composition relative to untreated plants.

[0009] Also disclosed are methods of manufacturing a fermented filtrate composition comprising steps of obtaining a material; and combining the material with a fermented filtrate, wherein the fermented filtrate is obtained by aerobically culturing at least one, e.g., two or more one gram (-) bacterial species. In some embodiments, the bacterial species is selected from Citrobacter sp., Comamonas sp., Enterobacter sp., Ensifer sp., Pseudomonas sp., and Rhizobium. In many embodiments, a supplement is added to the fermented filtrate, such as one or more of citric acid, betaine, and arginine. In many embodiments, the material is one or more of a worm tea, a fertilizer, a macronutrient, a micronutrient, a plant hormones a triglyceride, a fatty acid, a lipid, a carbohydrate, an amino acid, a protein, yeast, an organic material, fulvic acid, a humate or a vitamin.

[0010] In another aspect, a method of enhancing at least one characteristic of a plant is disclosed, wherein the method comprises steps of contacting soil with a fermented composition, wherein the composition may comprise a fermented filtrate and a material. In some embodiments, the fermented filtrate comprises a supplement. The fermented filtrate may be obtained by aerobically culturing at least one bacterial species. In some embodiments the bacterial species is selected from Citrobacter sp., Comamonas sp., Enterobacter sp., Ensifer sp., Pseudomonas sp., and Rhizobium sp. In some embodiments, the filtrate may be passed through a filter, for one example a 0.22 pm filter, a 0.45 pm filter and/or a 10 pm filter. In one embodiment, the material may be a worm casting tea having a pH from about 3.0 to 9.0, or a fertilizer, e.g., 10-4-5 or 20-10-20.

BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 is a graphic depiction of the dry weight of hemp flower yield in control plants and plants treated with worm castings tea and fermented filtrate composition. Plants had higher yield with Worm Castings Tea application and fermented filtrate composition application compared to control plants [One way Welch ANOVA (W2.000, 62.82 = 30.79, P < 0.0001 ) followed by Dunnett’s T3 multiple comparisons tests (Control vs. Worm Castings Tea, P = 0.0032; Control vs. fermented filtrate composition , P < 0.0001)].

[0012] Figures 2a and 2b are graphic depictions of the total terpene (FIG. 2a) and cannabinoid (FIG. 2b) content in hemp for control plants and plants treated with worm castings tea and fermented filtrate (Turbo prepared using alfalfa feedstock).

[0013] Figures 3a-e are graphic depictions of plant traits where plants treated with worm castings tea and 0.6 mL/gal fermented filtrate are significantly improved relative to control. Leaf area (FIG. 3a) showed a p-value of 0.04, leaf dry mass (FIG. 3b) showed a p-value of 0.06, shoot dry mass (FIG. 3c) showed a p-value of 0.07, total mass (FIG. 3d) showed a p-value of 0.06, and stem diameter (FIG. 3e) showed a p-value of 0.04.

[0014] Figures 4a-c are graphic depictions of plant traits where plants treated with worm castings tea and 5 mL/gal fermented filtrate are significantly improved relative to control. Canopy projected area (FIG. 4a) showed a p-value of 0.06, root length (FIG. 4b) showed a p-value of 0.01 , and root surface area (FIG. 4c) showed a p-value of 0.02.

[0015] Figure 5 is a graphic depiction of significantly higher root dry mass in plants treated with worm castings tea and 0.6 mL/gal fermented filtrate (p=0.05) or the worm castings tea and 25 mL/gal fermented filtrate (p=0.05).

[0016] Figure 6 is a graphic depiction of decrease of sodium levels in plants treated with Turbo as indicated by a decrease in the concentration of sodium per gram dried plant tissue (“percent rate”) in cucumber when Turbo is provided together with synthetic or organic fertilizer amendments. The samples analyzed included Turbo alone or T urbo plus: Grow A and Grow B (see below); 3-1 -1 (Natures Source); 3-2-1 (Natures Source); 10-4- 5 (Natures Source); UAN 32 (Yara);10-34-0 (Two Rivers Terminal); Micronutrients (Natures Source); Humic Acid (Live Earth Products); Kelp (Afrikelp); Mammoth Silica; Worm Casting Tea (Denali Biosolutions); 8-4-4 (Natures Source). The results are detailed in Example 6.

[0017] Figure 7 is a graphic depiction of the results of amino acid analyses of Turbo Lot #s 1584 and 1738 (prepared as described in Example 7, below), versus filtered alfalfa feedstock. The analysis was conducted by Creative Proteomics.

[0018] Figure 8 is a graphic depiction of the results of the fresh mass of tomato plants after 3 weeks of growth following treatment with Turbo plus and minus Fertilizer (10-4-5).

[0019] Figure 9 a graphic depiction of the results of the fresh mass of tomato plants after 4 weeks of growth following treatment with Turbo plus and minus Fertilizer (20-10-20).

[0020] Figure 10 is an image of the results from a seed bag assay which depicts the size, shape, and growth of an immature plant and roots resulting from seeds pre-germinated and inserted into the perforated upper fold of a “dispo” bag.

[0021] Figures 11 a-c provide a graphic depiction of the results from analysis of roots from the seed bag assay detailed in Example 11 , where “Reach” (R), the deepest distance the roots reached (FIG. 11 a), “Spread” (S) is the widest distance the roots spread out in each bag (FIG. 1 1 b), and the root growth area (A) root area is calculated by (RxS)/2 (FIG. 1 1 c). DETAILED DESCRIPTION

[0022] Disclosed herein are fermented filtrates that enhance plant growth and/or a plant characteristic alone or when combined with various materials. In some embodiments, the fermented filtrate is a bacterial culture that has been filtered to remove bacteria. The filtrate may comprise various compositions, compounds, molecules, proteins, peptides, amino acids, nucleic acids, and the like. In many embodiments, the bacteria include one or more of Citrobacter sp., Comamonas sp., Enterobacter sp., Ensifer sp., Pseudomonas sp., and Rhizobium sp.

[0023] The disclosed fermented filtrates, when provided to plants alone or when combined with a material, may result in enhanced plant growth and/or an improved plant characteristic.

DEFINITIONS.

[0024] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the disclosure pertains. Although other processes and materials similar, or equivalent, to those described herein can be used to produce concentrated microbial compositions, the system and processes are described herein provides significant advantages are further detailed herein. Other features and advantages of the disclosure will be apparent from the following detailed description, and from the claims.

[0025] In describing and claiming the present disclosure, the following terminology will be used in accordance with the definitions set out below.

[0026] As used in this specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise.

[0027] The term "aero hydraulic bioreactor" is used herein with reference to a bioreactor wherein the contents are mixed by infusion of air into the liquid in the bioreactor. This differs from traditional bioreactors which require mechanical means to mix bioreactor contents. See, US Provisional Application Serial No. 63/457,915, expressly incorporated by reference herein.

[0028] The term "CPU” is an abbreviation for “colony forming unit” and “CFU/mL” is the number of colony forming units per milliliter of a microbial solution.

[0029] The term "comprises" and grammatical equivalents thereof are used herein to mean that, in addition to the features specifically identified, other features are optionally present. For example, a composition or device "comprising" (or "which comprises") components A, B and C can contain only components A, B and C, or can contain components A, B and C and one or more other components. When a range is given as "(a first number) to (a second number)" or "(a first number) - (a second number)", this means a range whose lower limit is the first number and whose upper limit is the second number. The terms “plural”, “multiple”, “plurality” and “multiplicity” are used herein to denote two or more than two features. Parts, percentages and ratios given in this specification are by weight unless otherwise noted.

[0030] The term "concentrated bacterial culture" means a bacterial culture with a concentration of from 10⁴ CFU/mL to 10¹¹ CFU/mL. The lower limit is known to those of skill in the art in terms of what is considered concentrated enough to be a product and the upper limit is set by the practical physical threshold to which gram-negative (-) bacterial cells can historically be grown. For example, if 1 liter of a 10⁷ CFU/mL cell stock such as that produced using an aero-hydraulic bioreactor is added to 999 liters of 1 x media, it yields 1000 liters of a 10⁴ CFU/mL culture (the lower threshold of what is defined as a concentrated bacterial culture).

[0031] The term "consortium" as used herein means a mixture of two or more different microbes, e.g., bacteria of different strains or species.

[0032] The term “continuous-liquid-feed” or “CLF” as used herein means applying a treatment to plants, e.g., a fermented filtrate as a water supplement.

[0033] The term "culture” as used herein means any combination of bacterial species and feedstock incubated for any period of time with or without aeration.

[0034] The term “EC” is used herein with reference to electroconductivity.

[0035] The term “feedstock” as used herein means a plant-based extract, typically prepared by a heated liquid steep of specific plant matter, which serves as nutrient/culture medium for microbial cultures. In some embodiments, a 1 x feedstock also referred to as 1 x growth media includes about 90 grams of plant matter steeped for 1 hour at 70°C per gallon of tap water. In some embodiments, the ratio of plant material mass to water is from about 45 g/gal to 90 g/gal, and the plant matter may be steeped at temperature of from about 70°C to boiling.

