Endophyte combinations for improving plant and algae productivity and stress tolerance

Abstract

Endophyte combinations, compositions, and methods of using the endophyte combinations and compositions for improving plant and algae productivity and tolerance to environmental stresses.

Description

Endophyte Combinations for Improving Plant and Algae Productivity and Stress ToleranceAttorney Docket No.: S167659.001Endophyte Combinations for Improving Plant and Algae Productivity and Stress ToleranceCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 578,731, filed August 25, 2023, and U.S. Provisional Application No. 63 / 578,738, filed August 25, 2023, each of which is incorporated herein by reference.ACKNOWLEDGMENT OF GOVERNMENT SUPPORT

[0002] This invention was made with government support under 89233218CNA000001 awarded by the National Nuclear Security Administration. The government has certain rights in the invention.BACKGROUND

[0003] Global climate change is affecting agriculture worldwide owing to an increase in surface temperature that will reduce soil moisture. In light of this and declining natural resources, there is a pressing need to find solutions to sustain our global agriculture. However, in many situations, boosting agricultural productivity relies heavily on chemical fertilizers, pesticides, and herbicides, which cause adverse environmental impacts and affect soil fertility over time. Another adverse effect of climate change that is becoming a significant challenge is the increased salinity of agricultural soil. Salinity stress is one of the most devastating stresses that causes a major reduction in the land area suitable for agriculture.SU ARY

[0004] Described are synthetic combinations of endophytes comprising: at least one first endophyte that reduces ethylene concentration in a plant; at least one second endophyte that increases growth-promoting hormones levels in the plant; and at least one third endophyte that fixes nitrogen.

[0005] Also described are synthetic combinations of endophytes comprising: at least two different endophytes that reduce ethylene concentration in a plant, at least two different endophytes that increases growth-promoting hormones levels in the plant; and / or at least two different endophytes that fix nitrogen.

[0006] In some embodiments, the at least one first endophyte comprises a Bacillus species or a Pseudomonas species; the at least one second endophyte comprises aRhizobium species, a Pseudomonas species, or an Azotobacter species; and the at least one third endophytecomprises a Rhizobium species, a Mesorhizobium species, an Arthrobacter species, an Azospirillum species, an Azotobacter species, a Sinorhizobium species, or a Bradyrhizobium species.

[0007] In some embodiments, the at least one first endophyte comprises Bacillus licheniformis, Bacillus megaierium. Bacillus subtilis, Pseudomonas putida, or Pseudomonas fluorescens,' the at least one second endophyte comprises a Rhizobium leguminosarum, Rhizobium phaseoli. Pseudomonas putida, Pseudomonas fluorescens, or Azotobacter vinelandii'. and the at least one third endophyte comprises Rhizobium leguminosarum, Rhizobium phaseoli. Mesorhizobium amor phae. Arthrobacter oryzae, Azospirillum brasilense, Azotobacter chroococcum. Azotobacter vinelandii, or Sinorhizobium meliloti.

[0008] In some embodiments, the at least one first endophyte comprises Bacillus licheniformis Bacillus megaterium, or Bacillus subtilis,' the at least one second endophyte comprises a Pseudomonas putida or Pseudomonas fluorescens,' and the at least one third endophyte comprises Rhizobium leguminosarum, Rhizobium phaseoli, Mesorhizobium amorphae, Arthrobacter oryzae, Azospirillum brasilense. Azotobacter chroococcum, Azotobacter vinelandii, Sinorhizobium meliloti, or Bacillus subtilis.

[0009] Also described are synthetic combinations of endophytes comprising: at least one first endophyte that reduces ethylene concentration in a plant; and at least one second endophyte that increases growth-promoting hormones levels in the plant.

[0010] Also described are synthetic combinations of endophytes comprising: at least one first endophyte that reduces ethylene concentration in a plant; and at least one second endophyte that fixes nitrogen.

[0011] Also described are synthetic combinations of endophytes comprising: at least one first endophyte that increases growth-promoting hormones levels in the plant; and at least one second endophyte that fixes nitrogen.

[0012] In some embodiments, the at least one first endophyte that reduces ethylene concentration in a plant comprises aBacillus species or a Pseudomonas species; and the at least one second endophyte that increases growth-promoting hormones levels in the plant comprises a Rhizobium species, a Pseudomonas species, or an Azotobacter species.

[0013] In some embodiments, the at least one first endophyte that reduces ethylene concentration in a plant comprises aBacillus species or a Pseudomonas species; and the at least one second endophyte that fixes nitrogen comprises a Rhizobium species, a Mesorhizobium species, an Arthrobacter species, an Azospirillum species, an Azotobacter species, a Sinorhizobium species, or a Bradyrhizobium species.

[0014] In some embodiments, the at least one first endophyte that increases growthpromoting hormones levels in the plant comprises a Rhizobium species, a Pseudomonas species, or an Azotobacter species: and the at least one second endophyte that fixes nitrogen comprises a Rhizobium species, a Mesorhizobium species, an Arthrobacter species, an Azospirillum species, an Azotobacter species, a Sinorhizobium species, or a Bradyrhizobium species.

[0015] In some embodiments, the synthetic combination of endophytes comprises at least one endophyte selected from the group consisting of: Arthrobacter oryzae, Azospirillum brasilense, Azotobacter chroococcum. Azotobacter vinelandii, Bacillus licheniformis, Bacillus megaterium, Bacillus subtilis, Mesorhizobium amorphae. Pseudomonas putida, Pseudomonas lluorescens. Rhizobium leguminosarum. Rhizobium phaseoli, and Sinorhizobium meliloti.

[0016] In some embodiments, the synthetic combination of endophytes comprises at least 2, 3, 4, 5, 6, 7, 8, or more endophytes selected from the group consisting of: Arthrobacter oryzae. Azospirillum brasilense, Azotobacter chroococcum, Azotobacter vinelandii. Bacillus licheniformis, Bacillus megaterium, or Bacillus subtilis, Mesorhizobium amorphae, Pseudomonas putida. Pseudomonas fluorescens. Rhizobium leguminosarum, Rhizobium phaseoli, and Sinorhizobium meliloti.

[0017] In some embodiments, the synthetic combination of endophytes comprises 3, 4, 5,6, 7, 8, or more endophytes selected from the group consisting of: Arthrobacter oryzae, Azospirillum brasilense, Azotobacter chroococcum, Azotobacter vinelandii, Bacillus licheniformis. Bacillus megaterium, or Bacillus subtilis, Mesorhizobium amorphae, Pseudomonas putida, Pseudomonas fluorescens, Rhizobium leguminosarum, Rhizobium phaseoli, and Sinorhizobium meliloti.

[0018] In some embodiments, the synthetic combination of endophytes comprises 4. 5, 6,7, 8, or more endophytes selected from the group consisting of: Arthrobacter oryzae. Azospirillum brasilense, Azotobacter chroococcum, Azotobacter vinelandii, Bacillus licheniformis, Bacillus megaterium, or Bacillus subtilis, Mesorhizobium amorphae, Pseudomonas putida, Pseudomonas fluorescens, Rhizobium leguminosarum, Rhizobium phaseoli. and Sinorhizobium meliloti.

[0019] In some embodiments, any of the described synthetic combination of endophytes can be formulated with one or more excipients, one or more agrochemically active compounds, one or more biocontrol agents, or combinations thereof. The synthetic combinations of endophytes can be formulated as a liquid, a dispersion, a colloidal dispersion, a solution, a suspension, a colloidal suspension, an emulsion, a foam, a slurry, a lyophilized cake or powder,a spray-dried powder, a pellet, a biologically pure pellet, a coated pellet, a powder, a flowable powder, or a granule suitable for applying to a seed, plant, plant part, or environment of the seed, plant or plant part.

[0020] In some embodiments, the synthetic combination of endophytes is applied to a seed in an amount effective to colonize a plant germinated from the seed with the synthetic combination of endophytes; applied to a seedling, a plant, or a leaf of the plant in an amount effective to colonize the seedling or the plant with the synthetic combination of endophytes; or applied to an environment of a seed, plant or plant part in an amount effective to colonize a plant grown in the environment with the synthetic combination of endophytes. The plant can be a monocot plant, a dicot plant, or an algae. The seed can be a monocot plant seed or a dicot plant seed. In some embodiments, the plant is an agricultural crop plant. The agricultural crop plant can be, but is not limited to, wheat, barley, rice, millet, oats, triticale, rye, bamboo, sugarcane plant, soybean, canola, rapeseed, cotton, alfalfa, cassava, potato, tomato, pea, chickpea, lentil, flax, or pepper.

[0021] In some embodiments, the synthetic combination of endophytes is co-cultured with algae.

[0022] In some embodiments, a plant colonized with the synthetic combination of endophytes has one or more of: increased plant biomass production, increased root biomass, increased root length, increased yield, and increased flowering, relative to an untreated reference plant that has not been colonized with the endophyte population.

[0023] In some embodiments, a plant colonized with the synthetic combination of endophytes has increased synchronization of flow ering or decreased seed shattering relative to an untreated reference plant that has not been colonized with the endophyte population.

[0024] In some embodiments, a plant colonized with the endophyte population has one or more of: increased abiotic stress resistance, increased salt tolerance, increased pH tolerance, or increased heat tolerance, relative to an untreated reference plant that has not been colonized with the endophyte population.

[0025] Described are methods for preparing an agricultural seed composition comprising contacting the surface of a plurality of seeds with any of the described synthetic combination of endophytes. In some embodiments, the synthetic combination of endophytes is provided as a coating on the surface of the seed.

[0026] Described are methods of growing a plant in a controlled environment comprising contacting or applying any of the described synthetic combinations of endophytes to a seed, plant, plant part, or environment of the seed, plant or plant part and growing the plant in thecontrolled environment. The controlled environment can be, but is not limited to, a greenhouse or a hydroponic system. In some embodiments, the plant is colonized with a synthetic combination of endophytes as described herein and grown in the controlled environment. In some embodiments, the plant is grown in the controlled environment and the synthetic combination of endophytes is provided in the water or growing medium of the plant. In some embodiments, the controlled environment comprises a hydroponic system.

[0027] Also described are seed or plants or colonized with any of the described synthetic combinations of endophytes.

[0028] Described are methods for reducing seed shattering, increasing plant biomass production, increasing yield, or increasing abiotic stress resistance in a plant or algae comprising applying any of the described synthetic combinations of endophytes to a seed, plant, plant part, algae, or environment of the seed, plant, plant part, or algae in an amount effective to colonize the plant or algae or plant grown from the seed with the synthetic combination of endophytes.BRIEF DESCRIPTION OF THE FIGURES

[0029] FIG. 1. Graph illustrating increase biomass productivity in algae treated with endophytes.

[0030] FIG. 2. Image illustrating increase biomass productivity in algae treated with endophytes.

[0031] FIG. 3A-B. Images and graphs illustrating improved seedling growth in two different crops, Camelina (A), and Sorghum (B and C) following treatment of seeds with endophyte combinations. The average and standard deviation of 15 seedlings is reported, and significant differences (a Student’s t-test determined *) between Control and treated, P < 0.0001. Seedlings were grown on agar plates after seed treatment with endophyte combinations.

[0032] FIG. 3C. Photograph demonstrating of root colonization by endophytes.

[0033] FIG. 4. Photograph illustrating growth of Sorghum plants grown from endophyte- treated and untreated (control) Sorghum seed in non- sterile soil (20 days old plants).

[0034] FIG. 5. Graph illustrating increased plant grow th in Sorghum plants grown from seed treated with endophyte combination.

[0035] FIG. 6. Graph illustrating increased salt tolerance in Sorghum plants grown from seed treated with endophyte combination.

[0036] FIG. 7. Photo illustrating increased total plant biomass in Sorghum plants grown from seed treated with endophyte combination.

[0037] FIG. 8. Photo illustrating increased salt tolerance in Sorghum plants grown from seed treated with endophyte combination.

[0038] FIG. 9. Graph illustrating differential production of metabolites in tomato treated with Bacillus megaterium and Rhizohium phaseoli.

[0039] FIG. 10. Graph illustrating differential production of metabolites in tomato treated with Azotobacter vinelandii, Rhizohium leguminosarum, Arthrobacter oryzae. and Bacillus licheniformis .

[0040] FIG. 11. Graph illustrating differential production of metabolites in tomato treated with Rhizohium leguminosarum. Bacillus licheniformis, Azospirillum brasilense. and Azotobacter chroococcum.

[0041] FIG. 12. Graphs illustrating differential production of metabolites in algae cocultured with Bacillus megaterium + Rhizohium phaseoli.

[0042] FIG. 13. Photo and graph illustrating increased biomass in lettuce plants following endophyte combination treatment.

[0043] FIG. 14. Photo and graph illustrating increased biomass in tomato plants following endophyte combination treatment.

[0044] FIG. 15. Graph illustrating increased number of flow ers in tomato plants following endophyte combination treatment.DETAILED DESCRIPTIONI. Definitions

[0045] As used herein the singular forms "a". “and’; and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “an endophyte” includes a plurality of endophytes and reference to “a plant or seed” includes reference to one or more plants or seeds, and so forth. All technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure belongs unless clearly indicated otherwise.

[0046] As used herein, “apply” refers to the action of introducing or contacting an endophyte, combination of endophytes, or composition or formulation of endophytes to a seed, plant, plant part, algae or environment of a seed, plant, plant part, or algae to improve plant performance.

