Agricultural beneficial microorganisms, microbial compositions and consortia

Abstract

The present application relates to agriculturally beneficial microorganisms, microbial compositions and consortia. The present disclosure relates to isolated microorganisms, novel strains including microorganisms, microbial consortia, and agricultural compositions comprising the same. In addition, the present disclosure teaches methods of utilizing the microorganisms, microbial consortia, and agricultural compositions comprising the same in methods of conferring beneficial properties to target plant species. In particular aspects, the disclosure provides methods of enhancing desired plant traits in agronomically important crop species.

Description

[0001] This application is a divisional application. The parent application number is 202180048399.9, the application date is June 16, 2021, and the invention title is "Beneficial microorganisms, microbial compositions and aggregates for agriculture".

[0002] Cross-reference to related applications

[0003] This application is entitled to the benefit of U.S. Provisional Patent Application Serial No. 62 / 705,239, filed on June 17, 2020, pursuant to claim 35U.SC119(e), the entire contents of which are incorporated herein by reference.

[0004] References to sequence lists submitted electronically

[0005] An official copy of the sequence list is submitted electronically via EFS-Web as an ASCII format sequence list and is submitted with this specification. The file name of the sequence list is 20041USPCT_Microbes5_SequenceListing_ST25.TXT, created on June 3, 2021, and its size is 41.2KB. The sequence list contained in this ASCII format file is an integral part of this specification and is incorporated herein by reference in its entirety. Technical Field

[0006] This disclosure relates in general to the field of biology, and in particular to microorganisms and microbial compositions for plant improvement. Background Technology

[0007] According to the United Nations World Food Program, nearly 900 million people worldwide suffer from malnutrition. The malnutrition epidemic is particularly pronounced in developing countries, where one in six children is underweight. The lack of available food can be attributed to a variety of socioeconomic factors; however, regardless of the ultimate cause, the fact remains that there is a shortage of food to feed the growing world population, projected to reach 9 billion by 2050. The United Nations estimates that agricultural output must increase by 70% to 100% by 2050 to feed the projected global population.

[0008] These staggering figures on the world's population and malnutrition underscore the critical importance of agricultural efficiency and productivity in sustaining a growing global population. The technological advancements achieved through modern intercropping agriculture, resulting in unprecedented crop yields, are impressive. However, despite progress through technological innovations such as genetically engineered crops and novel insecticides and herbicides, improved crop performance is still needed to meet the demands of an exponentially increasing global population.

[0009] Scientists estimate that if the global agricultural yield gap (i.e., the difference between observed optimal yields and results elsewhere) could be narrowed, global crop production would increase by 45% to 70%. In other words, if all farmers (regardless of their location worldwide) could achieve the highest achievable yields expected in their respective regions, much of the global food production deficit could be resolved. However, addressing how to achieve higher yields across the heterogeneous global landscape presents a significant challenge.

[0010] Typically, yield disparities can be explained by insufficient water, substandard farming practices, inadequate fertilizer, and the inability to use herbicides and pesticides. However, drastically increasing the use of water, fertilizer, herbicides, and pesticides worldwide is not only economically unfeasible for most parts of the world, but would also have adverse environmental consequences.

[0011] Therefore, it is entirely impractical to meet global agricultural output expectations by simply scaling up the current high-input agricultural systems used by most developed countries.

[0012] Therefore, there is an urgent need in this field for improvement methods to enhance crop performance and confer beneficial traits to desired plant species. Summary of the Invention

[0013] This invention includes isolated and biologically pure microbial cells, particularly useful in agriculture. The disclosed microbial cells can be used in their isolated and biologically pure state or formulated into agriculturally acceptable compositions. Agriculturally beneficial microbial aggregates (comprising at least two members of the disclosed microbial cells) and methods for utilizing said aggregates in agricultural applications are also provided. In some aspects, genomic modifications of microorganisms (individuals, aggregates, and / or communities) are envisioned to improve microbial traits and the plants associated with the microorganisms.

[0014] This disclosure addresses the important question of how to improve crop performance, thereby narrowing the global yield gap, while providing ways to confer other beneficial traits onto plant species.

[0015] The solutions provided in this disclosure for improving crop performance and increasing yield are harmless to Earth's resources because they do not rely on increased water consumption or the input of synthetic chemicals into the system. Instead, this disclosure utilizes microorganisms to confer beneficial traits on desired plants, including increased yield.

[0016] Therefore, this disclosure provides an environmentally sustainable solution that enables farmers to increase the yield of important crops without relying on increased use of synthetic herbicides and pesticides.

[0017] In its implementation, this disclosure provides an effective and widely applicable agricultural platform that utilizes microorganisms and microbial aggregates (multiple microorganisms, in some respects, for improving the health of the associated plant or a desired phenotype, such as agronomic traits) to promote one or more desired plant characteristics.

[0018] The microorganisms disclosed herein improve the performance of plants (such as crop plants) through direct and indirect mechanisms. In some aspects, the microorganisms form a symbiotic relationship with the plant. In some aspects, the microorganisms produce compounds (e.g., metabolites) that confer benefits on the plant or can be used by the plant to improve its properties. In some aspects, the microorganisms increase the solubility of one or more compositions (such as nutrients), thereby benefiting the plant. In some aspects, the microorganisms confer tolerance to exogenous substances (such as herbicides or pesticides) on the plant. In some aspects, the microorganisms produce compositions that are harmful to plant pests (such as insects). In some aspects, the microorganisms fix nitrogen, thereby improving the nutritional status of the plant. Other aspects besides the exemplary and non-limiting aspects listed above are contemplated.

[0019] In some embodiments, a single microorganism is utilized. In some aspects, the single microorganism is isolated and purified. In some aspects, the single microorganism is a taxonomic species of bacteria. In some aspects, the single microorganism is an identifiable strain of a taxonomic species of bacteria. In some aspects, the single microorganism is a novel, recently discovered strain of a taxonomic species of bacteria.

[0020] In some aspects, a synthetic composition is provided comprising: (a) microbial cells, exudates derived therefrom, or culture broth derived therefrom, wherein the microbial cells are selected from the group consisting of: i. microbial cells comprising a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; ii. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and iii. microbial cells obtained or derived from microorganisms of any of the following taxa: *Arthrobotrys oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methylotrophicus*, *Bacillus pumilus*, *Bacillus tequilensis*, *Bacillus velezensis*, and *Lysinibacillus*. (a) at least one heterologous composition selected from the group consisting of: plant components, formulation components, agricultural compositions, and any combination thereof; wherein the microorganism is present at a concentration of at least about 10^2 CFU / mL in a liquid formulation or at least about 10^2 CFU / g in a non-liquid formulation.

[0021] In some aspects, a synthetic composition is provided comprising: (a) microbial cells, exudates derived therefrom, or culture broth derived therefrom, wherein the microbial cells are selected from the group consisting of: i. containing and selected from SEQ ID NO. ii. Microbial cells whose sequences NO:1-21 share at least 97% identity with a 16S or ITS sequence; and iii. Microbial cells obtained or derived from microorganisms in Table 1 or Table 1A; and iii. Microbial cells obtained or derived from any of the following taxa: *Oligospora oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesii*, *Bacillus fusiformis*, *Bacillus arabinogalactanophilus*, *Discozoella aurea*, *Bacillus alginate*, *Bacillus erythropoietinus*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; and (b) at least one heterologous composition selected from the group consisting of: plant components, formulation components, agricultural compositions, and any combination thereof; wherein the microorganism is present at a concentration of at least about 10^2 CFU / mL in a liquid formulation or at least about 10^2 CFU / g in a non-liquid formulation; the synthetic composition further comprises at least one additional microorganism.

[0022] In some aspects, a synthetic composition is provided comprising: (a) microbial cells, exudates derived therefrom, or culture broth derived therefrom, wherein the microbial cells are selected from the group consisting of: i. containing and selected from SEQ ID NO. ii. Microbial cells whose sequences NO:1-21 share at least 97% identity with a 16S or ITS sequence; and iii. Microbial cells obtained or derived from microorganisms in Table 1 or Table 1A; and iii. Microbial cells obtained or derived from any of the following taxa: Oligosporium, Bacillus amyloliquefaciens, Bacillus megaterium, Bacillus methyltrophicus, Bacillus pumilus, Bacillus tekira, Bacillus belesii, Bacillus fusiformis, Microbes arabinogalactanophilus, Discobacterium aureum, Bacillus alginate, Bacillus erythropoietinus, Bacillus illinoiense, Bacillus taichung, and Bacillus rubrum; and (b) at least one heterologous composition selected from the group consisting of: plant components, formulation components, agricultural compositions, and any combination thereof; wherein the microorganism is present at a concentration of at least about 10^2 CFU / mL in a liquid formulation or at least about 10^2 CFU / g in a non-liquid formulation; the synthetic composition further comprises at least one additional microorganism; wherein the at least one additional microorganism is selected from Table 2.

[0023] In some aspects, a synthetic composition is provided comprising: (a) microbial cells, exudates derived therefrom, or culture broth derived therefrom, wherein the microbial cells are selected from the group consisting of: i. containing and selected from SEQ ID NO. ii. Microbial cells whose sequences NO:1-21 share at least 97% identity with a 16S or ITS sequence; and iii. Microbial cells obtained or derived from microorganisms in Table 1 or Table 1A; and iii. Microbial cells obtained or derived from any of the following taxa: *Sclerotium oligosporum*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesii*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus erinaceus*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; and (b) at least one heterologous composition selected from the group consisting of: plant components, formulation components, agricultural compositions, and any combination thereof; wherein the microorganism is present at a concentration of at least about 10^2 CFU / mL in a liquid formulation or at least about 10^2 CFU / g in a non-liquid formulation, wherein the plant component is a seed.

[0024] In some aspects, a synthetic composition is provided comprising: (a) microbial cells, exudates derived therefrom, or culture broth derived therefrom, wherein the microbial cells are selected from the group consisting of: i. containing and selected from SEQ ID NO. ii. Microbial cells that share at least 97% identity with the 16S or ITS sequence of NO:1-21; and iii. Microbial cells obtained or derived from the microorganisms listed in Table 1 or Table 1A; and iii. Microbial cells obtained or derived from any of the following taxa: *Sclerotium oligosporum*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesii*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus erinaceus*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; and (b) at least one heterologous composition selected from the group consisting of: plant components, formulation components, agricultural compositions, and any combination thereof; wherein the microorganism is present at a concentration of at least about 10^2 CFU / mL in a liquid formulation or at least about 10^2 CFU / g in a non-liquid formulation, wherein the plant component is a seed; wherein the seed contains a transgene.

[0025] In some aspects, a synthetic composition is provided comprising: (a) microbial cells, exudates derived therefrom, or culture broth derived therefrom, wherein the microbial cells are selected from the group consisting of: i. containing and selected from SEQ ID NO. ii. Microbial cells whose sequences NO:1-21 share at least 97% identity with a 16S or ITS sequence; and iii. Microbial cells obtained or derived from the microorganisms listed in Table 1 or Table 1A; and iii. Microbial cells obtained or derived from any of the following taxa: *Sclerotium oligosporum*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesii*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus iridoides*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; and (b) at least one heterologous composition selected from the group consisting of: plant components, formulation components, agricultural compositions, and any combination thereof; wherein the microorganism is present at a concentration of at least about 10^2 CFU / mL in a liquid formulation or at least about 10^2 CFU / g in a non-liquid formulation; wherein the plant component is a leaf.

[0026] In some aspects, a synthetic composition is provided comprising: (a) microbial cells, exudates derived therefrom, or culture broth derived therefrom, wherein the microbial cells are selected from the group consisting of: i. containing and selected from SEQ ID NO. ii. Microbial cells whose sequences NO:1-21 share at least 97% identity with a 16S or ITS sequence; and iii. Microbial cells obtained or derived from the microorganisms listed in Table 1 or Table 1A; and iii. Microbial cells obtained or derived from any of the following taxa: *Sclerotium oligosporum*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesii*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus erinaceus*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; and (b) at least one heterologous composition selected from the group consisting of: plant components, formulation components, agricultural compositions, and any combination thereof; wherein the microorganism is present at a concentration of at least about 10^2 CFU / mL in a liquid formulation or at least about 10^2 CFU / g in a non-liquid formulation; wherein the plant component is a root.

[0027] In some aspects, a synthetic composition is provided comprising: (a) microbial cells, exudates derived therefrom, or culture broth derived therefrom, wherein the microbial cells are selected from the group consisting of: i. containing and selected from SEQ ID NO. ii. Microbial cells whose sequences NO:1-21 share at least 97% identity with a 16S or ITS sequence; and iii. Microbial cells obtained or derived from the microorganisms listed in Table 1 or Table 1A; and iii. Microbial cells obtained or derived from any of the following taxa: *Sclerotium oligosporum*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesii*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus erinaceus*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; and (b) at least one heterologous composition selected from the group consisting of: plant components, formulation components, agricultural compositions, and any combination thereof; wherein the microorganism is present at a concentration of at least about 10^2 CFU / mL in a liquid formulation or at least about 10^2 CFU / g in a non-liquid formulation; wherein the plant component is a whole plant.

[0028] In some aspects, a synthetic composition is provided comprising: (a) microbial cells, exudates derived therefrom, or culture broth derived therefrom, wherein the microbial cells are selected from the group consisting of: i. microbial cells comprising a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; ii. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and iii. microbial cells obtained or derived from microorganisms of any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belye*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophila*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus ehime*. (a) Bacillus, Bacillus ilignata, Bacillus taichungensis, Bacillus rubrum; and (b) at least one heterologous composition selected from the group consisting of: plant components, formulation components, agricultural compositions, and any combination thereof; wherein the microorganism is present at a concentration of at least about 10^2 CFU / mL in a liquid formulation or at least about 10^2 CFU / g in a non-liquid formulation; wherein the formulation component is selected from the group consisting of: compounds that improve the stability of the microorganism, preservatives, carriers, surfactants, anti-complexing agents, and any combination thereof.

[0029] In some aspects, a synthetic composition is provided comprising: (a) microbial cells, exudates derived therefrom, or culture broths derived therefrom, wherein the microbial cells are selected from the group consisting of: i. microbial cells comprising a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; ii. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and iii. microbial cells obtained or derived from microorganisms of any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belye*, *Bacillus fusiformis*, *Bacillus arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*. (a) Bacillus subtilis, Bacillus erythrophylloides, Bacillus ilignata, Bacillus taichungensis, Bacillus rubrum; and (b) at least one heterologous composition selected from the group consisting of: plant components, formulation components, agricultural compositions, and any combination thereof; wherein the microorganism is present at a concentration of at least about 10^2 CFU / mL in a liquid formulation or at least about 10^2 CFU / g in a non-liquid formulation; wherein the agricultural composition comprises a fungicide, a nematicide, a bactericide, an insecticide, a herbicide, or any combination thereof.

[0030] In some aspects, a variety of synthetic compositions are provided comprising: (a) microbial cells, exudates derived therefrom, or culture broths derived therefrom, wherein the microbial cells are selected from the group consisting of: i. microbial cells comprising a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; ii. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and iii. microbial cells obtained or derived from microorganisms of any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belye*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus ehime*, *Bacillus ilignosa*. (a) Bacterium, Bacillus taichungensis, Bacillus rubrum; and (b) at least one heterologous composition selected from the group consisting of: plant components, formulation components, agricultural compositions, and any combination thereof; wherein the microorganism is present at a concentration of at least about 10^2 CFU / mL in a liquid formulation or at least about 10^2 CFU / g in a non-liquid formulation; wherein the synthetic composition is substantially enclosed within an object selected from the group consisting of: tubes, bottles, wide-mouth bottles, ampoules, packaging, containers, bags, boxes, storage boxes, envelopes, cartons, containers, silos, shipping containers, carriages, and boxes.

[0031] In some aspects, a variety of synthetic compositions are provided, the various synthetic compositions comprising: (a) microbial cells, exudates derived therefrom, or culture broths derived therefrom, wherein the microbial cells are selected from the group consisting of: i. containing and selected from SEQ ID. ii. Microbial cells whose sequences NO:1-21 share at least 97% identity with a 16S or ITS sequence; and iii. Microbial cells obtained or derived from the microorganisms listed in Table 1 or Table 1A; and iii. Microbial cells obtained or derived from any of the following taxa: *Sclerotium oligosporum*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesii*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus erinaceus*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; and (b) at least one heterologous composition selected from the group consisting of: plant components, formulation components, agricultural compositions, and any combination thereof; wherein the microorganism is present at a concentration of at least about 10^2 CFU / mL in a liquid formulation or at least about 10^2 CFU / g in a non-liquid formulation; wherein the synthetic composition is at a temperature below zero degrees Celsius.

[0032] In some aspects, a synthetic composition is provided comprising: (a) microbial cells, exudates derived therefrom, or culture broth derived therefrom, wherein the microbial cells are selected from the group consisting of: i. microbial cells comprising a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; ii. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and iii. microbial cells obtained or derived from microorganisms of any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus bereaves*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus ehime*, *Bacillus illinoi*, *Bacillus taichung*, etc. (a) *Bryophyte rubrum*; and (b) at least one heterologous composition selected from the group consisting of: plant components, formulation components, agricultural compositions, and any combination thereof; wherein the microorganism is present at a concentration of at least about 10^2 CFU / mL in a liquid formulation or at least about 10^2 CFU / g in a non-liquid formulation; wherein the plant component is obtained from plants selected from the group consisting of: corn, soybean, wheat, cotton, cucumber, tomato, pepper, potato, strawberry, orange, lemon, lime, apple, green beans, zucchini, pea, lettuce, broccoli, celery, cauliflower, sorghum, and canola.

[0033] In some aspects, a synthetic composition is provided comprising: (a) microbial cells, exudates derived therefrom, or culture broth derived therefrom, wherein the microbial cells are selected from the group consisting of: i. containing and selected from SEQ ID NO. ii. Microbial cells whose sequences NO:1-21 share at least 97% identity with a 16S or ITS sequence; and iii. Microbial cells obtained or derived from the microorganisms listed in Table 1 or Table 1A; and iii. Microbial cells obtained or derived from any of the following taxa: *Oligospora spp.*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesii*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella aurea*, *Bacillus alginate*, *Bacillus erythropoietinus*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; and (b) at least one heterologous composition selected from the group consisting of: plant components, formulation components, agricultural compositions, and any combination thereof; wherein the microorganism is present at a concentration of at least about 10^2 CFU / mL in a liquid formulation or at least about 10^2 CFU / g in a non-liquid formulation; wherein the agricultural composition comprises a growth medium.

[0034] In some aspects, a synthetic composition is provided comprising: (a) microbial cells, exudates derived therefrom, or culture broth derived therefrom, wherein the microbial cells are selected from the group consisting of: i. containing and selected from SEQ ID NO. ii. Microbial cells whose sequences NO:1-21 share at least 97% identity with a 16S or ITS sequence; and iii. Microbial cells obtained or derived from the microorganisms listed in Table 1 or Table 1A; and iii. Microbial cells obtained or derived from any of the following taxa: *Sclerotium oligosporum*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesii*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus erinaceus*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; and (b) at least one heterologous composition selected from the group consisting of: plant components, formulation components, agricultural compositions, and any combination thereof; wherein the microorganism is present at a concentration of at least about 10^2 CFU / mL in a liquid formulation or at least about 10^2 CFU / g in a non-liquid formulation; wherein the agricultural composition comprises a growth medium; wherein the growth medium comprises soil.

[0035] In some aspects, a variety of synthetic compositions are provided comprising: (a) microbial cells, exudates derived therefrom, or culture broths derived therefrom, wherein the microbial cells are selected from the group consisting of: i. microbial cells comprising a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; ii. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and iii. microbial cells obtained or derived from microorganisms of any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belye*, *Bacillus fusiformis*, *Bacillus arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus erythropoietinus*, *Bacillus ilignata ... (a) Bacillus subtilis, Bacillus taichungensis, Bacillus rubrum; and (b) at least one heterologous composition selected from the group consisting of: plant components, formulation components, agricultural compositions, and any combination thereof; wherein the microorganism is present at a concentration of at least about 10^2 CFU / mL in a liquid formulation or at least about 10^2 CFU / g in a non-liquid formulation; wherein the agricultural composition comprises a growth medium; wherein the growth medium comprises soil; wherein the plurality of synthetic compositions are placed in the soil in a regular pattern with substantially equal spacing between each of the synthetic compositions.

[0036] In some aspects, a synthetic composition is provided comprising: (a) an exudate or culture broth of a plurality of cells, wherein these cells comprise at least one microbial cell selected from the group consisting of: i. microbial cells comprising a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; ii. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and iii. microbial cells obtained or derived from microorganisms of any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesia*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella aurea*, *Bacillus alginate*, *Bacillus erythropoietinus*, *Bacillus ilignata*, *Bacillus taichungensis*, *Bacillus rubrum*; and (b) at least one heterologous composition selected from the group consisting of: plant components, formulation components, agricultural compositions, and any combination thereof.

[0037] In some aspects, a method is provided for regulating agronomically important traits in plants obtained or derived from plant components, the method comprising treating said plant component with a formulation containing microbial cells, exudates derived therefrom, or culture broths derived therefrom, wherein the microbial cells are selected from the group consisting of: a. microbial cells containing a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; b. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and c. microbial cells obtained or derived from microorganisms of any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesia*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus ehime*, *Bacillus ilignosa*, *Bacillus taichung*, and *Brochiropterygium*.

[0038] In some aspects, a method is provided for regulating agronomically important traits in plants obtained or derived from plant components, the method comprising treating said plant component with a formulation containing microbial cells, exudates derived therefrom, or culture broths derived therefrom, wherein the microbial cells are selected from the group consisting of: a. containing and selected from SEQ ID NO. NO:1-21 are microbial cells that share at least 97% identity with a 16S or ITS sequence; b. microbial cells obtained or derived from the microorganisms listed in Table 1 or Table 1A; and c. microbial cells obtained or derived from any of the following taxa: *Sclerotium oligosporum*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesii*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus eigensis*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; wherein the agronomically important trait is selected from the group consisting of the following: disease resistance, drought tolerance, heat tolerance, cold tolerance, salt tolerance, metal tolerance, herbicide tolerance, chemical tolerance, improved water use efficiency, improved nitrogen use efficiency, improved nitrogen fixation, insect resistance, and herbivorous animal... Resistance to plant growth, pathogen resistance, increased yield, increased yield under water-limited conditions, enhanced health, improved vitality, improved growth, enhanced photosynthetic capacity, enhanced nutrition, changes in protein content, changes in oil content, increased biomass, increased shoot length, increased root length, improved root architecture, increased seed weight, changes in seed carbohydrate composition, changes in seed oil composition, increased radicle length, increased pod number, delayed senescence, greenness retention, changes in seed protein composition, increased dry weight of mature plant reproductive components, increased fresh weight of mature plant reproductive components, increased number of mature plant reproductive components per plant, increased chlorophyll content, increased number of pods per plant, increased pod length per plant, increased number of seeds per plant, increased seed weight per plant, reduced number of wilted leaves per plant, reduced number of severely wilted leaves per plant, increased number of non-wilted leaves per plant, or improved plant visual appearance.

[0039] In some aspects, a method is provided for regulating agronomically important traits in plants obtained or derived from plant components, the method comprising treating said plant component with a formulation containing microbial cells, exudates derived therefrom, or culture broths derived therefrom, wherein the microbial cells are selected from the group consisting of: a. containing and selected from SEQ ID NO. a. Microbial cells sharing at least 97% identity with the 16S or ITS sequence of NO:1-21; b. Microbial cells obtained or derived from the microorganisms listed in Table 1 or Table 1A; and c. Microbial cells obtained or derived from any of the following taxa: *Sclerotium oligosporum*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesii*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella aurea*, *Bacillus alginate*, *Bacillus erythropoietinus*, *Bacillus ilignosa*, *Bacillus taichungensis*, *Bacillus rubrum*; wherein the microbial cells, exudates, or broths derived therefrom are present in an amount capable of providing benefit to the plant derived from the plant component, compared to plants from plant components that have never been treated with the microbial cells or exudates derived therefrom.

[0040] In some aspects, a method of cultivating a plant is provided, the method comprising introducing microbial cells, exudates therefrom, or culture broths therefrom into a plant component of said plant, wherein the microbial cells are selected from the group consisting of: a. microbial cells containing a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; b. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and c. microbial cells obtained or derived from microorganisms of any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesia*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus ehime*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; and wherein said microbial cells are heterologously disposed into the plant component.