[0036] The term "fertilizer” is used herein with reference to a substance containing one or more recognized plant nutrients that is applied to plants or soil. Some exemplary fertilizers include but are not limited to, 10-4-5 (Two Rivers Terminal), 10-34-0 and 20-10-20 (Jacks Peat Lite). [0037] The term "improve plant growth and development” as used herein means the increased growth and/or more rapid development of one or more of roots, shoots, leaves, flowers, and fruits.

[0038] As used herein, the term “increase the growth of roots” is used with reference to characteristics such as root length and root tip number, or may be determined using a seed bag assay (exemplified in Example 11 ), where the results are presented as: “Reach” (R), which is the deepest distance the roots reached, and “Spread” (S) is the widest distance the roots spread out in each bag, and the growth area (A) is calculated by multiplying R by S, and the root area is calculated by (RxS)/2.

[0039] The term "microbial composition" is used herein with reference to a culture of microbes, such as a culture of bacteria, e.g., gram negative bacteria.

[0040] The terms, “osmolytes”, “osmoprotectants”, and “osmolytic compounds”, are used herein with reference to compounds that play a crucial role in helping plants adapt to various environmental stresses, particularly osmotic stress caused by changes in water availability. Osmotic stress occurs when there is an imbalance between the amount of water inside plant cells and the water in the surrounding soil, leading to dehydration and potential damage to cellular structures. Osmolytes help plants manage this stress. Osmolytes allow more water into the plant and thereby decrease salt stress. Fertilizers and water in aquafers are high in salt. Osmolytic compounds include but are not limited to, betaine, proline, trehalose, some sugars including polyols such as mannitol and sorbitol, peptides, potassium, some acidic molecules and amino acids. Osmolytic compounds have also been reported to help scavenge reactive oxygen species (ROS) and protect cells from oxidative damage. Environmental stressors such as drought, high salinity, and intense light can lead to the production of reactive oxygen species (ROS) within plant cells. ROS can cause cellular damage and oxidative stress.

[0041] The term "sp." as used herein means species.

[0042] The term "substantially sterile” is used herein with reference to a fermented broth that has been filtered, e.g., through a 10 pm filter, a 0.45 pm filter, and/or a 0.22 pm. In some cases, a supplement is added to the fermented filtrate.

[0043] The term "Turbo" is used herein with reference to fermented filtrates derived from the culture of bacteria, e.g. gram (-) bacteria. In some embodiments, Turbo is filtered through a 0.22 micron filter and supplemented with 1% citric acid. In some embodiments, Turbo is filtered through a 10 micron filter and supplemented with from about 0.5M to about 1 .5M citric acid and from about 0.5M to about 1 .5M betaine. In some embodiments, Turbo is supplemented with arginine, e.g., from about 0.5M to about 1 .5M arginine. [0044] The term "water activity" is used herein with reference to the availability of water molecules in a substance. Higher water activity (a_w) means more available water. Lower water activity minimizes bacterial growth. Exemplary substances that lower water activity include but are not limited to betaine, citrate salts, e.g., sodium citrate or potassium citrate, n-acetyl glutamate, and arginine. Water activity can also be decreased under acidic conditions, e.g., by lowering the pH. Water activity can be measured using a hydrometer or by putting a fixed volume in a container with head space and measuring the humidity in the head space.

Fermented Filtrates

[0045] Various bacterial species may be useful in creating the disclosed fermented filtrate, also referred to as “Turbo”. In some embodiments, the bacterial species include one or more of Citrobacter sp., Comamonas sp., Enterobacter sp., Ensifer sp., Pseudomonas sp., and Rhizobium sp. In some embodiments, the bacterial species include one or more of Citrobacter freundii, Enterobacter cloacae, Comamonas testosteroni, Pseudomonas putida, Pseudomonas fluorescens, Rhizobium tibeticum, and Ensifer adherens. In some embodiments, the bacterial species are Comamonas testosteroni and Pseudomonas putida.

[0046] Various media, also referred to as feedstocks, may be used to grow the disclosed bacterial species prior to filtration, plant material. In some embodiments, that plant material may be organically grown. In one embodiment, the plant material is alfalfa, and a sterilized infusion may be used to create the disclosed feedstock. In some embodiments, the alfalfa may be Medicago sativa. In some embodiments, the plant material is wheat bran. In some embodiments, the plant material is soybean hulls, beet pulp, spent hops, rice bran, grape skin, grass clippings or oat bran. A variety of techniques may be useful in creating the disclosed feedstocks. In one embodiment, the media is a plant material infusion. In these embodiments, dry plant material in various forms, such as pelletized plant material, may be combined with water to create a slurry/steeping mixture and heated to about 71 °C for about 1 hour to create a raw feedstock or infusion media.

[0047] The raw feedstock or infusion media may be filtered to remove solid matter, microbes, and/or contaminants, and thereby create a filtered feedstock or infusion media. The filtered feedstock or infusion media may also be further sterilized by autoclaving at about 121 °C at about 15 psig for about 1 hour.

[0048] Various methods may be used to prepare cultures of the disclosed bacterial species. In one embodiment, cultures are inoculated to achieve an initial bacterial density of between about 1 E+05 (1 x10^A5 or 1 x10⁵) and about 1 x10⁶ Colony Forming Units (CFU)/ml_. In many embodiments, the initial concentration of each bacterial species is between about 1 x10⁵ and about 1 x10⁶ CFU/mL. Thus, a culture containing 2 different bacterial species may have an initial concentration of between about 2x10⁵ and about 2x10^s CFU/mL. In many embodiments, the volume of initial bacterial inoculum added to the cultures will not exceed about 1 % v/v basis. Thus, the initial volume of a bacterial inoculum added to 300 L of sterile, filtered infusion media by be less than about 3 mL.

[0049] Bacterial cultures may be grown or fermented under various methods to create the presently claimed fermented filtrate. In most embodiments, the culture is allowed to grow/ferment in the filtered infusion media under aerobic conditions for at least 3 days at about 25°C to create a fermented broth. In some embodiments, the culture is prepared using an aero hydraulic bioreactor as described in US Provisional Application Serial No. 63/457,915, expressly incorporated by reference herein.

[0050] In some embodiments, an autoclaved 5x concentrated feedstock (as determined by absorption at 400nm UV-Vis) is diluted with filter-sterilized water to generate large volumes of sterile 1 x feedstock, which is used to fill a bioreactor vessel. In some embodiments, the bioreactor is inoculated with approximately 5 mL of inoculum with a concentration of 10⁴ to about 10⁶ CFU/mL. Progress of the incubation is accomplished by periodically measuring the CFU/mL and pH of the culture in the bioreactor vessel.

[0051] Microbes may be removed from the fermented broth by various methods to create a filtrate. In some embodiments, microbes may be removed from the fermented broth by filtering, for example filtering the fermented broth through a 0.22 pm filter, to create a fermented filtrate. In many embodiments, one or more pre-filter steps may be performed prior to the 0.22 pm filtration. For example, the fermented broth may be passed through an initial filter, e.g., a 0.45 pm filter, to remove large particles and/or aggregates, including biofilms. In such cases, a supplement may be added to the fermented filtrate.

Supplements

[0052] In many embodiments, supplement, e.g., citric acid, betaine, arginine, sorbic acid, nitrites, nitrates, sulfites, benzoate, a sorbate, EDTA, a gallate, a paraben, a propolis extract, tartaric acid, malic acid, and/or ascorbic acid are added to the fermented filtrate. In some embodiments, the fermented filtrate comprises 1 % citrate (v/v). In some embodiments, the fermented filtrate comprises 1 M citric acid and 1 M betaine. In some embodiments, the fermented filtrate further comprises 1 M arginine. In some embodiments, microbes may be removed from the fermented broth by filtering through a 10 pm filter to create a fermented filtrate, followed by addition of one or more of citric acid, betaine, arginine or another supplement.

[0053] The concentration of citrate in the fermented filtrate may be from about 0.5%, to about 3.0%, from about 0.8%, to about 2.2%, from about 1 .0%, to about 2.5%, from about 1 .3% to about 2.5%, from about 1 .6% to about 2.8%, from about 0.1 M to about 3.0M, from about 0.5M to about 2.5M, from about 0.8M to about 2.2M, or from about 1.0M to about 2.0M, for example, 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%, 2.0%, 2.1 %, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%,

3.0%, 0.1 M, 0.2M, 0.3M, 0.4M, 0.5M, 0.6M, 0.7M, 0.8M, 0.9M, 1 ,0M, 1.1 M, 1.2M, 1.3M,

1.4M, 1.5M, 1.6M, 1.7M, 1.8M, 1.9M, 2.0M, 2.1 M, 2.2M, 2.3M, 2.4M, 2.5M, 2.6M, 2.7M,

2.8M, 2.9M, or 3.0M.

[0054] The concentration of betaine in the fermented filtrate may be from about 0.1 M to about 3.0M, from about 0.5M to about 2.5M, from about 0.8M to about 2.2M, or from about 1 ,0M to about 2.0M, for example, 0.1 M, 0.2M, 0.3M, 0.4M, 0.5M, 0.6M, 0.7M, 0.8M, 0.9M, 1 ,0M, 1 .1 M, 1 ,2M, 1 ,3M, 1 ,4M, 1 ,5M, 1 ,6M, 1 ,7M, 1 ,8M, 1 ,9M, 2.0M, 2.1 M, 2.2M, 2.3M, 2.4M, 2.5M, 2.6M, 2.7M, 2.8M, 2.9M, or 3.0M.