[0047] An “effective amount’" refers to a quantity of endophytes sufficient to colonize a plant and / or improve plant performance.

[0048] The “environment of a seed, plant or plant part” comprises the area surrounding the seed, plant, or plant part (or algae), including but not limited to the soil, the air, or in-furrow. The environment of a seed, plant or plant part may be in proximity, touching, adjacent to, or in the same field as the seed, plant, or plant part. The compositions described herein may be applied to the environment of the seed, plant, or plant part as a seed treatment, as a foliar application, as a granular application, as a soil application, or as an encapsulated application. As used herein, “in-furrow” is intended to mean within or near the area where a seed is planted. The compositions disclosed herein may be applied in-furrow concurrently or simultaneously with a seed. In another embodiment, the compositions disclosed herein may be applied sequentially, either before or after a seed is planted.

[0049] A “plant” refers to all plants, plant parts, seed, and plant populations, such as desirable and undesirable wild plants, cultivars, transgenic plants, and plant varieties (whether or not protectable by plant variety or plant breeder’s rights). Cultivars and plant varieties can be plants obtained by conventional propagation and breeding methods that can be assisted or supplemented by one or more biotechnological methods such as by use of double haploids, protoplast fusion, random and directed mutagenesis, molecular or genetic markers or by bioengineering and genetic engineering methods.

[0050] An “endophyte” is an endosymbiont, such as a bacterium or fungus, that lives within aplant for at least part ofits life cycle without causing apparent disease. Endophytes are present is all or nearly all plant species. Endophytes may assist the plant in nutrient acquisition, improve the plant's tolerance abiotic stress, or increase the plant’s resistance to pathogens. Bacterial endophytes include, but are not limited to: Acidovorax, Agrobacterium, Arthrobacter, Azospirillum, Azotobacter, Bacillus, Burkholderia, Chryseobacterium. Curtobacterium, Enter obacter, Escherichia, Methylobacterium, Mesorhizobium, PaeniBacillus, Pantoea, Pseudomonas, Ralstonia, Rhizobium, Sinorhizobium, SacchariBacillus, Sphingomona , Sinorhizobium, and Stenotrophomonas species. Fungal endophytes in include, but are not limited to: Acremonium, Alternaria, Cladosporium, Cochliobolus, Embellisia, Epicoccum, Fusarium, Nigrospora, Phoma, and Podospora species.

[0051] “Heterologous,” in reference to an endophyte and a plant, indicates the endophyte is not normally present in the plant or was not present in the seed, plant, or plant part prior to applying the endophyte to the seed, plant, or plant part.

[0052] “Heterologous,” in reference to a combination of endophytes, indicates that the combination of endophytes does not normally occur together in nature, do not normally commonly colonize a plant or particular plant species, or was not present in the seed, plant, or plant part to applying the combination of endophytes to the seed, plant, or plant part.II. Synthetic endophyte combinations

[0053] Described are agricultural products comprising endophytes or synthetic combinations of endophytes that are able to improve plant (including algae) growth. In some embodiments the endophyte combinations comprise at least one endophyte species the reduces ethylene concentrations in the plant, at least one endophyte species that increases endogenous levels of growth-promoting hormones, and at least one endophyte species with nitrogen fixation capability. Endophyte species that reduce ethylene concentrations in the plant include endophyte species that produce 1 -aminocyclopropane- 1 -carboxylate (ACC) deaminase. ACC deaminase is an enzyme that degrades the ethylene precursor, ACC (1-aminocyclopropane-l- carboxylate). In some embodiments, an ACC deaminase-producing endophyte species is a Bacillus species (e.g. Bacillus licheniformis (ATCC #14580) Bacillus megaterium (ATCC #14581), or Bacillus subtilis (ATCC #6051) or a Pseudomonas species (e.g, Pseudomonas putida (ATCC #12633) or Pseudomonas fluorescens (ATCC #13525)). Endophyte species that increase endogenous levels of growth-promoting hormones include endophyte species that increase auxin, cytokinin, and gibberellin production in the plant. In some embodiments, an endophyte species that increases auxin, cytokinin, and gibberellin production in the plant is a Rhizobium species (e.g., Rhizobium leguminosarum (ATCC #10004) or Rhizobium phaseoli (ATCC #31236)), a Pseudomonas species (e.g.. Pseudomonas putida (ATCC #12633) or Pseudomonas fluorescens (ATCC #13525)). or an Azotobacter species (e.g.. Azotobacter vinelandii (ATCC #478)). In some embodiments, nitrogen fixing endophyte species is a Rhizobium species (e.g. , Rhizobium leguminosarum (ATCC #10004) o Rhizobium phaseoli (ATCC #31236)); a Mesorhizobium species (e.g, Mesorhizobium amorphae (ATCC #BAA- 114)); an Arthrobacter species (e.g., Arthrobacter oryzae (ATCC #42149)); an Azospirillum species (e.g, Azospirillum brasilense (ATCC #29729)); an Azotobacter species (e.g.. Azotobacter chroococcum (ATCC #9043) or Azotobacter vinelandii (ATCC #478)); a Sinorhizobium species (e.g, Sinorhizobium meliloti (ATCC #9930); or a Bradyrhizobium species.

[0054] In some embodiments, one or more of the endophytes in the endophyte combination is heterologous to the seed, seedling, plant, plant part, or algae. In some embodiments, theendophyte combination is heterologous to the seed, seedling, plant, plant part, or algae. An endophyte or endophyte combination is considered heterologous to a seed, seedling, plant, plant part, or algae if the seed, seedling, plant, plant part, or algae does not normally contain detectable levels of the endophyte or endophyte combination.

[0055] In some embodiments, a strain may be derived from Rhizobium species (e.g, Rhizobium leguminosarum (ATCC #10004) or Rhizobium phaseoli (ATCC #31236)), a Pseudomonas species (e.g, Pseudomonas putida (ATCC #12633) ox Pseudomonas fluorescens (ATCC #13525)), or an Azotobacter species (e.g., Azotobacter vinelandii (ATCC #478)). In some embodiments, nitrogen fixing endophyte species is derived from a Rhizobium species (e.g, Rhizobium leguminosarum (ATCC #10004) ox Rhizobium phaseoli (ATCC #31236)); a Mesorhizobium species (e.g. , Mesorhizobium amorphae (ATCC #BAA-114)); ax Arthrobacter species (e.g, Arthrobacter oryzae (ATCC #42149)); an Azospirillum species (e.g, Azospirillum brasilense (ATCC #29729)); an Azotobacter species (e.g, Azotobacter chroococcum (ATCC #9043) or Azotobacter vinelandii (ATCC #478)); a Sinorhizobium species (e.g, Sinorhizobium meliloti (ATCC #9930); or a Bradyrhizobium species and retains at least one function (e.g, nitrogen fixing function) of the original strain. A derived strain retains at least one function of the original strain (e.g, ACC deaminase-production, induction of increased endogenous levels of growth-promoting hormones, or nitrogen fixing) and may be created from directed mutagenesis, natural evolution, or genetic modification, including genetic editing.

[0056] In some embodiments, the synthetic combination of endophytes comprises: (a) at least one endophyte selected from the group consisting of: a Bacillus species and a Pseudomonas species; (b) at least one endophyte selected from the group consisting of: a Rhizobium species, a Pseudomonas species, and an Azotobacter species; and (c) at least one endophyte selected from the group consisting of: a Rhizobium species, a Mesorhizobium species, an Arthrobacter species, an Azospirillum species, an Azotobacter species, a Sinorhizobium species, and a Bradyrhizobium species.

[0057] In some embodiments, the synthetic combination of endophytes comprises: (a) a Bacillus species; (b) at least one endophyte selected from the group consisting of: a Rhizobium species, a Pseudomonas species, and an Azotobacter species; and (c) at least one endophyte selected from the group consisting of: a Rhizobium species, a Mesorhizobium species, an Arthrobacter species, an Azospirillum species, an Azotobacter species, a Sinorhizobium species, and a Bradyrhizobium species.

[0058] In some embodiments, the synthetic combination of endophytes comprises: (a) a Pseudomonas species; (b) at least one endophyte selected from the group consisting of: a Rhizobium species, a Pseudomonas species, and an Azotobacter species; and (c) at least one endophyte selected from the group consisting of: a Rhizobium species, a Mesorhizobium species, an Arthrobacter species, an Azospirillum species, an Azotobacter species, a Sinorhizobium species, and a Bradyrhizobium species.

[0059] In some embodiments, the synthetic combination of endophytes comprises: (a) a Bacillus species; (b) a first Rhizobium species; and (c) at least one endophyte selected from the group consisting of: a second Rhizobium species, a Mesorhizobium species, an Arthrobacter species, an Azospirillum species, an Azotobacter species, a Sinorhizobium species, and a Bradyrhizobium species.

[0060] In some embodiments, the synthetic combination of endophytes comprises: (a) a Pseudomonas species; (b) a first Rhizobium species; and (c) at least one endophyte selected from the group consisting of: a second Rhizobium species, a Mesorhizobium species, an Arthrobacter species, an Azospirillum species, an Azotobacter species, a Sinorhizobium species, and a Bradyrhizobium species.

[0061] In some embodiments, the synthetic combination of endophytes comprises: (a) a Bacillus species; (b) a Pseudomonas species; and (c) at least one endophyte selected from the group consisting of: a Rhizobium species, a Mesorhizobium species, an Arthrobacter species, an Azospirillum species, an Azotobacter species, a Sinorhizobium species, and a Bradyrhizobium species.

[0062] In some embodiments, the synthetic combination of endophytes comprises: (a) a first Pseudomonas species; (b) a second Pseudomonas species; and (c) at least one endophyte selected from the group consisting of: a Rhizobium species, a Mesorhizobium species, an Arthrobacter species, an Azospirillum species, an Azotobacter species, a Sinorhizobium species, and a Bradyrhizobium species.

[0063] In some embodiments, the synthetic combination of endophytes comprises: (a) a Bacillus species; (b) an Azotobacter species; and (c) at least one endophyte selected from the group consisting of: a Rhizobium species, a Mesorhizobium species, an Arthrobacter species, an Azospirillum species, an Azotobacter species, a Sinorhizobium species, and a Bradyrhizobium species.

[0064] In some embodiments, the synthetic combination of endophytes comprises: (a) a first Pseudomonas species; (b) an Azotobacter species; and (c) at least one endophyte selected from the group consisting of: a Rhizobium species, a Mesorhizobium species, an Arthrobacterspecies, an Azospirillum species, an Azotobacter species, a Sinorhizobium species, and a Bradyrhizobium species.

[0065] In some embodiments, the synthetic combination of endophytes comprises: (a) a Bacillus species; (b) a Pseudomonas species; and (c) aRhizobium species.

[0066] In some embodiments, the synthetic combination of endophytes comprises: (a) a Bacillus species; (b) a Pseudomonas species; and (c) aMesorhizobium species.

[0067] In some embodiments, the synthetic combination of endophytes comprises: (a) a Bacillus species; (b) a Pseudomonas species; and (c) an Arthrobacter species.

[0068] In some embodiments, the synthetic combination of endophytes comprises: (a) a Bacillus species; (b) a Pseudomonas species; and (c) an Azospirillum species.

[0069] In some embodiments, the synthetic combination of endophytes comprises: (a) a Bacillus species; (b) a Pseudomonas species; and (c) an Azotobacter species.

[0070] In some embodiments, the synthetic combination of endophytes comprises: (a) a Bacillus species; (b) a Pseudomonas species; and (c) a Sinorhizobium species.

[0071] In some embodiments, the synthetic combinations of endophytes comprise: (a) a Bacillus species; (b) a Pseudomonas species; and (c) a Bradyrhizobium species.

[0072] In some embodiments, the synthetic combinations of endophytes comprise: (a) a Azotobacter species; (b) aRhizobium species; and (c) a Arthrobacter species.

[0073] In some embodiments, the synthetic combinations of endophytes comprise: (a) a Sinorhizobium species; (b) a Azotobacter species; and (c) a Bacillus species.

[0074] In some embodiments, the synthetic combinations of endophytes comprise: (a) a Azotobacter species; (b) a Rhizobium species; (c) a Arthrobacter species, and (d) a Bacillus species.

[0075] In some embodiments, the synthetic combinations of endophytes comprise: (a) a Sinorhizobium species; (b) a Azotobacter species; and (c) aRhizobium species.

[0076] In some embodiments, the synthetic combinations of endophytes comprise: (a) a Arthrobacter species; (b) a Sinorhizobium species; and (c) a Azospirillum species.

[0077] In some embodiments, the synthetic combinations of endophytes comprise: (a) a Rhizobium species; (b) a Bacillus species; (c) a Azospirillum species, and (d) a Azotobacter species.

[0078] In some embodiments, the synthetic combinations of endophytes comprise: (a) a Sinorhizobium species; (b) a Rhizobium species; (c) a Mesorhizobium species; and (d) a Arthrobacter species.

[0079] In some embodiments, the synthetic combinations of endophytes comprise: (a) a first Azotobacter species; (b) a Rhizobium species; (c) aArthrobacter species; (d) a Bacillus species; (e) a Sinorhizobium species; (f) a second Azotobacter species; (g) a Azospirillum species; (h) aMesorhizobium species.

[0080] In some embodiments, the synthetic combinations of endophytes comprise: (a) a Bacillus species; and (b) a Pseudomonas species.

[0081] In some embodiments, the synthetic combinations of endophytes comprise: (a) a Bacillus species; and (b) a Rhizobium species.