[0041] In some aspects, a method of cultivating a plant is provided, the method comprising introducing microbial cells, exudates therefrom, or culture broths therefrom into a plant component of the plant, wherein the microbial cells are selected from the group consisting of: a. containing and selected from SEQ ID NO. a. Microbial cells sharing at least 97% identity with the 16S or ITS sequence of NO:1-21; b. Microbial cells obtained or derived from the microorganisms listed in Table 1 or Table 1A; and c. Microbial cells obtained or derived from any of the following taxa: *Sclerotium oligosporum*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesii*, *Bacillus fusiformis*, *Bacillus arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus ehime*, *Bacillus ilynoides*, *Bacillus taichung*, *Bacillus rubrum*; and wherein said microbial cells are heterologously disposed into plant components; wherein said introduction into plant components is accomplished by indirect methods selected from the group consisting of: furrow application, soil irrigation application, and lateral application.

[0042] In some aspects, a method of cultivating a plant is provided, the method comprising introducing microbial cells, exudates therefrom, or culture broths therefrom into a plant component of the plant, wherein the microbial cells are selected from the group consisting of: a. containing and selected from SEQ ID NO. a. Microbial cells sharing at least 97% identity of a 16S or ITS sequence with sequences NO:1-21; b. Microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and c. Microbial cells obtained or derived from any of the following taxa: *Saccharomyces oligosporus*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesii*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella aurea*, *Bacillus alginate*, *Bacillus erythropoietinus*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; and wherein said microbial cells are heterologously disposed into plant components; wherein said introduction into plant components is accomplished by coating said plant components with a liquid preparation of the microorganism or an exudate derived therefrom.

[0043] In some aspects, a method of cultivating a plant is provided, the method comprising introducing microbial cells, exudates therefrom, or culture broths therefrom into a plant component of the plant, wherein the microbial cells are selected from the group consisting of: a. containing and selected from SEQ ID NO. a. Microbial cells sharing at least 97% identity of a 16S or ITS sequence with sequences NO:1-21; b. Microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and c. Microbial cells obtained or derived from any of the following taxa: *Saccharomyces oligosporus*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesii*, *Bacillus fusiformis*, *Bacillus arabinogalactanophilus*, *Discozoella aurea*, *Bacillus alginate*, *Bacillus erythropoietinus*, *Bacillus ilignosa*, *Bacillus taichungensis*, *Bacillus rubrum*; and wherein said microbial cells are heterologously disposed into plant components; wherein said introduction into plant components is accomplished by coating said plant components with a substantially non-liquid formulation of the microorganism or an exudate derived therefrom.

[0044] In some aspects, a method of cultivating a plant is provided, the method comprising introducing microbial cells, exudates therefrom, or culture broths therefrom into a plant component of said plant, wherein the microbial cells are selected from the group consisting of: a. microbial cells containing a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; b. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and c. microbial cells obtained or derived from microorganisms of any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesia*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus ehime*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; and wherein said microbial cells are heterologously disposed into a plant component; wherein said plant component is a seed.

[0045] In some aspects, a method of cultivating a plant is provided, the method comprising introducing microbial cells, exudates therefrom, or culture broths therefrom into a plant component of said plant, wherein the microbial cells are selected from the group consisting of: a. microbial cells containing a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; b. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and c. microbial cells obtained or derived from microorganisms of any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesia*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus ehime*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; and wherein said microbial cells are heterologously disposed into a plant component; wherein said plant component is a leaf.

[0046] In some aspects, a method of cultivating a plant is provided, the method comprising introducing microbial cells, exudates therefrom, or culture broths therefrom into a plant component of said plant, wherein the microbial cells are selected from the group consisting of: a. microbial cells containing a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; b. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and c. microbial cells obtained or derived from microorganisms of any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesia*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus ehime*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; and wherein said microbial cells are heterologously disposed into a plant component; wherein said plant component is a root.

[0047] In some aspects, a method of cultivating a plant is provided, comprising introducing microbial cells, exudates therefrom, or culture broths therefrom into a plant component of said plant, wherein the microbial cells are selected from the group consisting of: a. microbial cells containing a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; b. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and c. microbial cells obtained or derived from microorganisms of any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesia*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus ehime*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; and wherein said microbial cells are heterologously disposed into a plant component; wherein said plant component is a whole plant.

[0048] In some aspects, a method is provided for regulating agronomically important traits in a harvested product, the method comprising introducing microbial cells, exudates derived therefrom, or culture broths derived therefrom into an organism from which the harvested product is obtained, wherein the microbial cells are selected from the group consisting of: a. microbial cells containing a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; b. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and c. microbial cells obtained or derived from microorganisms of any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus bereaves*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus ehime*, *Bacillus ilignosa*, *Bacillus taichung*, and *Brochiropterygium*.

[0049] In some aspects, a method is provided for regulating agronomically important traits in a harvested product, the method comprising introducing microbial cells, exudates therefrom, or culture broths therefrom into the harvested product, wherein the microbial cells are selected from the group consisting of: a. microbial cells containing a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; b. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and c. microbial cells obtained or derived from microorganisms of any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus bereaves*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus ehime*, *Bacillus ilignosa*, *Bacillus taichung*, and *Brochiropterygium*.

[0050] In some aspects, a method is provided for regulating agronomically important traits in a harvested product, the method comprising introducing microbial cells, exudates derived therefrom, or culture broths derived therefrom into an organism from which the harvested product is obtained, wherein the microbial cells are selected from the group consisting of: a. microbial cells containing a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; b. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and c. microbial cells obtained or derived from any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus bereaves*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus ehime*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; wherein the harvested product is a fruit.

[0051] In some aspects, a method is provided for regulating agronomically important traits in a harvested product, the method comprising introducing microbial cells, exudates therefrom, or culture broths therefrom into the harvested product, wherein the microbial cells are selected from the group consisting of: a. microbial cells containing a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; b. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and c. microbial cells obtained or derived from microorganisms of any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belleus*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus ehime*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; wherein the harvested product is a fruit.

[0052] In some aspects, a method is provided for regulating agronomically important traits in a harvested product, the method comprising introducing microbial cells, exudates derived therefrom, or culture broths derived therefrom into an organism from which the harvested product is obtained, wherein the microbial cells are selected from the group consisting of: a. microbial cells containing a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; b. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and c. microbial cells obtained or derived from any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesia*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus ehime*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; wherein the harvested product is a vegetable.

[0053] In some aspects, a method is provided for regulating agronomically important traits in a harvested product, the method comprising introducing microbial cells, exudates therefrom, or culture broths therefrom into the harvested product, wherein the microbial cells are selected from the group consisting of: a. microbial cells containing a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; b. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and c. microbial cells obtained or derived from any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belleus*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus ehime*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; wherein the harvested product is a vegetable.

[0054] In some aspects, a method is provided for regulating agronomically important traits in a harvested product, the method comprising introducing microbial cells, exudates derived therefrom, or culture broths derived therefrom into an organism from which the harvested product is obtained, wherein the microbial cells are selected from the group consisting of: a. microbial cells containing a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; b. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and c. microbial cells obtained or derived from microorganisms of any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus bereaves*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus ehime*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; wherein the harvested product is a seed.

[0055] In some aspects, a method is provided for regulating agronomically important traits in a harvested product, the method comprising introducing microbial cells, exudates therefrom, or culture broths therefrom into the harvested product, wherein the microbial cells are selected from the group consisting of: a. microbial cells containing a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; b. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and c. microbial cells obtained or derived from any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus bereaves*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus ehime*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; wherein the harvested product is a seed.

[0056] In some aspects, a method is provided for regulating agronomically important traits in a harvested product, the method comprising introducing microbial cells, exudates derived therefrom, or culture broths derived therefrom into an organism from which the harvested product is obtained, wherein the microbial cells are selected from the group consisting of: a. microbial cells containing a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; b. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and c. microbial cells obtained or derived from any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus bereaves*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella spp.*, *Bacillus alginate*, *Bacillus ehime*, *Bacillus ilignosa*, *Bacillus taichung*, *Bacillus rubrum*; wherein the harvested product is fiber.

[0057] In some aspects, a method is provided for regulating agronomically important traits in a harvested product, the method comprising introducing microbial cells, exudates therefrom, or culture broths therefrom into the harvested product, wherein the microbial cells are selected from the group consisting of: a. microbial cells containing a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; b. microbial cells obtained or derived from microorganisms listed in Table 1 or Table 1A; and c. microbial cells obtained or derived from any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesia*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella aurea*, *Bacillus alginate*, *Bacillus erythropoietinus*, *Bacillus ilignata*, *Bacillus taichungensis*, *Bacillus rubrum*; wherein the harvested product is fiber.

[0058] In some aspects, a substantially cell-free preparation obtained or derived from a microbial culture is provided, wherein the microorganism is selected from the group consisting of: a. microbial cells comprising a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; b. microbial cells obtained or derived from microorganisms in Table 1 or Table 1A; and c. microbial cells obtained or derived from microorganisms of any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesia*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella aurea*, *Bacillus alginate*, *Bacillus erythropoietinus*, *Bacillus ilignosa*, *Bacillus taichung*, and *Brochiropterygium*.

[0059] In some aspects, a purified composition is provided prepared from a substantially cell-free preparation obtained or derived from a microbial culture, wherein the microorganism is selected from the group consisting of: a. microbial cells comprising a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO: 1-21; b. microbial cells obtained or derived from microorganisms in Table 1 or Table 1A; and c. microbial cells obtained or derived from microorganisms of any of the following taxa: *Arthropoda oligospora*, *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus methyltrophicus*, *Bacillus pumilus*, *Bacillus tekira*, *Bacillus belesia*, *Bacillus fusiformis*, *Microbacterium arabinogalactanophilus*, *Discozoella aurea*, *Bacillus alginate*, *Bacillus erythropoietinus*, *Bacillus ilignata*, *Bacillus taichungensis*, and *Brochiropterygium*.

[0060] In some respects, a strain of isolated bacterial strains selected from Table 1 or Table 1A, or a strain of isolated bacterial strains having substantially similar morphological and physiological characteristics, substantially similar genetic characteristics, their offspring, mutants, or gene-edited, altered, or modified variants are provided.

[0061] In some aspects, an isolated bacterial strain is provided, which contains a polynucleotide sequence that shares at least 97% sequence identity with any of the strains in SEQ ID NO:1-21.

[0062] In some aspects, an agricultural composition is provided comprising: a) an isolated bacterial strain comprising a polynucleotide sequence sharing at least 97% sequence identity with any of SEQ ID NO:1-21; and b) an agriculturally acceptable vector; wherein the bacterial strain is present in the agricultural composition in an amount that effectively produces a modified phenotype in the associated plant.

[0063] In some aspects, an agricultural composition is provided comprising: a) an isolated strain of bacteria containing a polynucleotide sequence sharing at least 97% sequence identity with any of SEQ ID NO:1-21; and b) an agriculturally acceptable vector; wherein the strain of bacteria is present in the agricultural composition in an amount that effectively produces a modified phenotype in the associated plant; wherein the agricultural composition is formulated as a seed coating, foliar spray, soil irrigation, impregnation treatment, furrow treatment, soil conditioner, granules, broadcast treatment, or post-harvest disease control treatment.

[0064] In some aspects, a microbial cell and plant component are provided, the microbial cell comprising a 16S or ITS sequence sharing at least 97% identity with a sequence selected from SEQ ID NO:1-21, wherein the microbial cell is heterologously disposed into the plant component.

[0065] In some implementations, a single microorganism from Table 1 or Table 1A is used.

[0066] In some embodiments, microorganisms from the genus *Bacillus* are used. In some embodiments, a combination of one or more microorganisms from the genus *Bacillus* is used. In some embodiments, microorganisms from the genus *Paenibacillus* are used. In some embodiments, a combination of one or more microorganisms from the genus *Paenibacillus* is used. In some embodiments, microorganisms from the genus *Arthrobotrys* (teleomorph Orbilia) are used. In some embodiments, microorganisms from the genus *Lysinibacillus* are used. In some embodiments, microorganisms from the genus *Microbacterium* are used. In some embodiments, microorganisms from the genus *Talaromyces* are used.

[0067] In some respects, the single microorganism (whether a taxonomically identifiable species or strain) is combined with one or more other microorganisms of different species or strains. In some respects, the combination of two or more microorganisms forms a consortia or consortium. The terms consortia and consortium are used interchangeably.

[0068] In some aspects, this disclosure provides for the development of highly functional microbial aggregates that facilitate the development and expression of desired phenotypic or genotypic plant traits. In some embodiments, the aggregates of this disclosure possess functional properties that do not exist in nature when the individual microorganisms live alone. That is, in various embodiments, specific microbial species are combined into an aggregate such that the microbial ensemble possesses functional properties that any single member of the aggregate does not possess when considered individually.

[0069] In some implementations, the functional attribute possessed by the microbial aggregate is the ability to confer one or more beneficial characteristics on the plant species, such as: increased growth, increased yield, increased nutrient utilization (e.g., nitrogen, phosphate, etc.), increased nitrogen use efficiency, enhanced stress tolerance, enhanced drought tolerance, increased photosynthetic rate, enhanced water use efficiency, enhanced pathogen resistance, and modification of plant architecture (which does not necessarily affect plant yield, but rather addresses plant function), etc.

[0070] In some embodiments, individual microorganisms do not possess the ability to confer these beneficial properties to plants when they exist in nature. Instead, in some embodiments, these microorganisms are artificially combined into aggregates to form functional compositions that possess properties and functional characteristics not found in nature. In some embodiments, the aggregate may comprise microorganisms that have been gene-edited, altered, or modified by means of genetic material (including DNA, RNA, proteins, and / or combinations thereof) modified using techniques known to those skilled in the art.

[0071] However, in other embodiments, this disclosure provides isolated and biologically pure individual microorganisms that can confer beneficial properties to a desired plant species without requiring the microorganisms to be combined into aggregates.

[0072] In some embodiments, the microbial aggregate may be any combination of one or more individual microorganisms from Table 1 or Table 1A. In other embodiments, a single microorganism from Table 1 or Table 1A is used in combination with one or more microorganisms selected from Table 2. In other embodiments, one or more microorganisms from Table 1 and / or Table 1A are used in combination with another microorganism from said table or one or more microorganisms from Table 2. In some embodiments, the microbial aggregate comprises two, three, four, five, six, seven, eight, nine, ten, or more than ten microorganisms.

[0073] Another object of this disclosure relates to the use of isolated microorganisms and microbial aggregates as plant growth promoters. In other respects, the isolated microorganisms and microbial aggregates act as growth regulators, which can, for example, resist normal senescence, thereby causing an increase in biomass.

[0074] Another objective of this disclosure relates to the use of isolated microorganisms and microbial aggregates as soil health enhancers and plant health enhancers. In other respects, the isolated microorganisms and microbial aggregates act as biostimulants.

[0075] Additional objectives of this disclosure relate to the use of isolated microorganisms and microbial aggregates as insecticides. In other aspects, the isolated microorganisms and microbial aggregates act as biofungicides. In still other aspects, the isolated microorganisms and microbial aggregates act as bionematicides.

[0076] Another objective of this disclosure is to design microbial aggregates capable of performing synergistic, multidimensional activities. In some respects, the microorganisms constituting the aggregate act synergistically. In other respects, the effect of the microbial aggregate on a certain plant characteristic is greater than the effect observed when any single microbial member of the aggregate is used alone. That is, in some respects, the aggregate exhibits a greater-than-additive effect on the desired plant characteristic compared to the effect found when any single member of the aggregate is used alone.

[0077] In some respects, aggregates enable the establishment of other plant-microbe interactions, for example by acting as primary colonists or foundational populations that set the trajectory of future microbiome development.

[0078] In the implementation scheme, this disclosure relates to synergistic combinations (or mixtures) of microbial isolates.

[0079] In several respects, the aggregates taught in this paper offer a wide range of agricultural applications, including: increased yields of grains, fruits, and flowers; enhanced growth of plant parts; improved nutrient utilization (e.g., nitrogen, phosphate, etc.); increased disease resistance; biopesticide effects, including improved resistance to fungi, insects, and nematodes; increased survival rates in extreme climates; and modification of other desired plant phenotypic traits. Notably, these benefits to plants can be obtained without any harmful environmental side effects.

[0080] In some respects, the individual microorganisms of this disclosure or aggregates containing them can be combined to form agriculturally acceptable compositions.

[0081] In some embodiments, the agricultural compositions disclosed herein include, but are not limited to: wetting agents, compatibilizers, defoamers, detergents, chelating agents, drift reducers, neutralizers, buffers, corrosion inhibitors, dyes, odorants, spreading agents, penetration aids, adhesives, binders, dispersants, thickeners, stabilizers, emulsifiers, freezing point inhibitors, antimicrobial agents, fertilizers, pesticides, herbicides, inert carriers, polymers, etc.

[0082] In one embodiment of this disclosure, microorganisms (including isolated single species or strains, aggregates, or combinations thereof, such as metabolites) are supplied in the form of seed coatings or other seed applications. In some embodiments, seed coatings may be applied to naked and untreated seeds. In other embodiments, seed coatings may be applied to previously treated seeds. Therefore, in some embodiments, this disclosure teaches a method of treating seeds that includes applying isolated strains or microbial aggregates to the seeds. In some embodiments, isolated strains or microbial aggregates are applied in the form of an agricultural composition comprising an agriculturally acceptable carrier. In some embodiments, the agricultural composition may be formulated as: soil irrigation agents, foliar sprays, impregnation treatments, furrow treatments, soil conditioners, granules, broadcast treatments, post-harvest disease control treatments, or seed treatments. In some embodiments, the agricultural composition may be applied alone or in a rotary spraying procedure with other agricultural products. In some embodiments, the agricultural composition may be tank-compatible. In some embodiments, the agricultural composition may be tank-compatible with other agricultural products. In some implementations, the agricultural composition may be compatible with equipment used for ground, air, and irrigation application.

[0083] In some embodiments, the applied microorganisms may become endophytic and thus be present in the treated growing plants and their offspring. In other embodiments, the microorganisms may be applied simultaneously with the seed treatment agent in a co-treatment manner.

[0084] In one embodiment of this disclosure, the microorganisms are supplied in the form of granules or plugs applied to plant growth media or soil irrigation. In other embodiments, the microorganisms are supplied in the form of foliar application, such as foliar sprays or liquid compositions. Foliar sprays or liquid application can be applied to growing plants or growth media, such as soil.

[0085] In other embodiments, the microorganisms (including isolated single species, strains, or aggregates, or combinations thereof, such as metabolites) are supplied in the form of fertilizers, pesticides, or other amendments applicable to the soil. In some embodiments, the microorganisms are supplied in the form of fertilizers, pesticides, or other amendments applied to the soil prior to planting. In some embodiments, the microorganisms are supplied in the form of fertilizers, pesticides, or other amendments applied to the soil concurrently with planting. In some embodiments, the microorganisms are supplied in the form of fertilizers, pesticides, or other amendments applied to the soil after planting.

[0086] In other embodiments of this disclosure, microorganisms (including isolated single species, strains, or aggregates) and / or combinations thereof (e.g., metabolites) are supplied in the form of post-harvest disease control applications.

[0087] In embodiments, the agricultural compositions of this disclosure can be formulated as: (1) a solution; (2) a wettable powder; (3) a spreadable powder; (4) a soluble powder; (5) an emulsion or suspension concentrate; (6) a seed dressing agent; (7) a tablet; (8) a water-dispersible granule; (9) a water-soluble granule (slow-release or immediate-release); (10) a microencapsulated granule or suspension; (11) as an irrigation component; and (12) a component of fertilizers, pesticides, and other compatibility modifiers, etc. In some respects, the compositions can be diluted in an aqueous medium prior to conventional spray application. The compositions of this disclosure can be applied to soil, plants, seeds, rhizosphere, root sheath, or other areas where the application of the microbial composition will be beneficial.

[0088] Another object of this disclosure relates to agricultural compositions formulated to provide high colony-forming unit (CFU) bacterial communities or aggregates. In some aspects, the agricultural compositions have adjuvants that provide a relevant shelf life. In embodiments, the CFU concentration of the taught agricultural composition is higher than the concentration at which the microorganisms would naturally exist outside the disclosed methods. In another embodiment, the agricultural composition contains microbial cells at a concentration of 10^2-10^12 CFU / g carrier or 10^5-10^9 CFU / g carrier. In one aspect, the microbial cells are applied directly to seeds as a seed coating at a concentration of 10^5-10^9 CFU. In other aspects, the microbial cells are applied as an outer seed coating at a concentration of 10^5-10^9 CFU on top of another seed coating. In still other aspects, the microbial cells are applied together with another seed treatment agent as a co-treatment agent at a ratio of 10^5-10^9 CFU.

[0089] In all respects, this disclosure relates to agricultural microbial preparations for promoting plant growth. In all respects, this disclosure provides the taught isolated microorganisms and aggregates comprising them for formulation as agricultural biological inoculants. The taught biological inoculants can be applied to plants, seeds, or soil, or in combination with fertilizers, pesticides, and other compatibility modifiers. Suitable examples of formulating biological inoculants comprising isolated microorganisms can be found in U.S. Patent No. 7,097,830, which is incorporated herein by reference.

[0090] The disclosed microbial preparations can: reduce the need for nitrogen fertilizers, dissolve minerals, provide biological insecticidal protection for plants, protect plants from pathogens (e.g., fungi, insects, and nematodes), and enable plants to obtain valuable nutrients such as nitrogen and / or phosphates, thereby reducing and eliminating the need for the use of chemical pesticides and chemical fertilizers.

[0091] In some embodiments, isolated and biologically pure microorganisms of this disclosure may be used in methods of conferring one or more beneficial characteristics or traits upon a desired plant species.

[0092] In some embodiments, in methods of imparting one or more beneficial characteristics or traits to a desired plant species, an agriculturally acceptable composition comprising isolated and biologically pure microorganisms of the present disclosure may be used.

[0093] In some embodiments, the aggregates of this disclosure may be used in methods of imparting one or more beneficial characteristics or traits to a desired plant species.

[0094] In some embodiments, agriculturally acceptable compositions comprising the polymers of this disclosure may be used in methods of imparting one or more beneficial characteristics or traits to a desired plant species.

[0095] In some aspects, the isolated and biologically pure microorganisms and / or aggregates of this disclosure are derived from an accelerated microbial selection process (“AMS” process). The AMS process utilized in some aspects of this disclosure is described, for example, in the following documents: (1) International Patent Application No. PCT / NZ2012 / 000041, published on September 20, 2012, with International Publication No. WO 2012125050 A1, and (2) International Patent Application No. PCT / NZ2013 / 000171, published on March 27, 2014, with International Publication No. WO 2014046553 A1, each of which is incorporated herein by reference in its entirety for all purposes. The AMS process described in this disclosure is, for example, in the following documents: Figures 1 to 4 middle.

[0096] However, in other embodiments, the microorganisms disclosed herein are not derived from an accelerated microbial selection process. In some aspects, the microorganisms utilized in embodiments of this disclosure are selected from members of a database. In particular, the microorganisms utilized in embodiments of this disclosure are selected from microorganisms existing in a database based on specific characteristics of the microorganisms.

[0097] This disclosure provides a way to effectively enhance a plant component or part by coating it with isolated microorganisms or microbial aggregates in an amount that would normally not be found on a plant component or part.

[0098] Some embodiments described herein are methods for preparing agricultural seed compositions or seed coatings, the methods comprising: contacting the surface of a seed with a formulation containing a purified microbial community, the purified microbial community comprising at least one isolated microorganism that is heterologous to the seed or rarely present on the seed. Other embodiments require the preparation of agricultural plant compositions comprising: contacting the surface of a plant with a formulation containing a purified microbial community, the purified microbial community comprising at least one isolated microorganism that is heterologous to the plant. In other aspects, the formulation or microorganisms are introduced into the interior of the seed, for example, into the cotyledons or other seed tissues such as the plumule.