[0055] The concentration of arginine in the fermented filtrate may be from about 0.1 M to about 3.0M, from about 0.5M to about 2.5M, from about 0.8M to about 2.2M, or from about 1 ,0M to about 2.0M, for example, 0.1 M, 0.2M, 0.3M, 0.4M, 0.5M, 0.6M, 0.7M, 0.8M, 0.9M, 1 ,0M, 1 .1 M, 1 ,2M, 1 ,3M, 1 ,4M, 1 ,5M, 1 ,6M, 1 ,7M, 1 ,8M, 1 ,9M, 2.0M, 2.1 M, 2.2M, 2.3M, 2.4M, 2.5M, 2.6M, 2.7M, 2.8M, 2.9M, or 3.0M.

[0056] Some Turbo embodiments include but are not limited to:

Grow-20l0-A0: Mammoth P, where the cultured bacteria are Citrobacter freundii, Enterobacter cloacae, Comamonas testosteroni, and Pseudomonas putida, which are grown for 7 days in an alfalfa extract with shaking at 150 RPM and 25°C. On the 7th day of fermentation, 1 % v/v citric acid is added and the culture is centrifuged for 10 minutes at 10,00 RPM, followed by filtration of the supernatant is through a 0.45 pm filter then a 0.22 pm filter;

Grow-20l0-B1 : Mammoth P Canada, where the cultured bacteria are Comamonas testosteroni and Pseudomonas putida, which are grown for 7 days in a wheat bran extract with shaking at 150 RPM and 25°C. On the 7th day of fermentation, 1 % v/v citric acid is added, and the culture is centrifuged for 10 minutes at 10,00RPM, followed by filtration of the supernatant through a 0.22 pm filter; and

Grow-2113-B9: Mammoth P Canada plus Mammoth NFC1 where the cultured bacteria are Comamonas testosteroni, Pseudomonas putida, Pseudomonas fluorescens, Rhizobium tibeticum, and Ensifer adherens, which are grown for 7 days in a wheat bran extract with shaking at 150 RPM and 25°C. On the 7th day of fermentation, 1 % v/v citric acid is added, and the culture is centrifuged for 10 minutes at 10,00RPM, followed by filtration of the supernatant through a 0.22 pm filter.

Combination Materials.

[0057] The disclosed filtrates may be combined with various materials to create compositions with enhanced function. In various embodiments, combining the disclosed filtrate and materials may result in enhancement of one or more plant growth characteristic or plant characteristic. In various embodiments, ‘combination,’ as used herein may include applying the filtrate and material as a preprepared mixture, separately at or about the same time, or one after the other. In some embodiments, where the filtrate and material are not applied at the same time, the interval between applying the second component (i.e. filtrate, if material is applied first; or material, if filtrate is applied first) of the combination may be from minutes to days or weeks. For example greater than about 1 minute, 10 minutes, 30 minutes, 1 h, 2 h, 3 h, 4 h, 5 h, 6 h, 7 h, 8 h, 9 h, 10 h, 15 h, 24 h, 48 h, 72 h, 96 h, one week, 2 weeks, 2.5 weeks, 3 weeks, 3.5 weeks, 4 weeks, 4.5 weeks, 5 weeks, 6 weeks, 7 weeks, 2 months, 2.5 months, 3 months, 3.5 months, or 4 months, and less than about 5 months, 4.5 months, 4 months, 3.5 months, 3 months, 2.5 months, 2 months, 7 weeks, 7.5 weeks, 6.5 weeks, 6 weeks, 5.5 weeks, 5 weeks, 4.5 weeks, 4 weeks, 3.5 weeks, 3 weeks, 2.5 weeks, 2 weeks, 10 days, 9 days, 8 days, 7 days, 6 days, 5 days, 96 h, 72 h, 48 h, 24 h, 15 h, 10 h, 9 h, 8 h, 7 h, 6 h, 5 h, 4 h, 3 h, 2 h, 1 h, 30 min, 10 min, or 2 min.

[0058] In some embodiments the disclosed filtrates are provided to plants in water as a Continuous-Liquid-Feed (CLF) or water supplement alone or in combination with one or more materials. Various materials may be combined with the disclosed fermented filtrate. In many embodiments, the material may comprise one or more composition, compound, molecule, or element known to be useful in aiding plant growth and/or a plant characteristic. In one example the material is a worm casting. Other materials which may be combined with filtrate to achieve an effect on plant development and/or nutrient bioavailability include but are not limited to one or more of, fertilizers, macronutrients such as nitrogen (N), phosphate (P) and potassium (K), micronutrients such as boron (B), zinc (Zn), manganese (Mn), iron (Fe), copper (Cu), molybdenum (Mo), chloride (Cl), calcium (Ca), nickel (Ni), magnesium (Mg), silica (Si), plant hormones including rooting and growth hormones, triglycerides, fatty acids, lipids, carbohydrates including simple sugars, amino acids, proteins, yeast, organic materials, fulvic acids, humates (humic acid materials and solutions), and vitamins. [0059] The disclosed materials may be obtained from various sources. In some embodiments, the source of the material(s) may be chemical, biological, and/or organic matter. Exemplary biological or organic matter includes but is not limited to composted plant matter, composted manures and guano, bone and blood meals; seaweed/kelp, fish emulsion, worm casting tea, alfalfa tea, wood chips, mycorrhizae, yucca extract, willow extract, insect frass, azymite, dolomite, steep, protein hydrolysate and feather meal.

[0060] Worm castings are digested waste produced by worms. In most cases, the worms may be fed or grown in various forms of waste biomass. In most cases, worms defecate at or near the top of a soil, biomass, or castings pile, and then return into the pile for additional feeding. Worm castings may be in their natural granular form, or they may be processed, for example by pelletizing or liquefaction. Liquid worm castings (referred to as tea), may be made in various ways. In one embodiment, worm castings are combined with water and allowed to sit for about 24 hours. Liquid castings may also be obtained directly from a castings pile as they may be produced naturally by the worms while feeding. In this case, the liquid may be collected at or near the bottom of a pile or worm bed. In some cases, the tea may be aerated and/or a sugar source may be added to the water. The Worm casting tea may have a pH from about 3.0 to 9.0, for example, greater than about 3.0, greater than about 3.5, greater than about 4.0, greater than about 4.5, greater than about 5.0, greater than about 5.5, greater than about 6.0, or less than about 9.0, less than about 8.5, less than about 8.0, less than about 7.5, less than about 7.0, or less than about 6.5. In some cases, the castings may be placed in a bag or filter material before adding to the water, this may aid in removal of solid matter at the end of the process. In most embodiments, the water is not bactericidal, that is, the water does not kill, inhibit, or otherwise impede growth of bacteria. Thus, in many cases, for example where municipal water is used to produce the tea, the water may be filtered, treated, or otherwise altered to reduce or eliminate anti-bacterial compounds, molecules, and elements. In some embodiments, municipal water may be allowed to sit for days or weeks until chlorine is neutralized or off gassed. In other embodiments, untreated well water may be used.

Additives.

[0061] The disclosed fermented compositions may further comprise one or more additives. In one embodiment, the additive is one or more compound or composition for stabilizing, optimizing, or preserving the composition, aiding in its storage or application, and/or treating the plant or soil. In some embodiments, the additive is selected from one or more of wetting agents, dispersants, surfactants, water trapping agents, zeolites, enzymes, pest control agents, pesticides, acaracides, molluscicides, insecticides, fungicides, nematicides, and antifoam compounds.

Turbo Compositions.

[0062] The presently disclosed fermented filtrate may be combined with a material to create a composition for enhancing plant growth and/or a plant characteristic. As used herein, the term “fermented compositions” may be used with reference to combinations of the disclosed fermented filtrate and a material that when combined, enhance plant growth and/or a plant characteristic. In various embodiments, the fermented composition may be in various forms, for example, liquid, solid, powder, pellets, granules, etc. The filtrate and material may be combined in various ratios typically presented as mL/gal. or percentage (%) depending upon the material and method of application, e.g., CLF or soil drench. The disclosed filtrates, materials, additives, and fermented compositions may have a pH of between about 3.0 and about 1 1 .0, for example, greater than about 3.0, greater than about 3.5, greater than about 4.0, greater than about 4.5, greater than about 5.0, greater than about 5.5, greater than about 6.0, or less than about 11 .0, less than about 10.5, less than about 10.0, less than about 9.5, less than about 9.0, less than about 8.5, less than about 8.0, less than about 7.5, less than about 7.0, or less than about 6.5, for example pH 3.5.

Methods of using Fermented Filtrates to Enhance Plant growth.

[0063] Disclosed herein are various methods of using the disclosed filtrate to enrich soil and/or enhance plant growth and/or a plant characteristic in the presence of one or more materials. The disclosed fermented compositions may, in some cases, be applied to soil, seed, and seedling, or mature plants. The fermented compositions may be applied to the soil surface or mixed into the soil using methods known in the art, such as spraying, drenching, injecting, tilling, and/or plowing. The fermented composition may also be applied directly to the plant via foliar application using methods to the art such as spraying. The fermented composition may be applied directly to the soil or mixed with water and/or fertilizer prior to application. In some embodiments, the fermented filtrate may be applied to plant parts, plant roots, seeds, or growth media before, during, or after introduction or germination of the plant.