[0082] In some embodiments, the synthetic combinations of endophytes comprise: (a) a Pseudomonas species; and (b) a Rhizobium species.

[0083] In some embodiments, the synthetic combinations of endophytes comprise: (a) a first Bacillus species; (b) a second Bacillus species; (c) a first Pseudomonas species; and (d) a second Pseudomonas species.

[0084] In some embodiments, the synthetic combinations of endophytes comprise: (a) at least one first endophyte selected from the group consisting of: Bacillus licheniformis (ATCC #14580). Bacillus megaterium (ATCC #14581), Pseudomonas putida (ATCC #12633), and Pseudomonas fluorescens (ATCC #13525); (b) at least one second endophyte selected from the group consisting of: Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), Pseudomonas putida (ATCC #12633), Pseudomonas fluorescens (ATCC #13525). and Azotobacter vinelandii (ATCC #478); and (c) at least one third endophyte selected from the group consisting of: Rhizobium leguminosarum (ATCC # 10004), Rhizobium phaseoli (ATCC #31236), Mesorhizobium amorphae (ATCC #BAA-114), Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), and Sinorhizobium meliloti (ATCC #9930). In some embodiments, the first, second, and third endophytes are all different (e.g., if the first endophyte is Pseudomonas putida, then the second endophyte is not Pseudomonas putida).

[0085] In some embodiments, the synthetic combination of endophytes comprises: (a) Bacillus licheniformis (ATCC #14580) ox Bacillus megaterium (ATCC #14581); (b) at least one second endophyte selected from the group consisting of: Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), Pseudomonas putida (ATCC #12633), Pseudomonas fluorescens (ATCC #13525), av Azotobacter vinelandii (ATCC #478); and (c) at least one third endophyte selected from the group consisting of: Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), Mesorhizobium amorphae (ATCC#BAA-114), Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043). Azotobacter vinelandii (ATCC #478). and Sinorhizobium meliloti (ATCC #9930). In some embodiments, the first, second, and third endophytes are all different (e.g., if the second endophyte is Rhizobium leguminosarum, then the third endophyte is not Rhizobium leguminosarum).

[0086] In some embodiments, the synthetic combination of endophytes comprises: (a) Pseudomonas putida (ATCC #12633) ox Pseudomonas fluorescens (ATCC #13525); (b) at least one second endophyte selected from the group consisting of: Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), Pseudomonas putida (ATCC #12633), Pseudomonas fluorescens (ATCC #13525), and Azotobacter vinelandii (ATCC #478); and (c) at least one third endophyte selected from the group consisting of: Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), Mesorhizobium amorphae (ATCC #BAA-114), Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), and Sinorhizobium meliloti (ATCC #9930). In some embodiments, the first, second, and third endophytes are all different (e.g, if the second endophyte is Rhizobium leguminosarum, then the third endophyte is not Rhizobium leguminosarum).

[0087] In some embodiments, the synthetic combination of endophytes comprises: (a) Bacillus licheniformis (ATCC #14580) or Bacillus megaterium (ATCC #14581); (b) Rhizobium leguminosarum (ATCC #10004) or Rhizobium phaseoli (ATCC #31236); and (c) at least one third endophyte selected from the group consisting of: Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), Mesorhizobium amorphae (ATCC #BAA-114), Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478). and Sinorhizobium meliloti (ATCC #9930). In some embodiments, the first, second, and third endophytes are all different (e.g, if the second endophyte is Rhizobium leguminosarum, then the third endophyte is not Rhizobium leguminosarum).

[0088] In some embodiments, the synthetic combination of endophytes comprises: (a) Pseudomonas putida (ATCC #12633) or Pseudomonas fluorescens (ATCC #13525); (b) Rhizobium leguminosarum (ATCC #10004) or Rhizobium phaseoli (ATCC # 1236); and (c) at least one third endophyte selected from the group consisting of: Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), Mesorhizobium amorphae (ATCC #BAA-114), Arthrobacter oryzae (ATCC #42149). Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), andSinorhizobium meliloti (ATCC #9930). In some embodiments, the first, second, and third endophytes are all different (e g., if the second endophyte is Rhizobium leguminosarum, then the third endophyte is not Rhizobium leguminosarum).

[0089] In some embodiments, the synthetic combination of endophytes comprises: (a) Bacillus licheniformis (ATCC #14580) or Bacillus megaterium (ATCC #14581); (b) Pseudomonas putida (ATCC #12633) or Pseudomonas fluorescens (ATCC #13525); and (c) at least one third endophyte selected from the group consisting of: Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), Mesorhizobium amorphae (ATCC #BAA-114), Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), and Sinorhizobium meliloti (ATCC #9930).

[0090] In some embodiments, the synthetic combination of endophytes comprises: (a) Pseudomonas putida (ATCC #12633) or Pseudomonas fluorescens (ATCC #13525); (b) Pseudomonas putida (ATCC #12633) or Pseudomonas fluorescens (ATCC #13525); and (c) at least one third endophyte selected from the group consisting of: Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236). Mesorhizobium amorphae (ATCC #BAA-114), Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), and Sinorhizobium meliloti (ATCC #9930). In some embodiments, the first and second endophytes are different (e g, if the first endophyte is Pseudomonas putida, then the second endophyte is Pseudomonas fluorescens).

[0091] In some embodiments, the synthetic combination of endophytes comprises: (a) Bacillus licheniformis (ATCC #14580) or Bacillus megaterium (ATCC #14581); (b) Azotobacter vinelandii (ATCC #478); and (c) at least one third endophyte selected from the group consisting of: Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), Mesorhizobium amorphae (ATCC #BAA-114), Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), and Sinorhizobium meliloti (ATCC #9930). In some embodiments, the second and third endophytes are different (e.g. the third endophyte is not Azotobacter vinelandii).

[0092] In some embodiments, the synthetic combination of endophytes comprises: (a) Pseudomonas putida (ATCC #12633) or Pseudomonas fluorescens (ATCC #13525); (b) Azotobacter vinelandii (ATCC #478); and (c) at least one third endophyte selected from the group consisting of: Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC#31236), Mesorhizobium amorphae (ATCC #BAA-114), Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), and Sinorhizobium meliloti (ATCC #9930). In some embodiments, the second and third endophytes are different (e.g., the third endophyte is not Azotobacter vinelandii).

[0093] In some embodiments, the synthetic combination of endophytes comprises: (a) Bacillus licheniformis (ATCC #14580) or Bacillus megaterium (ATCC #14581); (b) Pseudomonas putida (ATCC #12633) or Pseudomonas fluorescens (ATCC #13525); and (c) Rhizobium leguminosarum (ATCC #10004) or Rhizobium phaseoli (ATCC #31236).

[0094] In some embodiments, the synthetic combination of endophytes comprises: (a) Bacillus licheniformis (ATCC #14580) or Bacillus megaterium (ATCC #14581); (b) Pseudomonas putida (ATCC #12633) ox Pseudomonas fluorescens (ATCC #13525); and (c) Mesorhizobium amorphae (ATCC #BAA-114).

[0095] In some embodiments, the synthetic combination of endophytes comprises: (a) Bacillus licheniformis (ATCC #14580) or Bacillus megaterium (ATCC #14581); (b) Pseudomonas putida (ATCC #12633) ox Pseudomonas fluorescens (ATCC #13525); and (c) Arthrobacter oryzae (ATCC #42149).

[0096] In some embodiments, the synthetic combination of endophytes comprises: (a) Bacillus licheniformis (ATCC #14580) or Bacillus megaterium (ATCC #14581); (b) Pseudomonas putida (ATCC #12633) ox Pseudomonas fluorescens (ATCC #13525); and (c) Azospirillum brasilense (ATCC #29729).

[0097] In some embodiments, the synthetic combination of endophytes comprises: (a) Bacillus licheniformis (ATCC #14580) or Bacillus megaterium (ATCC #14581); (b) Pseudomonas putida (ATCC #12633) ox Pseudomonas fluorescens (ATCC #13525); and (c) Azotobacter chroococcum (ATCC #9043) ox Azotobacter vinelandii (ATCC #478).

[0098] In some embodiments, the synthetic combination of endophytes comprises: (a) Bacillus licheniformis (ATCC #14580) or Bacillus megaterium (ATCC #14581); (b) Pseudomonas putida (ATCC #12633) ox Pseudomonas fluorescens (ATCC #13525); and (c) Sinorhizobium meliloti (ATCC #9930).

[0099] In some embodiments, the synthetic combination of endophytes comprises at least one endophyte selected from the group consisting of: Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), Bacillus licheniformis (ATCC #14580), Bacillus megaterium (ATCC #14581), Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonasputida (ATCC #12633), Pseudomonas fluorescens (ATCC #13525), Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), and Sinorhizobium meliloti (ATCC #9930).

[0100] In some embodiments, the synthetic combination of endophytes comprises at least two endophytes selected from the group consisting of: Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), Bacillus licheniformis (ATCC #14580). Bacillus megaterium (ATCC #14581), Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonas putida (ATCC #12633), Pseudomonas fluorescens (ATCC #13525), Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), and Sinorhizobium meliloti (ATCC #9930).

[0101] In some embodiments, the synthetic combination of endophytes comprises at least three endophytes selected from the group consisting of: Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), Bacillus licheniformis (ATCC #14580), Bacillus megaterium (ATCC #14581). Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonas putida (ATCC #12633), Pseudomonas fluorescens (ATCC #13525), Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), and Sinorhizobium meliloti (ATCC #9930).

[0102] In some embodiments, the synthetic combination of endophytes comprises at least four endophytes selected from the group consisting of: Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), Bacillus licheniformis (ATCC #14580), Bacillus megaterium (ATCC #14581), Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonas putida (ATCC #12633), Pseudomonas fluorescens (ATCC #13525), Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), and Sinorhizobium meliloti (ATCC #9930).

[0103] In some embodiments, the synthetic combination of endophytes comprises at least five endophytes selected from the group consisting of: Arthrobacter oryzae (ATCC #42149). Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), Bacillus licheniformis (ATCC #14580), Bacillus megaterium (ATCC #14581), Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonas putida (ATCC #12633), Pseudomonas fluorescens (ATCC #13525), Rhizobiumleguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), and Sinorhizobium meliloti (ATCC #9930).

[0104] In some embodiments, the synthetic combination of endophytes comprises at least six endophytes selected from the group consisting of: Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), Bacillus licheniformis (ATCC #14580), Bacillus megaterium (ATCC #14581), Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonas putida (ATCC #12633), Pseudomonas fluorescens (ATCC #13525), Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), and Sinorhizobium meliloti (ATCC #9930).

[0105] In some embodiments, the synthetic combination of endophytes comprises at least seven endophytes selected from the group consisting of: Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), Bacillus licheniformis (ATCC #14580), Bacillus megaterium (ATCC #14581), Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonas putida (ATCC #12633). Pseudomonas fluorescens (ATCC #13525). Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), and Sinorhizobium meliloti (ATCC #9930).

[0106] In some embodiments, the synthetic combination of endophytes comprises at least eight endophytes selected from the group consisting of: Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), Bacillus licheniformis (ATCC #14580), Bacillus megaterium (ATCC #14581), Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonas putida (ATCC #12633), Pseudomonas fluorescens (ATCC #13525), Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), and Sinorhizobium meliloti (ATCC #9930).

[0107] In some embodiments, the synthetic combination of endophytes comprises at least nine endophytes selected from the group consisting of: Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043). Azotobacter vinelandii (ATCC #478), Bacillus licheniformis (ATCC #14580), Bacillus megaterium (ATCC #14581), Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonas putida (ATCC #12633), Pseudomonas fluorescens (ATCC #13525), Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), and Sinorhizobium meliloti (ATCC #9930).

[0108] In some embodiments, the synthetic combination of endophytes comprises at least ten endophytes selected from the group consisting of Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), Bacillus licheniformis (ATCC #14580), Bacillus megaterium (ATCC #14581), Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonas putida (ATCC #12633), Pseudomonas fluorescens (ATCC #13525), Rhizobium leguminosarum (ATCC #10004). Rhizobium phaseoli (ATCC #31236), and Sinorhizobium meliloti (ATCC #9930).

[0109] In some embodiments, the synthetic combination of endophytes comprises at least eleven endophytes selected from the group consisting of: Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), Bacillus licheniformis (ATCC #14580), Bacillus megaterium (ATCC #14581), Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonas putida (ATCC #12633), Pseudomonas fluorescens (ATCC #13525), Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), and Sinorhizobium meliloti (ATCC #9930).

[0110] In some embodiments, the synthetic combination of endophytes comprises Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043). Azotobacter vinelandii (ATCC #478), Bacillus licheniformis (ATCC #14580), Bacillus megaterium (ATCC #14581), Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonas putida (ATCC #12633), Pseudomonas fluorescens (ATCC #13525), Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), and Sinorhizobium meliloti (ATCC #9930).[OHl] In some embodiments, the endophyte combination comprises Azotobacter chroococcum, Rhizobium leguminosarum, and Arthrobacter oryzae.

[0112] In some embodiments, the endophyte combination comprises Sinorhizobium meliloti , Azotobacter vinelandii, and Bacillus licheniformis .

[0113] In some embodiments, the endophyte combination comprises Azotobacter vinelandii, Rhizobium leguminosarum, Arthrobacter oryzae, and Bacillus licheniformis .

[0114] In some embodiments, the endophyte combination comprises Sinorhizobium meliloti, Azotobacter chroococcum, and Rhizobium leguminosarum.

[0115] In some embodiments, the endophyte combination comprises Arthrobacter oryzae, Sinorhizobium meliloti, and Azospinllum brasilense.