[0099] In some respects, applying the isolated microorganisms, microbial aggregates, exudates, metabolites, and / or agricultural compositions of this disclosure to seeds or plants can modulate agronomically important traits. Agronomically important traits may include, for example, disease resistance, drought tolerance, heat tolerance, cold tolerance, salt tolerance, metal tolerance, herbicide tolerance, chemical tolerance, improved water use efficiency, improved nitrogen use efficiency, improved nitrogen stress resistance, improved nitrogen fixation, improved nutrient (e.g., phosphate, potassium, etc.) utilization, insect resistance, herbivore resistance, pathogen resistance, reduced pathogen levels (e.g., via the secretion of metabolites that affect pathogen survival), increased yield, increased yield under water-limited conditions, enhanced health, improved vitality, improved growth, improved photosynthetic capacity, enhanced nutrition, altered protein content, altered oil content, increased biomass, increased shoot length, increased root length, improved root architecture, increased seed weight, faster seed germination, altered seed carbohydrate composition, altered seed oil composition, pod number, delayed senescence, greenness retention, and altered seed protein composition. In some respects, the regulation modulates at least two, three, four, or more agronomically important traits. In some respects, the regulation has a positive effect on one of the aforementioned agronomic traits.

[0100] In some respects, the isolated microorganisms, aggregates, and / or agricultural compositions of this disclosure can be applied to plants to modulate or alter plant characteristics, such as changes in oil content, protein content, seed carbohydrate composition, seed oil composition, seed protein composition, chemical resistance, cold tolerance, delayed senescence, disease resistance, drought tolerance, ear weight, improved growth, enhanced health, heat tolerance, herbicide tolerance, herbivore resistance, improved nitrogen fixation, improved nitrogen use efficiency, improved root architecture, improved water use efficiency, increased biomass, decreased biomass, increased root length, decreased root length, increased seed weight, increased shoot length, decreased shoot length, and increased yield. Increased yield under water-limited conditions, grain quality, grain moisture content, metal tolerance, number of spikes, number of grains per spike, number of pods, enhanced nutrition, pathogen resistance, insect resistance, improved photosynthetic capacity, salt tolerance, greenness retention, improved vigor, increased dry weight of mature seeds, increased fresh weight of mature seeds, increased number of mature seeds per plant, increased chlorophyll content, increased number of pods per plant, increased pod length per plant, decreased number of wilted leaves per plant, decreased number of severely wilted leaves per plant and increased number of non-wilted leaves per plant, detectable regulation of metabolite levels, detectable regulation of transcript levels, and detectable regulation of the proteome.

[0101] In some implementations, the agricultural formulations taught herein comprise at least one member selected from the group consisting of: agriculturally compatible carriers, thickeners, microbial stabilizers, fungicides, antimicrobial agents, herbicides, nematicides, insecticides, plant growth regulators, rodenticides, and nutrients.

[0102] The methods described herein may include contacting seeds or plants with at least 100 CFU or spores, at least 300 CFU or spores, at least 1,000 CFU or spores, at least 3,000 CFU or spores, at least 10,000 CFU or spores, at least 30,000 CFU or spores, at least 100,000 CFU or spores, at least 300,000 CFU or spores, at least 1,000,000 CFU or spores or more of the microorganisms taught herein.

[0103] The methods described herein may include contacting seeds or plants with a composition comprising metabolites produced by a single microorganism or microbial aggregate disclosed herein. In some aspects, the method includes contacting seeds or plants with a composition comprising at least 1 mg of metabolites produced by a single microorganism or microbial aggregate disclosed herein. In some aspects, the method includes contacting seeds or plants with a composition comprising at least 10 mg of metabolites produced by a single microorganism or microbial aggregate disclosed herein. In some aspects, the method includes contacting seeds or plants with a composition comprising at least 100 mg of metabolites produced by a single microorganism or microbial aggregate disclosed herein. In some aspects, the method includes contacting seeds or plants with a composition comprising at least 1 g of metabolites produced by a single microorganism or microbial aggregate disclosed herein. In some aspects, the method includes contacting seeds or plants with a composition comprising at least 10 g of metabolites produced by a single microorganism or microbial aggregate disclosed herein. In some aspects, the method includes contacting seeds or plants with a composition comprising at least 100 g of metabolites produced by a single microorganism or microbial aggregate disclosed herein. In some aspects, the method includes contacting seeds or plants with a composition comprising at least 1 kg of metabolites produced by a single microorganism or microbial aggregate disclosed herein. In some aspects, the method includes contacting seeds or plants with a composition comprising more than 1 kg of metabolites produced by a single microorganism or microbial aggregate disclosed herein.

[0104] In some embodiments of the methods described herein, the isolated microorganisms of this disclosure are present in the formulation in an amount that is effectively detectable within and / or on the target tissue of an agricultural plant. For example, at least 100 CFU or spores, at least 300 CFU or spores, at least 1,000 CFU or spores, at least 3,000 CFU or spores, at least 10,000 CFU or spores, at least 30,000 CFU or spores, at least 100,000 CFU or spores, at least 300,000 CFU or spores, at least 1,000,000 CFU or spores, or more of microorganisms are detected within and / or on the target tissue of the plant. Alternatively or additionally, the microorganisms of this disclosure may be present in the formulation in an amount that effectively increases the biomass and / or yield of the plant to which such formulation is applied by at least 1%, at least 2%, at least 3%, at least 5%, at least 10%, at least 15%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100% or more when compared with a reference agricultural plant to which the formulation of this disclosure is not applied. Alternatively or additionally, the microorganisms of this disclosure may be present in the formulation in an amount of at least 1%, at least 2%, at least 3%, at least 5%, at least 10%, at least 15%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100% or more, when compared with a reference agricultural plant that has not been treated with the formulation of this disclosure.

[0105] In some embodiments of the methods described herein, one or more metabolites isolated from the microorganisms or aggregates of this disclosure are present in the formulation in an amount that is effectively detectable within and / or on the target tissue of the agricultural plant. For example, metabolites in amounts of at least 1 mg, at least 10 mg, at least 50 mg, at least 100 mg, at least 200 mg, at least 400 mg, at least 600 mg, at least 800 mg, at least 1 g, or more are detected in and / or on the target tissue of the plant. Alternatively or additionally, metabolites isolated from the microorganisms and aggregates of this disclosure may be present in the formulation in an amount that effectively increases the biomass and / or yield of the plant to which such formulation is applied by at least 1%, at least 2%, at least 3%, at least 5%, at least 10%, at least 15%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, or more when compared with a reference agricultural plant to which the formulation of this disclosure is not applied. Alternatively or additionally, metabolites isolated from the microorganisms and aggregates of this disclosure may be present in the formulation in an amount of at least 1%, at least 2%, at least 3%, at least 5%, at least 10%, at least 15%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100% or more, when compared with a reference agricultural plant to which the formulation of this disclosure has not been applied, to effectively detectably adjust the agronomic traits of interest of the plant to which such formulation has been applied.

[0106] In some embodiments, the agricultural compositions taught herein are shelf-stable. In some aspects, the microorganisms taught herein are freeze-dried. In some aspects, the microorganisms taught herein are spray-dried. In some aspects, the microorganisms taught herein are placed in a liquid formulation. In some aspects, the microorganisms taught herein are present on granules.

[0107] This article also describes a variety of isolated microorganisms enclosed in objects selected from the group consisting of: bottles, wide-mouth bottles, ampoules, packaging, utensils, bags, boxes, storage boxes, envelopes, cartons, containers, silos, shipping containers, carriages, and crates.

[0108] In some aspects, combining a selected plant species with the disclosed microorganisms (operational taxonomic units (OTUs), strains, or compositions comprising any of the foregoing) results in increased crop yields and the generation of their products. Therefore, in one aspect, this disclosure provides a synthetic combination of a seed of a first plant and a microbial preparation coated onto the surface of the seed of the first plant, such that the microorganisms are present at a higher level on the surface of the seed compared to those present on the surface of an uncoated reference seed. In another aspect, this disclosure provides a synthetic combination of a portion of a first plant and a microbial preparation coated onto the surface of that portion of the first plant, such that the microorganisms are present at a higher level on the surface of that portion of the first plant compared to those present on the surface of an uncoated reference plant portion. The above methods can be used alone or in conjunction with plant breeding and transgenic technologies.

[0109] In some embodiments, the isolated bacterial strains may be selected from the group consisting of: Bacillus tekirae deposited as NRRL accession number B-67810, Bacillus methyltrophicus deposited as NRRL accession number B-67812, Bacillus amyloliquefaciens deposited as NRRL accession numbers B-67815, B-67947 or B-67947, Bacillus alginate-like bacteria deposited as NRRL accession number B-67813 or B-67811, Agaricus aureus / Oligospora spp. deposited as NRRL accession number 67879 (the sexual and asexual forms of the same species of microorganism, respectively), Bacillus pumilus deposited as NRRL accession number B-67878, and Bacillus fusiformis deposited as NRRL accession number B-67871.

[0110] In some embodiments, the bioconfluent may comprise bacterial strains isolated from *Bacillus alginate* deposited as NRRL accession numbers B-67813 and B-67811. In some embodiments, the bioconfluent may comprise bacterial strains isolated from *Bacillus amyloliquefaciens* deposited as NRRL accession numbers B-67947 and B-67813. In some embodiments, the bioconfluent may comprise bacterial strains isolated from *Bacillus belyss* deposited as NRRL accession number B-50614 and *Bacillus pumilus* deposited as NRRL accession number B-67878.

[0111] In some embodiments, the isolated bacterial strains have morphological and physiological characteristics substantially similar to those of the bacterial strains isolated in this disclosure. In some embodiments, the isolated bacterial strains have genetic characteristics substantially similar to those of the bacterial strains isolated in this disclosure. In some embodiments, the isolated bacterial strains are naturally occurring or artificially generated mutants of the bacterial strains isolated in this disclosure. In some embodiments, the isolated bacterial strains are gene-edited, altered, or modified bacterial strains. In some embodiments, the bacterial strains isolated in this disclosure are located in substantially pure cultures. In some embodiments, the bacterial strains isolated in this disclosure are located in pure cultures. In some embodiments, the bacterial strains isolated in this disclosure are located in cell fractions, extracts, or supernatants.

[0112] In some embodiments, progeny and / or mutants of the isolated bacterial strains of this disclosure are envisioned. In some embodiments, progeny, mutants, and / or genetically modified forms of the isolated bacterial strains of this disclosure are envisioned. In some embodiments, the isolated bacterial strains of this disclosure comprise a polynucleotide sequence sharing at least 97% sequence identity with any of SEQ ID NO:1-12.

[0113] In some embodiments, cell-free or inactivated preparations of the isolated bacterial strains of this disclosure, or mutants of said isolated bacterial strains, are contemplated. In some embodiments, cell-free or inactivated preparations of the isolated bacterial strains of this disclosure, or mutants of said isolated bacterial strains, or gene-edited, altered, or modified variants, are contemplated. In some embodiments, metabolites produced from the isolated bacterial strains of this disclosure, or mutants of said isolated bacterial strains, are contemplated. In some embodiments, metabolites produced from the isolated bacterial strains of this disclosure, or mutants of said isolated bacterial strains, or genetically modified variants, are contemplated.

[0114] In some embodiments, the agricultural composition comprises an isolated bacterial strain and an agriculturally acceptable vector. The isolated bacterial strain may be present in the composition at a concentration of 1 × 10^2 to 1 × 10^12 CFU / g. The agricultural composition may be formulated as a seed coating.

[0115] In some embodiments, a method of conferring at least one beneficial trait to a plant species includes applying an isolated strain of bacterial strain to the plant or the growth medium in which the plant is situated. In some embodiments, a method of conferring at least one beneficial trait to a plant species includes applying the agricultural composition of this disclosure to the plant or the growth medium in which the plant is situated.

[0116] In some embodiments, this disclosure teaches a method for cultivating a plant having at least one beneficial trait. In some embodiments, the method includes applying an isolated strain of bacterial strains or microbial aggregates to seeds of the plant; sowing or planting the seeds; and planting the plant. In some embodiments, the isolated strain of bacterial strains or microbial aggregates are applied in the form of an agricultural composition that also comprises an agriculturally acceptable carrier.

[0117] In some embodiments, the microbial aggregate comprises at least two microorganisms selected from the group consisting of: A) *Bacillus tekirae*, *Bacillus methyltrophicus*, *Bacillus amyloliquefaciens*, *Bacillus alginate*, *Discocele aureosulcata* / *Arthrozoa oligospora* (sexual and asexual forms, respectively), *Bacillus pumilus*, and *Bacillus fusiformis*; and

[0118] B) *Arthrobacter cupressi*, *Arthrobactermysorens*, *Arthrobacter nicotinovorans*, *Arthrobacter pascens*, *Bacillus megaterium*, *Bacillus subtilis*, *Bacillus thuringiensis*, *Bacillus belesii*, *Brevibacterium frigoritolerans*, *Herbaspirillum chlorophenolicum*, *Kosakonia radicincitans*, *Bacillus fusiformis*, *Massilia kyonggiensis*, *Massilia niastensis*, *Novosphingobium sediminicola*, *Paenibacillus* amylolyticus), Paenibacillus glycanilyticus, Paenibacillus polymyxa, Pseudomonas fluorescens, Pseudomonas jinjuensis, Pseudomonas oryzihabitans, Pseudomonas putida, Rahnella aquatilis, and Tumebacillus permanentifrigoris; and combinations thereof, wherein at least one microorganism is selected from A).

[0119] In some embodiments, the microbial aggregate comprises at least two isolated bacterial strains selected from the group consisting of: A) *Bacillus tekirae* deposited as NRRL accession number B-67810, *Bacillus methyltrophus* deposited as NRRL accession number B-67812, *Bacillus amyloliquefaciens* deposited as NRRL accession number B-67815, *Bacillus amyloliquefaciens* deposited as NRRL accession number B-67947, and *Bacillus amyloliquefaciens* deposited as NRRL accession number B-67949. Bacillus, Bacillus alginate-containing Bacillus deposited under NRRL accession number B-67813, Bacillus alginate-containing Bacillus, Bacillus alginate-containing Bacillus, Bacillus alginate-containing Bacillus deposited under NRRL accession number B-67811, Aureobasidium aureum / Oligospora spp. deposited under NRRL accession number 67879 (sexual and asexual forms, respectively), Bacillus pumilus deposited under NRRL accession number B-67878, and Bacillus fusiformis deposited under NRRL accession number B-67871;And B) *Arthrobacter cylindrica* deposited under NRRL accession number B-67183, *Arthrobacter cylindrica* and *Arthrobacter mysore* deposited under NRRL accession number B-67289, *Arthrobacter trophoblastus*, *Bacillus megaterium*, *Bacillus megaterium*, *Bacillus megaterium*, *Bacillus megaterium*, *Bacillus subtilis*, *Bacillus subtilis*, *Bacillus thuringiensis*, and *Bacillus vesalis* deposited under NRRL accession number B-67370, and *Bacillus vesalis* deposited under NRRL accession number B-50614. Bacteria, including: *Aeromonas frostridae* (NRRL accession number B-67360), *Cyclopyralidus* (NRRL accession number B-67236), *Cyclopyralidus* (NRRL accession number B-67197), *Sacchariformis* (NRRL accession number B-67171), *Sacchariformis* (NRRL accession number B-67946), *Bacillus fusiformis*, *Morphus gypseus* (NRRL accession number B-67198), and *Bacillus fusiformis* (NRRL accession number B-67235). Niasmacaster, Niasmacaster deposited under NRRL accession number B-67199, Niasmacaster, Neosphingomonas sphingosine mononitrate deposited under NRRL accession number B-67945, Bacillus amyloliquefaciens, Bacillus polymyxa degenerates deposited under NRRL accession number B-67204, Bacillus polymyxa, Pseudomonas fluorescens, Pseudomonas fluorescens, Pseudomonas fluorescens, Pseudomonas fluorescens, Pseudomonas fluorescens, Pseudomonas fluorescens, Pseudomonas fluorescens, Pseudomonas jinjuensis deposited under NRRL accession number B-67207, as NRRL accession The following are deposited under accession number B-67225: *Pseudomonas aeruginosa*, *Pseudomonas aeruginosa*, *Pseudomonas aeruginosa*, *Pseudomonas aeruginosa*, *Pseudomonas aeruginosa*, *Pseudomonas aeruginosa*, *Pseudomonas aeruginosa*, *Pseudomonas aeruginosa*, *Pseudomonas aeruginosa*, *Pseudomonas aeruginosa*, *Pseudomonas aeruginosa*, *Bacillus aquaticus*, *Bacillus aeruginosa* permanently frozen and expanded as NRRL accession number B-67301, and *Bacillus aeruginosa* permanently frozen and expanded as NRRL accession number B-67302; and combinations thereof, wherein at least one microorganism is selected from A).

[0120] In some embodiments, the microbial aggregates have morphological and physiological characteristics substantially similar to those of the microbial aggregates disclosed herein. In some embodiments, the microbial aggregates have genetic characteristics substantially similar to those of the microbial aggregates disclosed herein. In some embodiments, the microbial aggregates are located in substantially pure cultures. In some embodiments, subsequent generations of any microorganisms within the microbial aggregate are envisioned. In some embodiments, mutants of any microorganisms within the microbial aggregate are envisioned. In some embodiments, gene-edited, altered, or modified variants of any microorganisms within the microbial aggregate are envisioned. In some embodiments, cell-free or inactivated preparations of the microbial aggregate or mutants or gene-edited, altered, or modified variants of any microorganism within the microbial aggregate are envisioned. In some embodiments, metabolites produced by the microbial aggregate or mutants or gene-edited, altered, or modified variants of any microorganism within the microbial aggregate are envisioned.

[0121] In some embodiments, the agricultural composition comprises a microbial aggregate and an agriculturally acceptable carrier. The microbial aggregate of the agricultural composition may be present in the composition at a concentration of 1 × 10^3 to 1 × 10^12 bacterial cells / gram. In some embodiments, the agricultural composition is formulated as a seed coating. In some embodiments, a method of conferring at least one beneficial trait to a plant species includes applying the microbial aggregate to the plant or the growth medium in which the plant is situated. In some embodiments, a method of conferring at least one beneficial trait to a plant species includes applying the agricultural composition to the plant or the growth medium in which the plant is situated.

[0122] In some embodiments, the microbial aggregate comprises microorganisms selected from the group consisting of: *Bacillus tekirae* deposited under NRRL accession number B-67810, *Methyltrophic Bacillus* deposited under NRRL accession number B-67812, *Bacillus amyloliquefaciens* deposited under NRRL accession number B-67815, *Bacillus amyloliquefaciens* deposited under NRRL accession number B-67947, *Bacillus amyloliquefaciens* deposited under NRRL accession number B-67949, and so on. The following are Bacillus alginate, Bacillus alginate, and Bacillus alginate deposited under NRRL accession number B-67813; Bacillus alginate deposited under NRRL accession number B-67811; Aureobacterium glomeratum / Oligospora spp. (sexual and asexual forms, respectively) deposited under NRRL accession number 67879; Bacillus pumilus deposited under NRRL accession number B-67878; and Bacillus fusiformis deposited under NRRL accession number B-67871.

[0123] In some embodiments, a method of conferring at least one beneficial trait to a plant species includes applying at least one isolated bacterial species to the plant or the growth medium in which the plant is situated, wherein the at least one isolated bacterial species is selected from the group consisting of: Bacillus tekirae, Bacillus methyltrophicus, Bacillus amyloliquefaciens, Bacillus alginate, Discocele aureus / Oligospora spp. (sexual and asexual forms, respectively), Bacillus pumilus, Bacillus fusiformis, and combinations thereof.

[0124] In another embodiment, at least one isolated bacterial species is a strain selected from the group consisting of: *Bacillus tekirae* deposited as NRRL accession number B-67810, *Bacillus methyltrophicus* deposited as NRRL accession number B-67812, *Bacillus amyloliquefaciens* deposited as NRRL accession number B-67815, *Bacillus amyloliquefaciens* deposited as NRRL accession number B-67947, *Bacillus amyloliquefaciens* deposited as NRRL accession number B-67949, and so on. Bacillus alginate, Bacillus alginate, Bacillus alginate, Bacillus alginate deposited under NRRL accession number B-67813, Bacillus alginate deposited under NRRL accession number B-67811, Aureobasidium auriculatum / Oligospora spp. (sexual and asexual forms, respectively) deposited under NRRL accession number 67879, Bacillus pumilus deposited under NRRL accession number B-67878, and Bacillus fusiformis deposited under NRRL accession number B-67871, as well as combinations thereof.

[0125] In some embodiments, the isolated bacterial strains are selected from Table 1 or Table 1A. In some embodiments, isolated bacterial strains with substantially similar morphological and physiological characteristics to those selected from Table 1 or Table 1A are envisioned. In some embodiments, isolated bacterial strains with substantially similar genetic characteristics to those from Table 1 or Table 1A are envisioned. In some embodiments, substantially pure cultures of isolated bacterial strains from Table 1 or Table 1A are envisioned. In some embodiments, progeny or mutants of isolated bacterial strains from Table 1 or Table 1A are envisioned. In some embodiments, cell-free or inactivated preparations derived from isolated bacterial strains from Table 1 or Table 1A or their mutants are envisioned. In some embodiments, metabolites produced by isolated bacterial strains from Table 1 or Table 1A or their mutants are envisioned.

[0126] In some embodiments, the agricultural composition comprises isolated bacterial strains from Table 1 or Table 1A and an agriculturally acceptable vector. In some embodiments, the isolated bacterial strains are present in the agricultural composition at a concentration of 1 × 10^2 to 1 × 10^12 CFU / g. In some embodiments, the agricultural composition is formulated as a seed coating. In some embodiments, a method of conferring at least one beneficial trait to a plant species includes applying isolated bacterial strains from Table 1 or Table 1A to a plant or the growth medium in which said plant is situated. In some embodiments, a method of conferring at least one beneficial trait to a plant species includes applying the agricultural composition of this disclosure to a plant or the growth medium in which said plant is situated.

[0127] In some embodiments, the microbial aggregate comprises at least two microorganisms selected from those listed in Table 1 or Table 1A. In some embodiments, the microbial aggregate comprises at least two microorganisms, wherein at least one microorganism is selected from Table 1 or Table 1A, and the other microorganisms are selected from Table 2.

[0128] In some embodiments, plant seeds enhanced with microbial seed coating comprise plant seeds and a seed coating applied to said plant seeds, wherein the seed coating comprises at least two microorganisms listed in Tables 1, 1A, and 2, and at least one microorganism is selected from Table 1 or Table 1A. In another embodiment, the seed coating comprises a combination of microorganisms. In yet another embodiment, the seed coating comprises at least one microorganism listed in Table 1 or Table 1A at a concentration of 1 × 10^2 to 1 × 10^9 CFU per seed. In some embodiments, the microorganisms selected from Table 1 are used in agriculture. In some embodiments, the synthetic combination of plant and microorganism comprises at least one plant and at least one microorganism selected from Table 1 or Table 1A.

[0129] In some embodiments, methods for enhancing or promoting desired phenotypic traits of a plant species include applying at least one microorganism selected from Table 1 or Table 1A to the plant or the growth medium in which the plant is situated. In another embodiment, the application of the at least one bacterium is performed by: coating plant seeds with the bacterium, coating plant parts with the bacterium, spraying the bacterium onto plant parts, spraying the bacterium into furrows where the plant or seeds will be placed, watering the bacterium onto plant parts or the area where the plant will be placed, dispersing the bacterium onto plant parts or the area where the plant will be placed, seeding the bacterium onto plant parts or the area where the plant will be placed, and combinations thereof.

[0130] In any of the methods described, the microorganism may comprise a 16S rRNA nucleic acid sequence having at least 97% sequence identity with a 16S rRNA nucleic acid sequence of a bacterium selected from the genera or species provided in Table 1 or Table 1A. Attached Figure Description

[0131] This disclosure will be more fully understood through the following detailed description and the accompanying drawings and sequence listings that form part of this application.

[0132] Figure 1 A schematic diagram of a generalized process of the disclosed accelerated microbial selection (AMS) (also referred to herein as directed microbial selection) method is shown. When viewed in the context of a microbial aggregate, the diagram illustrates the directed evolution of the microbial aggregate. This process is one method for obtaining the beneficial microorganisms of this disclosure.

[0133] Figure 2 A generalized process flow diagram of the implementation scheme for obtaining the beneficial microorganisms of this disclosure is shown.

[0134] Figure 3 A graphical representation and associated flowchart of an embodiment of the beneficial microorganisms of this disclosure are shown.

[0135] Figure 4 A graphical representation and associated flowchart of an embodiment of the beneficial microorganisms of this disclosure are shown.

[0136] Figure 5 This shows Bacillus tekirae BEC80 colonizing the root (a GFP-tagged microorganism on cauliflower roots).