[0064] The disclosed filtrates, methods, and fermented compositions (i.e. containing the disclosed filtrate and a material) may be used to enhance growth and/or a characteristic of plants cultured by various methods, including hydroponic, aeroponic systems, which is then delivered to the soil and/or plant in liquid or dry form. In another aspect, the fermented composition may be used to enrich seeds prior to planting. In various embodiments, the plant or seed may be grown in various environments, including fields and containers. Plants, for use with the presently disclosed filtrates, materials, fermented compositions, and methods may be grown in aeroponic, hydroponic, aquaponic, in-vitro, and traditional horticulture techniques.

[0065] When Turbo is provided in a continuous liquid feed environment, the concentration of Turbo in the water is from about 0.1 mL/gallon to about 0.6ml_/gallon, about 0.2mL/gallon to about 0.5ml_/gallon, about 0.1 mL/gallon, 0.2mL/gallon, 0.3mL/gallon, 0.4mL/gallon, 0.5mL/gallon or 0.6mL/gallon.

[0066] When Turbo is provided as a soil application, the concentration of Turbo is from about 0.2mL/gallon to about 2.5mL/gallon, about 0.4mL/gallon to about 2.2mL/gallon, about 0.5mL/gallon to about 2. OmL/gallon, 0.2ml_/gallon, 0.3mL/gallon, 0.4mL/gallon,

0.5mL/gallon, 0.6mL/gallon, 0.7mL/gallon, 0.8mL/gallon, 0.9mL/gallon, 1 .OmL/gallon,

1 .1 mL/gallon, 1 .2mL/gallon, 1 .3mL/gallon, 1.4mUgallon, 1 .5mL/gallon, 1 ,6mL/gallon,

1 .7mL/gallon, 1 .8ml_/gallon, 1 .9ml_/gallon, 2. OmL/gallon, 2.1 mL/gallon, or 2.2mL/gallon on a weekly basis.

[0067] Fertilizers are provided consistent with manufacturers recommendations.

[0068] Various soil or soil media may be used to grow plants according to the present methods and with the presently disclosed filtrates, materials, and fermented compositions. In some embodiments, the soil media may be natural soil, synthetic soil, or a combination thereof. In some embodiments, the soil may comprise rockwool, coco fiber (coir) or chips (croutons), and/or other suitable media well known in the art. In some embodiments, seeds may be soaked in or coated with the presently disclosed fermented compositions prior to planting.

[0069] Soil media may include one or more additional media such as, without limitation, perlite, vermiculite, diatomaceous earth, rock wool, clay pebbles or balls, growstones, peat (for example sphagnum peat moss), sand, woodchips, sawdust, pebbles, gravel, and other media. In many cases, soilless grow media may be comprised of organic or inorganic materials, and may be mixed together. Generally, soil amendments in combination with soilless media may provide support for the growth of the plant, especially for the root.

[0070] Plant growth and/or plant characteristics may be assessed by measuring one or more plant characteristics. In some embodiments plant number, germination time, plant height, stem diameter, branch diameter, biomass production, root weight, total fresh weight, total dry weight, fruit number, fruit size, fruit weight, bud number, flower number, flower size, etc. may be measured. [0071] Various plant types (e.g. food plants, fiber plants, dye plants, edible plants, medicinal plants, oil-producing and oilseed plants, ceremonial plants, ornamental plants, flowering and nonflowering plants, decorative plants, commercial plants, etc. as well as seeds and parts thereof) may benefit from the disclosed compositions and methods, and these plant types may be useful in determining enhanced plant growth and/or plant characteristics. In one embodiment, the plant may be a vegetable, fruit, tree, turf, grass, weed, etc. In some embodiments, the plant is Cannabis sativa, tomato, a cucurbit, jalapeno pepper, hemp, etc. Growth and/or plant characteristics may be measured in plants grown under a controlled environment, for example in a greenhouse. In some embodiments, plants may be grown in perlite media in a hydroponic system. In various embodiments, a greenhouse containing the plants may be kept at a temperature between 75-85 °F. Plants may be grown under 16 hours of light for the first four weeks followed by ambient light conditions (13-15 hours of daylight).

[0072] The disclosed fermented compositions, filtrates, and methods may enhance growth and/or plant characteristics to varying degrees. In some embodiments, the measured characteristic may increase more than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 110%, 120%, 130%, 140%, 150%, 200%, 300%, 400%, 500%, or more, and less than about 1000%, 500%, 400%, 300%, 200%, 150%, 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, or 2% over a given interval of time. As one example, if the disclosed filtrate, added alone, is able to enhance one or more of plant growth or a plant characteristic by about 10 percent, and a second composition may enhance one or more of plant growth or a plant characteristic by about 10 percent - in these cases, combining the disclosed filtrate and material enhance one or more of plant growth or a plant characteristic by 20% or more that is the effect of combining the material with the disclosed filtrate may be greater than additive. In many embodiments, the disclosed combination of material and filtrate may result in enhancing a characteristic by more than about 1-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, or 10-fold, and less than about 20-fold, 10-fold, 9-fold, 8-fold, 7-fold, 6-fold, 5-fold, 4-fold, 3-fold, or 2-fold compared to the material enhancement and filtrate enhancement separately. In some embodiments, the enhanced plant growth and/or changes in a plant characteristics may result from an additive combination of the material enhancement and filtrate enhancement. In some embodiments, the enhanced plant growth and/or changes in a plant characteristics may result from the ability to use less of the material or the filtrate to obtain a similar, same, or greater measured growth or characteristic. [0073] In some embodiments, the interval of time of measured plant growth may be more than about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 2 weeks, 2.5 weeks, 3 weeks, 3.5 weeks, 4 weeks, 4.5 weeks,

5 weeks, 6 weeks, 7 weeks, 2 months, 2.5 months, 3 months, 3.5 months, 4 months, 4.5 months, 5 months, 5.5 months, 6 months, 6.5 months, 7 months, 7.5 months, or 8 months, and less than about 9 months, 8.5 months, 8 months, 7.5 months, 7 months, 6.5 months,

6 months, 5.5 months, 5 months, 4.5 months, 4 months, 3.5 months, 3 months, 2.5 months, 2 months, 7 weeks, 7.5 weeks, 6.5 weeks, 6 weeks, 5.5 weeks, 5 weeks, 4.5 weeks, 4 weeks, 3.5 weeks, 3 weeks, 2.5 weeks, 2 weeks, 13 days, 12 days, 11 days, 10 days, 9 days, 8 days, 7 days, 6 days, 5 days, 4 days, 3 days, or 2 days.

Plant characteristics.

[0074] The presently disclosed methods and compositions may be useful in enhancing one or more characteristics of a plant. In some embodiments, the characteristics may include one or more of growth rate (for example as measured by height, weight, canopy, or root area), micro-nutrient content, macronutrient content, active compound content, etc., number of fruit, flowers, leaves, buds, etc. Plant growth may be assessed by measuring one or more plant characteristics. In some embodiments, one or more of seed germination rate, seed germination percentage, germination time, root length, root density, root biomass, enzyme activity, plant height, stem diameter, fresh or dry biomass (e.g., total biomass, leaf biomass, flower biomass, seed biomass, etc.), leaf number, leaf area, fruit number, fruit size, fruit weight, bud number, timing of budding, flower number, flower size, biochemical and nutrient concentrations in all or some of the different plant components, and/or the rate of change of these or other characteristics.

[0075] In some embodiments, the disclosed fermented filtrates (Turbo) mitigate the effects of salt stress or drought. A high sodium (Na+) concentration in fertilizers, water sources, and soil can cause salt stress which may have detrimental effects on plants. When the soil has a high sodium concentration, the difference in water potential makes it more difficult for plants to take up water from the soil. A reduction in water availability to plant cells due to salt stress or drought can result in dehydration of plants and impair nutrient uptake. In addition, a high sodium concentration can directly damage plant roots, affecting plant growth and development. Turbo appears to be most effective at enhancing plant characteristics in a high salt environment, as exemplified by the application of Turbo to plants in the presence of high salt fertilizers.

[0076] With respect to cannabis plants, in some embodiment the active content may be one or more of a cannabinoid such as CBDA (Cannabidiolic acid), cannabidiol (CBD), THCA (A⁹-tetrahydrocannabinolic acid), A⁹-tetrahydrocannabinol (THC), CBCA (Cannabichromenenic acid), CBGVA (Cannabigerovarinic acid), THCVA (Tetrahydrocanabivarinic acid), CBDVA (Cannabidivarinic acid), CBCVA (Cannabichromevarinic acid) and CBGA (Cannabigerolic acid), CBC (Cannabichromene), CBGV (Cannabigerivarin), THCV (Tetrahydrocannabivarin), CBDV (Cannabidivarin), CBCV (Cannabichromevarin), and CBG (Cannabigerol), or terpene, including but not limited to myrcene, limonene, pinene, linalool, caryophyllene, humulene, guaiol, camphene and linalool, and any derivatives thereof.