[0116] In some embodiments, the endophyte combination comprises Rhizobium leguminosarum, Bacillus licheniformis. Azospirillum brasilense, and Azotobacter chroococcum.

[0117] In some embodiments, the endophyte combination comprises Sinorhizobium meliloti , Rhizobium leguminosarum, Mesorhizobium amorphae, and Arthrobacter oryzae.

[0118] In some embodiments, the endophyte combination comprises Azotobacter chroococcum, Rhizobium leguminosarum, Arthrobacter oryzae, Bacillus licheniformis. Sinorhizobium meliloti, Azotobacter chroococcum. Azospirillum brasilense, and Mesorhizobium amorphae.

[0119] In some embodiments, the endophyte combination comprises Bacillus subtilis and Pseudomonas fluorescens .

[0120] In some embodiments, the endophyte combination comprises Bacillus subtilis and Rhizobium phaseoli.

[0121] In some embodiments, the endophyte combination comprises Pseudomonas fluorescens and Rhizobium phaseoli.

[0122] In some embodiments, the endophyte combination comprises Bacillus subtilis. Pseudomonas fluorescens, and Rhizobium phaseoli.

[0123] In some embodiments, the endophyte combination comprises Bacillus Megaterium, Bacillus subtilis, Pseudomonas fluorescens, and Pseudomonas putida.

[0124] In some embodiments, the endophyte combination comprises Bacillus Megaterium and Rhizobium phaseoli.

[0125] The described synthetic combinations of endophytes (endophyte combinations) can be used to confer one or more improved agronomic traits to a plant. The endophyte combinations colonize within plants, resulting in one or more of: decreased ethylene production, increased abiotic stress resistance (e.g, increased salinity tolerance), increased endogenous production of growth-promoting hormones (e.g., auxin, cytokinin, and gibberellin), increased nitrogen fixation, increase flowering, increased synchronization of flowering, reduced seed shattering, increased plant resilience, increased plant quality, increased yield quality, and increased yield (i.e., plant growth). Increased yield can include increasing biomass and / or increasing seed harvest.

[0126] In some embodiments, the described endophyte combinations increase nitrogen fixation and production of other plant hormones in plants to which they are applied. Increased nitrogen fixation and production of hormones in plants leads to increased plant growth and increased biomass yield.

[0127] In some embodiments, the described endophyte combinations increase synchronous flowing and reduce sees shattering in plants. In many crops, such as oilseed (e.g., camelina, soybean, canola, and flax) and grain (rice and sorghum) crops, seed shattering is a significant cause of crop loss. While seed shattering is a natural process essential for seed dispersal in wild plants, it creates a significant harvest loss in oilseed and grain crops, which can suffer a 30- 50% annual harvest loss due to an extended asynchronous flowering period and uncontrolled seed shattering. Further, the severity of seed shattering loss is increasing due to rising temperatures and global climate change. Irregular increases in temperature due to global warming increase asynchronous flowering period and uncontrolled seed shattering. Annual seed yield for oilseed and grain crops heavily depends on the timing of harvest. Timing of harvest is complicated by irregular flowering and irregular growth habit. Ethylene is a plant hormone than mediates plant stress response and interacts with other plant hormones to regulate plant development. When ethylene is over-produced, it initiates a stress response that hinders plant grow th. Ethylene reduction is known to increase plant growth and reduce fruit dropping. By decreasing ethylene production and improving the overall growth of the plant, the described synthetic combinations of endophytes enhance synchronized flowering in plants and reduce seed shattering. Reducing seed shattering increases overall plant seed production and eases harvest. Increased synchronized flowering and reduced seed shattering facilitates single-step harvesting.

[0128] In some embodiments, the described endophyte combinations increase nutrient availability and modulate phytohormones by increasing the concentrations of auxin, gibberellin, cytokinin, and ACC-deaminase. Increasing nutrient availability, increasing concentrations of plant growth hormones, and / or decreasing ethylene concentration can lead to increased resistance to abiotic stress, such as soil salinity stress.

[0129] In some embodiments, the described endophyte combinations can be used as biofertilizers, biostimulants, or biocontrol agents. The endophyte combinations can be used to reduce the use of chemical fertilizers, pesticides, and herbicides.

[0130] The described endophyte combinations can be used with any plant. The plant can be a monocot or a dicot. The described endophyte combinations can also be used with algae species.

[0131] In some embodiments, the described endophyte combinations can be used to confer one or more improved agronomic traits in a food crop, an oil seed crop, a grain crop, or an algae species. The oil seed crop can be. but is not limited to: camelina, soybean, cottonseed, sunflower seed, canola, rapeseed, flax (linseed), castor beans, safflow er, sorghum, peanut, andBrassica species. The grain crop can be, but is not limited to, a cereal grain, a pseudocereal grain, a pulse grain, com. rice, rye. sorghum, millet, wheat, soybean, oats, and barley. The agronomic traits can be, but are not limited to, decreased ethylene production, increased abiotic stress resistance (e.g., increased salinity, pH, or heat tolerance), increased endogenous production of grow th-promoting hormones (e g., auxin, cytokinin, and gibberellin), increased nitrogen fixation, increased synchronization of flowering, reduced seed shattering, increase flowering, and increased yield (i.e., plant growth). Increased yield can include increased biomass, increased carbon storage, and / or increased seed or fruit harvest.

[0132] The described endophyte combinations also increase survival under adverse outdoor environmental conditions.

[0133] The described endophyte combinations are able to be applied to a seed, plant, plant part, or soil, by simple inoculation.

[0134] In some embodiments, one or more endophytes in an endophyte combination are tagged with a marker to facilitate identification. The tag can be, but is not limited to, a genetic tag, or a marker gene that encodes a detectable protein. The detectable protein can be, but is not limited to, a fluorescent protein (e.g., GFP or sfGFP).III. Compositions

[0135] Any of the described endophyte combinations can be provided in a composition or formulation. Any of the described endophyte combinations can be combined with one or more excipients, one or more agrochemically active compounds, one or more biocontrol agents, and combinations thereof.

[0136] An excipient is any ingredient that is intentionally added to the formulation but is not itself expected to exert an effect on plant growth or health. Excipients may act to (a) aid in manufacture, (b) protect, support, or enhance stability, plant delivery, or bioavailability, (c) assist in product identification, and / or (d) enhance any other attribute of the overall safety, effectiveness, or deliver}' or the composition during storage or use. An excipient may or may not be an inert substance. In some embodiments, the excipient is an agriculturally acceptable excipient. An excipient can be. but is not limited to, a carrier, an adjuvant, a solubilizing agent, a suspending agent, a diluent, an oxygen scavenger, an antioxidant, a food material, an anticontaminant agent, or combinations thereof. Excipients include, but are not limited to: absorption enhancers, adhesives, anti-foaming agents, anti-oxidants, binders, buffering agents, carriers, coating agents, colors, delivery enhancers, dextran, dextrose, diluents, disintegrants, dispersants, dust control agents, emulsifiers, extenders, fillers, flavors, glidants, humectants,lubricants, oils, polymers, preservatives, saline, salts, solvents, sugars, suspending agents, sustained release matrices, sweeteners, thickening agents, tonicity agents, vehicles, waterrepelling agents, and wetting agents. Excipients also include, cellulose, microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, dibasic calcium phosphate, glycine, polysaccharides, starch, milk sugar, and high molecular weight polyethylene gly cols, and the like. A carrier can be, but is not limited to, a solvent or dispersion medium containing, for example, water, saline, phosphate buffered saline, gel. solvent, solubilizer, alcohol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol), silicon, waxes, petroleum jelly, vegetable oil, sugar, gelatin, amylose, talc, surfactants, and suitable mixtures thereof. A carrier may also contain adjuvants or additives such as preservatives, wetting agents, emulsifying agents, and dispersing agents. In some embodiments, the carrier is an agriculturally acceptable carrier. Agriculturally acceptable refers to those properties and / or substances which are acceptable for use in agriculture.

[0137] An agrochemically active compound is a substance that may be used for treating a seed, a plant, plant part, or the environment of the seed or plant or plant part. Agrochemically active compounds include, but are not limited to. fungicides, bactericides, insecticides, acaricides, nematicides, molluscicides, safeners, plant growth regulators, and plant nutrients.

[0138] A biocontrol agent is a biological organism or component thereof, other than an endophyte or the endophyte composition, that reduces the incidence or severity of diseases caused by plant pathogens. A biocontrol agent can be. but is not limited to, microorganisms or modified organisms.

[0139] The described endophyte combinations can be provided (formulated) as a liquid, a dispersion, a colloidal dispersion, a solution, a suspension, a colloidal suspension, an emulsion, a foam, a slurry’, a lyophilized (freeze-dried) cake or powder, a spray-dried powder, a pellet, a biologically pure pellet, a coated pellet, a powder, a flowable powder, or a granule. A liquid formulation or composition may contain one or more of: a buffer, salt, sorbitol, and / or glycerol. The endophyte combination may be formulated as a heterogeneous mixture or a homogeneous mixture. Powders, pellets, and granules may be formulated using encapsulation technologies known in the art.

[0140] The described endophyte combinations can be provided (formulated) as heterogeneous mixture or a homogeneous mixture.

[0141] The compositions and formulations are prepared using methods available in the art for formulating bacterial and fungal compositions.

[0142] In some embodiments, a synthetic combination of endophytes is provided in a growth media or fertilizer. The growth media can be a solid growth media or a liquid growth media (e.g. a hydroponic liquid growth media). Solid and liquid growth media and fertilizers known in the art can be amended to contain any of the described synthetic combination of endophytes.

[0143] In some embodiments, the endophytes are grown in one or more cultures prior to formulation or applying the endophytes to a seed, plant, plant part, or environment of the seed, plant, or plant part. The endophytes in the combination can be grown in individual cultures (e.g. , a separate culture for each endophyte) and combined prior to formulation or applying the endophytes to a seed, plant, plant part, or environment of the seed, plant, or plant part. Alternatively, the endophytes in the combination can be grown together in a single culture prior to formulation or applying the endophytes to a seed, plant, plant part, or environment of the seed, plant, or plant part. The endophytes in the combination can also be grown in two or more separate cultures, each containing one or more of the endophytes in the culture, and combined prior to formulation or applying the endophytes to a seed, plant, plant part, or environment of the seed, plant, or plant part. If grown in two of more cultures, the endophytes may be combined prior to formulation or applying the endophytes to a seed, plant, plant part, or environment of the seed, plant or plant part or the endophytes may be formulated or applied separately to the seed, plant, plant part, or the environment of the seed, plant, or plant part.

[0144] In some embodiments, an endophyte combination is formulated for use with a seed. In some embodiments, an endophyte combination is formulated for use as a seed treatment. In some embodiments, an endophyte combination is formulated for application to a plant or plant part (e.g., a leaf). In some embodiments, an endophyte combination is formulated for application to soil.

[0145] An endophyte combination formulated as a seed treatment may be applied to a seed (or plurality of seeds) using methods available in the art for treating seeds. Such methods include, but are not limited to, rotary coaters, drum coaters, fluidized bed techniques, spouted beds, rotary’ mists, and combinations thereof. The compositions may be applied to seeds using continuous or batch coating processes.

[0146] In some embodiments, an endophyte combination is formulated to contain the endophytes in an amount effective to colonize a plant. The compositions can be formulated for one-time treatment of a seed, plant, plant part, or soil or the compositions can be formulated for repeat application, such as to a plant, plant part, or soil.

[0147] The endophytes may be purified from a culture prior to formulation of application to a seed, plant, plant part, or environment of the seed, plant, or plant part.

[0148] In some embodiments, the described endophyte combinations are biologically pure cultures of the endophytes.

[0149] In some embodiments, a synthetic combination of endophytes is provided or formulated as a liquid, a dispersion, a colloidal dispersion, a solution, a suspension, a colloidal suspension, an emulsion, or a slurry at a concentration of about l x IO4to about l x IO9colony forming units (cfu) per mL, about 1 x 1 CP to about 1 x 109cfu per mL, about 1 x 106to about 1 x 109cfu per mL, about 1 x 107to about 1 x 109cfu per mL, or about 1 x 108to about 1 x 109cfu per mL.IV. Plants

[0150] The described synthetic combinations of endophytes may be use with or applied to any plant seed, plant seedling, plant, or plant part. The plant can be, but is not limited to, a naturally occurring plant, a cultivated plant (e.g., a cultivar), a genetically modified plant, or an agricultural crop plant. Cultivars and plant varieties can be plants obtained by conventional propagation and breeding methods that can be assisted or supplemented by one or more biotechnological methods such as by use of double haploids, protoplast fusion, random and directed mutagenesis, molecular or genetic markers or by bioengineering and genetic engineering methods. The described synthetic combinations of endophytes may be used with or applied to algae species. The plant can be, but is not limited to. a plant grown in a field, a plant grown in a greenhouse, or a plant grown in a hydroponic system.

[0151] The plant can be a dicotyledon (dicot) or a monocotyledon (monocot). In some embodiments, the plant is an oil seed crop or a grain crop. The oil seed crop can be, but is not limited to: camelina, soybean, cottonseed, sunflower seed, canola, rapeseed, flax (linseed), castor beans, safflower, sorghum, peanut, and Brassica species. The grain crop can be, but is not limited to, a cereal grain, a pseudocereal grain, a pulse grain, com, rice, rye, sorghum, millet, wheat, soybean, oats, and barley. The plant can also be a food crop. Food crops include, but are not limited to, vegetable crops. Vegetable crops include, but are not limited to, tomato and lettuce.