[0137] Figure 6 The activity status of Bacillus tekiria BEC80 across a certain temperature and pH range is shown at 0, 2, 3 and 7 days.

[0138] Figure 7 The relative yields of iturin in Bacillus tekirae BEC80 and a reference Bacillus strain are shown.

[0139] Figure 8 The relative yields of nutrient-rich elements in Bacillus tekirae BEC80 and the reference Bacillus strain are shown.

[0140] Figure 9 A photograph shows the plant in situ biostimulant activity of Bacillus methyltrophicus BEC60.

[0141] Figure 10 The image shows Bacillus methyltrophicus BEC60 colonizing roots (fluorescently labeled microorganisms in wheat roots).

[0142] Figure 11 The images show untreated, post-harvest apples treated with 100 ppm of a commercially available treatment agent and with Bacillus tekirae BEC80.

[0143] Figure 12A The population counts of *C. elegans* over time are shown. OP50 bacterial (-) control, Bt (+) control, and Bacillus (-) control are included.

[0144] Figure 12B This study illustrates a plant in situ action model test (tomato). Counts of root-knot nematode larvae and adults at different stages of infection in tomato roots after inoculation.

[0145] Figure 13 The growth of Bacillus amyloliquefaciens BEC69 across a wide range of temperature and pH conditions is shown.

[0146] Figure 14 The colonization of plant components of Bacillus amyloliquefaciens BEC69 (a fluorescently labeled microorganism) in tomatoes, soybeans, wheat, and corn is shown.

[0147] Figure 15 This study illustrates the colonization of plant components of Bacillus alginate BEC68 (a fluorescently labeled microorganism) in the rhizosphere tissue of maize.

[0148] Figure 16 The image shows BEC77 colonizing the surface of aboveground and underground plants (cauliflower leaf and rhizosphere) (fluorescently labeled microorganism).

[0149] Figure 17 The image shows BEC68 and BEC78 co-colonizing in plant roots (fluorescently labeled microorganisms).

[0150] Figure 18A This paper demonstrates the enhanced phosphate rock solubilizing activity of some of the biostimulant microorganisms disclosed herein.

[0151] Figure 18B This paper demonstrates the enhanced magnesium silicate solubilizing activity of some of the biostimulant microorganisms disclosed herein.

[0152] Figure 19 The growth of *Microbacterium arabinogalactanus* BEC102 across a wide range of temperature and pH conditions is shown.

[0153] The sequence description and accompanying sequence listing conform to the rules governing the disclosure of nucleotide and amino acid sequences in regulatory patent applications as set forth in 37 C. FR § § 1.821 and 1.825. The sequence description includes the three-letter codes of the amino acids as defined in 37 C. FR § § 1.821 and 1.825, which are incorporated herein by reference.

[0154] The microorganisms described in this application are deposited at the Agricultural Research Culture Collection (NRRL), an international depository located at 1815 North University Street, Peoria, IL 61604, USA.

[0155] These deposits were prepared in accordance with the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure.

[0156] These deposits are prepared in accordance with and meet the standards set forth in 37 C. FR § § 1.801-1.809 and the Manual of Patent Examining Procedure § § 2402-2411.05.

[0157] The NRRL registration numbers, deposit dates, and descriptions of the aforementioned Budapest Treaty deposits are provided in Tables 1 and 2.

[0158] Table 1: Microorganisms

[0159]

[0160] Table 1A: Microorganisms

[0161] Table 2: Microorganisms

[0162]

[0163] Detailed Implementation

[0164] While the following terms are believed to be well understood by those skilled in the art, they are set forth in order to explain the subject matter disclosed herein.

[0165] The term "a" or "an" refers to one or more of the entity, that is, multiple referents. Therefore, the terms "a" or "an," "one or more," and "at least one" are used interchangeably herein. Furthermore, the reference to "an element" by the indefinite article "a" or "an" does not preclude the possibility of more than one element, unless the context explicitly requires the existence of exactly one element.

[0166] As used herein, the terms “microorganism” or “microbe” should be interpreted broadly. These terms are used interchangeably and include, but are not limited to, the two prokaryotic domains (i.e., bacteria and archaea) and eukaryotic fungi and protists. In some embodiments, this disclosure refers to “microbes” in Tables 1-2, or in the various other tables or paragraphs present in this disclosure. This characterization may refer not only to the taxonomic genera of bacteria identified in the tables, but also to the taxonomic species identified, as well as the various novel and recently identified bacterial strains in the tables.

[0167] As used herein, the term "microorganism" or "microbial body" refers to any species or taxonomic unit of a microbial body, including but not limited to archaea, bacteria, microalgae, fungi (including molds and yeasts), mycoplasma, microspores, nanobacteria, oomycetes, and protozoa. In some embodiments, a microorganism or microbial body encompasses a single cell (e.g., a single-celled microorganism) or more than one cell (e.g., a multicellular microorganism). Thus, a "microbial body community" can refer to multiple cells of a single microorganism that share a common genetic derivation.

[0168] As used herein, the term “bacteria” generally refers to any prokaryotic organism and may include organisms from the kingdoms Eubacteria (bacteria), Archaea (archaea), or both. In some cases, bacterial genera or other taxonomic classifications have been redesignated for other reasons (such as, but not limited to, evolutionary fields of whole-genome sequencing), and it should be understood that such redesignations fall within the scope of any taxonomic claim. For example, certain species of the genus *Erwinia* have been described in the literature as belonging to the genus *Pantoea* (Zhang, Y., Qiu, S. Examining phylogenetic relationships of *Erwinia* and *Pantoea* species using whole genome sequence data. Antonie van Leeuwenhoek 108, 1037–1046 (2015)).

[0169] The term "16S" refers to the DNA sequence of the 16S ribosomal RNA (rRNA) sequence of bacteria. 16S rRNA gene sequencing is an established method for studying bacterial phylogeny and taxonomy.

[00166] As used herein, the term "fungus" generally refers to any organism from the fungal kingdom. Historically, fungi have been taxonomically classified based on their morphological characteristics. Since the mid-19th century, it has been recognized that some fungi have pleomorphic life cycles, and different names are used for different forms of the same fungus. In 1981, the Sydney Congress of the International Mycological Association established rules for naming fungi asexual, sexual, or holotype based on their state (Taylor, JW One Fungus = One Name: DNA and fungal nomenclature twenty years after PCR. IMA Fungus 2, 113–120 (2011)). With the development of genome sequencing, it has become clear that molecular phylogenetic taxonomy does not align with morphological nomenclature (Shenoy, BD; Jeewon, R.; and Hyde, KD (2007). Impact of DNA sequence-data on the taxonomy of anamorphic fungi. Fungal Diversity 26:1-54). Therefore, in 2011, the International Botanical Congress adopted a resolution approving the International Nomenclature Code for Algae, Fungi and Plants (Melbourne Code) (2012), which stipulates that “one fungus = one name” (Hawksworth, DL Managing and coping with names of pleomorphic fungiin aperiod of transition. IMA Fungus 3, 15–24 (2012)).

[0170] The term "internal transcribed spacer" ("ITS") refers to the spacer DNA (non-coding DNA) located between the small subunit ribosomal RNA (rRNA) and the large subunit (LSU) rRNA gene in the corresponding transcribed region of a chromosome or polycistronic rRNA precursor transcript. ITS gene sequencing is an established method for studying fungal phylogeny and taxonomy. In some cases, the "large subunit" ("LSU") sequence is used to identify fungi. LSU gene sequencing is an established method for studying fungal phylogeny and taxonomy. Some fungal microorganisms of the present invention can be described by ITS sequences, and some fungal microorganisms can be described by LSU sequences. It should be understood that both are equally descriptive and accurate for determining taxonomy.

[0171] The term "microbial consortia" or "microbial consortium" refers to a subset of a microbial community of a single species, or a strain of a species, that can be described as performing a common function, or as participating in or contributing to or associated with identifiable parameters or plant phenotypic traits. This community may contain one or more species of microorganisms, or strains of a species. In some cases, these microorganisms coexist symbiotically within the community.

[0172] The term "microbial community" refers to a group of microorganisms comprising two or more species or strains. Unlike microbial aggregates, microbial communities do not necessarily perform a common function, nor do they necessarily participate in or cause or relate to identifiable parameters or plant phenotypic traits.

[0173] The terms “accelerated microbial selection” or “AMS” are used interchangeably with the terms “directed microbial selection” or “DMS” and refer to an iterative selection method used in some embodiments of this disclosure to obtain a claimed microbial species or an aggregate of said species.

[0174] As used herein, the terms “isolated strain,” “isolated,” “isolated microorganism,” and similar terms are intended to mean one or more microorganisms that have been isolated from at least one material associated with a particular environment (e.g., soil, water, plant tissue).

[0175] Therefore, the “isolated microorganism” does not exist in its natural environment; rather, it is removed from its natural environment and placed in a non-natural state of existence through the various techniques described herein. Thus, the isolated strain can exist, for example, in the form of a biologically pure culture or spores (or other forms of the strain) combined with an agricultural carrier.

[0176] In some aspects of this disclosure, the isolated microorganisms exist in the form of isolated and biologically pure cultures. Those skilled in the art will understand that an isolated and biologically pure culture of a particular microorganism means that the culture is substantially free of (to the extent scientifically reasonable) other living organisms and contains only the single microorganism in question. The culture may contain varying concentrations of the microorganism. This disclosure states that isolated and biologically pure microorganisms are generally “certainly different from less pure or impure material.” See, for example, In re Bergstrom, 427F.2d 1394, (CCPA 1970) (discussing purified prostaglandins), also see Inre Bergy, 596F.2d 952 (CCPA 1979) (discussing purified microorganisms), also see Parke-Davis & Co. v HKMulford & Co., 189F.95 (SDNY 1911) (Learned Hand, discussing purified adrenaline), Partial Maintenance, Partial Withdrawal, 196F.496 (2d Cir. 1912), each of which is incorporated herein by reference. Furthermore, in some aspects, this disclosure provides certain quantitative measures of concentration or purity limits that must be found in isolated and biologically pure microbial cultures. In some embodiments, the presence of these purity values ​​is another property distinguishing the microorganisms disclosed in this invention from those that exist in their natural state. See, for example, Merck & Co. v. Olin Mathieson Chemical Corp., 253F.2d 156 (4th Cir. 1958) (discussing purity limits for vitamin B12 produced by microorganisms), which is incorporated herein by reference.

[0177] As used herein, “isolated isolate” should be considered as meaning a composition or culture that, after being isolated from one or more other microorganisms, primarily comprises a single genus, species, or strain of microorganisms. This phrase should not be considered as an indication of the degree of isolation or purification of the microorganisms. However, “isolated isolate” may essentially contain only one genus, species, or strain of microorganisms.

[0178] As used herein, the term "growth medium" is any culture medium suitable for supporting plant growth. As examples, media can be natural or artificial, including but not limited to: soil, potting mixes, bark, vermiculite, hydroponic solutions applied alone to solid plant support systems, and tissue culture gels. It should be understood that media can be used alone or in combination with one or more other media. They can also be used with or without the addition of exogenous nutrients and physical support systems for roots and leaves.

[0179] In one embodiment, the growth medium is a naturally occurring culture medium, such as soil, sand, mud, clay, humus, topsoil, rock, or water. In another embodiment, the growth medium is artificial. Such an artificial growth medium may be configured to simulate the conditions of a naturally occurring culture medium; however, this is not required. The artificial growth medium may be made from one or more of any amount and combination of materials, including sand, minerals, glass, rock, water, metals, salts, nutrients, and water. In one embodiment, the growth medium is sterile. In another embodiment, the growth medium is not sterile.

[0180] Culture media can be modified or enriched with additional compounds or components, such as components that can facilitate the interaction and / or selection of specific microbial communities with and between plants and each other. For example, antibiotics (such as penicillin) or sterilizing agents (e.g., quaternary ammonium salts and oxidants) may be present, and / or physical conditions (such as salinity, phytonutrients (e.g., organic and inorganic minerals (such as phosphorus, nitrogen salts, ammonia, potassium, and micronutrients such as cobalt and magnesium), pH, and / or temperature) may be modified.

[0181] The term "plant" generally includes the whole plant, plant organs, plant tissues, seeds, plant cells, and their offspring. Plant cells include, but are not limited to, cells derived from seeds, suspension cultures, plumules, meristematic zones, callus, leaves, roots, buds, gametophytes, sporophytes, pollen, and microspores. "Plant component" is intended to refer to the whole plant or plant part, which may include differentiated and / or undifferentiated tissues, such as, but not limited to, plant tissues, parts, and cell types. In one embodiment, a plant component is one of the following: whole plant, seedling, meristematic tissue, ground tissue, vascular tissue, cortex, seed, leaf, root, bud, stem, flower, fruit, stolons, bulb, tuber, corm, bud, shoot, bud, bract, nodule tissue, and various forms of cells and cultures (e.g., single cells, protoplasts, plumules, callus). The term "plant organ" refers to plant tissues or groups of tissues that constitute different morphological and functional parts of a plant. As used herein, “plant part” is synonymous with “part” of a plant and refers to any part of a plant, which may include different tissues and / or organs, and is used interchangeably with the term “tissue” throughout the text.

[0182] "Offspring" includes any subsequent generations of an organism produced through sexual or asexual reproduction.

[0183] As used herein, the term "plant component" refers to intact plant callus, plant masses, and plant cells that can be derived from plant cells, plant protoplasts, plant cell tissue cultures, or from plant or plant parts such as embryos, pollen, ovules, seeds, leaves, flowers, branches, fruits, grains, spikes, rachis, bark, stems, roots, root tips, anthers, etc., and their own parts. Grain is intended to refer to mature seeds produced by commercial growers for purposes other than the growth or reproduction of a species. Progeny, variants, and mutants of regenerated plants are also included within the scope of this invention, provided that these parts contain introduced polynucleotides.

[0184] Similarly, "plant reproductive component" is intended to generally refer to any part of a plant that can create other plants through the sexual or asexual reproduction of that plant, such as, but not limited to: seeds, seedlings, roots, buds, cuttings, scions, grafted seedlings, stolons, bulbs, tubers, corms, buds, or bracts. Plant components may be located in the plant or in plant organs, tissue cultures, or cell cultures.

[0185] The term "monocotyledonous" or "monocotyledonous plant" refers to the subclass Angiosperms, also known as the "monocotyledonous plant class," whose seeds typically contain only one embryonic leaf or cotyledon. The term includes references to the whole plant, plant components, plant organs (e.g., leaves, stems, roots, etc.), seeds, plant cells, and their offspring.

[0186] The term "dicotyledonous" or "dicotyledonous plant" refers to the subclass Angiosperms, also known as the "dicotyledonous plant class," whose seeds typically contain two embryonic leaves or cotyledons. The term includes references to the whole plant, plant components, plant organs (e.g., leaves, stems, roots, etc.), seeds, plant cells, and their offspring.

[0187] As used in this article, the term "cultivar" refers to a plant variety, strain, or family that has been produced through horticultural or agronomic techniques and is not typically found in wild-type populations.

[0188] As used herein, “improved” should be interpreted broadly to encompass improvements in plant characteristics compared to control plants or to known average quantities associated with the characteristics in question. For example, “improved” plant biomass associated with the application of the beneficial microorganisms or aggregates of this disclosure can be demonstrated by comparing the biomass of plants treated with the microorganisms taught herein with the biomass of untreated control plants. Alternatively, the biomass of plants treated with the microorganisms taught herein can be compared with the average biomass typically achieved by a given plant (as expressed in scientific or agricultural publications known to those skilled in the art). In this disclosure, “improved” does not necessarily require that the data be statistically significant (e.g., p < 0.05); rather, any quantifiable difference indicating that one value (e.g., average treatment value) differs from another value (e.g., average control value) can be considered “improved.”

[0189] As used herein, “suppression and inhibition” and similar terms should not be interpreted as requiring complete suppression or inhibition, but may be necessary in some implementations.

[0190] As used herein, the term “genotype” refers to the genetic composition of an individual cell, cell culture, tissue, organism (e.g., plant) or population of organisms.

[0191] The compositions and methods described herein can provide plants with modified "agronomical traits," "agronomically important traits," or "traits of agronomic interest," which may include, but are not limited to, the following: disease resistance, drought tolerance, heat tolerance, cold tolerance, salt tolerance, metal tolerance, herbicide tolerance, improved water use efficiency, improved nitrogen use efficiency, improved nitrogen fixation, insect resistance, herbivore resistance, pathogen resistance, increased yield, enhanced health, improved vitality, improved growth, improved photosynthetic capacity, enhanced nutrition, altered protein content, altered oil content, increased biomass, increased shoot length, increased root length, improved root architecture, metabolite regulation, proteome regulation, increased seed weight, altered seed carbohydrate composition, altered seed oil composition, altered seed protein composition, and altered seed nutrient composition compared to isoline plants that do not contain modifications derived from the methods or compositions described herein.

[0192] "Agronomic trait potential" is intended to refer to the ability of a plant component to exhibit a phenotype (preferably, a modified agronomic trait) at some point in its life cycle or to transfer said phenotype to another plant component it is associated with in the same plant.

[0193] As used herein, the terms “molecular marker,” “marker,” or “genetic marker” refer to an indicator used in methods for observing differences in the characteristics of nucleic acid sequences. Examples of such indicators are restriction fragment length polymorphism (RFLP) markers, amplified fragment length polymorphism (AFLP) markers, single nucleotide polymorphisms (SNPs), insertion mutations, microsatellite markers (SSRs), sequence-characterized amplified regions (SCARs), enzyme digestion amplified polymorphic sequences (CAPS) markers, or isoenzyme markers, or combinations of markers described herein (which define specific gene and chromosomal locations). The localization of molecular markers near alleles is a procedure that can be performed by a person with experience in molecular biotechnology.

[0194] As used herein, the term "trait" refers to a characteristic or phenotype. For example, in the context of some embodiments of this disclosure, crop yield refers to the amount of marketable biomass (e.g., fruit, fiber, cereal) produced by the plant. Desired traits may also include other plant characteristics, including but not limited to: water use efficiency, nutrient use efficiency, productivity, mechanical harvestability, fruit maturity, shelf life, insect / disease resistance, early maturity, stress tolerance, etc. Traits can be inherited in a dominant or recessive manner, or in a partially or incompletely dominant manner. Traits can be monogenic (i.e., determined by a single locus) or polygenic (i.e., determined by more than one locus), or can be the result of the interaction of one or more genes with the environment.

[0195] As used herein, the term “phenotype” refers to an observable characteristic of an individual cell, cell culture, organism (e.g., plant), or population of organisms, which arises from the interaction between an individual’s genetic makeup (i.e., genotype) and its environment.

[0196] As used herein, a “synthetic nucleotide sequence” or “synthetic polynucleotide sequence” is a nucleotide sequence that is known not to exist in nature or is not naturally occurring. Generally, when compared with any other naturally occurring nucleotide sequence, such a synthetic nucleotide sequence will contain at least one nucleotide difference.

[0197] As used herein, the term "nucleic acid" refers to a polymer of nucleotides (ribonucleotides or deoxyribonucleotides) of any length or similar. The term refers to the primary structure of a molecule and therefore includes double-stranded and single-stranded DNA, as well as double-stranded and single-stranded RNA. It also includes modified nucleic acids, such as methylated and / or terminally capped nucleic acids, nucleic acids containing modified bases, nucleic acids with modified backbones, etc. The terms "nucleic acid" and "nucleotide sequence" are used interchangeably.

[0198] As used herein, the term "gene" refers to any segment of DNA associated with a biological function. Therefore, genes include, but are not limited to, coding sequences and / or regulatory sequences required for their expression. Genes may also include unexpressed segments of DNA, such as recognition sequences that form other proteins. Genes can be obtained from a variety of sources, including cloning from sources of interest or synthesis from known or predicted sequence information, and may include sequences designed to have desired parameters.

[0199] As used herein, the terms “homologous” or “homology” or “orthologous” are known in the art and refer to related sequences that share a common ancestor or family member and are determined based on the degree of sequence identity. The terms “homology,” “homology,” “substantially similar,” and “substantially corresponding” are used interchangeably herein. They refer to nucleic acid fragments in which changes to one or more nucleotide bases do not affect the ability of the nucleic acid fragment to mediate gene expression or produce a particular phenotype. These terms also refer to modifications of nucleic acid fragments of this disclosure, such as the deletion or insertion of one or more nucleotides that substantially do not alter the functional characteristics of the resulting nucleic acid fragment relative to the initial, unmodified fragment. Therefore, it should be understood that, as those skilled in the art will appreciate, this disclosure covers more than the specific exemplary sequences. These terms describe the relationship between a gene found in one species, subspecies, variety, cultivar, or strain and a corresponding or equivalent gene in another species, subspecies, variety, cultivar, or strain. For the purposes of this disclosure, homologous sequences are compared. “Homologous sequences” or “homology” or “orthologous” are considered, believed, or known to be functionally related. Functional relationships can be indicated in any of a variety of ways, including but not limited to: (a) degree of sequence identity and / or (b) identical or similar biological functions. Preferably, both (a) and (b) are indicated. Homology can be determined using software programs readily available in the art, such as those discussed in Current Protocols in Molecular Biology (edited by FMAusubel et al., 1987), Supplement 30, Section 7.718, Table 7.71. Some alignment programs are MacVector (Oxford Molecular Ltd, Oxford, UK), ALIGN Plus (Scientific and Educational Software, Pennsylvania), and AlignX (Vector NTI, Invitrogen, Carlsbad, CA). Another alignment program is Sequencher (Gene Codes, Ann Arbor, Michigan), which uses default parameters.

[0200] As used herein, the term "nucleotide change" refers to, for example, nucleotide substitution, deletion, insertion, chemical alteration, or any of the foregoing, as is well understood in the art.

[0201] As used herein, the term “protein modification” refers to, for example, amino acid substitution, amino acid modification, deletion and / or insertion, as well as is well understood in the art.

[0202] As used herein, the terms “at least a portion” or “fragment” for nucleic acid or polypeptide mean a portion having the smallest size characteristic of such a sequence, or any larger fragment (at most and including the full-length molecule). Fragments of polynucleotides disclosed herein may encode the biologically active portion of a gene regulatory element. The biologically active portion of a gene regulatory element may be prepared by isolating a portion of one of the polynucleotides disclosed herein that contains the gene regulatory element and assessing its activity as described herein. Similarly, a portion of a polypeptide may be 4 amino acids, 5 amino acids, 6 amino acids, 7 amino acids, etc., up to a full-length polypeptide. The length of the portion to be used will depend on the specific application. A portion of nucleic acid that can be used as a hybridization probe may be as short as 12 nucleotides; in some embodiments, it is 20 nucleotides. A portion of a polypeptide that can be used as an epitope may be as short as 4 amino acids. The portion of a polypeptide that functions as a full-length polypeptide will typically be longer than 4 amino acids.

[0203] As used herein, the term "primer" refers to an oligonucleotide that can anneal to the amplification target to allow DNA polymerase to ligate it, thus acting as the starting point for DNA synthesis when under conditions that induce primer extension product synthesis (i.e., in the presence of nucleotides and reagents for polymerization such as DNA polymerase and at suitable temperature and pH). Primers are preferably single-stranded to obtain maximum amplification efficiency. Preferably, primers are oligodeoxyribonucleotides. Primers must be long enough to initiate the synthesis of extension products in the presence of reagents for polymerization. The precise length of the primer will depend on many factors, including temperature and primer composition (A / T versus G / C content). A bidirectional primer pair consists of a forward primer and a reverse primer, as commonly used in the field of DNA amplification (such as PCR amplification).