[0077] The compositions and methods described herein find utility in agriculture as well as other fields where cultures of gram (-) bacteria are a part of manufacturing processes utilizing biological systems to produce commercially important biomaterials. Examples include pharmaceuticals, textiles, food ingredients, fuel enzymes, and more.

EXAMPLES

Example 1 - Determining Effect Of Fermentation Broth On Plant Properties.

[0078] A randomized block design was used to test the effect of the presently disclosed compositions and methods on various plant properties. Each block consisted of 15 plants (planted in a single row). The present test was performed on clones from the hemp strain Cherry Uno. Each treatment arm included three replicated blocks, for a total of 45 (i.e. 3x15) plants per treatment. All plants were grown in sterile coco coir cubes and were fed a standard fertilizer regime each week. Electrical Conductivity (EC) was monitored and kept between 1 .8 and 2.3. Greenhouse temperatures were set at 75 °F during the day and 60-65 °F at night. Natural lighting was used to maintain an 18:6 light cycle during vegetative stage (4 weeks) and a 12:12 light cycle during flowering period (8 weeks). Nutrients were applied at the rates recommended by manufacturer, and delivered through a custom constructed drip irrigation system. Nutrients were delivered approximately three times per week for the first twelve weeks, with only tap water (5.8 pH) being applied the last week to flush the growth medium. Watering events occurred as needed (growers’ discretion) with frequency increasing over time as plant biomass increased. The experimental design was a randomized complete design.

[0079] Treatments solutions consisted of a (1) control (liquid fertilizer only), (2) worm castings tea (3:1 dilution in tap water), and (3) fermented composition (3:1 dilution tea into tap water with 0.6ml/gal fermented alfalfa filtrate added to dilution). After treatment solutions were prepared, they were buffered to a pH range accepted for hydroponic production (6.0 - 6.5). The buffered treatment solution was then applied manually to the substrate surface at 250ml during vegetative stage and 500ml during the flowering period per plant for each treatment. [0080] At harvest, plants were cut at the base of the stem and the intact plants were air dried, in the dark, at 27°C (80°F) or ambient temperature with airflow for one week. Potency and terpene values were also collected at the time of harvest from five randomly selected plants per treatment. After drying, dry flower was then removed or “shucked” from the stem and weighed separately from the main stem and ancillary branches. The flowers were trimmed from the main shoot inflorescence (apical bud), dried in a forced air oven at 38°C (100°F) for 12-24hrs, homogenized into one sample per treatment, and then sent to Phytatech Laboratories (Denver, CO) to be analyzed for terpenes and cannabinoids.

[0081] All statistical analyses were conducted using GraphPad Prism Version 9.1.2 for macOS (GraphPad Software, San Diego, California USA). Normality assumptions were met based on a Shapiro-Wilk test (alpha = 0.05) and observation of a quantile-quantile plot. Yield means were compared using Welch’s ANOVA test for unequal variances followed by Dunnett’s T3 multiple comparisons test.

[0082] As shown in Figures 1 and 2, enhanced flower yield and trends of increased quality were observed in plants treated with Worm Casting Tea and fermented composition (3:1 dilution tea into tap water with 0.6ml/gal fermented alfalfa filtrate added to dilution). Plants treated with Worm Casting Tea produced 19% greater yield compared to control (P = 0.0032) and plants treated with Grow Me (Turbo + worm casting tea) produced 34% greater yield compared to control (P < 0.0001 ) (Figure 1 ). Terpene and cannabinoid content was higher in both the Worm Casting Tea and Grow Me treated plants compared to the control (Figure 2). Plants treated with Worm Casting Tea produced ten different terpenes, while Grow Me treated plants produced eleven terpenes in comparison to seven terpenes produced in control plants (Figure 2, left panel).

Example 2 - Rhizotron Trial for Determining Plant Development.

[0083] Tomato variety HM1823 was analyzed using a randomized block design to test various development characteristics. In these experiments, various amounts of the present filtrate material was combined with worm casting tea and applied to the plants, and compared to a Control of water + grower’s standard fertilizer alone. The addition of the presently disclosed filtrate composition greatly enhanced the analyzed characteristics.

Materials and Methods.

[0084] Tomato seedlings were grown at the University of Florida’s Gulf Coast Research Center (GCREC) in an environmentally controlled greenhouse. At five weeks, the seedlings were transplanted into rhizotrons using soil collected from an onsite agricultural field. The rhizotrons were then inclined at 30° to promote root growth on the bottom side and were covered to keep roots in darkness for the remainder of the two-week trial.

[0085] Greenhouse temperatures remained at ambient and natural lighting was used throughout the trial. Soil used in the rhizotrons was unfumigated and packed to its original bulk density, the field from which it was collected was consistently fertilized at 50lbs/ac N, 100lbs/ac P2O5, 100lbs/ac K₂O, 37lbs/ac Ca, 29lbs/ac. Watering events occurred as needed (growers’ discretion). The experimental design was a randomized complete block design, six blocks with six plants per block (one plant per treatment per block including a control).

[0086] A single treatment event occurred at the time of transplant where the treatments were applied manually (1 OOml/plant) to the surface of the substrate. Treatments consisted of a control (tap water), Filtrate at various application rates, and worm casting tea with various rates of Filtrate added into the solution.

Data Collection.

[0087] All data were collected at the end of the 14-day trial. Stem width (mm) data were collected using calipers at the base of the plant 1 cm above the top of the rhizotron. Leaf area (cm2/plant) and canopy projected area (cm2/plant) were measured using Imaged software. For canopy area, a single overhead image was taken approximately 45cm from the top of each intact plants’ canopy using a compact digital camera (SONY DSC-RX100). For leaf area, photos of freshly harvested leaves were taken individually. Photos were then uploaded into the Imaged software where pixels were converted from RGB to binary, which were then calculated into area measurements. Leaf dry weight (g), shoot dry weight (g), and root dry weight (g) were collected using destructive sampling methods. Roots were manually separated from the substrate and then gently washed by hand using tap water. Roots were destructively spread to minimize root overlapping and then imaged using a high-resolution scanner (EPSON V850) and the software WinRhizo to calculate root length (cm) and root surface area (cm²). The fresh roots, shoots, and leaves were then dried separately in a forced air oven at 65°C for 48h until constant weight was reached, and then weighed for individual and total dry mass (g) metrics.

Statistical Analysis.

[0088] All statistical analyses were conducted using GraphPad Prism (Version 9.3.1 ; GraphPad Software, San Diego, California USA). Outliers were detected and removed using Grubb’s (a = 0.05) from the root length WCT + Filtrate dataset and the shoot dry mass and total mass Filtrate datasets. Normality and variance assumptions were met, and means were compared using unpaired Student’s t tests. Results.

[0089] Results are presented in Figure 3, Panels a-e, and Figure 4, Panels a-c, and summarized, briefly, below. Briefly, where worm castings tea was supplemented with 0.6 mL/gal Filtrate, the plant traits are significantly higher than the control plants. Specifically, leaf area was enhanced (with a p-value vs. Control of 0.04), leaf dry mass was enhanced (p-value vs. Control of 0.06), shoot dry mass was enhanced (p-value of 0.07), total mass was enhanced (p-value of 0.06), and stem diameter was enhanced (p-value of 0.04). Worm castings tea supplemented with 5 mL/gal Filtrate also resulted in significantly higher measurements for several traits versus Control: for example canopy projected area (p- value of 0.06), root length (p-value of 0.01 ), and root surface area (p-value of 0.02). Finally, the root dry mass characteristic was significantly higher in the worm castings tea supplemented with either 0.6 mL/gal Filtrate (p value of 0.05) or 25 mL/gal Filtrate (p value of 0.05).

[0090] WCT+0.6 mL Filtrate/qal application rate: canopy projected area increased by 23.6%, root projected area increased by 29.9%, plant height increased by 3.2%, stem diameter increased by 8.5%, leaf area increased by 19.9%, leaf dry weight increased by 17.6%, stem dry weight increased by 9.3%, shoot dry weight increased by 15.7%, root dry weight increased by 18.2%, root to shoot ratio increased by 3%, root length increased by 24.5%, root surface area increased by 30.3%, and root tip number increased by 14.4%. (Figs. 4 and 5).

[0091] WCT+5mL Filtrate/qal application rate: canopy projected area increased by 38.0%, root projected area increased by 20.3%, plant height increased by 3.5%, stem diameter increased by 4.7%, leaf area increased by 14.5%, leaf dry weight increased by 11 .6%, stem dry weight increased by 10.8%, shoot dry weight increased by 11 .4%, root dry weight increased by 14.3%, root to shoot ratio increased by 2.9%, root length increased by 50.6%, root surface area increased by 59.9%, and root tip number increased by 20.7%. (Figs. 4 and 5).

[0092] WCT+25mL Filtrate/qal application rate: canopy projected area increased by 29.5%, root projected area increased by 15.9%, plant height increased by 5.8%, stem diameter increased by 7%, leaf area increased by 14.1%, leaf dry weight increased by 13.0%, stem dry weight increased by 10.7%, shoot dry weight increased by 12.5%, root dry weight increased by 17.9%, root to shoot ratio increased by 6%, root length increased by 31 .9%, root surface area increased by 43.4%, and root tip number increased by 5.5%. (Figs. 4 and 5). Example 3 - Testing of Fermented Filtrate and Worm castings Plus Fermented Filtrate on Industrial Hemp.