[0152] Also described are seeds treated with any of the described synthetic combinations of endophytes. The seed can be any plant seed treated with any of the described endophyte combinations or any of the described compositions or formulations thereof. The seed can be a dicotyledon (dicot) seed or a monocotyledon (monocot) seed. In some embodiments, the seed is an oil crop seed or a grain crop seed. The oil crop seed can be, but is not limited to: camelinaseed, soybean seed, cotonseed seed, sunflower seed, canola seed, rapeseed seed, flax (linseed) seed, castor bean seed, safflower seed, sorghum seed, peanut seed, and Brassica species seed. The grain crop seed can be, but is not limited to, a cereal grain seed, a pseudocereal grain seed, a pulse grain seed, acom seed, a rice seed, a rye seed, a sorghum seed, a millet seed, a wheat seed, a soybean seed, an oat seed, or a barley seed. The plant can also be a food crop. Food crops include, but are not limited to, vegetable crops. Vegetable crops include, but are not limited to, tomato and letuce.

[0153] Also described are seedlings, plants, and plant parts treated with any of the described synthetic combinations of endophytes. The seedling, plant, and plant part can be any seedling, plant, or plant part treated with any of the described endophyte combinations or any of the described compositions or formulations thereof. The seedling or plant can also be a seedling or plant grown from a seed or plant part treated with any of the described endophyte combinations or any of the described compositions or formulations thereof. The plant part can also be a plant part of a seedling or plant treated with any of the described endophyte combinations or any of the described compositions or formulations thereof. The plant part can also be a plant part of a plant grown from a seedling or seed treated with any of the described endophyte combinations or any of the described compositions or formulations thereof. The plant part can also be a plant part derived from a seedling or plant treated with any of the described endophyte combinations or any of the described compositions or formulations thereof. The plant can be any plant. The plant can be a dicotyledon (dicot) or a monocotyledon (monocot). Tn some embodiments, the plant is an algae. In some embodiments, the plant is a food crop, an oil seed crop, or a grain crop. The oil seed crop can be, but is not limited to: camelina, soybean (Glycine max'), cotonseed, sunflower (Helianthus annuus), canola, rapeseed, flax (linseed), castor beans, safflower (Carthamus tinctorius), sorghum (Sorghum bicolor, Sorghum vulgar e), peanut (Arachis hypogaea), and Brassica species. The grain crop can be, but is not limited to, a cereal grain, a pseudocereal grain, a pulse grain, com, rice (Oryza sativa), rye (Secale cereale), sorghum (Sorghum bicolor, Sorghum vulgare), millet (Pennisetum glaucum), wheat (Triticum aestivum), soybean (Glycine max), oats, or barley. The food crop can be, but is not limited to, a vegetable crop. A vegetable crop can be, but is not limited to, tomato and letuce.

[0154] A plant part refers to any above ground or below ground part or organ of a plant. A plant part can be, but are not limited to, a shoot, a leaf, a blossom, a root, a needle, a stem, a branch, a fruiting body, a fruit, a tuber, a corm, or a rhizome. A plant part can also be avegetative or generative propagating material, such as, but not limited to a cutting, a corm, a rhizome, a tuber, or a runner.

[0155] Additional exemplary plants include, but are not limited to, B. napus. B. rapa. B. juncea, alfalfa (Medicago sativ ), proso millet (Panicum miliaceum), foxtail millet Setaria italica), finger millet (Eleusine coracana), tobacco (Nicotiana tabacum), potato (Solanum tuberosum), cotton (Gossypium barbadense, Gossypium hirsiitum). sweet potato (Ipomoea batatus). cassava (Manihot esculenta), coffee (Coffea spp.), coconut (Cocos nucifera). pineapple (Ananas comosus), citrus trees (Citrus spp.), cocoa (Theobroma cacao), tea (Camellia sinensis), banana (Musa spp.), avocado (Persea americana), fig (Ficus casica), guava (Psidium guajava), mango (Mangifera indica), olive (Olea europaea), papaya (Carica papaya), cashew (Anacardium occidental), macadamia (Macadamia integrifolia), almond (Prunus amygdalus), sugar beets (Beta vulgaris), sugarcane (Saccharum spp.), vegetables (e.g., tomato or lettuce), ornamentals, and conifers.

[0156] Also described are algae species treated with any of the described synthetic combinations of endophytes. The algae species can be any algae species treated with any of the described endophyte combinations or any of the described compositions or formulations thereof.V. Method of use

[0157] Described are methods for increasing plant or algae health and / or yield comprising applying any of the described endophyte combinations or any of the described compositions or formulations thereof to a seed, a plant, a plant part, soil, or an algae. The endophyte combination is applied to the seed, a plant, plant part, soil, or algae in an amount effective to colonize a seed (or a plant grown from the seed), a plant, plant part, soil, or algae. Colonization of a plant with any of the synthetic combination of endophytes as described herein can be carried out before seed germination or at any stage of plant growth. Colonization of a plant indicates the endophytes of the endophyte combination are present inside the plant and in an endophytic relationship with the plant. Increasing plant health and / or yield comprises: decreasing ethylene production, increasing abiotic stress resistance (e.g, increasing salinity tolerance), increasing endogenous production of growth-promoting hormones (e.g., auxin, cytokinin, and gibberellin), increasing nitrogen fixation, increasing root formation, increasing root mass, increasing root function, increasing flowering, increasing, synchronization of flowering, reducing seed shattering, and increasing yield (i.e., plant growth). Increasing yield can include increasing biomass (plant fresh weight or plant dry weight, included fruit weightor number) and / or increasing seed harvest relative to an untreated reference plant that has not been treated with the endophyte population. Increasing algae health and / or yield comprises: decreasing ethylene production, increasing abiotic stress resistance (e.g., increasing salinity tolerance), increasing endogenous production of growth-promoting hormones (e.g., auxin, cytokinin, and gibberellin), increasing nitrogen fixation, and / or increasing yield (i.e., algae growth). Increasing yield can include increasing biomass (algae fresh weight or algae dry weight) relative to an untreated reference algae that has not been treated with the endophyte population.

[0158] Also described are methods for increasing algae health and / or grow th comprising co-culture of the algae with any of the described endophyte combinations.

[0159] In some embodiments, an endophyte combination or a composition or formulation comprising the endophyte combination is applied a seed. The endophyte combination is applied to the seed in an amount effective to colonize a plant grown from the seed.

[0160] In some embodiments, the seeds are contacted with the synthetic combination of endophytes. In some embodiments the seeds are incubated in a solution containing the synthetic combination of endophytes. The seeds can be contacted with the synthetic combination of endophytes or incubated in a solution containing the synthetic combination of endophytes for sufficient time to allow' colonization of the seed. The seeds can be contacted w ith the synthetic combination of endophytes or incubated in a solution containing the synthetic combination of endophytes for about 5 to about 120 minutes. In some embodiments, the seeds are contacted with the synthetic combination of endophytes or incubated in a solution containing the synthetic combination of endophytes for about 30 minutes. In some embodiments, seeds are contacted with the synthetic combination of endophytes for a period of time and washed prior to planting.

[0161] In some embodiments, an endophyte combination or a composition or formulation comprising the endophyte combination is applied a seedling. The endophyte combination is applied to the seedling in an amount effective to colonize the seedling or a plant grown from the seedling.

[0162] In some embodiments, an endophyte combination or a composition or formulation comprising the endophyte combination is applied a plant part. The endophyte combination is applied to the plant part in an amount effective to colonize the plant part of a plant grow n from the plant part.

[0163] In some embodiments, an endophyte combination or a composition or formulation comprising the endophyte combination is applied a plant. The endophyte combination is applied to the plant in an amount effective to colonize the plant.

[0164] In some embodiments, an endophyte combination or a composition or formulation comprising the endophyte combination is applied to soil in which a seed is to be planted, a seed has been planted, or a plant is growing. The endophyte combination is applied to the soil in an amount effective to colonize a plant grown in the soil. In some embodiments, the endophyte combination is applied to the soil prior to planting a seed. In some embodiments, the endophyte combination is applied to the soil after to planting a seed. In some embodiments, the endophyte combination is applied to the soil concomitantly with planting a seed. In some embodiments, the endophyte combination is applied to the soil after a seed planted in the soil has germinated. In some embodiments, the endophyte combination is applied to the soil after a seed planted in the soil has sprouted.

[0165] Also described are methods of growing a plant in a controlled environment comprising contacting or applying any of the described synthetic combinations of endophytes to a seed, plant, plant part, or environment of the seed, plant or plant part and growing the plant in the controlled environment. In some embodiments, the plant is colonized with a synthetic combination of endophytes as described herein and grown in the controlled environment. In some embodiments, the plant is grown in the controlled environment and the synthetic combination of endophytes is provided in the water or a growing medium of the plant or otherwise applied to the plant in the controlled environment. The controlled environment can be, but is not limited to, a greenhouse or a hydroponic system.

[0166] A controlled environment can include one or more of: a means for controlling and / or providing nutrients and / or to the plants, a means for controlling oxygen levels in a growth media (e.g, solid or liquid media), a means for controlling oxygen levels in the air surrounding the plant, a means for controlling and / or adjusting temperature of growth media, water, and / or air surrounding the plants, a means for circulating water, and a means for providing and / or controlling light to the plants.

[0167] In some embodiments, the controlled environment comprises a hydroponic system. In some embodiments, in a hydroponic system, plant roots are submerged in a liquid medium that provides water, oxygen, and nutrients. The hydroponic system can be any hydroponic system known in the art. A hydroponic system can include one or more of: a means for controlling and / or providing nutrients (e.g. , in a liquid growth media) and / or water to the plants, a means for controlling oxygen levels in a liquid growth media and / or water, a means forcontrolling oxygen levels in the air surrounding the plant, a means for controlling and / or adjusting temperature of liquid growth media, water and / or air surrounding the plants, a means for circulating water, and a means for providing and / or controlling light to the plants.

[0168] In some embodiments, a plant is colonized with a synthetic combination of endophytes as described herein prior to growth in the controlled environment. In some embodiments, a plant growing in a controlled environment is contacted with a synthetic combination of endophytes as described herein. In some embodiments, the plant is contracted with the endophyte combination via a liquid growth media.

[0169] In some embodiments, an endophyte combination or a composition or formulation comprising the endophyte combination is applied an algae species. The endophyte combination is applied to the algae species in an amount effective to colonize the algae species.

[0170] The seed, seedling, plant, plant part, or algae can be treated with any of the described synthetic combinations of endophytes. Treating includes any method of applying the endophyte combination, or a composition or formulation comprising the endophyte combination, to the seed, seedling, plant, plant part, soil, or algae. Various methods of applying the endophyte combination are available in the art. Such methods include, but are not limited to, spraying, coating, misting, or in the case of algae, co-culture. The endophyte combination can be applied wet, partially dried (or desiccated), are as a dry powders, pellet, and granule.

[0171] In some embodiments, an endophyte combination is applied to a seed, seedling, plant, plant part, soil, or algae in combination with one or more agrochemically active compounds, one or more biocontrol agents, or combinations thereof. The agrochemically active compound can be, but it not limited to, a fungicide, a bactericide, an insecticide, an acaricide, a nematicide, a molluscicide, a safener, a plant growth regulator, or a fertilizer. The endophyte combination can be applied to the seed, seedling, plant, plant part, soil, or algae prior to an agrochemically active compound and / or biocontrol agent, after an agrochemically active compound and / or biocontrol agent, or concomitantly with an agrochemically active compound and / or biocontrol agent.

[0172] In some embodiments, the endophyte combinations applied to a seed, seedling, plant, plant part, soil, or algae at a rate of about I x lO4to about I x lO9colony forming units (cfu) per mL. In some embodiments, the endophyte combinations applied to a seed, seedling, plant, plant part, soil, or algae at a rate of about I xlO5to about I x lO9colony forming units (cfu) per mL. In some embodiments, the endophyte combinations applied to a seed, seedling, plant, plant part, soil, or algae at a rate of about I xlO6to about I x lO9colony forming units (cfu) per mL. In some embodiments, the endophyte combinations applied to a seed, seedling,plant, plant part, soil, or algae at a rate of about I xlO7to about I x lO9colony forming units (cfu) per mL. In some embodiments, the endophyte combinations applied to a seed, seedling, plant, plant part, soil, or algae at a rate of about I xlO8to about I x lO9colony forming units (cfu) per mL.

[0173] In some embodiments, plants treated with a described endophyte combination (either by treating the plants, seedlings of the plants or parts of the plants), have an increase in biomass of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, when compared with similar plants that were not treated with the endophyte combinations.

[0174] In some embodiments, plants grown from seeds treated with a described endophyte combination, have an increase in biomass of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, when compared with similar plants grown from seed that were not treated with the endophyte combination.

[0175] In some embodiments, algae treated or co-cultured with a described endophyte combination, have an increase in biomass of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, when compared with similar algae that were not treated or co-cultured with the endophyte combinations.

[0176] In some embodiments, plants treated with a described endophyte combination (either by treating the seeds, plants, seedlings of the plants or parts of the plants), have an increase in seed or fruit production of at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, when compared with similar plants that were not treated with the endophyte combinations.