[0204] The term "stringency" or "stringent hybridization conditions" refers to hybridization conditions that affect the stability of the hybrid, such as temperature, salt concentration, pH, formamide concentration, etc. These conditions are empirically optimized to maximize specific binding of primers or probes to their target nucleic acid sequences and minimize non-specific binding. The terminology used includes references to conditions under which probes or primers will hybridize with their target sequences to a much greater extent than other sequences (e.g., at least 2 times greater than background). Stringency conditions are sequence-dependent and vary under different conditions. Longer sequences hybridize specifically at higher temperatures. Generally, stringency conditions are chosen to be approximately 5°C lower than the thermal melting point (Tm) of the specific sequence at defined ionic strengths and pH. Tm is the temperature at which 50% of the complementary target sequence hybridizes with a perfectly matched probe or primer (at defined ionic strengths and pH). Typically, stringent conditions are those where the salt concentration is less than about 1.0 M Na+ ions at pH 7.0 to 8.3, typically about 0.01 to 1.0 M Na+ ion concentration (or other salts), and the temperature is at least about 30 °C (for short probes or primers (e.g., 10 to 50 nucleotides)) and at least about 60 °C (for long probes or primers (e.g., greater than 50 nucleotides)). Stringent conditions can also be achieved by adding a destabilizing agent such as formamide. Exemplary low-stringent conditions, or “reduced stringency conditions,” involve hybridization at 37 °C with a buffer of 30% formamide, 1 M NaCl, and 1% SDS, followed by washing at 40 °C in 2 × SSC. Exemplary high-stringent conditions involve hybridization at 37 °C in 50% formamide, 1 M NaCl, and 1% SDS, followed by washing at 60 °C in 0.1 × SSC. Hybridization procedures are well known in the art and have been described, for example, by Ausubel et al., 1998 and Sambrook et al., 2001. In some implementations, the stringent conditions are hybridization at 45°C in 0.25M Na2HPO4 buffer (pH 7.2) containing 1 mM Na2EDTA and 0.5% to 20% sodium dodecyl sulfate (such as 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20%), followed by washing in 5×SSC containing 0.1% (w / v) sodium dodecyl sulfate.

[0205] In some embodiments, the cell or organism has at least one heterologous trait. As used herein, the term "heterologous trait" refers to a phenotype of a cell or organism conferred by a foreign molecule or other organism (e.g., a microorganism), a fragment of DNA, a heterologous polynucleotide, or a heterologous nucleic acid.

[0206] Various phenotypic changes are of interest in this disclosure, including but not limited to modifications of fatty acid composition in plants, alterations in amino acid content, changes in pathogen defense mechanisms in plants, and increases in yields of economically important traits (e.g., cereal yields, forage yields, etc.). These results can be achieved by using the methods and compositions of this disclosure to provide the expression of heterologous products or the increased expression of endogenous products in plants.

[0207] "Synthetic assemblies" can include combinations of plants and microorganisms disclosed herein. Such assemblies can be achieved, for example, by coating the surface of seeds of plants (such as agricultural plants) or host plant tissues (roots, stems, leaves, etc.) with the microorganisms disclosed herein. Furthermore, "synthetic assemblies" can include combinations of microorganisms of various strains or species. A synthetic assembly has at least one variable that distinguishes it from any assembly existing in nature. This variable can, in particular, be the concentration of microorganisms on seeds or plant tissues that do not exist naturally, or a combination of microorganisms and plants that do not exist naturally, or a combination of microorganisms or strains that do not coexist naturally. In each of these cases, the synthetic assembly shows traces of artificiality and possesses structural and / or functional properties that do not exist when considering the individual elements of the assembly in isolation.

[0208] In some embodiments, the microorganism may be "endogenous" to the seed or plant. As used herein, a microorganism is considered "endogenous" to the plant or seed if it originates from a plant sample from which it is derived. That is, in cases where the microorganism is found to be associated with the plant in nature. In embodiments where the endogenous microorganism is applied to the plant, the endogenous microorganism is applied in an amount different from the level found on the plant in nature. Thus, if an endogenous microorganism for a given plant is present on the plant at a level not found in nature, the microorganism can still form a synthetic combination with the plant.

[0209] In some embodiments, a composition (such as a microorganism) may be "heterogeneous" (also referred to as "exogenous") to another composition (such as a seed or plant), and in some respects, is referred to herein as a "heterogeneous composition." As used herein, a microorganism is considered "heterogeneous" to a plant or seed if it does not originate from a plant sample from which it is derived. That is, a situation where the microorganism is found not to be associated with said plant in nature. For example, a microorganism typically associated with leaf tissue of a maize plant is considered exogenous to leaf tissue of another maize plant in which said microorganism is not found in nature. In another example, a microorganism typically associated with a maize plant is considered exogenous to a wheat plant in which said microorganism is not found in nature.

[0210] When a composition is applied, inoculated, associated with, or disposed of mechanically or manually onto or in a plant component, seedling, plant, plant growth medium, or treatment formulation, such that the treatment is present on or in the plant component, seedling, plant, plant growth medium, or formulation in a manner not found in nature prior to application of the treatment, the composition is "heterogeneously disposed of," for example, in a manner not found in nature in the plant variety, at that stage of plant development, in the plant tissue, in terms of abundance, or in the growth environment (e.g., drought). In some embodiments, this manner is contemplated as being selected from the group consisting of: the presence of microorganisms; the presence of microorganisms of varying cell numbers, concentrations, or amounts; the presence of microorganisms in or at other physical locations in or on different plant components, tissues, cell types, or plants; and the presence of microorganisms at different time periods, such as the developmental stage of the plant or plant component, time of day, time of season, and combinations thereof. In some embodiments, "heterogeneously disposed of" means that the microorganisms are applied to a tissue or cell type of a plant component that is different from where the microorganisms are naturally present. In some embodiments, "heterogeneously treated" means that a microorganism is applied to a plant component, seedling, or a specific developmental stage of the plant that is not associated in nature at that stage but may be associated at other stages. For example, if a microorganism is typically found during the flowering stage of a plant but not at other stages, then a microorganism applied during the seedling stage can be considered heterogeneously treated. In some embodiments, if a microorganism is typically found in the root tissue of a plant component but not in the leaf tissue, and the microorganism is applied to the leaf, then the microorganism is heterogeneously treated. In another non-limiting example, if a microorganism is naturally present in the mesophyll layer of a leaf tissue but is applied to the epidermis, then the microorganism will be considered heterogeneously treated. In some embodiments, "heterogeneously treated" means that the natural plant component, seedling, or plant does not contain detectable levels of the microorganism in that same plant component, seedling, or plant. In some implementations, "heterogeneously treated" means that the microorganisms are applied to the plant component, seedling, or plant at a greater concentration, quantity, or amount than they would be present in nature in the plant. For example, the microorganisms are heterogeneously treated when they are present at a quantity, quantity, or concentration that is at least 1.5 times, between 1.5 and 2 times, 2 times, between 2 and 3 times, 3 times, between 3 and 5 times, 5 times, between 5 and 7 times, 7 times, between 7 and 10 times, 10 times, or even more than 10 times greater than the concentration present before treatment. In another non-limiting example, microorganisms present in cypress (cupressaceous tree) tissue in nature would be considered heterogeneous to tissues of corn, wheat, cotton, or soybean plants.In another instance, microorganisms present in the leaf tissues of maize, spring wheat, cotton, and soybean plants in nature are considered heterologous to leaf tissues of another maize, spring wheat, cotton, or soybean plant that do not contain said microorganisms or contain different amounts of said microorganisms in nature.

[0211] Microorganisms can also be “heterogeneously disposed of” onto a given plant tissue. This means that the microorganism is placed on plant tissue on which it is not found in nature. For example, if a given microorganism is naturally present only on the roots of a given plant, then the microorganism can be exogenously applied to the aboveground tissue of the plant and will thus be “heterogeneously disposed of” onto said plant tissue. Therefore, when applied to a plant on which the microorganism does not naturally exist or does not naturally possess the amount of microorganism applied, the microorganism is considered to be heterogeneously disposed of.

[0212] The compositions and methods described herein can provide host plants with "modified" "agronomical traits" or "agronomically important traits," which may include, but are not limited to, the following: changes in oil content, protein content, seed carbohydrate composition, seed oil composition, and seed protein composition compared to isoline plants grown from seeds that do not contain the seed treatment preparations; chemical tolerance; cold tolerance; delayed senescence; disease resistance; drought tolerance; panicle weight; improved growth; enhanced health; heat tolerance; herbicide tolerance; herbivore resistance; improved nitrogen fixation; improved nitrogen use efficiency; improved root architecture; improved water use efficiency; increased biomass; increased root length; increased seed weight; and increased shoot length. Increased yield, increased yield under water-limited conditions, grain quality, grain moisture content, metal tolerance, number of ears, number of grains per ear, number of pods, enhanced nutrition, pathogen resistance, insect resistance, improved photosynthetic capacity, salt tolerance, greenness retention, increased vigor, increased dry weight of mature seeds, increased fresh weight of mature seeds, increased number of mature seeds per plant, increased chlorophyll content, increased number of pods per plant, increased pod length per plant, decreased number of wilted leaves per plant, decreased number of severely wilted leaves per plant and increased number of non-wilted leaves per plant, detectable regulation of metabolite levels, detectable regulation of transcriptomic levels, and detectable regulation of the proteome. The term "regulatory" refers to changes in agronomic traits altered by the presence of microorganisms, exudates, broths, metabolites, etc. In all respects, this regulation provides the conferment of beneficial traits.

[0213] Microorganisms and microbial bodies

[0214] As used herein, the term "microorganism" should be interpreted broadly. It includes, but is not limited to, prokaryotic bacteria and archaea, as well as eukaryotic fungi and protists.

[0215] As examples, microorganisms can include: Proteobacteria (such as Pseudomonas, Enterobacter, Stenotrophomonas, Burkholderia, Rhizobium, Herbaspirillum, Pantotheca, Serratia, Rahnella, Azospirillum, Azorhiz). The phylum Firmicutes includes bacteria such as *Izobium*, *Azotobacter*, *Duganella*, *Delftia*, *Bradyrhizobium*, *Sinorhizobium*, *Variovorax*, and *Halomonas*. Other phyla include *Bacillus*, *Bacillus-like*, *Lactobacillus*, *Mycoplasma*, and *Acetobacter*. m), Actinobacteria (such as *Brevibacterium*, *Janibacter*, *Streptomyces*, *Rhodococcus*, *Microbacterium*, *Curtobacterium*, *Cellulomonas*, and *Nocardioides*) and Ascomycota (such as *Trichoderma*, *Ampelomycetes*) s), genera such as Coniothyrium, Paecoelomyces, Penicillium, Cladosporium, Hypocrea, Beauveria, Metarhizium, Verticulum, Cordyceps, Pichia, and Candida, Basidiomycota (such as Coprinus)Genus *Corticium* and *Agaricus*, phylum Oomycota (such as *Pythium*), and phylum Mucoromycota (such as *Mucor* and *Mortierella*); as well as genera *Orbilia* / *Arthrobotrys*, *Lysinibacillus*, *Microbacterium*, *Basilobacter*, *Arthrobacter*, *Kosakonia*, *Masillia*, *Novosphingobium*, and *Tumebacillus*.

[0216] In a particular embodiment, the microorganisms are endophytic bacteria, epiphytes, or microorganisms residing in the rhizosphere or root sheath of a plant. That is, the microorganisms can be found in soil material attached to the roots of a plant or in areas immediately adjacent to the roots.

[0217] In one implementation, the microorganisms are endophytes. Endophytes are beneficial to the host plant by preventing pathogenic organisms from colonizing them. Endophytes create a "barrier effect" on the widespread colonization of plant tissues, where localized endophytes prevail and prevent the survival of pathogenic organisms. Endophytes may also produce chemicals that inhibit the growth of competitors, including pathogenic organisms.

[0218] In some implementations, the microorganisms are unculturable. This should be understood to mean that it is not yet known whether the microorganism is culturable or difficult to culture using methods known to those skilled in the art.

[0219] The microorganisms disclosed herein may be collected or obtained from any source, or contained in and / or associated with materials collected from any source.

[0220] In one implementation, the microorganisms are derived from any general terrestrial environment, including its soil, plants, fungi, animals (including invertebrates), and other biomes, including sediments, water, and biomes of lakes and rivers; from marine environments, their biomes, and sediments (e.g., seawater, marine mud, marine plants, marine invertebrates (e.g., sponges), marine vertebrates (e.g., fish)); terrestrial and marine lithosphere (topsoil and rocks, such as compressed subsurface rock, sand, and clay); cryosphere and its meltwater; atmosphere (e.g., filtered airborne dust, clouds, and raindrops); and urban, industrial, and other man-made environments (e.g., organic and mineral deposits on concrete, roadside drains, roof surfaces, and road surfaces).

[0221] In another implementation, microbial cells are collected from sources that may favor the selection of suitable microorganisms. As an example, this source could be a specific environment where other plants are suitable for growth or considered terroir-related. In another example, the source could be a plant possessing one or more desired traits, such as a plant that grows naturally in a specific environment or under certain conditions of interest. As an example, a plant may grow naturally in sandy or highly saline sand, or at extreme temperatures, or in conditions with very little water, or it may be resistant to certain pests or diseases in its environment, and it may be desirable for cash crops to grow under such conditions, especially when these are, for example, the only conditions available in a particular geographic location. As another example, microbial cells could be collected from cash crops grown in such environments, or more specifically, individual crop plants that best exhibit the traits of interest among crops grown in any particular environment, such as the fastest-growing plant among crops grown in saline-limited soils, or the least damaged plant among crops exposed to severe insect infestations or disease outbreaks, or plants with desired amounts of certain metabolites and other compounds (including fiber content, oil content, etc.), or plants exhibiting desired color, taste, or odor. Microorganisms can be collected from any material present in the plant or environment of interest, including fungi and other animal and plant biomes, soil, water, sediments, and other environmental elements as previously mentioned. In some embodiments, microorganisms are individual isolates from different environments.

[0222] In one embodiment, the microorganisms or combinations of microorganisms used in the methods of this disclosure may be selected from a pre-existing collection of individual microbial species or strains based on some knowledge of their potential or predicted benefits to the plant. For example, the microorganisms may be predicted to: enhance nitrogen fixation; release phosphate from soil organic matter; release phosphate from inorganic forms of phosphate (e.g., rock phosphate); “fix carbon” in root microspheres; live in the rhizosphere of the plant, thereby helping the plant absorb nutrients from the surrounding soil and then more easily provide those nutrients to the plant; increase the number of nodules on the plant roots, thereby increasing the number of symbiotic nitrogen-fixing bacteria (e.g., rhizobium species) per plant and the amount of nitrogen fixed by the plant; trigger plant defense responses such as ISR (inducible systemic resistance) or SAR (systemically acquired resistance), which help the plant resist the invasion and spread of pathogenic microorganisms; compete with microorganisms harmful to plant growth or health through antagonism or competitive use of resources (such as nutrients or space); alter the color of one or more parts of the plant, or change the plant's chemical status, odor, taste, or one or more other characteristics.

[0223] In one implementation, the microorganisms or combinations of microorganisms are selected from a collection of pre-existing individual microbial species or strains whose potential or predicted benefits to plants are unknown. For example, a collection of unidentified microorganisms isolated from plant tissues without prior knowledge of their ability to improve plant growth or health, or a collection of microorganisms collected to explore their potential for producing compounds that could lead to drug development.

[0224] In one embodiment, the microorganisms are obtained from materials from which they naturally reside (e.g., soil, rock, water, air, dust, plants, or other organisms). They may be provided in any suitable form, taking into account their intended use in the methods of this disclosure. However, by way of example only, the microorganisms may be provided in the form of aqueous suspensions, gels, homogenates, particles, powders, slurries, live organisms, or dried materials.

[0225] The microorganisms disclosed herein can be isolated from substantially pure or mixed cultures. They can be concentrated, diluted, or provided at their natural concentrations present in the source material. For example, microorganisms from salt sediments can be isolated for use in this disclosure by suspending the sediments in fresh water and allowing the sediments to settle to the bottom. The water containing most of the microorganisms can be removed by decantation after a suitable settling period and applied directly to the plant growth medium, or concentrated by filtration or centrifugation, diluted to an appropriate concentration, and applied to the plant growth medium along with the removed majority of the salt. As another example, microorganisms from mineralized or toxic sources can be similarly treated to recover the microorganisms for application to plant growth material, thereby minimizing the possibility of plant damage.

[0226] In another embodiment, the microorganisms are used in crude form, wherein they are not separated from the source material in which they naturally reside. For example, the microorganisms are provided in combination with the source material in which they reside; for example, in soil form, or from the roots, seeds, or leaves of a plant. In this embodiment, the source material may include one or more species of microorganisms.

[0227] In some implementations, the methods of this disclosure use mixed populations of microorganisms.

[0228] In embodiments of this disclosure, in which microorganisms are isolated from source materials (e.g., materials in which microorganisms naturally reside), any one or a combination of many standard techniques readily known to those skilled in the art can be used. However, by way of example, these techniques generally employ processes suitable for obtaining solid or liquid cultures of single microorganisms in substantially pure form, typically by physical separation on the surface of a solid microbial growth medium or by volumetric dilution in a liquid microbial growth medium. These processes may include separation from dried materials, liquid suspensions, slurries, or homogenates (wherein the material is spread in thin layers on a suitable solid gel growth medium), or by continuous dilution of the material in a sterile medium and inoculation into liquid or solid media.

[0229] Although not strictly necessary, in one embodiment, the material containing microorganisms may be pretreated prior to the separation process to multiply all microorganisms in the material, or to select portions of the microbial community by: enriching the material containing microbial nutrients (e.g., by pasteurizing the sample to select for heat-resistant microorganisms (e.g., bacilli)), or exposing the sample to low concentrations of organic solvents or sterilizing agents (e.g., household bleach) to enhance the survival rate of spore-forming or solvent-resistant microorganisms. As described above, the microorganisms can then be separated from the enriched material or the material treated to achieve selective viability.

[0230] In one embodiment of this disclosure, endophytic or epiphytic microorganisms are isolated from plant material. Many standard techniques known in the art can be used and microorganisms can be isolated from any suitable tissue of a plant, including, for example, roots, stems and leaves, as well as plant reproductive tissues. As an example, conventional methods for isolation from plants typically involve aseptic removal of the plant material of interest (e.g., root or stem length, leaves), surface sterilization with a suitable solution (e.g., 2% sodium hypochlorite), followed by placement of the plant material on a nutrient medium for microbial growth (see, for example, Strobel G and Daisy B (2003) Microbiology and Molecular Biology Reviews 67(4):491-502; Zinniel DK et al., (2002) Applied and Environmental Microbiology 68(5):2198-2208).

[0231] In one embodiment of this disclosure, microorganisms are isolated from root tissue. Another method for isolating microorganisms from plant material is detailed below.

[0232] In one implementation, the microbial community is exposed to selection pressure (before or at any stage of the method). For example, exposing the microorganisms to pasteurization before they are added to a plant growth medium (preferably sterile) may increase the probability that plants selected for the desired trait will be associated with sporoforming microorganisms that are more likely to survive under adverse conditions, during commercial storage, or when applied to seeds in a coating form in adverse environments.

[0233] In some embodiments, as mentioned above, the microorganisms can be used in crude form and do not need to be isolated from the plant or culture medium. For example, plant material or growth medium comprising microorganisms identified as beneficial to the selected plant can be obtained, and this plant material or growth medium can be used as a source of crude microorganisms for the next round of the method, or as a source of crude microorganisms at the end of the method. For example, whole plant material can be obtained and optionally treated, such as mulching or crushing. Alternatively, individual tissues or parts (such as leaves, stems, roots, and seeds) of the selected plant can be isolated from the plant and optionally treated, such as mulching or crushing. In some embodiments, one or more parts of the plant associated with one or more of the second group of microorganisms can be removed from one or more of the selected plants and, in the event of any successive repetition of the method, grafted onto one or more plants used in any step of the plant breeding method.

[0234] Exemplary microorganisms

[0235] In all respects, this disclosure provides novel strains of isolated microorganisms, including the identified microbial species presented in Table 1 or Table 1A.

[0236] In other respects, this disclosure provides complete microbial cultures isolated from the species and strains identified in Table 1 or Table 1A. These cultures may contain various concentrations of microorganisms.

[0237] In all respects, this disclosure provides for the use of microorganisms selected from Table 1 or Table 1A in agriculture.

[0238] In some embodiments, this disclosure provides isolated microbial species belonging to the following genera: Bacillus, Bacillus-like Bacillus, Discobacterium, Arthrozoa, Lysine Bacillus, Microbacterium, and / or Basilaria.

[0239] In some implementations, microorganisms from the genus Bacillus are used in agriculture to confer one or more beneficial properties on plant species.

[0240] In some implementations, microorganisms from the genus Bacillus are used in agriculture to confer one or more beneficial properties on plant species.

[0241] In some implementations, microorganisms from the genus *Discocephalum* are used in agriculture to confer one or more beneficial properties on plant species.

[0242] In some implementations, microorganisms from the genus Arthrozoa are used in agriculture to confer one or more beneficial properties on plant species.

[0243] In some implementations, microorganisms from the genus *Bacillus lysine* are used in agriculture to confer one or more beneficial properties on plant species.

[0244] In some implementations, microorganisms from the genus Microbacteria are used in agriculture to confer one or more beneficial properties on plant species.

[0245] In some implementations, microorganisms from the genus *Basilaria* are used in agriculture to confer one or more beneficial properties on plant species.

[0246] In some embodiments, this disclosure provides isolated microorganisms belonging to the group consisting of: Bacillus tekirae, Bacillus methyltrophicus, Bacillus amyloliquefaciens, Bacillus alginate, Discocele aureus / Oligospora spp. (sexual and asexual forms, respectively), Bacillus pumilus, Bacillus fusiformis, Bacillus megaterium, Bacillus taichungii, Bacillus erinaceus, Bacillus ilignosa, Microbacterium arabinogalactanophilus, and Bacillus rubrum.

[0247] In some embodiments, this disclosure provides isolated microorganisms belonging to the group consisting of: *Bacillus tekirae*, *Bacillus methyltrophicus*, *Bacillus amyloliquefaciens*, *Bacillus alginate*, *Discozoella aureae* / *Oligospora spp.* (sexual and asexual forms, respectively), *Bacillus pumilus*, *Bacillus fusiformis*, *Bacillus megaterium*, *Bacillus taichungensis*, *Bacillus erinaceus*, *Bacillus ilignosa*, *Microbacterium arabinogalactanophilus*, and *Brochytosporum*. Specific novel strains of these aforementioned species are shown in Table 1 or Table 1A.

[0248] Furthermore, this disclosure relates to microorganisms having properties substantially similar to those of the microorganisms identified in Table 1 or Table 1A.

[0249] The isolated microbial species identified in this disclosure and novel strains of said species can confer beneficial characteristics or traits, such as agronomically important traits, onto target plant species.

[0250] For example, the isolated microorganisms or aggregates of said microorganisms described in Table 1 or Table 1A can improve plant health and vigor. The improved plant health and vigor can be quantitatively measured, for example, by measuring the effect of said microbial application on plant phenotypic or genotypic traits.

[0251] Sources of microorganisms

[0252] The microorganisms disclosed herein were obtained in various regions of New Zealand and the United States, as well as elsewhere.

[0253] Isolation and culture of microorganisms

[0254] The microorganisms in Tables 1 and 1A were identified by using standard microscopy techniques to characterize their phenotypes, which were then used to identify the microorganisms as taxonomically recognized species.

[0255] The isolation, identification, and culture of the microorganisms disclosed herein can be achieved using standard microbiological techniques. Examples of such techniques can be found in Gerhardt, P. (ed.) Methods for General and Molecular Microbiology. American Society for Microbiology, Washington, DC (1994) and Lennette, EH (ed.) Manual of Clinical Microbiology, 3rd Edition, American Society for Microbiology, Washington, DC (1980), each of which is incorporated herein by reference.

[0256] Isolation can be achieved by streaking a sample on a solid culture medium (e.g., nutrient agar plate) to obtain a single colony (characterized by the phenotypic traits described above (e.g., Gram-positive / negative, ability to form spores in an aerobic / anaerobic manner, cell morphology, carbon source metabolism, acid / base production, enzyme secretion, metabolic secretions, etc.)) and reducing the likelihood of manipulation with contaminated cultures.

[0257] For example, with respect to the bacteria isolated in this disclosure, biologically pure isolates can be obtained through repeated subculturing of biological samples, streaking onto solid medium after each subculturing to obtain individual colonies. Methods for preparing lyophilized bacteria, thawing them, and growing them are well known, for example, Gherna, RL, and CAReddy. 2007. Culture Preservation, pp. 1019-1033, in CAReddy, TJ Beveridge, JA Breznak, GA Marzluf, TMS Schmidt, and LRSnyder (eds.), American Society for Microbiology, Washington, DC, p. 1033; this document is incorporated herein by reference. Therefore, it is contemplated that lyophilized liquid formulations and cultures stored long-term in a glycerol-containing solution at -70°C may be used to provide the formulations of this invention.