[0093] The ability of the presently disclosed compositions to affect growth of industrial hemp was also evaluated.

Materials And Methods.

[0094] The hemp (cultivar ‘Lifter’) was grown to evaluate the impact of worm castings tea with 0.6ml/gal of added Filtrate (WCT + 0.6 ml/gal Filtrate), and Filtrate at 0.6 mUgal as a drench and as a foliar application. Plants were grown from feminized seed of variety Lifter, started mid-May, and transplanted into prepared field in June. The experimental design was a randomized complete block with four replicates.

[0095] The WCT +0.6 ml/gal Filtrate treatment was applied once weekly via soil drench at 37.9ml/gal throughout the growing season. The Filtrate drench treatment was applied as a soil drench weekly at 0.6 mL/gal and the Filtrate foliar treatment was applied as a foliar spray weekly at 0.6 mL/gal. Control plots received baseline fertilizer applications that were applied to all plots in addition to unadulterated water at same rates to applied treatments. Plots consisted of five plants spaced 5’ apart in the row and between rows. Irrigation was applied on a weekly basis with adjustments made based on weekly rainfall amounts.

[0096] In early June, all plots were fertilized with 10Olbs N ac-1 , 60lbs P ac-1 , 80lbs K ac- 1 , using Pro-Gro (5-3-4) (North Country Organics; Bradford, VT). The fertility amendments were based on soil test results (University of Vermont Agricultural and Environmental Testing Laboratory, Burlington, VT). All soil fertilizer applications were products approved for use in certified organic systems.

[0097] Plants were monitored for flowering date and all plants within the trial had initiated flower formation in early August.

Data Collection.

[0098] Total plant weight, as well as dry flower weight measurements were taken. At the end of September, aggregate flower samples were taken, post-harvest, and sent to Bia Diagnostics (Colchester, VT) to be analyzed for terpenes and cannabinoids.

Statistical Analysis.

[0099] All statistical analyses were conducted using GraphPad Prism (Version 9.3.1 ; GraphPad Software, San Diego, California). Normality and variance assumptions were met, and means were compared using unpaired Student’s t tests to determine treatment effect. Results.

[0100] Seasonal precipitation and temperature were recorded with a Davis Instrument Vantage Pro2 weather station, equipped with a WeatherLink data logger at Borderview Research Farm in Alburgh, VT. The growing season was be characterized as hot and dry. Temperatures during the growing period were 5.97 degrees higher than the 30-year average. The month of the July was the exception with much of the month experiencing unseasonably cool but still dry conditions.

[0101] The results of this study are presented at FIGS. 9 and 10. Harvest quality was higher in plants treated with WCT + Filtrate compared to control plants. Total CBD% was 11 .75% (SD = 1.12) in WCT + Filtrate treated plants, while control plants had an average of 9.90% CBD (SD = 1.14). Though not statistically significant, total plant weight and flower weight showed positive trends in all treatments.

[0102] From this one year of study in receiving these distinct treatments, it appeared as if there was some noticeable impact on the chemical profiles of the Lifter cultivar in addition to the proportions of plant parts (flower, leaf, stem). However, no noticeable impacts were recorded on yields. Additional years and growing conditions could be further indicative of the impact of these products while applied throughout the growing season and could help to evaluate and document the potential benefits of these various soil amendment and foliar spray products.

Example 4 - Effect of Turbo on Tomato Plants.

[0103] A greenhouse in San Luis Obispo, CA was used for this study. Tomatoes were grown in artificial media following a standard commercial system. Each treatment was randomly distributed with 5 replicates and 10 plants per plot, for a total of 50 plants per treatment. All plants were treated with the grower’s standard fertilizer and all treatments occurred as a soil drench weekly throughout the growing cycle, beginning at transplant.

[0104] Filtrate 5 mL/gal application rate: Average plant vigor increased by 3.13%, average BRIX increased by 0.71 %, average fruit width increased by 0.33%, average fruit height increased by 1 .05%, average fruit color increased by 1 .89%, total marketable fruit count increased by 3.21 %, total marketable weight decreased by 0.87%, total unmarketable fruit count decreased by 8.33%, total unmarketable fruit weight decreased by 31 .52%, total number of plants harvested decreased by 4.21%, and total marketable weight per plant increased by 11 .49%

[0105] Filtrate 25 mL/gal application rate: Average plant vigor increased by 4.51 %, average BRIX did not change, average fruit width decreased by 0.01 %, average fruit height decreased by 0.76%, average fruit color decreased by 0.94%, total marketable fruit count increased by 7.22%, total marketable weight increased by 8.83%, total unmarketable fruit count increased by 25%, total unmarketable fruit weight increased by 34.69%, total number of plants harvested increased by 4.21%, and total marketable weight per plant increased by 1 1 .93%

[0106] Filtrate 50 mL/gal application rate: Average plant vigor increased by 2.78%, average BRIX increased by 4.29%, average fruit width increased by 12.09%, average fruit height decreased by 1 .94%, average fruit color decreased by 2.83%, total marketable fruit count decreased by 3.44%, total marketable weight decreased by 4.37%, total unmarketable fruit count decreased by 8.33%, total unmarketable fruit weight increased by 14.29%, total number of plants harvested decreased by 6.31 %, and total marketable weight per plant increased by 12.06%

[0107] Although the number of replicates were small and the data was not statistically significant, this data shows that certain application rates of Filtrate can decrease unmarketable fruit count and weight, and increase total marketable weight, as well as the total marketable weight per plant. All application rates also show no negative impacts in fruit quality.

Example 5 - Effect of Turbo on Cucumber.

Materials and Methods.

[0108] Cucumber seeds were planted in indoor soil mix with no charge in standard 1801 trays (18 wells/tray). Each treatment was applied to one entire tray twice weekly starting 10 days after planting for a total of 4 applications by both soil drench and foliar application. No additional nutrients were applied. Plants were harvested about one month after planting, then dried and weighed.

[0109] The treatments were a control that was treated with either water (control), Nature’s Source Micronutrients, or Turbo at 5% plus Nature’s Source Micronutrients.

[0110] Although no statistical analysis was completed, dry mass was increased for Turbo +micronutrient-treated plants compared to plants where micronutrients were applied alone and as compared to the water control.

Example 6. Mitigation of Salt Stress.

[0111] In this study, the effect of Turbo alone and Turbo in combination with various fertilizers, on sodium (Na+) concentration was evaluated in cucumber (Cucumis sativus) plants in a grow room in 50 cell flats with a temperature range of 23-25°C during the day and approximately 18°C at night with a photo period of approximately 18 hours on and 6 hours off. The soil was Lamberts LM-HP. Two flats (100 plants) were assigned per treatment. Industry standard water soluble 20-10-20 fertilizer (Jacks Peat Lite) was used at an EC of 0.79 (1.81 g/gal) applied at 10 mL/cell once per day. On day 7 and day 14 fertilizer was applied at an EC of 1.3 (3.62g/gal). Turbo treatments were applied weekly at an assigned application rate for a total of 3 applications. Cucumbers were harvested on day 21 . Each plant was then cut at the base, weighed and recorded as fresh mass. The aboveground fresh mass samples were sent to Midwest labs for drying and tissue nutrient testing.

[0112] The Materials evaluated alone and in combination with Turbo included: Grow A and Grow B (see below); 3-1 -1 (Natures Source); 3-2-1 (Natures Source); 10-4-5 (Natures Source); UAN 32 (Yara); 10-34-0 (Two Rivers Terminal); Micronutrients (Natures Source); Humic Acid (Live Earth Products); Kelp (Afrikelp); Mammoth Silica; Worm Casting Tea (Denali Biosolutions); and 8-4-4 (Natures Source).

[0113] The contents of Grow A and Grow B, are, Grow A: Total Nitrogen, 4%; Nitrate Nitrogen, 3.93%; Ammoniacal Nitrogen, 0.07%; Soluble Potash, 1.00%; Calcium, 4.20%; Magnesium, 0.19%; Boron, 0.01%; Iron, 0.06%; Manganese, 0.013%; Molybdenum, 0.0007%; Zinc, 0.0045%. Grow B: Total Nitrogen, 1.07%; Nitrate Nitrogen, 0.20%; Ammoniacal Nitrogen, 0.87%; Soluble Potash, 5.00%; Phosphate, 3.00%; Magnesium, 0.89%; Sulfur, 1.30%; Copper, 0.005%.

[0114] The results shown in Figure 6 illustrate a decrease in the concentration of sodium in plants treated with Turbo as indicated by a decrease in the concentration of sodium per gram of dried plant tissue (“percent rate”) in cucumber when Turbo was included in synthetic or organic fertilizer amendments as compared to plants treated with fertilizer alone. The results suggest that Turbo can protect plants from salt stress and that the effect is greater for “high salt” fertilizers.

Example 7. Evaluation of Amino Acid Content in Turbo as Compared to Alfalfa Feedstock.