[0177] In some embodiments, treating plants with a described endophyte combination (either by treating the seeds, plants, seedlings of the plants or parts of the plants), results in decreased seed shattering. The decrease in seed shattering can be at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, or at least 80%, when compared with similar plants that were not treated with the endophyte combinations.

[0178] In some embodiments, treating plants with a described endophyte combination (either by treating the seeds, plants, seedlings of the plants or parts of the plants), results in decreased heterogeneity of seed ripening in a field. Heterogeneity of seed ripening can bereduced by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%. at least 70%, or at least 80%. when compared with similar plants that were not treated with the endophyte combinations.

[0179] Reducing seed shattering or reducing heterogeneity of seed ripening in a field can improve harvesting efficiency and or costs, by eliminating or reducing steps typically taken to account for seed shattering or heterogeneity of seed ripening. Such steps include, but are not limited to, swathing, and use of chemicals to reduce shattering.

[0180] Various aspects of the invention are further described and illustrated by way of the several examples which follow, none of which are intended to limit the scope of the invention.EXAMPLES

[0181] Example 1. Endophyte combinations improve seedling growth in Camelina and Sorghum.

[0182] Various endophyte combinations, included a Bacillus species, a Pseudomonas species and an Bradyrhizobium species, were analyzed for their effect on improving seedling growth in Camelina sativa and Sorghum. Camelina w as used as a multipurpose model oilseed crop. Camelina seed or plants were treated with 16 individual endophytes known to produce ACC deaminase (an ethylene reducing enzyme) and other plant hormones. Endophytes that improved the Camelina root phenotype were selected and tested in various combinations as listed in Table 1. Camelina and Sorghum treated with the four combinations exhibited dramatic improvements in seedling growth (FIG. 3A-B). The treated Camelina showed significantly improved root phenotype, seedling fresh w eight, and had higher number of leaves per seedling (FIG. 3A). Soaking the seeds with the endophyte combinations led to root colonization (FIG. 3C). Root colonization was shown using a Pseudomonas endophyte tagged with a sfGFP. The data demonstrate the described endophyte combination strongly improve plant grow th.Example 2. Endophyte combinations improve growth of algae.

[0183] Algal strains T. striata and P. celeri were treated with six endophyte strains screened for growth promotion under 1% CO2 supplementation and grown in an emersed photobioreactor (ePBR). Effect of the endophyte combinations on algal biomass productivity, quality, and resilience was analyzed. Three beneficial endophyte strains and endophyte combinations were identified that improved growth in the two algal strains. The results indicated that algae treated with the endophyte combinations exhibited a clear increase inbiomass productivity (as much as 80%, FIGs. 1 and 2). The increase in biomass indicates an increase in productivity and carbon storage (e.g, lipid and carbohydrate storage).

[0184] Because the endophyte combinations colonize and improve plant health and productivity in both Camelina and algae, it is expected the endophyte combinations will improve plant growth in any plant.Example 3. Testing of endophyte combinations.

[0185] Various endophyte combinations were analyzed to determine their effect on plant grow th. Endophyte combinations as shown in Table 1 were applied to the seeds and the seeds were planted in pots (soil) or germinated on plates. Plants were then monitored for germination and growth under a variety of conditions.Table 1. Endophyte combinations.X = complete experiment ** = experiment in progress

[0186] For plants grown in soil from treated seeds, an increase in growth and biomass was observed. Improvements in plant height of 15-20% was observed relative to plants grown from seed not treated with the endophyte combinations (untreated seeds). Increases in tap root size and branching of roots was also observed. Root fresh weight was improved on average 1.7-fold relative to plants grown from seed not treated with the endophyte combinations.

[0187] For seeds germinated on plates, germination, and growth under salinity' stress (100 rnM salt) was measured. Plants germinated and grown from treated seeds exhibited higher germination levels and increased plant growth in 100 mM salt relative to plants germinated and grown from untreated seeds under the same conditions.Example 4. Growth of seed treated with endophytes.

[0188] Forage sorghum (Sorghum vulgare) or Camelina (Camelina sativa) seed were sanitized and then inoculated with combination of bacterial endophytes as shown in Table 2. Seeds were submerged in 70% ethanol for one minute, then soaked in 10% bleach solution for 10 minutes to sanitize the seed surface. Following sanitization, seeds were rinsed five times with sterile water to wash away residual bleach, then soaked in mixtures of bacterial cultures (endophytes) or sterile LB bacterial growth medium for 30 minutes. Inoculated seedswere then rinsed once with sterile water to remove residual bacterial culture and growth medium. Combinations of bacterial cultures as listed in Table 2 were prepared with equal volumes of culture. For all combinations, bacterial cultures used were at optical densities (X=600nm) ranging from 0.4-0.9 and cell density ranging from about l * 108to 1 *109cells mL ’.Table 2. Endophyte combinations

[0189] Inoculated seeds were planted into one of two experimental systems: petri plates or potting soil. Petri plates used for short-term growth and germination assays were prepared by adding 7 mL of sterile water or 100 mM sodium chloride solution and an autoclaved paper disk or the agar media (prepared with half strength Murashige and Skoog Media) to the plate. For germination assays, ten seeds were placed onto the saturated paper in the plate. For growth on agar plates, seeds were placed vertically on agar surface after germination. Plates were sealed with parafilm and incubated under illumination at 25°C. Following 7-10 days of growth, seedlings were photographed and seedling size, development, and germination rate were compared. For medium-term growth experiments in the laboratory', small pots were filled with nonsterile potting soil (Magik-Moss Brand) to grow control and inoculated seeds to evaluate the effect of the indicated endophyte combinations on plant growth under natural environment (as the potting soil contained different microbial populations and w as not sterilized). Prior to planting, soil was hydrated to near saturation with water, or 100 mM sodium chloride solution to load salt into the growth medium. Three inoculated seeds were planted per pot, approximately 1 cm deep. Pots were covered with plastic wrap until emergence. Followingemergence, each pot was watered with an equal volume of water 2 to 3 times per week, so as to prevent saturation of the soil and flushing of the salt from the pot. Plants were grown under 150 pE light intensity on a 14 / 10 (hours) light / dark cycle at room temperature for about one month. Following the grow th period, plants were photographed (FIG. 4), and plant height, fresh shoot and root biomass, dry shoot and root biomass were measured. Root morphology' and length was measured and calculated from root scans using the WinRhizo program.Example 5. Growth of seed treated with endophytes.

[0190] Sorghum seeds w ere sanitized with 70% ethanol and 10% bleach, rinsed, then added to one of three different endophyte combinations of Table 3 and allowed to soak for 30 minutes. Each endophyte culture was grown in LB broth for 48 h at 25°C, shaking at 160 rpm then diluted in fresh LB to the indicated density. For all combinations, bacterial cultures used were at optical densities (1 = 600 nm) ranging from 0.4-0.9 and cell density ranging from about I x lO8to I x lO9cells mL1. Inoculated seeds were planted in commercial potting soil (non- sterile) and 100 mL water was applied after planting. Plants were watered 3x weekly with 50 mL water. Plants were grown in the laboratory under 110-140 pE light, with the LED lights powered on 16 h per day. Plants were measured and destructively sampled 22 days after planting.

[0191] Results are shown in FIG. 4 and FIG. 5. Endophyte treatment of seeds resulted in increased root fresh weight, increased shoot fresh w eight, increase plant height, and increased dry shoot weight. Treatments were statistically compared to the control using t-tests. Stars indicate significance level (* <0.05; ** <0.01).Table 3.Example 6. Endophyte treatment increases salt tolerance in Sorghum.

[0192] Sorghum seeds were sanitized with 70% ethanol and 10% bleach, rinsed, then added to one of three different endophyte combinations of Table 3 and allowed to soak for 30 minutes.Each endophyte culture was grown in LB broth for 48 h at 25°C, shaking at 160 rpm then diluted in fresh LB to the indicated density. Inoculated seeds were planted in commercial potting soil (non-sterile) and 100 mL of 0. IM sodium chloride solution was applied for salinity to both treated and untreated sorghum after planting. Plants were watered 3x weekly with 50 mL water. Plants were grown in the laboratory under 110-140 pE light, with the LED lights powered on 16 h per day. Plants were measured and destructively sampled 22 days after planting.

[0193] Results are shown in FIG. 6. Endophyte treatment of seeds with combinations 1 and 3 resulted in increased root fresh weight, increased shoot fresh weight, increased plant height, and increased dry' shoot weight. Endophyte treatment of seeds with combination 2 resulted in increased root fresh weight and increased shoot fresh weight. Treatments were statistically compared to the control using t-tests. Stars indicate significance level (* <0.05; ** <0.01).Example 7. Sorghum greenhouse experiments.

[0194] Sorghum seeds were sanitized with 70% ethanol and 10% bleach, rinsed, then added to one of three different endophyte combinations and allowed to soak for 30 minutes. Each endophyte culture was grown in LB broth for 48 h at 25°C, shaking at 160 rpm then diluted in fresh LB to the indicated density. Inoculated seeds were planted in commercial potting soil (non-sterile) and 600 mL of either fresh water or lOOmM NaCl solution was applied after planting to setup the standard and high-salinity treatments respectively. Plants were grown in the greenhouse for 10 weeks. No extra fertilization was used.

[0195] Results for plants grown under normal conditions are shown in FIG. 7 for endophyte combination #2. Endophyte treatment of seeds resulted in plants having increased biomass.

[0196] Results for plants grown under salt stress conditions are shown in FIG. 8 for endophyte combination #2. Endophyte treatment of seeds resulted in plants having increased biomass, indicating increased salt tolerance.Example 8. Plant-endophyte metabolite data.

[0197] We performed metabolite analysis in control and endophyte treated tomato seedlings. Several metabolites were identified showing significantly different expression in tomato treated with endophyte combinations (FIGs. 9-11). The identified differentially expressed metabolites included biocontrol agents. The biocontrol agents included bioactive compounds (succinimide), antimicrobials (quercetin, toxopyrimidine, timonacic. isorhamnetin. and isoquercetin), antioxidants (sinapine) and pest and weed control agents (thiophanate, terpin,and pseudopelletierine) and insect control agents (precocene II). Interestingly, we found some compounds specific to either root or stem with pesticide, herbicide, and insecticide properties along with many antimicrobial compounds and these results show endophytes' potential as a broad-spectrum biocontrol product.Example 9. Algae co-culture metabolite data.

[0198] We performed metabolite analysis in control and endophyte treated algae co-cultures. Picochlorum and Tetraselmis were separately co-cultured with Bacillus megaterium + Rhizobium phaseoli (BM+RP). Several metabolites were identified showing significantly different expression in algae co-cultured with endophyte combinations. Results are shown in Table 4 and FIG. 12.Table 4. Algal metabolites:Example 10. Hydroponics growth data

[0199] Tomato and letuce seeds were sanitized with 70% ethanol and 10% bleach, rinsed, then added to one of three different endophyte combinations as in Table 3 and allowed to soak for 30 minutes. Each endophyte culture was grown in LB broth for 48 h at 25°C, shaking at 160 rpm then diluted in fresh LB to the indicated density. Inoculated seeds were planted in small germination plugs. After seed germination, seedlings were transferred to hydroponics systems. Plants were grown for 8 weeks in hydroponics systems.

[0200] Results for letuce plants grown hydroponically are shown in FIG. 13 for endophyte combination (AV+RL+AO+BL). Endophyte treatment of seeds resulted in plants having increased biomass. Treatments were statistically compared to the control using t-tests. Stars indicate significance level (* <0.05).

[0201] Results for tomato plants grown hydroponically are shown in FIG. 14 for endophyte combination (AV+RL+AO+BL). Endophyte treatment of seeds resulted in plants having increased biomass. Treatments were statistically compared to the control using t-tests. Stars indicate significance level (* <0.05).

[0202] As shown in FIG. 15, endophyte treatment of tomato seeds also increased flowering in tomato plants.

Claims

Claims:

1. A synthetic combination of endophytes for improving plant or algae grow th comprising two or more endophyte species selected from:(a) at least one first endophyte species the reduces ethylene concentration in the plant;(b) at least one second endophyte species that increases endogenous levels of growth-promoting hormones; and(c) at least one third endophyte species with nitrogen fixation capability.

2. The synthetic combination of endophytes of claim 1, wherein(a) the at least one first endophyte species is selected from the group consisting of: a Bacillus species and a Pseudomonas species;(b) the at least one second endophyte species is selected from the group consisting of: aRhizobium species, a Pseudomonas species, and an Azotobacter species; and(c) the at least one third endophyte species is selected from the group consisting of: aRhizobium species, aMesorhizobium species, an Arthrobacter species, an Azospirillum species, an Azotobacter species, a Sinorhizobium species, and a Bradyrhizobium species.

3. The synthetic combination of endophytes of claim 2, wherein (a) the at least one first endophyte comprises Bacillus licheniformis, Bacillus megaterium, Bacillus subtilis, Pseudomonas puiida. or Pseudomonas fluorescens,' (b) the at least one second endophyte comprises a Rhizobium leguminosarum, Rhizobium phaseoli, Pseudomonas putida. Pseudomonas fluorescens , Azotobacter chroococcum, or Azotobacter vinelandii,' and (c) the at least one third endophyte comprises Rhizobium leguminosarum, Rhizobium phaseoil. Mesorhizobium amorphae, Arthrobacter oryzae, Azospirillum brasilcnse. Azotobacter chroococcum, Azotobacter vinelandii, or Sinorhizobium meliloti.