[0258] The bacteria of this disclosure can multiply in liquid culture medium under aerobic conditions. The culture medium used to grow the bacterial strains of this disclosure includes a carbon source, a nitrogen source, and inorganic salts, as well as specific required substances such as vitamins, amino acids, and nucleic acids. Examples of suitable carbon sources for growing bacterial strains include, but are not limited to, starch, peptone, yeast extract, amino acids, sugars such as glucose, arabinose, mannose, glucosamine, and maltose; salts of organic acids such as acetic acid, fumaric acid, adipic acid, propionic acid, citric acid, gluconic acid, malic acid, pyruvic acid, and malonic acid; alcohols such as ethanol and glycerol; and oils or fats such as soybean oil, rice bran oil, olive oil, corn oil, and sesame oil. The amount of carbon source added varies depending on the type of carbon source and is generally between 1 and 100 grams per liter of culture medium. Preferably, the culture medium contains 0.1% to 5% (w / v) of glucose, starch, and / or peptone as the primary carbon source. Examples of suitable nitrogen sources for the growth of bacterial strains of the present invention include, but are not limited to, amino acids, yeast extracts, tryptone, beef extract, peptone, potassium nitrate, ammonium nitrate, ammonium chloride, ammonium sulfate, ammonium phosphate, ammonia, or combinations thereof. The amount of nitrogen source varies depending on the type of nitrogen source and is generally between 0.1 and 30 grams per liter of culture medium. Inorganic salts, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, magnesium sulfate, magnesium chloride, ferric sulfate, ferrous sulfate, ferric chloride, ferrous chloride, manganese sulfate, manganese chloride, zinc sulfate, zinc chloride, copper sulfate, calcium chloride, sodium chloride, calcium carbonate, and sodium carbonate may be used alone or in combination. The amount of inorganic acids varies depending on the type of inorganic salt and is generally between 0.001 and 10 grams per liter of culture medium. Examples of substances with specific requirements include, but are not limited to, vitamins, nucleic acids, yeast extracts, peptone, meat extract, malt extract, dried yeast, and combinations thereof. Cultivation can be carried out at temperatures that allow the growth of bacterial strains (generally between 20°C and 46°C). In some embodiments, the temperature range is 30°C to 37°C. For optimal growth, in some implementations, the culture medium may be adjusted to a pH of 7.0 to 7.4. It should be understood that commercially available culture media, such as nutrient broth or nutrient agar, available from Difco, Detroit, MI, can also be used to culture bacterial strains. It should be understood that the culture time may vary depending on the type of culture medium used and the concentration of sugar as the primary carbon source.

[0259] In all respects, the incubation period is 24 to 96 hours. The bacterial cells thus obtained are isolated using methods well known in the art. Examples include, but are not limited to, membrane filtration and centrifugation. The pH may be adjusted using sodium hydroxide or similar methods, and the culture may be dried using a freeze dryer until the water content becomes 4% or less. Microbial co-cultures can be obtained by propagating the individual strains as described above. It should be understood that these microbial strains can be co-cultured when compatible culture conditions are available.

[0260] Identification of microorganisms

[0261] Microorganisms can be classified into a genus based on polyphasic taxonomy, which integrates all available phenotypic and genotypic data into a consensus classification (Vandamme et al., 1996. Polyphasic taxonomy, a consensus approach to bacterial systematics. Microbiol Rev 1996, 60:407-438). A generally accepted method for defining species by genotypic association is based on overall genome correlation, such that strains sharing approximately 70% or higher correlation when performing DNA-DNA hybridization at 5°C or lower (ΔTm, the difference in melting temperature between homologous and heterologous hybrids) under standard conditions are considered members of the same species. Therefore, populations sharing more than the aforementioned 70% threshold can be considered variants of the same species.

[0262] For bacteria and microorganisms, 16S rRNA sequences are typically used to determine classification and distinguish species because if the 16S rRNA sequence shares less than the specified sequence identity percentage with the reference sequence, the two organisms from which the sequence is obtained are considered to be different species.

[0263] Therefore, microorganisms can be considered the same species if they share at least 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity in their 16S or 16S rRNA or rDNA sequences. In some respects, microorganisms can only be considered the same species if they share at least 95% identity.

[0264] In addition, microbial strains of a species can be defined, such as strains that share at least 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequence identity in their 16S rRNA sequences.

[0265] Comparisons can also be made between a reference sequence and a 23S rRNA sequence. In some respects, microorganisms can only be considered the same strain if they share at least 95% identity. In some implementations, "substantially similar genetic characteristics" means microorganisms that share at least 95% identity.

[0266] For fungal microorganisms, ITS (internal transcribed sequence) is commonly used for classification and identification. Within the ribosomal cistron region, the internal transcribed spacer (ITS) has the highest probability of successfully identifying the widest range of fungi and has the most well-defined barcode gap between interspecific and intraspecific variation, and has been proposed as a formal fungal identification sequence (Schoch et al., PNAS, April 17, 2012, 109(16)6241-6246).

[0267] In one embodiment, the microbial strains of this disclosure include those containing a polynucleotide sequence that shares at least 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of SEQ ID NO:1-21.

[0268] In one embodiment, the microorganisms of this disclosure include those containing a polynucleotide sequence that shares at least 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of SEQ ID NO:1-21.

[0269] In one embodiment, the microbial aggregates of this disclosure comprise two or more microorganisms containing a polynucleotide sequence that shares at least 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of SEQ ID NO:1-21.

[0270] In one embodiment, the microbial aggregate of the present disclosure comprises two or more microbial strains, wherein at least one of these microbial strains contains a polynucleotide sequence sharing at least 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of SEQ ID NO:1-21.

[0271] In one embodiment, the microbial aggregate of the present disclosure comprises two or more microbial strains, wherein at least one of these microbial strains contains a polynucleotide sequence sharing at least 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with any of SEQ ID NO:1-21, and wherein at least one of these microorganisms is optionally selected from Table 2.

[0272] Unculturable microorganisms cannot usually be assigned to a specific species without a defined phenotype. Instead, a provisional name within a genus can be given to the microorganism, as long as its 16S rRNA sequence conforms to the principle of identity with a known species.

[0273] One approach is to observe the distribution of a large number of lineages of closely related species in sequence space and identify clusters of lineages that are well separated from other clusters. This method has been developed by assessing clustering patterns using tandem sequences of multinucleate (housekeeper) genes and has been called multiple-site sequence analysis (MLSA) or multiple-site sequence phylogenetic analysis. MLSA has been successfully used to explore clustering patterns in a large number of lineages assigned to very closely related species by current taxonomic methods, to examine relationships between a few lineages within a genus or a broader taxonomic group, and to solve specific taxonomic problems. More generally, this method can be used to explore the presence of bacterial species, i.e., to observe whether large groups of similar lineages always belong to well-separated clusters, or, in some cases, whether there is genetic continuity in which no obvious separation into clusters is observed.

[0274] To more accurately determine the genus, phenotypic traits (such as morphology, biochemistry, and physiological characteristics) are determined for comparison with a reference genus prototype. Colony morphology may include color, shape, pigmentation, slime production, etc. Cell characteristics are described in terms of shape, size, Gram reaction, extracellular material, presence of endospores, presence and location of flagella, motility, and inclusion bodies. Biochemical and physiological characteristics describe the organism's growth under varying temperature, pH, salinity, and atmospheric conditions, and under different single carbon and nitrogen sources. Those skilled in the art will reasonably understand the phenotypic traits defining the genera of this disclosure. For example, species of the genus *Rhizobium* are identified using the color, form, and texture of colonies on a specific agar (e.g., YMA).

[0275] In one embodiment, the bacterial microorganisms taught herein are identified using 16S rRNA gene sequences. It is known in the art that 16S rRNA contains hypervariable regions, which can provide species / strain-specific characteristic sequences that can be used for bacterial identification. In this disclosure, many microorganisms are identified via partial (500 bp to 1200 bp) 16S rRNA sequence characteristics. In each embodiment, each strain represents a pure colony isolate selected from an agar plate. Selection is based on any defined morphological characteristics of the colonies on the agar medium to represent the diversity of organisms present. In embodiments, the medium used is R2A, PDA, nitrogen-free semi-solid medium, or MRS agar. After 24 hours of growth, colony descriptions are performed for each “picked” isolate, and the colony descriptions are then entered into our database. Sequence data for each isolate are subsequently obtained.

[0276] Phylogenetic analysis of the 16S rRNA gene was used to define “substantially similar” species belonging to a common genus, and also to define “substantially similar” lines of a given taxonomic species. Furthermore, we documented the physiological and / or biochemical characteristics of the isolates, which could be used to highlight subtle and significant differences between lines that elicit favorable plant behaviors.

[0277] Microbial aggregates

[0278] In all respects, this disclosure provides microbial aggregates comprising a combination of at least two microorganisms selected from those identified in Table 1 and / or Table 1A.

[0279] In other respects, this disclosure provides microbial aggregates comprising a combination of at least two microorganisms, wherein at least one microorganism is selected from the microorganisms identified in Table 1 or Table 1A and additional microorganisms may optionally be selected from the microorganisms identified in Table 2.

[0280] In some embodiments, the aggregates of this disclosure comprise two, three, four, five, six, seven, eight, nine, ten, or more microorganisms. The microorganisms in the aggregate are different microbial species or different strains of microbial species.

[0281] In some embodiments, this disclosure provides an aggregate comprising at least one isolated microbial species belonging to the genera Bacillus, Bacillus-like Bacillus, Discobacterium, Arthrozoa, Lysine Bacillus, Microbacterium, or Basilaria.

[0282] In some embodiments, this disclosure provides an aggregate comprising at least one isolated microbial species belonging to the genera Bacillus, Bacillus subtilis, Discobacterium, Arthrozoonium, Bacillus lysineus, Microbacterium, or Basilaria, and optionally further comprising at least one isolated microbial species belonging to the genera Arthrozoonium, Bacillus, Breozoa, Spirochetes, Cossackella, Bacillus lysineus, Masséria, Neosphingolipidella, Bacillus subtilis, Pseudomonas, Rahn's, or Bacillus bloatosa.

[0283] In some embodiments, this disclosure provides an aggregate comprising at least one isolated microbial species belonging to the genera Bacillus, Bacillus subtilis, Discobacterium, Arthrozoonium, Bacillus lysineus, Microbacterium, or Basilaria, and the aggregate further comprising at least one isolated microbial species belonging to the genera Arthrozoonium, Bacillus, Breozoa, Spirochetes, Cossackella, Bacillus lysineus, Masséria, Neosphingolipids, Bacillus subtilis, Pseudomonas, Rahn's bacillus, and Bacillus bloatosa.

[0284] In some embodiments, this disclosure provides an aggregate comprising at least one isolated microbial species selected from the group consisting of: Bacillus tekirae, Bacillus methyltrophicus, Bacillus amyloliquefaciens, Bacillus alginate, Discocele aureus / Oligospora spp. (sexual and asexual forms, respectively), Bacillus pumilus, Bacillus fusiformis, Bacillus megaterium, Bacillus taichungii, Bacillus erinaceus, Bacillus ilignosa, Microbacterium arabinogalactanophilus, and Bacillus rubrum.

[0285] In some embodiments, this disclosure provides an aggregate comprising: at least one novel isolated microbial strain of species selected from the group consisting of: *Bacillus tekirae*, *Bacillus methyltrophicus*, *Bacillus amyloliquefaciens*, *Bacillus alginate*, *Discocele aureus* / *Arthrozoa oligosporus* (sexual and asexual forms, respectively), *Bacillus pumilus*, *Bacillus fusiformis*, *Bacillus megaterium*, *Bacillus taichungii*, *Bacillus erinaceus*, *Bacillus ilignosa*, *Microbacterium arabinogalactanum*, and *Bruchwegianella*, and optionally further comprising selected from... At least one isolated microbial strain of species comprising the following groups: *Arthrobacter cylindrica*, *Arthrobacter mysore*, *Arthrobacter nicotinate*, *Arthrobacter trophoblast*, *Bacillus megaterium*, *Bacillus subtilis*, *Bacillus thuringiensis*, *Brevibacterium frostridae*, *Clonolactone*, *Sacchariformis*, *Bacillus fusiformis*, *Masses gypseum*, *Masses niastragali*, *Neosphingosine monosporum*, *Bacillus amyloliquefaciens*, *Bacillus polymyxa*, *Pseudomonas fluorescens*, *Pseudomonas jinjuensis*, *Pseudomonas oryzae*, *Pseudomonas putidae*, *Laenia aquaticus*, and *Bacillus perfringens*.

[0286] In some embodiments, this disclosure provides an aggregate comprising: at least one novel isolated microbial strain of species selected from the group consisting of: *Bacillus tekirae*, *Bacillus methyltrophicus*, *Bacillus amyloliquefaciens*, *Bacillus alginate*, *Discocele aureus* / *Schizophyllum commune* (sexual and asexual forms, respectively), *Bacillus pumilus*, *Bacillus fusiformis*, *Bacillus megaterium*, *Bacillus taichungii*, *Bacillus erinaceus*, *Bacillus ilignosa*, *Microbacterium arabinogalactanum*, and *Bruchwegianella*, and the aggregate further comprising: At least one isolated microbial strain of the species comprising each group: *Arthrobacter cylindrica*, *Arthrobacter mysore*, *Arthrobacter nicotinate*, *Arthrobacter trophoblast*, *Bacillus megaterium*, *Bacillus subtilis*, *Bacillus thuringiensis*, *Brevibacterium frostridae*, *Clonolactone-ladenella*, *Sacchariformis sacchariformis*, *Bacillus fusiformis*, *Masses niastragali*, *Neosphingosine monosporus*, *Bacillus amyloliquefaciens*, *Bacillus polymyxa*, *Pseudomonas fluorescens*, *Pseudomonas jinjuensis*, *Pseudomonas oryzae*, *Pseudomonas putidae*, *Laenia aquaticus*, and *Bacillus perfringens*.

[0287] In some embodiments, this disclosure provides an aggregate comprising isolated microbial strains of Bacillus pumilus and isolated microbial strains of Bacillus belyssus.

[0288] The specific novel strains of these aforementioned species can be found in Tables 1, 1A and 2.

[0289] The microbial components of an aggregate may be selected from any microorganisms identified in Tables 1, 1A and 2, provided that at least one microorganism in any aggregate includes at least one microorganism selected from Table 1 or Table 1A.

[0290] Microbial-produced compositions

[0291] In some cases, the microorganisms disclosed herein may produce one or more compounds and / or have one or more activities, such as producing one or more of the following: producing metabolites, producing plant hormones (such as auxins), producing acetoin, producing antimicrobial compounds, producing siderophores, producing polyketides, producing phenazines, producing cellulase, producing pectinase, producing chitinase, producing glucanase, producing xylanase, fixing nitrogen or dissolving mineral phosphates.

[0292] For example, the microorganisms disclosed herein can produce plant hormones selected from the group consisting of: auxins, cytokinins, gibberellins, ethylene, brassinolide, and abscisic acid.

[0293] Therefore, "metabolites produced by the microorganisms of this disclosure" is intended to represent any molecule (small molecule, vitamin, mineral, protein, nucleic acid, lipid, fat, carbohydrate, etc.) produced by the microorganisms of this disclosure. Typically, the precise mechanism by which the microorganisms of this disclosure confer beneficial traits on a given plant species is unknown. It is presumed that, in some cases, the microorganisms produce metabolites beneficial to the plant. Therefore, in some respects, cell-free or inactivated preparations of the microorganisms are beneficial to the plant, because the microorganisms need not be alive to confer beneficial traits on a given plant species, as long as the preparation contains metabolites produced by said microorganisms that are beneficial to the plant.

[0294] In one embodiment, the microorganisms of this disclosure can produce auxin (e.g., indole-3-acetic acid (IAA)). Auxin production can be analyzed. Many of the microorganisms described herein are capable of producing the plant hormone auxin indole-3-acetic acid (IAA) when grown in a culture. Auxins play a crucial role in altering plant physiology, including the extent of root growth.

[0295] Therefore, in one embodiment, the microorganisms of this disclosure exist in the form of a community disposed on the surface or within the tissues of a given plant species. The microorganisms can effectively produce compositions, such as metabolites, by generating a detectably increased amount of the composition found on or within the plant when compared to a reference plant not treated with the microorganisms of this disclosure or with a cell-free or inactive preparation. The compositions produced by the microbial community can be beneficial to the plant species.

[0296] Compositions produced by such microorganisms may be present in cell culture broths or media in which the microorganisms grow, or may encompass exudates produced by the microorganisms. As used herein, “exudate” refers to one or more compositions secreted by or extracted from one or more microbial cells. As used herein, “broth” refers to the common composition of cell culture media after the microbial cells have been placed in the culture medium. The composition of the broth may change over time, altering during different stages of microbial growth and / or development. Broth and / or exudates may improve the traits of the plant to which they become associated.

[0297] Microbial-induced traits in plants

[0298] This disclosure utilizes microorganisms to confer beneficial characteristics (or traits) on desired plant species (such as agronomic species of interest). In this disclosure, the terms "beneficial characteristic" or "beneficial trait" are used interchangeably and refer to the regulation of a desired plant phenotypic or genetic characteristic of interest by applying microorganisms or microbial aggregates as described herein. As previously mentioned, in some respects, it may be highly desirable that metabolites produced by a given microorganism ultimately play a role in regulating or conferring beneficial traits on a given plant.

[0299] There are many beneficial traits that can be modulated by applying the microorganisms disclosed herein. For example, microorganisms may have the ability to confer one or more beneficial traits on plant species, such as increased growth, increased yield, increased nitrogen use efficiency, enhanced stress tolerance, enhanced drought tolerance, increased photosynthetic rate, enhanced water use efficiency, enhanced pathogen resistance, and modification of plant architecture (which does not necessarily affect plant yield, but rather addresses plant function and causes the plant to increase the production of metabolites of interest), etc.

[0300] In all respects, the microorganisms taught in this paper offer a wide range of agricultural applications, including: increasing the yield of grains, fruits, and flowers; improving the growth of plant parts; enhancing the ability to utilize nutrients (e.g., nitrogen, phosphate, etc.); enhancing disease resistance; providing bio-insectic effects (including enhancing resistance to fungi and nematodes); improving survival rates in extreme climates; and modifying other desired plant phenotypic traits.

[0301] In some respects, the isolated microorganisms, aggregates, and / or agricultural compositions of this disclosure can be applied to plants to modulate or alter plant characteristics, such as changes in oil content, protein content, seed carbohydrate composition, seed oil composition, seed protein composition, chemical resistance, cold tolerance, delayed senescence, disease resistance, drought tolerance, ear weight, improved growth, enhanced health, heat tolerance, herbicide tolerance, herbivore resistance, improved nitrogen fixation, improved nitrogen use efficiency, improved nutrient (e.g., phosphate, potassium, etc.) utilization, improved root architecture, improved water use efficiency, increased biomass, increased root length, increased seed weight, increased shoot length, increased yield, and improved yield under water-limited conditions. Increased quantity, grain quality, grain moisture content, metal tolerance, number of spikes, number of grains per spike, number of pods, enhanced nutrition, pathogen resistance, reduced pathogen levels (e.g., through the secretion of metabolites that affect pathogen survival), insect resistance, improved photosynthetic capacity, salt tolerance, greenness retention, increased vigor, increased dry weight of mature seeds, increased fresh weight of mature seeds, increased number of mature seeds per plant, increased chlorophyll content, increased number of pods per plant, increased pod length per plant, reduced number of wilted leaves per plant, reduced number of severely wilted leaves per plant and increased number of non-wilted leaves per plant, detectable regulation of metabolite levels, detectable regulation of transcript levels, and detectable regulation of the proteome.

[0302] In some aspects, the isolated microorganisms, aggregates, and / or agricultural compositions of this disclosure can be applied to plants to negatively modulate specific plant traits. For example, in some aspects, the microorganisms of this disclosure can reduce phenotypic traits of interest, as such functionality may be desired in some applications. For example, the microorganisms of this disclosure may have the ability to reduce root growth or root length. Or the microorganisms may have the ability to reduce shoot growth or plant growth rate, as such regulation of plant traits may be desired in some applications.

[0303] In some embodiments, the isolated microorganisms, aggregates, and / or agricultural compositions of this disclosure can be applied to plants to confer nematode stress tolerance. Suitably, in such embodiments, the microorganisms may be selected from the group consisting of: *Discocephalus aureus* / *Oligospora*, *Bacillus fusiformis*, and *Bacillus velenzensis*, or the aggregate may comprise *Bacillus velenzensis* and *Bacillus pumilus* or composed of them. Suitably, the microorganisms may be selected from the group consisting of: *Discocephalus aureus* / *Oligospora* BEC93, *Bacillus fusiformis* BEC91, and *Bacillus velenzensis* BEC89A, or the aggregate may comprise *Bacillus velenzensis* BEC89A and *Bacillus pumilus* BEC89B or composed of them.

[0304] In some embodiments, the isolated microorganisms, aggregates, and / or agricultural compositions of this disclosure may be applied to plants to provide biostimulation (biostimulant effect). Suitably, in such embodiments, the microorganisms may be selected from the group consisting of: *Bacillus amyloliquefaciens*, *Bacillus megaterium*, *Bacillus tekira*, *Microbacterium arabinogalactanophilus*, *Bacillus alginate*, *Bacillus erythropoietinus*, *Bacillus ilignosa*, *Bacillus taichungensis*, and *Rhodotorula rubrum*. Appropriately, the microorganisms may be selected from the group consisting of: Bacillus amyloliquefaciens BEC69, Bacillus alginate BEC68A, Bacillus alginate BEC68B, Bacillus alginate BEC68C, Bacillus alginate BEC68D, Bacillus amyloliquefaciens BEC77A, Bacillus amyloliquefaciens BEC77B, Bacillus amyloliquefaciens BEC69, Bacillus tegira BEC78, Bacillus megaterium BEC71, Bacillus taichung BEC110, Bacillus erinaceus BEC120, Bacillus ilignonii BEC108, Microbes arabinogalactanophilus BEC102, and Bacillus rubrum BEC101.

[0305] In some embodiments, the isolated microorganisms, aggregates, and / or agricultural compositions of this disclosure may be applied to plants to impart disease resistance. Suitably, in such embodiments, the microorganisms may be selected from the group consisting of *Bacillus tekirae* and *Bacillus methyltrophicus*. Suitably, the microorganisms may be selected from the group consisting of *Bacillus tekirae* BEC80 and *Bacillus methyltrophicus* BEC60 and BEC56.

[0306] Agricultural Composition

[0307] In some embodiments, the microorganisms of this disclosure are combined with agricultural compositions. Agricultural compositions generally refer to organic and inorganic compounds that may include compositions that promote the cultivation of microorganisms and / or plant components; compositions involved in the formulation of microorganisms to be applied to plant components (e.g., but not limited to: wetting agents, compatibilizers (also called "compatibilizers"), defoamers, detergents, chelating agents, drift reducers, neutralizers and buffers, corrosion inhibitors, dyes, odorants, spreaders (also called "dispersants"), penetration aids (also called "penetrators"), adhesives (also called "binders" or "binding agents"), dispersants, thickeners (also called "thickeners"), stabilizers, emulsifiers, freezing point inhibitors, antimicrobial agents, etc.); compositions involved in conferring protection to plant components or plants (e.g., but not limited to: insecticides, nematicides, fungicides, bactericides, herbicides, etc.); and other compositions that may be of interest for a particular application.

[0308] In some embodiments, the agricultural compositions of this disclosure are solid. When using solid compositions, it may be desirable to include one or more carrier materials with isolated active microorganisms or aggregates. In some embodiments, this disclosure teaches the use of carriers, including but not limited to: mineral soils such as silica, silica gel, silicates, talc, kaolin, activated clay, limestone, chalk, loess, clay, dolomite, diatomaceous earth, calcium sulfate, magnesium sulfate, magnesium oxide, milled synthetic materials, fertilizers such as ammonium sulfate, ammonium phosphate, ammonium nitrate, thiourea and urea, plant-derived products such as grain flour, bark powder, wood flour and nutmeg powder, cellulose powder, palygorskite, montmorillonite, mica, vermiculite, synthetic silica and synthetic calcium silicate, or combinations of these substances.