[0115] In this study, the amino acid content of 2 lots of Turbo (1584 and 1738) prepared by a 7 day fermentation of Citrobacter freundii, Enterobacter cloacae, Comamonas testosteroni, and Pseudomonas putida in a 180 gallon fermenter containing filtered alfalfa medium. On the 7th day 1 % v/v of citric acid was added to the fermenter and mixed for 10 minutes. After mixing, the fermented culture was passed through a depth filter, a 0.45 micron filter, then a 0.22 micron filter was compared to the amino acid content of the alfalfa feedstock used to make the Turbo. Microbial filtrate and alfalfa feedstock samples were submitted to Creative Proteomics for free amino acids according to the following protocol. [0116] 1 OOpL of each sample was extracted for amino acids using a methanol: chloroform extraction. The sample was run using a LC-MS/MS method for amino acids detection and quantification. An external standard curve was run using known concentrations of Ala, Arg, Asn, Asp, Gin, Glu, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, and Vai. This method does not detect Gly and Cys. The standard curves were used to calculate the concentration of all amino acids in the samples from the peak area detected. The results shown in Figure 7 (ng/mL) show an enrichment of nearly all amino acids when in Turbo as compared to alfalfa feedstock.

Example 8. Treatment of Cucumber plants with of Turbo with and without Synthetic Fertilizer.

[0117] Previous studies were carried out using cucumber and tomato plants. The effect of Turbo prepared by a 7 day fermentation of Pseudomonas putida, Enterobacter freundii, Comamonas testosteroni and Citrobacter cloacae using an alfalfa feedstock, followed by centrifugation n for 10 minutes at 10,00RPM and filtration though a 0.2p filter, or autoclaving at 121 °C, at about 15 psig for about 1 hour. The studies included treatments with Turbo alone versus addition of 1 % citric acid or 1 M citrate and 1 M betaine, fertilizer (20-10-20), and a lower Turbo application rate was evaluated. The treatments were applied weekly at a drench rate of 0.6 ml/gal. For each treatment, there were 18 replicates (1 tray/18 x 3” wells). In these studies, fresh weights were not measured and data was reported as “% rate” for nutrients based on dry plant material on a tray-by-tray basis, not individual plants. This resulted in few replicates and the evaluation of dry plant material was determined not to reflect nutrient uptake because it fails to take into account that healthier plants are often larger. In light of this observation, it was determined that fresh weight (mass) of plant material would be more accurate reflection of plant growth and health.

[0118] In this study, the effect of eleven treatments on the fresh mass of Cucumber plants after 4 weeks of growth was evaluated. The plants grown in a grow room with a temperature range of 23-25°C during the day and approximately 18°C at night with a photo period of approximately 18 hours on and 6 hours off. The soil was Lamberts LM-HP. The treatments are described in Table 1 , below.

[0119] Turbo was either Grow-20l0-B1 or Grow-20AE0-B1 prepared by either, (a) culture of Comamonas testosteroni and Pseudomonas putida for 7 days in a 1X wheat bran extract with shaking at 150 RPM and 25°C, followed by addition of 1 mol/L (1 M) Betaine and 1 M Citrate and filtration though a 10p filter (Grow-20AE0-A1 ). [0120] Turbo was applied every 4 days over the 4 week period of the experiment at a treatment rate of 2%. 50 cell trays were employed in this study so each treatment had 200 plants associated with it. The treatments were provided until “run through” such that the soil was fully wetted after each application.

[0121] Turbo+Betaine+Citrate for 10-34-0 at 2.8mL/gal (2ml_ of turbo to 100 mL of stock fertilizer solution).

[0122] Turbo+Betaine+Citrate for 10-4-5 at 3.31 mL/gal (2mL of Turbo to 100 mL of stock fertilizer solution).

[0123] 10-34-0 includes: 10% ammoniacal nitrogen, available phosphate: 34.00%. (Two Rivers Terminal).

[0124] 10-4-5 includes: (total nitrogen: 10%: 2.10% ammoniacal nitrogen, 1.80% nitrate nitrogen, 5.15% urea nitrogen, 0.75% other water soluble nitrogen, 0.20% water insoluble nitrogen);available phosphate: 4%; soluble potash: 5%; calcium: 0.01 %; magnesium: 0.05% (0.05% water soluble magnesium); sulfur: 0.2% (0.2% combined sulfur); boron: 0.002%; copper: 0.001% (0.001% water soluble copper); iron: 0.002% (0.002% water soluble iron); manganese: 0.014% (0.014% water soluble manganese); molybdenum: 0.0001% (0.0001% water soluble molybdenum); zinc: 0.005% (water soluble zinc). (Nature’s Source).

[0125] Table 1. Treatments.

[0126] Table 2. Fresh Mass (g).

[0127] Conclusions. The only observed significant differences between treatment groups are that treatment with betaine and citrate had no effect and all the other treatment groups resulted in higher fresh weights than water or water plus betaine and citrate. While not wishing to be bound by theory, humic acid (which is a rich source of organic matter) was used as a carrier for nutrients. The results suggest that it is not a good carrier to test the effect of Turbo on plants.

Example 9. Effect of Turbo Plus and Minus Fertilizer (10-4-5) on Tomato.

[0128] In this study, the effect of four treatments on the fresh mass of tomato plants after 3 weeks of growth was evaluated. Tomatoes (n=100 per treatment) were grown for three weeks and underwent two applications of treatments at fixed application rates and fertilization rates. The plants grown in a grow room with a plants in a grow room with a temperature range of 23-25°C during the day and approximately 18°C at night with a photo period of approximately 18 hours on and 6 hours off. The soil was Lamberts LM-HP. After three weeks of growth, all plants were harvested and weighed and the fresh mass (grams; g) was evaluated. The treatments provided until “run through” such that the soil is fully wetted after each application, e.g., in an amount of approximately 150-200 ml/well, are described in Table 3, below.

[0129] Table 3. Treatments.

[0130] Grow-20G0-A0 Turbo was prepared by culturing Citrobacter freundii, Enterobacter cloacae, Comamonas testosteroni, and Pseudomonas putida for 7 days in a 180 gallon fermenter containing filtered alfalfa feedstock. On the 7th day 1 % v/v of citric acid is added to the fermenter and mixed for 10 minutes. After mixing, the fermented culture was passed through a depth filter, a 0.45 micron filter, then a 0.22 micron filter

[0131] Grow-20l0-B1 Turbo was prepared by culturing Comamonas testosteroni and Pseudomonas putida in the lab for 7 days in 1 x wheat bran extract with shaking at 150 RPM and 25°C. On the 7th day of fermentation, 1% v/v citric acid was added. The culture was then centrifuged for 10 minutes at 10,00RPM, and the supernatant was passed through a 0.45 pm filter then a 0.22 pm filter.

[0132] Grow-20l0-A1 Turbo was prepared by culturing Comamonas testosteroni and Pseudomonas putida in the lab for 7 days in 1 x alfalfa feedstock with shaking at 150 RPM and 25°C. On the 7th day of fermentation, 1 % v/v citric acid was added. The culture was then centrifuged for 10 minutes at 10,00RPM, and the supernatant was passed through a 0.45 pm filter then a 0.22 pm filter.

[0133] The results are provided in Figure 8 and the results of statistical analysis comparing treatment groups is provided in Table 4, below.

[0134] Table 4. Pairwise Comparisons of Fresh Mass (g) Between Treatment Groups.

[0135] Adjusted p-values include a Bonferroni correction which is a multiple-comparison correction used when several dependent or independent statistical tests are being performed simultaneously.

[0136] In summary, treatment of tomato plants with 10-4-5+Grow-20l0-B1 (Turbo) resulted in plants with fresh weights that were was 22% greater than the control, and among the treatments this was the effect that was the most significantly different from the control (p«0.05).

Example 10. Effect of Bacterial Cultures or Turbo Plus and Minus Fertilizer (20-10-20) on Tomato.

[0137] In this study, the effect of eight treatments on the fresh mass of tomato plants after 4 weeks of growth was evaluated. Tomatoes (n= 100 per treatment) were grown for four weeks and underwent weekly applications of treatments at fixed application rates and fertilization rates that mimic continuous liquid feed. The plants grown in a grow room with a plants in a grow room with a temperature range of 23-25°C during the day and approximately 18°C at night with a photo period of approximately 18 hours on and 6 hours off. The soil was Lamberts LM-HP. After three weeks of growth, all plants were harvested and weighed and the fresh mass (grams; g) was evaluated. The treatments are described in Table 5, below.

[0138] Table 5. Treatments.

[0139] Grow-20l0-A0 Turbo was prepared by culturing Citrobacter freundii, Enterobacter cloacae, Comamonas testosteroni, and Pseudomonas putida for 7 days in 1x alfalfa extract with shaking at 150 RPM and 25°C. On the 7th day of fermentation, 1 % v/v citric acid was added. The culture was then centrifuged for 10 minutes at 10,00RPM, and the supernatant was passed through a through a 0.45 pm filter then a 0.22 pm filter.

[0140] Grow-21 C3-B9 was prepared by culturing Comamonas testosteroni, Pseudomonas putida, Pseudomonas fluorescens, Rhizobium tibeticum, and Ensifer adherens in the lab for 7 days in 1 x wheat bran extract with shaking at 150 RPM and 25°C.