4. A synthetic combination of endophytes for improving plant or algae grow th comprising two or more endophyte species selected from the group comprising: a Arthrobacter oryzae, a Azospirillum brasilense, aAzotobacter chroococcum, aAzotobacter vinelandii, a Bacillus licheniformis, a Bacillus megaterium, a Bacillus subtilis, a Mesorhizobium amorphae, a Pseudomonas putida, a Pseudomonas fluorescens, a Rhizobium leguminosarum, aRhizobium phaseoli, and a Sinorhizobium meliloti.

5. The synthetic combination of endophytes of claim 4, wherein the synthetic combination of endophytes comprises:(a) at least one Azotobacter species, at least one Rhizobium species, at least one Arthrobacter species, and at least one Bacillus species;(b) at least one Rhizobium species, at least one Bacillus species, at least one Azospirillum species, and at least one Azotobacter species;(c) at least one a Bacillus species or Pseudomonas species and at least one Rhizobium species or Azotobacter species;(d) at least one a Bacillus species and at least one Pseudomonas species;(e) at least one Rhizobium species and at least Azotobacter species;(!) at least one Arthrobacter species and at least one Sinorhizobium species; or(g) at least one Azotobacter species, at least one Rhizobium species, and at least one Arthrobacter species.

6. The synthetic combination of endophytes of claim 4, wherein the synthetic combination of endophytes comprises at least two, at least three, at least four, at least five, at least six, at least seven, or at least eight endophytes selected from the group consisting of: Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), Bacillus licheniformis (ATCC #14580). Bacillus megaterium (ATCC #14581), Bacillus subtilis (ATCC #6051), Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonas putida (ATCC #12633), Pseudomonas fluorescens (ATCC #13525), Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), Sinorhizobium meliloti (ATCC #9930), and a species derived from any of Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), Bacillus licheniformis (ATCC #14580), Bacillus megaterium (ATCC #14581), Bacillus subtilis (ATCC #6051), Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonas putida (ATCC #12633). Pseudomonas fluorescens (ATCC #13525), Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), or Sinorhizobium meliloti (ATCC #9930) wherein the derived species retains at least one function of the original strain.

7. The synthetic combination of endophytes of claim 4, wherein the synthetic combination of endophytes comprises:(a) Azotobacter vinelandii, Rhizobium leguminosarum, Arthrobacter oryzae, and Bacillus licheniformis',(b) Rhizobium leguminosarum. Bacillus licheniformis, Azospirillum brasilense. and Azotobacter chroococcum;(c) Azotobacter chroococcum. Rhizobium leguminosarum, and Arthrobacter oryzae,'(d) Sinorhizobium meliloti. Azotobacter vinelandii, and Bacillus licheniformis(e) Sinorhizobium meliloti, Azotobacter chroococcum, and Rhizobium leguminosarum.'(f) Arthrobacter oryzae, Sinorhizobium meliloti, and Azospirillum brasilense,(g) Sinorhizobium meliloti, Rhizobium leguminosarum, Mesorhizobium amorphae, and Arthrobacter oryzae;(h) Azotobacter chroococcum, Rhizobium leguminosarum, Arthrobacter oryzae, Bacillus licheniformis, Sinorhizobium meliloti, Azotobacter vinelandii, Azospirillum brasilense, and Mesorhizobium amorphae;(i) Bacillus subtilis and Pseudomonas fluorescens,'(j) Bacillus subtilis and Rhizobium phaseoli;(k) Pseudomonas fluorescens and Rhizobium phaseoli;(l) Bacillus subtilis, Pseudomonas fluorescens, and Rhizobium phaseoli;(m) Bacillus Megaterium. Bacillus subtilis, Pseudomonas fluorescens, and Pseudomonas putida; or(n) Bacillus Megaterium and Rhizobium phaseoli.

8. The synthetic combination of endophytes of claim 7, wherein the synthetic combination of endophytes comprises:(a) Azotobacter vinelandii (ATCC #478), Rhizobium leguminosarum (ATCC #10004), Arthrobacter oryzae (ATCC #42149), and Bacillus licheniformis (ATCC #14580);(b) Rhizobium leguminosarum (ATCC #10004), Bacillus licheniformis (ATCC #14580), Azospirillum brasilense (ATCC #29729), and Azotobacter chroococcum (ATCC #9043);(c) Azotobacter chroococcum (ATCC #9043), Rhizobium leguminosarum (ATCC #10004), and Arthrobacter oryzae (ATCC #42149);(d) Sinorhizobium meliloti (ATCC #9930). Azotobacter vinelandii (ATCC #478), and Bacillus licheniformis (ATCC #14580);(e) Sinorhizobium meliloti (ATCC #9930), Azotobacter chroococcum (ATCC #9043). and Rhizobium leguminosarum (ATCC #10004).(f) Arthrobacter oryzae (ATCC #42149), Sinorhizobium meliloti (ATCC #9930), and Azospirillum brasilense (ATCC #29729);(g) Sinorhizobium meliloti (ATCC #9930), Rhizobium leguminosarum (ATCC #10004), Mesorhizobium amorphae (ATCC #BAA-114), and Arthrobacter oryzae (ATCC #42149);(h) Azotobacter chroococcum (ATCC #9043), Rhizobium leguminosarum (ATCC #10004), Arthrobacter oryzae (ATCC #42149), Bacillus licheniformis (ATCC #14580), Sinorhizobium meliloti (ATCC #9930), Azotobacter vinelandii (ATCC #478), Azospirillum brasilense (ATCC #29729), and Mesorhizobium amorphae (ATCC #BAA-114);(i) Bacillus subtilis (ATCC #6051) and Pseudomonas fluorescens (ATCC #13525);(j) Bacillus subtilis (ATCC #6051) and Rhizobium phaseoli (ATCC #31236);(k) Pseudomonas fluorescens (ATCC #13525) and Rhizobium phaseoli (ATCC #31236);(l) Bacillus subtilis (ATCC #6051), Pseudomonas fluorescens (ATCC #13525), and Rhizobium phaseoli (ATCC #31236);(m) Bacillus Megaterium (ATCC #14581), Bacillus subtilis (ATCC #6051), Pseudomonas fluorescens (ATCC #13525), and Pseudomonas putida (ATCC #12633); or(n) Bacillus Megaterium (ATCC #14581) and Rhizobium phaseoli (ATCC #31236).

9. The synthetic combination of endophytes of any one of claims 1-8, wherein the synthetic combination of endophytes further comprises one or more of: an excipient, an agrochemically active compound, and a biocontrol agent.

10. The synthetic combination of endophytes of any one of claims 1-8, wherein the synthetic combination of endophytes is formulated as a liquid, a dispersion, a colloidal dispersion, a solution, a suspension, a colloidal suspension, an emulsion, a foam, a slurry', a lyophilized cake or powder, a spray-dried powder, a pellet, a biologically pure pellet, a coated pellet, a powder, a flowable powder, or a granule.

11. The synthetic combination of endophytes of claim 10, wherein synthetic combination of endophytes is formulated as a liquid, a dispersion, a colloidal dispersion, a solution, asuspension, a colloidal suspension, an emulsion, or a slurry at a concentration of about 1 x 104to about 1 x 109colony forming units (cfu) per mL, about 1 x 105to about 1 x 109cfu per mL, about 1 x 106to about 1 x 109cfu per mL, about 1 x 107to about 1 x 109cfu per mL, or about 1 x 108to about 1 x 109cfu per mL.

12. The synthetic combination of endophytes of any one of claims 1-11, wherein synthetic combination of endophytes is formulated for:(a) application to a seed in an amount effective to colonize a plant germinated from the seed with the synthetic combination of endophytes;(b) application to a seedling, a plant, or a leaf of the plant in an amount effective to colonize the seedling or the plant with the synthetic combination of endophytes;(c) application to an environment of a seed, plant, or plant part in an amount effective to colonize a plant grown in the environment with the synthetic combination of endophytes; or(d) coating the surface of the seed; wherein the seed is a monocot seed or a dicot seed or the plant is a monocot plant or a dicot plant.

13. The synthetic combination of endophytes of any one of claims 1-12, wherein synthetic combination of endophytes:(a) increases plant biomass production, increases root biomass, increases root length, or increases yield in a plant colonized by the synthetic combination of endophytes relative to an untreated reference plant that has not been colonized by synthetic combination of endophytes;(b) increases synchronization of flowering or decreases seed shattering in a plant colonized by the synthetic combination of endophytes relative to an untreated reference plant that has not been colonized by the endophyte population;(c) increases flowing in a plant colonized by the synthetic combination of endophytes relative to an untreated reference plant that has not been colonized by the endophyte population and / or(d) increases abiotic stress resistance, increases salt tolerance, increases pH tolerance, or increases heat tolerance in a plant colonized by the synthetic combination of endophytes relative to an untreated reference plant that has not been colonized by the endophyte population.

14. The method of claim 13, wherein the plant is an agricultural crop plant, optionally wherein the agricultural crop plant is selected from the group consisting of: wheat, barley, rice, millet, oats, triticale, rye, bamboo, sugarcane plant, soybean, canola, rapeseed, cotton, alfalfa, cassava, potato, tomato, pea, chickpea, lentil, flax, and pepper.

15. The synthetic combination of endophytes any one of claims 1-1 1, wherein synthetic combination of endophytes is formulated for application to algae or co-culture with algae.

16. The synthetic combination of endophytes of claim 16. wherein synthetic combination of endophytes:(a) increases biomass production or increases yield of algae colonized by the synthetic combination of endophytes relative to an untreated reference algae that has not been colonized by the endophyte population; and / or(b) increases abiotic stress resistance, increases salt tolerance, increases pH tolerance, and / or increases heat tolerance of algae colonized by the synthetic combination of endophytes relative to an untreated reference algae that has not been colonized by the endophyte population.

17. A synthetic combination of endophytes for improving plant or algae growth comprising tw o or more endophyte species selected from the group comprising: Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelardii (ATCC #478), Bacillus licheniformis (ATCC #14580), Bacillus Megaterium (ATCC #14581), Bacillus subtilis (ATCC #6051), Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonas putida (ATCC #12633), Pseudomonas fluorescens (ATCC #13525), Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), Sinorhizobium meliloti (ATCC #9930), and a species derived from any of Arthrobacter oryzae (ATCC #42149), Azospinllum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), Bacillus licheniformis (ATCC #14580), Bacillus megaterium (ATCC #14581), Bacillus subtilis (ATCC #6051), Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonas putida (ATCC #12633). Pseudomonas fluorescens (ATCC #13525), Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), ox Sinorhizobium meliloti (ATCC #9930) wherein the derived species retains at least one function of the original strain.

18. The synthetic combination of endophytes of claim 17, wherein the synthetic combination of endophytes comprises:(a) at least one of the Bacillus species or the Pseudomonas species and at least one of the Rhlzobium species or the Azotobacter species;(b) at least one of the Bacillus species and at least one of the Pseudomonas species;(c) at least one of the Rhizobium species and at least one of the Azotobacter species:(d) the Arthrobacter species and the Sinorhizobium species;(e) at least one Azotobacter species, at least one Rhizobium species, and at least one Arthrobacter species;(1) at least one Rhizobium species, at least one Bacillus species, at least one Azospirillum species, and at least one Azotobacter species;(g) at least one Azotobacter species, at least one Rhizobium species, at least one Arthrobacter species, and at least one Bacillus species; or(h) at least one first endophyte comprising one or more of Bacillus licheniformis, Bacillus Megaterium, Bacillus subtilis, Pseudomonas fluorescens, and Pseudomonas putida.’ at least one second endophyte comprising one or more of Rhizobium leguminosarum. Rhizobium phaseoli, Pseudomonas fluorescens, Pseudomonas putida, Azotobacter chroococcum, and Azotobacter vinelandii,’ and at least one third endophyte comprising one or more of Rhizobium leguminosarum, Rhizobium phaseoli, Mesorhizobium amorphae, Arthrobacter oryzae. Azospirillum brasilense. Azotobacter chroococcum. Azotobacter vinelandii, and Sinorhizobium meliloti.

19. The synthetic combination of endophytes of claim 17 or 18, wherein the synthetic combination of endophytes is formulated at a concentration of about 1 * 104to about 1 x 109cfu per mL, about 1 x 105to about 1 x 109cfu per mL, about 1 x 106to about 1 x 109cfu per mL, about 1 x 107to about 1 x 109cfu per mL, or about 1 x 108to about 1 x 109cfu per mL.

20. A method of improving plant or algae growth comprising applying a synthetic combination of endophytes to a seed, a plant, a plant part, the algae, or an environment of the seed, plant, plant or the algae in an amount effective to colonize the plant, a plant grown from the seed, or the algae, wherein the synthetic combination of endophytes comprises the synthetic combination of endophytes of any one of claims 1-19.

21. A method of improving plant or algae growth comprising applying a synthetic combination of endophytes to a seed, a plant, a plant part, the algae, or an environment of theseed, plant, plant or the algae in an amount effective to colonize the plant, a plant grown from the seed, or the algae, wherein the synthetic combination of endophytes comprises two or more endophyte species selected from the group comprising: a Arthrobacter oryzae. a Azospirillum brasilense, a Azotobacter chroococcum. a Azotobacter vinelandii, a Bacillus licheniformis, a Bacillus megaterium, a Bacillus subtilis, aMesorhizobium amorphae. a Pseudomonas put Ida. a Pseudomonas fluorescens, aRhizobium leguminosarum, aRhizobium phaseoli. and a Sinorhizobium meliloti.