[0309] Growth composition

[0310] In some embodiments, compositions that promote growth and development are provided to microorganisms and / or plant components. Exemplary compositions include liquids (such as broth, culture media) and / or solids (such as soil, nutrients). Various organic or inorganic compounds may be added to the growth composition, alone or in combination with plant components, to benefit the health of the microorganisms, such as, but not limited to, amino acids, vitamins, minerals, carbohydrates, monosaccharides, and lipids.

[0311] Pharmaceutical composition

[0312] One or more compositions other than microorganisms or microbial-derived compositions may be combined for various applications, stability, activity and / or storage reasons. Additional compositions may be referred to as "formulation components".

[0313] In some embodiments, the agricultural compositions of this disclosure are liquids. Therefore, in some embodiments, this disclosure teaches that the agricultural compositions disclosed herein may comprise compounds or salts such as monoethanolamine salts, sodium sulfate, potassium sulfate, sodium chloride, potassium chloride, sodium acetate, ammonium bisulfate, ammonium chloride, ammonium acetate, ammonium formate, ammonium oxalate, ammonium carbonate, ammonium bicarbonate, ammonium thiosulfate, ammonium diphosphate, ammonium monophosphate, sodium ammonium hydrogen phosphate, ammonium thiocyanate, ammonium aminosulfonate, or ammonium carbamate.

[0314] In some embodiments, this disclosure teaches that agricultural compositions may comprise binders such as: polyvinylpyrrolidone, polyvinyl alcohol, partially hydrolyzed polyvinyl acetate, carboxymethyl cellulose, starch, ethylene pyrrolidone / vinyl acetate copolymer, and polyvinyl acetate, or combinations thereof; lubricants such as magnesium stearate, sodium stearate, talc, or polyethylene glycol, or combinations thereof; defoamers such as silicone emulsions, long-chain alcohols, phosphate esters, ethynyl glycol, fatty acids, or organofluorine compounds; and complexing agents such as salts of ethylenediaminetetraacetic acid (EDTA), salts of triazinotriacetic acid, or salts of polyphosphate, or combinations thereof.

[0315] In some embodiments, the agricultural composition comprises a surfactant. In some embodiments, the surfactant is added to a liquid agricultural composition. In other embodiments, the surfactant is added to a solid formulation, particularly those designed to be diluted with a carrier prior to application. Thus, in some embodiments, the agricultural composition comprises a surfactant. Surfactants are sometimes used alone or together with other additives (such as minerals or vegetable oils) as adjuvants to spray can mixtures to improve the biocompatibility of microorganisms with a target. The type of surfactant used in bioenhancers generally depends on the nature and mode of action of the microorganisms. Surfactants can be anionic, cationic, or nonionic and can be used as emulsifiers, wetting agents, suspending agents, or for other purposes. In some embodiments, the surfactant is a nonionic surfactant, such as alkyl ethoxylates, linear fatty alcohol ethoxylates, and fatty amine ethoxylates. Surfactants commonly used in the field of pharmaceutical formulations and also applicable to the formulations of this invention are described in McCutcheon's Detergents and Emulsifiers Annual, MC Publishing Corp., Ridgewood, NJ, 1998 and Encyclopedia of Surfactants, Volumes I-III, Chemical Publishing Co., New York, 1980-81. In some embodiments, this disclosure teaches the use of surfactants comprising alkali metal, alkaline earth metal, or ammonium salts of fatty acids, including aromatic sulfonic acids (e.g., lignin sulfonic acid, phenol sulfonic acid, naphthalene sulfonic acid, and dibutylnaphthalene sulfonic acid), alkyl ethers, dodecyl ethers, fatty alcohol sulfates, and fatty alcohol glycol ether sulfates; condensates of sulfonated naphthalene and its derivatives with formaldehyde; condensates of naphthalene or naphthalene sulfonic acid with phenol and formaldehyde; condensates of phenol or phenol sulfonic acid with formaldehyde; condensates of phenol with formaldehyde and sodium sulfite; polyoxyethylene octylphenyl ether; ethoxylated isooctylphenol, ethoxylated octylphenol, or ethoxylated nonylphenol; tributylphenyl polyethylene glycol ether; alkyl aryl polyether alcohol; isotetrazol; ethoxylated castor oil; ethoxylated triarylphenol; salts of phosphorylated triarylphenol ethoxylates; dodecyl alcohol polyethylene glycol ether acetate; sorbitol esters; lignin-sulfite waste liquid or methylcellulose; or combinations thereof.

[0316] In some embodiments, this disclosure teaches other suitable surfactants, including salts of alkyl sulfates, such as diethanolammonium dodecyl sulfate; alkyl aryl sulfonates, such as calcium dodecylbenzenesulfonate; alkylphenol-epoxide addition products, such as nonylphenol-C18 ethoxylate; ethanol-epoxide addition products, such as tridecyl alcohol-C16 ethoxylate; soaps, such as sodium stearate; alkyl naphthalene-sulfonates, such as sodium dibutylnaphthalenesulfonate; and dialkyl esters of sulfosuccinates, such as di(2-ethylhexyl)sulfonate. Sodium succinate; sorbitol esters, such as sorbitol oleate; quaternary ammonium, such as dodecyltrimethylammonium chloride; polyethylene glycol esters of fatty acids, such as polyethylene glycol stearate; block copolymers of ethylene oxide and propylene oxide; salts of monoalkyl and dialkyl phosphates; vegetable oils, such as soybean oil, rapeseed / canola oil, olive oil, castor oil, sunflower oil, coconut oil, corn oil, cottonseed oil, linseed oil, palm oil, peanut oil, safflower oil, sesame oil, tung oil, etc.; and esters of the above vegetable oils, especially methyl esters.

[0317] In some embodiments, the agricultural composition includes a wetting agent. A wetting agent is a substance that, when added to a liquid, improves the spreading or penetrating ability of the liquid by reducing the interfacial tension between the liquid and the surface over which it spreads. Wetting agents serve two main functions in agricultural chemical formulations: improving the wetting rate of powders in water during processing and manufacturing to prepare soluble liquid concentrates or suspension concentrates; and shortening the wetting time of wettable powders and improving the penetration rate of water into water-dispersible particles during mixing of the product with water in a spray can or other container. Examples of wetting agents used in the agricultural compositions of this disclosure (including wettable powders, suspension concentrates, and water-dispersible particle formulations) in some embodiments are: sodium dodecyl sulfate; sodium dioctyl sulfosuccinate; alkylphenol ethoxylates; and fatty alcohol ethoxylates.

[0318] In some embodiments, the agricultural compositions of this disclosure include a dispersant. A dispersant is a substance that adsorbs onto the surface of particles and helps maintain the dispersed state of the particles and prevents them from re-aggregating. In some embodiments, a dispersant is added to the agricultural compositions of this disclosure to promote dispersion and suspension during manufacturing and to ensure that the particles are redispersed in water in a spray can. In some embodiments, the dispersant is used in wettable powders, suspension concentrates, and water-dispersible particles. Surfactants used as dispersants have the ability to strongly adsorb onto the particle surface and provide charged or steric barriers that prevent particle re-aggregation. In some embodiments, the most commonly used surfactants are anionic, nonionic, or a mixture of both.

[0319] In some embodiments, sodium lignosulfonate is the most commonly used dispersant for wettable powder formulations. In some embodiments, polyelectrolytes (such as sodium naphthalenesulfonate formaldehyde condensates) are used for suspension concentrates to provide excellent adsorption and stabilization. In some embodiments, tristyrene-phenol ethoxylate esters are also used. In some embodiments, alkylaryl ethylene oxide condensates and EO-PO block copolymers are sometimes combined with anionic surfactants as dispersants for suspension concentrates.

[0320] In some embodiments, the agricultural compositions of this disclosure comprise polymeric surfactants. In some embodiments, the polymeric surfactants have extremely long hydrophobic "backbone" and numerous ethylene oxide chains that form the "teeth" of a "comb"-like surfactant. In some embodiments, these high molecular weight polymers can give the suspension concentrate excellent long-term stability because the hydrophobic backbone has numerous points anchored to the particle surface. Examples of dispersants used in the agricultural compositions of this disclosure in some embodiments are: sodium lignin sulfonate; sodium naphthalene sulfonate formaldehyde condensate; tristyrene-phenol ethoxylate phosphate; fatty alcohol ethoxylates; alkyl ethoxylates; EO-PO block copolymers; and graft copolymers.

[0321] In some embodiments, the agricultural compositions of this disclosure include an emulsifier. An emulsifier is a substance that stabilizes a suspension of droplets in one liquid phase in another liquid phase. Without an emulsifier, the two liquids would separate into two immiscible liquid phases. In some embodiments, the most commonly used emulsifier blends comprise alkylphenols or fatty alcohols having 12 or more ethylene oxide units and oil-soluble calcium salts of dodecylbenzenesulfonic acid. A hydrophilic-lipophilic balance (“HLB”) value in the range of 8 to 18 will generally provide a well-stabilized emulsion. In some embodiments, emulsion stability can sometimes be improved by adding a small amount of EO-PO block copolymer surfactant.

[0322] In some embodiments, the agricultural compositions of this disclosure include a solubilizer. The solubilizer is a surfactant that forms micelles in water at concentrations exceeding a critical micelle concentration. The micelles are then capable of dissolving or solubilizing water-insoluble materials within the hydrophobic portion of the micelles. Commonly used surfactants for dissolution are nonionic surfactants: sorbitan monooleate; sorbitan monooleate ethoxylate; and methyl oleate.

[0323] In some embodiments, the agricultural compositions of this disclosure comprise organic solvents. Organic solvents are primarily used in formulations of emulsifiable concentrates, ULV formulations, and to a lesser extent in granule formulations. Sometimes mixtures of solvents are used. In some embodiments, this disclosure teaches the use of solvents including aliphatic paraffin oils, such as kerosene or refined paraffin. In other embodiments, this disclosure teaches the use of aromatic solvents, such as xylene and higher molecular weight fractions of C9 and C10 aromatic solvents. In some embodiments, chlorinated hydrocarbons may be used as co-solvents to prevent pesticide crystallization when the formulation is emulsified in water. Sometimes alcohols are used as co-solvents to improve solubility.

[0324] In some embodiments, the agricultural composition includes a gelling agent. Thickeners or gelling agents are primarily used in the formulation of suspension concentrates, emulsions, and suspension emulsions to alter the rheology or flowability of the liquid and prevent the separation and sedimentation of dispersed particles or droplets. Thickeners, gelling agents, and anti-settling agents generally fall into two categories: water-insoluble particles and water-soluble polymers. It is possible to use clay and silica to produce suspension concentrate formulations. In some embodiments, the agricultural composition includes one or more thickeners, including but not limited to: montmorillonite, such as bentonite; magnesium aluminum silicate; and palygorskite. In some embodiments, this disclosure teaches the use of polysaccharides as thickeners. The most commonly used types of polysaccharides are natural extracts of seeds and seaweed or synthetic derivatives of cellulose. Some embodiments utilize xanthan gum and some embodiments utilize cellulose. In some embodiments, this disclosure teaches the use of thickeners, including but not limited to: guar gum; locust bean gum; carrageenan; alginate; methylcellulose; sodium carboxymethyl cellulose (SCMC); and hydroxyethyl cellulose (HEC). In some embodiments, this disclosure teaches the use of other types of antisettling agents, such as modified starch, polyacrylates, polyvinyl alcohol, and polyethylene oxide. Another good antisettling agent is xanthan gum.

[0325] In some embodiments, the presence of surfactants (which reduce interfacial tension) can cause foaming in water-based formulations during preparation and mixing operations during application via spray cans. Therefore, in some embodiments, to reduce the tendency to foam, defoamers are typically added during the preparation stage or before filling into the bottle / spray can. Generally, there are two types of defoamers: silicone and non-silicone. Silicone defoamers are typically aqueous emulsions of dimethylpolysiloxane, while non-silicone defoamers are water-insoluble oils such as octanol and nonanol, or silica. In both cases, the function of the defoamer is to transfer the surfactant from the air-water interface.

[0326] In some embodiments, the agricultural composition contains a preservative.

[0327] In some embodiments, the agricultural composition may be formulated as: soil irrigation agent, foliar spray, impregnation treatment agent, furrow treatment agent, soil conditioner, granules, broadcast treatment agent, post-harvest disease control treatment agent, or seed treatment agent. In some embodiments, the agricultural composition may be applied alone or in combination with other agricultural products according to a rotary spraying procedure.

[0328] In some embodiments, the agricultural composition is tank-compatible. In some embodiments, the agricultural composition is tank-compatible with other agricultural products. In some embodiments, the agricultural composition is compatible with equipment used for ground, air, and irrigation application.

[0329] In some embodiments, the agricultural composition may be applied to genetically modified seeds or plants.

[0330] Protective composition

[0331] Furthermore, individual microorganisms, microbial aggregates, or microbial communities developed according to the disclosed methods can be combined with known active agents available in the agricultural field, such as: insecticides, herbicides, fungicides, cytotoxic agents, viricides, acaricides, nematicides, scabicides, plant growth regulators, rodenticides, anti-algae agents, biocontrol agents, or beneficial reagents. Additionally, microorganisms, microbial aggregates, or microbial communities developed according to the disclosed methods can be combined with known fertilizers. Such combinations can exhibit synergistic properties. Furthermore, individual microorganisms, microbial aggregates, or microbial communities developed according to the disclosed methods can be combined with inert ingredients. Additionally, in some aspects, the disclosed microorganisms are combined with bioactive agents.

[0332] In some embodiments, individual microorganisms or microbial aggregates or communities developed according to the disclosed methods may be combined with biopesticides that act as herbicides, fungicides, insecticides, viricides, acaricides, nematicides, scabicides, rodenticides, and / or antialgae agents. Such biopesticides may be, but are not limited to, macroorganisms (e.g., beneficial nematodes, etc.), microbial organisms (e.g., Serenade, Bt, etc.), plant extracts (e.g., Timorex Gold, etc.), biochemical substances (e.g., insect pheromones, etc.), and / or minerals and oils (e.g., canola oil).

[0333] Pesticides and biological pesticides

[0334] In some embodiments, the agricultural compositions of this disclosure comprise an insecticide used in combination with the taught microorganisms. In some embodiments, the agricultural compositions of this disclosure comprise a bio-insecticide used in combination with the taught microorganisms.

[0335] In some implementations, individual microorganisms or microbial aggregates or communities developed according to the disclosed methods can be combined with known pesticides in the agricultural field, such as pesticides that act as herbicides, fungicides, insecticides, viricides, acaricides, nematicides, scabicides, rodenticides and / or antialgae agents.

[0336] In some implementations, individual microorganisms or microbial aggregates or communities developed according to the disclosed methods can be combined with known biopesticides in the agricultural field, such as biopesticides that act as herbicides, fungicides, insecticides, viricides, acaricides, nematicides, scabicides, rodenticides and / or antialgae agents.

[0337] For example, in some embodiments, this disclosure teaches agricultural compositions comprising one or more of the following active ingredients: macroorganisms (e.g., beneficial nematodes, etc.), microorganisms (e.g., Serenade, Bt, etc.), plant extracts (e.g., Timorex Gold, etc.), biochemical substances (e.g., insect pheromones, etc.), and / or minerals and oils (e.g., canola oil).

[0338] In some embodiments, the individual microorganisms, microbial aggregates, or microbial communities developed according to the disclosed methods may be combined with herbicides selected from the group consisting of: acetamides selected from the group consisting of: acetochlor, metolachlor, butachlor, succinyl-methyl, fenfluroxychlor, fluthiamethoxam, bensulfuron-methyl, metolachlor, pyrazosulfuron, chlorpyrifos, clodinafop-propionate, clethodim, pretilachlor, thiamethoxam, and thifensulfuron-methyl; amino acid derivatives selected from the group consisting of: bisphosphonium, glufosinate, and glyphosate; aryloxyphenoxypropionates selected from the group consisting of: clodinafop-propionate, cyhalofop-butyl, quizalofop-p-ethyl, haloxyfop-P-ethyl, oxadiazon, quizalofop-P-ethyl, quizalofop-P-ethyl, quizalofop-P-ethyl, quizalofop-P-ethyl, and quizalofop-P-ethyl; and dichlorvos. Herbicides and paraquat; (thiolated) carbamates selected from the following groups: chlorpyrifos, butachlor, carbaryl, betaine, piperazine, EPTC, quizalofop-p-ethyl, chlorpyrifos, berberine, betaine, bensulfuron-methyl, barnyardgrass, quizalofop-p-ethyl, and oxadiazon; Cyclohexane selected from the following groups: butylbenzyl, clethodim, thiamethoxam, cyclobenzyl, haloxyfop-methyl, pyrazosulfuron, and oxadiazon; Dinitroanilines selected from the following groups: flurbichlor, ethylbutadiene, azoxystrobin, pendimethalin, ambroxol, and trifluralin; Diphenyl ethers selected from the following groups: trifluralin, bensulfuron-methyl, chlorpyrifos, chlorpyrifos, flufenoxuron, quizalofop-p-ethyl, and oxyfluorfen. Hydroxybenzonitrile selected from the following groups: bromobenzonitrile, chlorpyrifos, and iodobenzonitrile; imidazoline ketones selected from the following groups: imazalil, methoxypromethazine, methyl imazalazine, metribuzin, metribuzin, and imazalazine; phenoxyacetic acids selected from the following groups: chlorfenapyr, 2,4-dichlorophenoxyacetic acid (2,4-D), 2,4-DB, doxypropionic acid, MCPA, MCPA-thioethyl ester, MCPB, and 2,4-methylchloropropionic acid; pyrazines selected from the following groups: chlorpyrifos, flupyridaben, cyhalofop-butyl, flumethrin, and pyrazosulfuron; pyridines selected from the following groups: chlorpyrifos, dichlorpyrifos, pyrfluthrin, flusulfanilamide, flupyrazosulfuron, and chlorpyrifos. Flupyrazole and thiamethoxam; sulfonylureas selected from the group consisting of: sulfadiazine, tetrazolium sulfadiazine, bensulfadiazine, chlorpyrifos, chlorsulfuron, ethersulfuron, cypromethazine, ethoxysulfuron, pyrimisulfuron, flupyrsulfuron, flupyrsulfuron, formamidesulfuron, chlorpyrifos, azoxysulfuron, iodosulfuron, mesosulfuron, mesosulfuron, nicosulfuron, epoxysulfuron, flupyrsulfuron, flusulfuron, pyrimisulfuron, sulfadiazine, mesosulfuron, sulfonylsulfuron, thifensulfuron, etherbensulfuron, benzylsulfuron, trifluridinesulfuron, flumethanil, trifluridinesulfuron and 14(2-chloro-6-propyl-imidazol[1,2-b]pyridazin-3-yl)sulfonyl)-3-(4,6-dimethoxy-pyrimidin-2-yl)urea;Triazine compounds selected from the group consisting of the following: atrazine, atrazine, cyprodinil, isoamyl, etaziclomefone, cyclomethonium, benzoate, cyprodinil, promethazine, simazine, terbufenozide, decazine, and triazine fluroxypyr; urea compounds selected from the group consisting of the following: chlormequat, chlorfluazuron, diuron, fenfluroxypyr, isoproturon, linuron, methylbenzylthiazoline, and butyrazoline; acetolactate synthase inhibitors selected from the group consisting of the following: bispyribac-sodium, chlorpyrifos-sulfuron-methyl, dichlorvos-sulfuron-methyl, fluroxypyr, pyrazopyr-sulfuron-methyl, sulfadiazine-sulfuron-methyl, penoxsulam, propanil-sulfuron-methyl, propyltriazine. Glufosinate, pyrimisulfuron, cyclopyrimisulfuron, pyrimisulfuron, pyrimisulfuron, sulfonylpyrazosulfuron, and pyrazosulfuron; and compounds selected from the group consisting of: azoxystrobin, aminotriazole, sparphos, flubutyrazole, glufosinate, bencarbazone, benfluresate, pyrazosulfuron, bentazon, dicyclosulfuron, chlorpyrifos, brobutyrazole, flupropyrazosulfuron, phosmet, pyrazosulfuron-methyl, pyrazosulfuron-methyl, pyrazosulfuron-methyl, pyrazosulfuron-methyl, indole-methyl, diflubenzuron, cyclohexane, isoxaflutole, bensulfuron-methyl, propargylsulfonamide, dicamba, fenvalerate, flupyrazole, and *Drechslera*. Monoceras), Herbicides, Ethoxybenzamide, Tetracycline, Fluroxypyr, Propyrimethanil, Flumetsulam, Fluroxypyr, Fluroxypyr, Furazolidone, Indoxime, Isoxazolidone, Isoxazolidone, Cyclopyralid, Propanil, Pendimethalin, Quinolinic acid, Chlormethalin, Mesotrione, Methylarsic acid, Herbicides, Propyrimethanil, Oxychlor, Oxychlor, Cyclopentachlor, Cyclopyralid, Bisoxazolidin, Pyrazosulfuron, Pyrasuloftole, Pyrazosulfuron ... Cyclo[3.2.1]oct-3-en-2-one, (3-[2-chloro-4-fluoro-5-(3-methyl-2,6-dioxo-4-trifluoromethyl-3,6-dihydro-2H-pyrimidin-1-yl)-phenoxy]-pyridin-2-yloxy)-ethyl acetate, methyl 6-amino-5-chloro-2-cyclopropyl-pyrimidin-4-carboxylate, 6-chloro-3-(2-cyclopropyl-6-methyl-phenoxy)-pyridazin-4-ol, 4-amino-3-chloro-6-(4-chloro-phenyl)-5-fluoro-pyridin-2-carboxylic acid, methyl 4-amino-3-chloro-6-(4-chloro-2-fluoro-3-methoxy-phenyl)-pyridin-2-carboxylate, and methyl 4-amino-3-chloro-6-(4-chloro-3-dimethylamino-2-fluoro-phenyl)-pyridin-2-carboxylate.

[0339] In some embodiments, the individual microorganisms, microbial aggregates, or microbial communities developed according to the disclosed methods may be combined with insecticides selected from the group consisting of: organo(thio)phosphates selected from the group consisting of: acephate, methyl pyrazophos, phosmet, chlorpyrifos, methyl chlorpyrifos, chlorpyrifos, diazinon, dichlorvos, chlorpyrifos, dimethoate, phorate, ethion, fenitrothion, fenthion, isoxazophos, malathion, methamidophos, chlorpyrifos, methyl parathion, phosmet, phosmet, phosmet, parathion, parathion, parathion, phosmet, phosmet, phosmet, phorate, phorate, phorate, phosmet, phorate, phorate, phosmet, phorate, phosmet, phorate, phosmet, phorate, phosmet, terbufos, phosmet Phosphorus, triazophos, and trichlorfon; carbamates selected from the group consisting of: carbaryl, aldicarb, cypermethrin, carbofuran, carbofuran, thiocarb, fenoxycarb, furazolidone, methiocarb, methomyl, chlorpyrifos, propoxur, thiamethoxam, and pymetrozine; pyrethroids selected from the group consisting of: allethrin, bifenthrin, cypermethrin, deltamethrin, cypermethrin, α-cypermethrin, β-cypermethrin, ζ-cypermethrin, deltamethrin, fenvalerate, fenvalerate, cypermethrin, cypermethrin, cypermethrin, lambda-cyhalothrin, permethrin, pyrethrin I and II, benzalkonium chloride, flusilazole, and deltamethrin. Heptafluthrin, methamidophos, tetrabromopyrethrin, tetrafluorobenzyl, propofol, and tetrafluoromethrin; insect growth regulators selected from the group consisting of: a) chitin synthesis inhibitors, wherein the chitin synthesis inhibitors are benzoylureas selected from the group consisting of: flufenoxuron, cyramazin, diflubenzuron, flufenoxuron, flufenoxuron, flufenoxuron, lufenuron, flufenoxuron, flufenoxuron, flufenoxuron, chlorfenapyr; thiamethoxam, bensulfuron, thiamethoxam, etoxazole, and tetradifon; b) ecdysone antagonists selected from the group consisting of: chlorfenapyr, methoxyfenozide, tebufenozide, and azadirachtin; c) juvenile hormone analogs selected from the group consisting of: pyriproxyfen, tebufenozide, and phenoxyfenozide. d) Lipid biosynthesis inhibitors selected from the group consisting of: spirodiclofen, spirodiclofen, and spirotetramat; Nicotinic receptor agonists / antagonists selected from the group consisting of: thiamethoxam, fipronil, imidacloprid, thiamethoxam, acetamiprid, acetamiprid, thiamethoxam, and 1-(2-chloro-thiazo-5-ylmethyl)-2-nitromimino-3,5-dimethyl-[1,3,5]triazine; GABA antagonists selected from the group consisting of: endosulfan, acetamiprid, fipronil, flupyradifurone, pyrazinoflurane, pyrazolium, and 5-amino-1-(2,6-dichloro-4-methyl-phenyl)-4-sulfinylamino-1H-pyrazole-3-thiocarbamate;Macrolide insecticides selected from the following groups: abamectin, emamectin, mibamectin, rapamycin, spinosad, and ethyl spinosad; mitochondrial electron transport inhibitor (METI) I scabies insecticides selected from the following groups: quinfenoxam, pyridaben, pymetrozine, azoxystrobin, and pyrimethanil; METI insecticides selected from the following groups... Compounds II and III: acaricide, fluacyprim, and flufenoxuron; brofenoxuron; oxidative phosphorylation inhibitors selected from the group consisting of: tricyclic tin, bufenozide, fenbutatin, and chlorfenapyr; cryomazine; synergistic ethers; sodium channel blockers selected from the group consisting of: indoxacarb and cyfluthrin; and compounds selected from the group consisting of: benclothiazide, bifenazate, batan, flonicamid, acetamiprid, pymetrozine, sulfur, chlorfenapyr, flufenoxuron, chlorantraniliprole, brofenoxuron (HGW86), pyridaben, pyrimethanil, dicofol, sulfadiazine, imicyafos, bispyribac-triflufenoxuron, and pyrifluquinazon.