[0141] Grow-20C0-B1 was prepared by culturing Comamonas testosteroni and Pseudomonas putida in the lab. An AHB was set up containing 1x wheat bran extract and the bacteria were added to the AHB at a target of 10⁵-10⁷ CFU/ml. The culture was mixed in the AHB for approximately 10 minutes then bottled. The entire process happens on the same day, so no fermentation time.

[0142] Grow-20l0-B1 Turbo was prepared by culturing Comamonas testosteroni and Pseudomonas putida in the lab for 7 days in 1 x wheat bran extract with shaking at 150 RPM and 25°C. On the 7th day of fermentation, 1% v/v citric acid was added. The culture was then centrifuged for 10 minutes at 10,00RPM, and the supernatant was passed through a 0.22 pm filter.

[0143] Grow-20B0-B1 was prepared by culturing Comamonas and Pseudomonas putida in the lab. The consortium is grown for 7 days in 1 x wheat bran extract with shaking at 150 RPM and 25°C.

[0144] 20-10-20, which includes: (total nitrogen: 20%, 8.00% ammoniacal nitrogen, 12.00% nitrate nitrogen); available phosphate: 10%; soluble potash: 20%; magnesium: 0.1500% (0.1500% water soluble magnesium); boron: 0.0200%; copper: 0.0100% (0.0100% chelated copper); iron: 0.1000% (0.1000% chelated iron); manganese: 0.0500% (0.0500% chelated manganese); molybdenum: 0.0100%; zinc: 0.0500% (0.0500% chelated zinc). (Jack’s Peat Lite). [0145] The results are provided in Figure 9 and the results of statistical analysis comparing treatment groups is provided in Tables 6 and 7, below.

[0146] Table 6. Pairwise Comparisons of Fresh Mass (g) Between Treatment Groups After 4 Weeks of Growth.

[0147] There was a significant difference in fresh weight (g) versus controls for all Turbo embodiments except Grow-20C0-B1 .

[0148] Table 7. Fresh Weights of Plants treated with Turbo (Percent Difference Relative to Controls).

[0149]

[0150] Example 1 1 . Seed-Bag Bioassay - Root Growth From Seeds Germinated in Media Containing Turbo With And Without Materials.

[0151] In this study, a seed bag bioassay was used to evaluate the effect of fermented filtrates on plant growth at the germination stage (from germination to initial root-growth). The “seed bag or “seed pouch” is a plastic sleeve into which a durable papery sheet is inserted and small hooks are attached to hang along suspension wires. Figure 10 depicts the size, shape, and growth of plants pre-germinated and inserted into the perforated upper fold of a“dispo” bag. See, Preiser FA, et al., J Nematol. 1981 Oct;13(4):535-7. Seed bags were hung along suspension wires in a high-humidity environment with controlled light on a grow-room’s germination shelves. The results are presented as: “Reach” (R), which is the deepest distance the roots reached, and “Spread” (S) is the widest distance the roots spread out in each bag. The growth area (A) was calculated by multiplying R by S, and the root area is calculated by (RxS)/2.

[0152] Up to 5 seeds were placed in the upper folds of each seed bag in a manner that they contacted perforations. Each bag was wetted with 16 mL of water or liquid product, and 200 mL of water was added to the bottom of the container, then the container lid was closed, where the holes used to create the suspensions allow a small amount of gas exchange. Containers were maintained at approximately 72°F for up to 10 days. If drying was observed 2 mL of water alone or water including treatments was added to all bags. Growth was monitored and the root growth profile was periodically photographed.

[0153] Preparation of MPCDN Turbo is described in Example 8.

[0154] Feedstocks used in this study were: WBE (wheat bran extract) and AAE (alfalfa extract). Treatments are described in Table 8, below.

[0155] Table 8. Treatments.

[0156] After conservative p-value correction (Bonferroni) a significant difference was observed (all increasing in size) between Water Control and the following treatments: 10- 4-5 Control, 3 Day AAE MPCDN, 3 Day WBE MPCDN, 7 Day AAE MPCDN, 7 Day WBE MPCDN, Humic Acid+WBE, and Humic Acid+WBE 3 Day MPCDN. Reach was the dominant factor more so than spread in the determination of root area, and roots often were hindered in the lateral growth by the edges of the bag itself. See, Figures 11 a-1 1 c.

[0157] While multiple embodiments are disclosed, still other embodiments of the present invention will become apparent to those skilled in the art from the following detailed description. As will be apparent, the invention is capable of modifications in various obvious aspects, all without departing from the spirit and scope of the present invention. Accordingly, the detailed description is to be regarded as illustrative in nature and not restrictive.

[0158] All references disclosed herein, whether patent or non-patent, are hereby incorporated by reference as if each was included at its citation, in its entirety. In case of conflict between reference and specification, the present specification, including definitions, will control.

[0159] Although the present disclosure has been described with a certain degree of particularity, it is understood the disclosure has been made by way of example, and changes in detail or structure may be made without departing from the spirit of the disclosure as defined in the appended claims.

Claims

We claim: . A composition comprising: a fermented filtrate from a bacterial culture comprising at least one gram negative (-) bacterial species and a feedstock, wherein the fermented filtrate is substantially sterile and free of microbes. . The composition of claim 1 , wherein the gram negative (-) bacterial species are selected from Citrobacter sp., Comamonas sp., Enterobacter sp., Ensifer sp., Pseudomonas sp., and Rhizobium sp. . The composition of claim 2, wherein the gram (-) bacterial species are two or more of Citrobacter freundii, Enterobacter cloacae, Comamonas testosteroni, Pseudomonas putida, Pseudomonas fluorescens, Rhizobium tibeticum, and Ensifer adherens. . The composition of claim 2, further comprising one or more supplements selected from citric acid, betaine, and arginine. . The composition of claim 2 or 4, further comprising one or more materials selected from the group consisting of a fertilizer, worm casting tea, rooting hormone, growth hormone, triglycerides, fatty acids, lipids, carbohydrates, simple sugars, amino acids, proteins, yeast, organic materials, fulvic acids, humates, and a vitamin.. The composition of claim 5, wherein the material is a fertilizer. . The composition of claim 6, wherein the fertilizer is 10-34-0, 10-4-5 or 20-10-20.. The composition of claim 5, wherein one material is worm casting tea with a pH of from about 3.0 to 9.0. . The composition of claim 4, wherein the citric acid in the fermented filtrate has a concentration of from 0.5% to 2.0% or from 0.5M to 1 .5M. 0. The composition of claim 9, further comprising betaine at a concentration of from 0.5M to 1.5M. 1. The composition of claim 10, further comprising arginine at a concentration of about from 0.5M to 1 .5M. 2. The composition of claim 5, wherein the feedstock is a plant-based extract derived from alfalfa, soybean hulls, wheat bran, beet pulp, spent hops, rice bran, grape skin, grass clippings or oat bran. The composition of claim 5, wherein the composition is osmolytic and is effective to decrease the concentration of sodium in plants treated with the composition. The composition of claim 5, wherein the composition is effective to increase fresh weight in plants treated with the composition relative to untreated plants. The composition of claim 5, wherein the composition is effective to increase the growth of roots in plants treated with the composition relative to untreated plants. The composition of claim 5 prepared using alfalfa feedstock, wherein the composition comprises a higher concentration of amino acids than the alfalfa feedstock. A method of manufacturing a sterile fermented filtrate from a gram negative (-) bacterial culture, comprising: a. inoculating a feedstock with at least one gram (-) bacterial species to produce a fermented culture; b. culturing the bacterial species for 3 or more days to generate a fermented filtrate; c. filtering the fermented culture through at least one of a 0.22 pm filter, a 0.45 pm filter and a 10 pm filter to produce a fermented filtrate; d. adding a supplement to the fermented filtrate and storing it in a sterile container. The method of claim 17, wherein the gram (-) bacteria are selected from the group consisting of Citrobacter sp., Comamonas sp., Enterobacter sp., Ensifer sp., Pseudomonas sp., and Rhizobium sp. The method of claim 18, wherein the gram (-) bacterial species are two or more of Citrobacter freundii, Enterobacter cloacae, Comamonas testosteroni, Pseudomonas putida, Pseudomonas fluorescens, Rhizobium tibeticum, and Ensifer adherens. The method of claim 17, wherein the supplement is one or more of citric acid at a concentration of from 0.5% to 2.0% or from 0.5M to 1.5M, betaine at a concentration of from 0.5M to 1 .5M, and arginine at a concentration of from 0.5M to 1.5M. The method of claim 17, wherein the feedstock is a plant-based extract derived from alfalfa, soybean hulls, wheat bran, beet pulp, spent hops, rice bran, grape skin, grass clippings or oat bran. A method of treating plants with a fermented filtrate composition according to claim 4, using continuous-liquid-feed (”CLF”) or soil application. The method according to claim 23, wherein the fermented filtrate composition comprises a fertilizer and plants treated with the fermented filtrate composition have a lower salt (sodium) concentration than plants treated with fertilizer alone. The method according to claim 23, wherein the fermented filtrate composition is effective to increase fresh weight in plants treated with the composition relative to untreated plants or plants treated with fertilizer alone. The composition of claim 5, wherein the fermented filtrate composition is effective to increase the growth of roots in plants treated with the composition relative to untreated plants.