22. The method of claim 21, wherein the synthetic combination of endophytes comprises:(a) at least one Azotobacter species, at least one Rhizobium species, at least one Arthrobacter species, and at least one Bacillus species;(b) at least one Rhizobium species, at least one Bacillus species, at least one Azospirillum species, and at least one Azotobacter species;(c) at least one a Bacillus species or Pseudomonas species and at least one Rhizobium species or Azotobacter species;(d) at least one a Bacillus species and at least one Pseudomonas species;(e) at least one Rhizobium species and at least Azotobacter species;(f) at least one Arthrobacter species and at least one Sinorhizobium species; or(g) at least one Azotobacter species, at least one Rhizobium species, and at least one Arthrobacter species.

23. The method of claim 21, wherein (a) the at least one first endophyte comprises Bacillus licheniformis, Bacillus megaterium, Bacillus subtilis. Pseudomonas putida, or Pseudomonas fluorescens: (b) the at least one second endophyte comprises aRhizobium leguminosarum, Rhizobium phaseoli. Pseudomonas putida, Pseudomonas fluorescens, Azotobacter chroococcum, or Azotobacter vinelandii,' and (c) the at least one third endophyte comprises Rhizobium leguminosarum, Rhizobium phaseoli, Mesorhizobium amorphae. Arthrobacter oryzae. Azospirillum brasilense. Azotobacter chroococcum. Azotobacter vinelandii, or Sinorhizobium meliloti.

24. The method of claim 21, wherein the synthetic combination of endophytes comprises at least two, at least three, at least four, at least five, at least six. at least seven, or at least eight endophytes selected from the group consisting of: Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), Bacillus licheniformis (ATCC #14580),Bacillus megaterium (ATCC #14581). Bacillus subtilis (ATCC #6051), Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonas putida (ATCC #12633), Pseudomonas fluorescen (ATCC #13525), Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), Sinorhizobium meliloti (ATCC #9930), and a species derived from any of Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), Bacillus licheniformis (ATCC #14580), Bacillus megaterium (ATCC #14581), Bacillus subtilis (ATCC #6051), Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonas putida (ATCC #12633), Pseudomonas fluorescens (ATCC #13525), Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), Sinorhizobium meliloti (ATCC #9930) wherein the derived species retains at least one function of the original strain.

25. The method of claim 21, wherein the synthetic combination of endophytes comprises:(a) Azotobacter vinelandii, Rhizobium leguminosarum, Arthrobacter oryzae, and Bacillus licheniformis,'(b) Rhizobium leguminosarum. Bacillus licheniformis. Azospirillum brasilense, and Azotobacter chroococcum,'(c) Azotobacter chroococcum, Rhizobium leguminosarum, and Arthrobacter oryzae,'(d) Sinorhizobium meliloti. Azotobacter vinelandii, and Bacillus licheniformis,'(e) Sinorhizobium meliloti, Azotobacter chroococcum. and Rhizobium leguminosarum,'(1) Arthrobacter oryzae, Sinorhizobium meliloti, and Azospirilliim brasilense,(g) Sinorhizobium meliloti, Rhizobium leguminosarum, Mesorhizobium amorphae, and Arthrobacter oryzae.'(h) Azotobacter chroococcum, Rhizobium leguminosarum, Arthrobacter oryzae, Bacillus licheniformis, Sinorhizobium meliloti, Azotobacter vinelandii, Azospirillum brasilense, and Mesorhizobium amorphae,(i) Bacillus subtilis and Pseudomonas fluorescens,'(j) Bacillus subtilis and Rhizobium phaseoli,'(k) Pseudomonas fluorescens and Rhizobium phaseoli,'(l) Bacillus subtilis, Pseudomonas fluorescens, and Rhizobium phaseoli,'(m) Bacillus Megaterium. Bacillus subtilis, Pseudomonas fluorescens, and Pseudomonas putida,' or(n) Bacillus Megaterium and Rhizobium phaseoli.

6. The method of claim 25, wherein the synthetic combination of endophytes comprises:(a) Azotobacter vinelandii (ATCC #478), Rhizobium leguminosarum (ATCC #10004), Arthrobacter oryzae (ATCC #42149), and Bacillus licheniformis (ATCC #14580);(b) Rhizobium leguminosarum (ATCC #10004), Bacillus licheniformis (ATCC #14580), Azospirillum brasilense (ATCC #29729), and Azotobacter chroococcum (ATCC #9043);(c) Azotobacter chroococcum (ATCC #9043), Rhizobium leguminosarum (ATCC #10004), and Arthrobacter oryzae (ATCC #42149);(d) Sinorhizobium meliloti (ATCC #9930). Azotobacter vinelandii (ATCC #478), and Bacillus licheniformis (ATCC #14580);(e) Sinorhizobium meliloti (ATCC #9930), Azotobacter chroococcum (ATCC #9043), mA Rhizobium leguminosarum (ATCC #10004).(f) Arthrobacter oryzae (ATCC #42149), Sinorhizobium meliloti (ATCC #9930), mA Azospirillum brasilense (ATCC #29729);(g) Sinorhizobium meliloti (ATCC #9930). Rhizobium leguminosarum (ATCC #10004), Mesorhizobium amorphae (ATCC #BAA-114), and Arthrobacter oryzae (ATCC #42149);(h) Azotobacter chroococcum (ATCC #9043), Rhizobium leguminosarum (ATCC #10004), Arthrobacter oryzae (ATCC #42149), Bacillus licheniformis (ATCC #14580), Sinorhizobium meliloti (ATCC #9930), Azotobacter vinelandii (ATCC #478), Azospirillum brasilense (ATCC #29729), and Mesorhizobium amorphae (ATCC #BAA-114);(i) Bacillus subtilis (ATCC #6051) and Pseudomonas fluorescens (ATCC #13525);(j) Bacillus subtilis (ATCC #6051) mARhizobium phaseoli (ATCC #31236);(k) Pseudomonas fluorescens (ATCC #13525) and Rhizobium phaseoli (ATCC #31236);(l) Bacillus subtilis (ATCC #6051), Pseudomonas fluorescens (ATCC #13525), and Rhizobium phaseoli (ATCC #31236);(m) Bacillus Megaterium (ATCC #14581), Bacillus subtilis (ATCC #6051), Pseudomonas fluorescens (ATCC #13525), and Pseudomonas putida (ATCC #12633); or(n) Bacillus Megaterium (ATCC #14581) and Rhizobium phaseoli (ATCC #31236).

27. The method of any one of claims 21-25, wherein the endophyte combination is applied to the seed, the seedling, the plant, the plant part, the algae, or the environment of the seed, plant, plant or algae at a rate of about 1 x 104to about 1 x 109colony forming units (cfu) per mL, about 1 x 105to about 1 x 109cfu per mL, about 1 x 106to about 1 x 109cfu per mL, about 1 x 107to about 1 x 109cfu per mL, or about 1 x 108to about 1 x 109cfu per mL.

28. The method of any one of claims 21-27, wherein the synthetic combination of endophytes is:(a) formulated with one or more excipients, one or more agrochemically active compounds, one or more biocontrol agents, or combinations thereof prior to applying the synthetic combination of endophytes to the seed, plant, plant part, algae, or the environment of the seed, plant, plant part, or algae; and / or(b) provided as a liquid, a dispersion, a colloidal dispersion, a solution, a suspension, a colloidal suspension, an emulsion, a foam, a slurry, a lyophilized cake or powder, a spray- dried powder, a pellet, a biologically pure pellet, a coated pellet, a powder, a flowable powder, a granule, or a hydroponic growth media, prior to applying the synthetic combination of endophytes to the seed, plant, plant part, algae, or the environment of the seed, plant, plant part, or algae.

29. The method of claim 28, wherein liquid, the dispersion, the colloidal dispersion, the solution, the suspension, the colloidal suspension, the emulsion, the foam, or the slurry contains the synthetic combination of endophytes at a concentration of about 1 x 104to about 1 x 109colony forming units (cfu) per mL, about 1 x 1 (p to about 1 x 109cfu per mL, about1 x 106to about 1 x 109cfu per mL, about 1 x 107to about 1 x 109cfu per mL, or about 1 x 108to about 1 x 109cfu per mL.

30. The method of claim 21, wherein the plant or algae comprises the plant, and the method comprises:(a) applying the synthetic combination of endophytes to a seed in an amount effective to colonize a plant germinated from the seed with the synthetic combination of endophytes;(b) applying the synthetic combination of endophytes to a seedling, a plant, or a leaf of the plant in an amount effective to colonize the seedling or the plant with the synthetic combination of endophytes;(c) applying the synthetic combination of endophytes to an environment of a seed, plant, or plant part in an amount effective to colonize a plant grown in the environment with the synthetic combination of endophytes: or(d) coating the surface of a seed with a formulation comprising the combination of endophytes.

31. The method of any one of claims 21-30, wherein the plant colonized with the synthetic combination of endophytes has:(a) increased plant biomass production, increased root biomass, increased root length, or increased yield, relative to an untreated reference plant that has not been colonized with the endophyte population;(b) increased synchronization of flow ering or decreased seed shattering relative to an untreated reference plant that has not been colonized with the endophyte population; and / or(c) increased abiotic stress resistance, increased salt tolerance, increased pH tolerance, or increased heat tolerance, relative to an untreated reference plant that has not been colonized with the endophyte population.

32. The method of claim 30 or 31, wherein the plant is a monocot plant or a dicot plant.

33. The method of claim 32, wherein the plant is an agricultural crop plant, optionally wherein the agricultural crop plant is selected from the group consisting of: wheat, barley, rice, millet, oats, triticale, rye, bamboo, sugarcane plant, soybean, canola, rapeseed, cotton, alfalfa, cassava, potato, tomato, pea, chickpea, lentil, flax, and pepper.

34. The method of 32 or 33, wherein the plant is grown in a field or a controlled environment, optionally wherein the controlled environment comprises a hydroponic system.

35. The method of any one of claims 21-29, wherein the plant or algae comprises the algae.

36. The method of claim 35, wherein the algae colonized with the synthetic combination of endophytes has:(a) increased biomass production or increased yield relative to an untreated reference algae that has not been colonized with the endophyte population; and / or(b) increased abiotic stress resistance, increased salt tolerance, increased pH tolerance, and / or increased heat tolerance, relative to an untreated reference algae that has not been colonized with the endophyte population.

37. A method of improving plant or algae grow th comprising applying a synthetic combination of endophytes to a seed, a plant, a plant part, the algae, or an environment of the seed, plant, plant or the algae in an amount effective to colonize the plant, a plant grown from the seed, or the algae, wherein the synthetic combination of endophytes comprises two or more endophyte species selected from the group comprising: Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478). Bacillus licheniformis (ATCC #14580). Bacillus Megaterium (ATCC #14581), Bacillus subtilis (ATCC #6051), Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonas putida (ATCC #12633), Pseudomonas fluorescens (ATCC #13525), Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), Sinorhizobium meliloti (ATCC #9930), and a species derived from any of Arthrobacter oryzae (ATCC #42149), Azospirillum brasilense (ATCC #29729), Azotobacter chroococcum (ATCC #9043), Azotobacter vinelandii (ATCC #478), Bacillus licheniformis (ATCC #14580), Bacillus megaterium (ATCC #14581), Bacillus subtilis (ATCC #6051), Mesorhizobium amorphae (ATCC #BAA-114), Pseudomonas putida (ATCC #12633). Pseudomonas fluorescens (ATCC #13525), Rhizobium leguminosarum (ATCC #10004), Rhizobium phaseoli (ATCC #31236), Sinorhizobium meliloti (ATCC #9930) wherein the derived species retains at least one function of the original strain.

38. The method of claim 37. wherein the synthetic combination of endophytes comprises:(a) at least one of the Bacillus species or the Pseudomonas species and at least one of the Rhizobium species or the Azotobacter species;(b) at least one of the Bacillus species and at least one of the Pseudomonas species;(c) at least one of the Rhizobium species and at least one of the Azotobacter species:(d) the Arthrobacter species and the Sinorhizobium species; or(e) at least one Azotobacter species, at least one Rhizobium species, and at least one Arthrobacter species;(1) at least one Rhizobium species, at least one Bacillus species, at least one Azospirillum species, and at least one Azotobacter species;(g) at least one Azotobacter species, at least one Rhizobium species, at least one Arthrobacter species, and at least one Bacillus species; or(h) at least one first endophyte comprising one or more of Bacillus licheniformis, Bacillus Megaterium, Bacillus subtilis, Pseudomonas fluorescens, and Pseudomonas putidcr, at least one second endophyte comprising one or more of Rhizobium leguminosarum, Rhizobium phaseoli, Pseudomonas fluorescens, Pseudomonas putida, Azotobacter chroococcum, and Azotobacter vinelandiv. and at least one third endophyte comprising one or more of Rhizobium leguminosarum, Rhizobium phaseoli, Mesorhizobium amorphae, Arthrobacter oryzae, Azospirillum brasilense, Azotobacter chroococcum, Azotobacter vinelandii, and Sinorhizobium meliloti.

39. The method of claim 37 or 38, wherein the endophyte combination is applied to the seed, the seedling, the plant, the plant part, the algae, or the environment of the seed, plant, plant or algae at a rate of about 1 * 104to about 1 x 109cfu per mL, about 1 * 105to about 1 x 109cfu per mL, about 1 x 106to about 1 x 109cfu per mL, about 1 x 107to about 1 x 109cfu per mL, or about 1 x 108to about 1 x 109cfu per mL.

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