[0340] In some embodiments, the present invention teaches the synergistic use of the microorganisms or microbial aggregates disclosed herein with known pesticides in the agricultural field, such as pesticides that act as herbicides, fungicides, insecticides, viricides, acaricides, nematicides, scabicides, rodenticides, and / or antialgae agents.

[0341] In some embodiments, the present invention teaches the synergistic use of the microorganisms or microbial aggregates disclosed herein with known biopesticides in the agricultural field, such as biopesticides that act as herbicides, fungicides, insecticides, viricides, acaricides, nematicides, scabicides, rodenticides, and / or antialgae agents.

[0342] In some embodiments, when microorganisms or microbial aggregates identified according to the taught methods are combined with insecticides, an additive effect on the plant phenotypic trait of interest is observed. In other embodiments, when microorganisms or microbial aggregates identified according to the taught methods are combined with insecticides, a synergistic effect on the plant phenotypic trait of interest is observed.

[0343] In some embodiments, when microorganisms or microbial aggregates identified according to the taught methods are combined with biopesticides, an additive effect on the plant phenotypic trait of interest is observed. In other embodiments, when microorganisms or microbial aggregates identified according to the taught methods are combined with biopesticides, a synergistic effect on the plant phenotypic trait of interest is observed.

[0344] The synergistic effect obtained through the taught method can be quantified according to the Colby formula (i.e., (E) = X + Y - (X*Y / 100)). See Colby, RS, “Calculating Synergistic and Antagonistic Responses of Herbicide Combinations,” 1967 Weeds, Vol. 15, pp. 20-22, the full text of which is incorporated herein by reference. Therefore, the term “synergistic” refers to the component that, in its presence, increases the expected effect beyond the cumulative amount.

[0345] The isolated microorganisms and aggregates disclosed herein can synergistically enhance the efficacy of agricultural active pesticide compounds and agricultural auxiliary pesticide compounds.

[0346] The isolated microorganisms and aggregates disclosed herein can synergistically enhance the efficacy of agricultural active biological pesticide compounds and agricultural auxiliary biological pesticide compounds.

[0347] Plant growth regulators and biostimulants

[0348] In some embodiments, the agricultural compositions disclosed herein comprise plant growth regulators and / or biostimulants used in combination with the taught microorganisms.

[0349] In some implementations, individual microorganisms or microbial aggregates or communities developed according to the disclosed methods can be combined with known plant growth regulators in the agricultural field, such as auxins, gibberellins, cytokinins, ethylene-producing agents, growth inhibitors, and growth retardants.

[0350] For example, in some embodiments, this disclosure teaches agricultural compositions comprising one or more of the following active ingredients, including: cyclopropionylpyridinol, sec-butylamine, alcohol, chlormequat chloride, cytokinin, butyrylhydrazine, ethephon, furazolidone, gibberellic acid, gibberellin mixture, indole-3-butyric acid (IBA), maleic hydrazine, mefludide, mepiquat amine, mepiquat pentaborate, naphthaleneacetic acid (NAA), 1-naphthylacetamide (NAD), n-decyl alcohol, placlobutrazol, calcium cyclohexane, anti-buckwheat ester, uniconazole, salicylic acid, abscisic acid, ethylene, brassinolide, jasmonic acid, polyamine, nitric oxide, strigolactone, or karrikins, etc.

[0351] In some embodiments, individual microorganisms, microbial aggregates, or microbial communities developed according to the disclosed methods can be combined with seed inoculants known in the agricultural field, such as: In some embodiments, the Bradyrhizobium inoculum is used in combination with any single microorganism or microbial aggregate disclosed herein. In certain aspects, when one of the aforementioned inoculums (e.g., Synergistic effects were observed when *Or slow-growing rhizobia* were combined with microorganisms or microbial aggregates as taught herein.

[0352] In some embodiments, the agricultural compositions disclosed herein comprise plant growth regulators comprising: kinetin, gibberellic acid, and indolebutyric acid, as well as copper, manganese, and zinc.

[0353] In some embodiments, this disclosure teaches agricultural compositions comprising one or more commercially available plant growth regulators, including but not limited to: Royaltac

[0354] Early Boll Cotton Super Early Green PGR Sucker DipN Turf Armor

[0355] and

[0356] In some embodiments, the present invention teaches the synergistic use of the microorganisms or microbial aggregates disclosed herein with plant growth regulators and / or stimulants (such as plant hormones or chemicals that affect the production or destruction of plant growth regulators).

[0357] In some embodiments, the present invention teaches plant hormones, which may include: auxins (e.g., indoleacetic acid IAA), gibberellins, cytokinins (e.g., kinetin), abscisic acid, ethylene (and their production, such as being regulated by ACC synthase and destroyed by ACC deaminase).

[0358] In some implementations, individual microorganisms, microbial aggregates, or microbial communities developed according to the disclosed methods may be combined with biostimulants. Such biostimulants may be, but are not limited to, microbial organisms, plant extracts, algae, acids, biochar, etc.

[0359] In some embodiments, individual microorganisms, microbial aggregates, or microbial communities developed according to the disclosed methods can be combined with fertilizers, which can be organic (e.g., manure, blood meal, fish meal, etc.), nitrogen-based (e.g., nitrates, ammonium, urea, etc.), phosphates, and potash fertilizers. Such fertilizers may also contain micronutrients, including but not limited to sulfur, iron, and zinc.

[0360] In some embodiments, the present invention teaches additional plant growth-promoting chemicals that can work synergistically with the microorganisms and microbial aggregates disclosed herein, such as humic acid, fulvic acid, amino acids, polyphenols, and protein hydrolysates.

[0361] Therefore, in some embodiments, this disclosure provides for combining the taught microorganism with... The combination of these microorganisms can be applied to any crop. Furthermore, this disclosure provides for the application of the taught microorganisms with... It can be applied in combination to any crop and using any method or amount of application.

[0362] In some embodiments, this disclosure teaches agricultural compositions containing biostimulants.

[0363] As used herein, the term "biostimulant" refers to any substance used to stimulate the growth of microorganisms that may be present in soil or other plant growth media.

[0364] The level of microorganisms in soil or growth media is directly related to plant health. Microorganisms feed on biodegradable carbon sources, and therefore plant health is also related to the amount of organic matter in the soil. While fertilizers provide nutrients to nourish and grow plants, in some embodiments, biostimulants provide biodegradable carbon (e.g., molasses, carbohydrates (e.g., sugars)) to nourish and grow microorganisms. Unless otherwise explicitly stated, biostimulants may contain a single ingredient or a combination of several different ingredients that, due to the effects of one or more of these ingredients (acting alone or in combination), can enhance microbial activity or plant growth and development.

[0365] In some embodiments, biostimulants are compounds that produce non-nutritive plant growth responses. In some embodiments, the many important benefits of biostimulants are based on their ability to influence hormone activity. Hormones in plants (plant hormones) are chemical messengers that regulate normal plant development and responses to the environment. Root and shoot growth, as well as other growth responses, are regulated by plant hormones. In some embodiments, compounds in biostimulants can alter the hormonal state of plants and have a significant impact on their growth and health. Therefore, in some embodiments, this disclosure teaches kelp, humic acid, fulvic acid, and vitamin B as common components of biostimulants. In some embodiments, the biostimulants of this disclosure enhance antioxidant activity, thereby enhancing the plant's defense system. In some embodiments, vitamin C, vitamin E, and amino acids (such as glycine) are antioxidants contained in the biostimulant.

[0366] In other embodiments, biostimulants can be used to stimulate the growth of microorganisms present in soil or other plant growth media. Previous studies have shown that when certain biostimulants containing specific organic seed extracts (e.g., soybean) are used in combination with microbial inoculants, the biostimulants can stimulate the growth of microorganisms contained in the microbial inoculum. Therefore, in some embodiments, this disclosure teaches one or more biostimulants that, when used with microbial inoculants, can enhance the populations of native microorganisms and inoculum microorganisms. For a review of some popular uses of biostimulants, see Calvo et al., 2014, Plant Soil 383:3-41.

[0367] A combination of plant components, microorganisms, and agricultural compositions

[0368] In some embodiments, this disclosure teaches that individual microorganisms or microbial aggregates or communities or any combination thereof (e.g., including any one or more microorganisms disclosed in Table 1 or Table 1A) can be applied to plant components (optionally in combination with any agricultural composition) to improve plant phenotype.

[0369] Isolated microorganisms, communities, or aggregates (often interchangeably referred to as "microbes" or "microbes") can be applied to heterologous plant components to form a synthetic assemblage. Microorganisms are considered heterologous to plant components if they are not normally associated with plant components in nature or are applied in quantities different from those found in nature. In some embodiments, microorganisms may be present in one part of a plant but not in another, and the introduction of microorganisms into another part of the plant is considered a heterologous association.

[0370] It is further envisioned that isolated microorganisms, or those in combination with plants or plant components, could be further associated with one or more agricultural compositions (such as those described above).

[0371] The concept of synthetic combinations of microorganisms with plant components, microorganisms with agricultural compositions, and microorganisms with plant components and agricultural compositions (often referred to as "synthetic compositions," i.e., compositions containing components that are not typically found in nature) is envisioned.

[0372] Plant component treatment

[0373] In some embodiments, this disclosure also relates to the finding that treating plant components with one or more of the microorganisms or agricultural compositions of this disclosure prior to sowing or planting can enhance desired plant traits, such as plant growth, plant health, and / or plant insect resistance.

[0374] Therefore, in some embodiments, this disclosure teaches the use of one or more microorganisms or microbial aggregates as plant component treatment agents. Plant component treatment agents can be plant component coatings applied directly to untreated and "naked" plant components. However, plant component treatment agents can be plant component coatings applied to plant components that have already been coated with one or more previous plant component coatings or plant component treatment agents. Previous plant component treatment agents may contain one or more active compounds (chemical or biological) and one or more inert components.

[0375] The term "plant component treatment agent" generally refers to the application of material to plant components before or during planting in soil. Plant component treatment agents having the microbial and other agricultural compositions disclosed herein have the advantage of delivering the treatment agent to areas where plant components are planted shortly before germination and emergence.

[0376] In other embodiments, this disclosure also teaches that the use of plant component treatment agents can minimize the amount of microorganisms or agricultural compositions required for successful plant treatment and further limit worker exposure to microorganisms and compositions compared to application techniques such as spraying over soil or over germinating plant components.

[0377] Furthermore, in some embodiments, this disclosure teaches that the microorganisms disclosed herein are important for enhancing plant life in the early stages (e.g., within the first thirty days after the emergence of plant components). Therefore, in some embodiments, delivering the microorganisms and / or compositions of this disclosure in the form of a plant component treatment agent allows them to be placed in the area of ​​action at a time when microbial activity is critical.

[0378] In some embodiments, the microbial compositions of this disclosure are formulated as plant component treatment agents. In some embodiments, it is envisioned that one or more layers of the microorganisms and / or agricultural compositions disclosed herein can be substantially uniformly coated onto plant components using treatment agent application equipment specially designed and manufactured to apply the plant component treatment product precisely, safely, and effectively, using conventional mixing, spraying methods, or combinations thereof. Such equipment utilizes various types of coating technologies, such as rotary coaters, drum coaters, fluidized bed technology, fountain beds, rotary sprays, or combinations thereof. Liquid plant component treatment agents (such as those disclosed herein) can be applied via a spinning "atomizer" disc or nozzle, which distributes the plant component treatment agent uniformly onto the plant component as it moves in a spray mode. In various embodiments, the plant component is subsequently mixed or tumbled for a period of time to achieve additional treatment agent distribution and drying.

[0379] Prior to coating with the microbial composition, the plant components may be induced or uninitiated to improve the uniformity of germination and emergence. In an alternative embodiment, the dry powder formulation may be metered onto the mobile plant components and allowed to mix until fully distributed.

[0380] In some embodiments, at least a portion of the surface area of ​​the plant component is coated with the microbial composition according to the present disclosure. In some embodiments, a plant component coating containing the microbial composition is applied directly to a bare plant component. In some embodiments, a plant component coating containing the microbial composition is applied to a plant component that has already been coated with a plant component. In some aspects, the plant component may have a plant component coating containing, for example, thiamethoxam and / or Bacillus firmus-I-1582, on which the composition of the present invention will be applied as a plant component coating. In some aspects, the taught microbial composition is applied as a plant component coating to a plant component that has already been coated with PONCHO. TM VOTiVO TM The plant component being treated. In some aspects, the plant component may have a plant component coating containing, for example, metalaxyl and / or thiamethoxam and / or Bacillus thuringiensis-I-1582, on which the composition of the present invention will be applied as a plant component coating. In some aspects, the taught microbial composition is applied as a plant component coating to a product treated with ACCELERON. TM The components of the plant being treated.

[0381] In some embodiments, the plant components treated with microorganisms have the following microbial spore concentrations or microbial cell concentrations: approximately 10^2 to 10^12, 10^2 to 10^11, 10^2 to 10^10, 10^2 to 10^9, 10^2 to 10^8, 10^2 to 10^7, 10^2 to 10^6, 10^2 to 10^5, 10^2 to 10^4, or 10^2 to 10^3 per plant component.

[0382] In some embodiments, the plant components treated with microorganisms have the following microbial spore concentrations or microbial cell concentrations: approximately 10^3 to 10^12, 10^3 to 10^11, 10^3 to 10^10, 10^3 to 10^9, 10^3 to 10^8, 10^3 to 10^7, 10^3 to 10^6, 10^3 to 10^5, or 10^3 to 10^4 per plant component.

[0383] In some embodiments, the plant components treated with microorganisms have the following microbial spore concentrations or microbial cell concentrations: approximately 10^4 to 10^12, 10^4 to 10^11, 10^4 to 10^10, 10^4 to 10^9, 10^4 to 10^8, 10^4 to 10^7, 10^4 to 10^6, or 10^4 to 10^5 per plant component.

[0384] In some embodiments, the plant components treated with microorganisms have the following microbial spore concentrations or microbial cell concentrations: about 10^5 to 10^12, 10^5 to 10^11, 10^5 to 10^10, 10^5 to 10^9, 10^5 to 10^8, 10^5 to 10^7, or 10^5 to 10^6 per plant component.

[0385] In some implementations, the plant components treated with microorganisms have a microbial spore concentration or microbial cell concentration of approximately 10⁵ to 10⁹ per plant component.

[0386] In some embodiments, the plant components treated with microbial bodies have the following microbial spore concentrations or microbial cell concentrations: at least about 1×10^3 or 1×10^4 or 1×10^5 or 1×10^6 or 1×10^7 or 1×10^8 or 1×10^9 per plant component.

[0387] In some embodiments, the amount of one or more of the microorganisms and / or agricultural composition applied to the plant component depends on the final formulation and the size or type of the plant or plant component utilized. In some embodiments, one or more of the microorganisms are present at about 2% w / w to about 80% w / w of the total formulation. In some embodiments, by weight, one or more of the microorganisms used in the composition are about 5% w / w to about 65% w / w, or 10% w / w to about 60% w / w of the total formulation.

[0388] In some embodiments, the plant component may also have more spores or microbial cells per plant component, for example, about 10^2, 10^3, 10^4, 10^5, 10^6, 10^7, 10^8, 10^9, 10^10, 10^11, 10^12, 10^13, 10^14, 10^15, 10^16 or 10^17 spores or cells per plant component.

[0389] In some embodiments, the thickness of the plant component of this disclosure may be at most 10μm, 20μm, 30μm, 40μm, 50μm, 60μm, 70μm, 80μm, 90μm, 100μm, 110μm, 120μm, 130μm, 140μm, 150μm, 160μm, 170μm, 18 0μm、190μm、200μm、210μm、220μm、230μm、240μm、250μm、260μm、270μm、280μm m、290μm、300μm、310μm、320μm、330μm、340μm、350μm、360μm、370μm、380μm、3 90μm, 400μm, 410μm, 420μm, 430μm, 440μm, 450μm, 460μm, 470μm, 480μm, 490μm, 500μm, 510μm, 520μm, 530μm, 540μm, 550μm, 560μm, 570μm, 580μm, 590μm, 600μm, 610μm, 620μm, 630μm, 640μm, 650μm, 660μm, 670μm, 680μm, 690μm, 700μm, 710μm, 720μm, 730μm, 740μm, 750μm, 760μm, 770μm, 780μm, 790μm, 800μm ,810μm,820μm,830μm,840μm,850μm,860μm,870μm,880μm,890μm,900μm,910μm,920μm,930μm,940μm,950μm,960μm,970μm,980μm,990μm,1000μm,101 0μm, 1020μm, 1030μm, 1040μm, 1050μm, 1060μm, 1070μm, 1080μm, 1090μm, 1100μm, 1110μm, 1120μm, 1130μm, 1140μm, 1150μm, 1160μm, 1170μm, 1180μm, 11 90μm, 1200μm, 1210μm, 1220μm, 1230μm, 1240μm, 1250μm, 1260μm, 1270μm, 1280μm, 1290μm, 1300μm, 1310μm, 1320μm, 1330μm, 1340μm, 1350μm, 1360μm, 1 370μm, 1380μm, 1390μm, 1400μm, 1410μm, 1420μm, 1430μm, 1440μm, 1450μm, 1460μm, 1470μm, 1480μm, 1490μm, 1500μm, 1510μm, 1520μm, 1530μm, 1540μm,1550μm, 1560μm, 1570μm, 1580μm, 1590μm, 1600μm, 1610μm, 1620μm, 1630μm, 1640μm, 1650μm, 1660μm, 1670μm, 1680μm, 1690μm, 1700μm, 1710μm, 1720μm ,1730μm,1740μm,1750μm,1760μm,1770μm,1780μm,1790μm,1800μm,1810μm,1820μm,1830μm,1840μm,1850μm,1860μm,1870μm,1880μm,1890μm,1900μm m, 1910μm, 1920μm, 1930μm, 1940μm, 1950μm, 1960μm, 1970μm, 1980μm, 1990μm, 2000μm, 2010μm, 2020μm, 2030μm, 2040μm, 2050μm, 2060μm, 2070μm, 2080 μm, 2090μm, 2100μm, 2110μm, 2120μm, 2130μm, 2140μm, 2150μm, 2160μm, 2170μm, 2180μm, 2190μm, 2200μm, 2210μm, 2220μm, 2230μm, 2240μm, 2250μm, 226 0μm, 2270μm, 2280μm, 2290μm, 2300μm, 2310μm, 2320μm, 2330μm, 2340μm, 2350μm, 2360μm, 2370μm, 2380μm, 2390μm, 2400μm, 2410μm, 2420μm, 2430μm, 24 40μm, 2450μm, 2460μm, 2470μm, 2480μm, 2490μm, 2500μm, 2510μm, 2520μm, 2530μm, 2540μm, 2550μm, 2560μm, 2570μm, 2580μm, 2590μm, 2600μm, 2610μm, 2 620μm, 2630μm, 2640μm, 2650μm, 2660μm, 2670μm, 2680μm, 2690μm, 2700μm, 2710μm, 2720μm, 2730μm, 2740μm, 2750μm, 2760μm, 2770μm, 2780μm, 2790μm, 2800μm, 2810μm, 2820μm, 2830μm, 2840μm, 2850μm, 2860μm, 2870μm, 2880μm, 2890μm, 2900μm, 2910μm, 2920μm, 2930μm, 2940μm, 2950μm, 2960μm, 2970μm,2980μm, 2990μm, or 3000μm.

[0390] In some embodiments, the thickness of the plant component coating disclosed herein may be 0.5 mm, 1 mm, 1.5 mm, 2 mm, 2.5 mm, 3 mm, 3.5 mm, 4 mm, 4.5 mm, or 5 mm.

[0391] In some embodiments, the plant component coating of this disclosure may be at least 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 10.5%, 11%, 11.5%, 12%, 12.5%, 13%, 13.5%, 14%, 14.5%, 15%, 15.5%, 16%, 16.5%, 17%, 17.5%, 18%, 18.5%, 19%, 19.5%, 20%, 20.5%, 21%, 21.5%, 22%, 22.5%, 23%, 23.5%, or 24% of the weight of the uncoated plant component. 24.5%, 25%, 25.5%, 26%, 26.5%, 27%, 27.5%, 28%, 28.5%, 29%, 29.5%, 30%, 30.5%, 31%, 31.5%, 32%, 32.5%, 33%, 33.5%, 34%, 34.5%, 35%, 35.5%, 36%, 36.5%, 37% 37.5%, 38%, 38.5%, 39%, 39.5%, 40%, 40.5%, 41%, 41.5%, 42%, 42.5%, 43%, 43.5%, 44%, 44.5%, 45%, 45.5%, 46%, 46.5%, 47%, 47.5%, 48%, 48.5%, 49%, 49.5%, or 50%.

[0392] In some embodiments, microbial spores and / or cells may be freely coated onto plant components or may be formulated in a liquid or solid composition prior to coating onto plant components. For example, a solid composition containing microorganisms may be prepared by mixing a solid carrier with a suspension of spores until the solid carrier is impregnated with the spore or cell suspension. The mixture may then be dried to obtain the desired particles.

[0393] In some other embodiments, the solid or liquid microbial compositions of this disclosure are envisioned to also contain functional agents, such as activated carbon, nutrients (fertilizers), and other agents capable of improving the germination and quality of the product or combinations thereof.

[0394] Plant component coating methods and compositions known in the art can be particularly useful when modified by adding one of the embodiments of this disclosure. Such coating methods and their application devices are disclosed, for example, in U.S. Patent Nos. 5,916,029, 5,918,413, 5,554,445, 5,389,399, 4,759,945, 4,465,017 and U.S. Patent Application No. 13 / 260,310, each of which is incorporated herein by reference.

[0395] Plant component coating compositions are disclosed in, for example, U.S. Patent Nos. 5,939,356, 5,876,739, 5,849,320, 5,791,084, 5,661,103, 5,580,544, 5,328,942, 4,735,015, 4,634,587, 4,372,080, 4,339,456, and 4,245,432, each of which is incorporated herein by reference.

[0396] In some embodiments, a variety of additives may be added to the plant component treatment formulation comprising the composition of the present invention. A binder may be added, and said binder comprises a natural or synthetic adhesive polymer that is non-phytotoxic to the plant component being coated. The binder may be selected from polyvinyl acetate; polyvinyl acetate copolymers; ethylene-vinyl acetate (EVA) copolymers; polyvinyl alcohol; polyvinyl alcohol copolymers; cellulose, including ethyl cellulose, methyl cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose, and carboxymethyl cellulose; polyvinylpyrrolidone; polysaccharides, including starch, modified starch, dextrin, maltodextrin, alginate, and deacetylated chitosan; fats; oils; proteins, including gelatin and corn gluten; gum arabic; shellac; vinylidene chloride and vinylidene chloride copolymers; calcium lignosulfonate; acrylic acid copolymers; polyvinyl acrylate; polyethylene oxide; acrylamide polymers and copolymers; hydroxyethyl polyacrylate, methacrylamide monomers; and polychloroprene.

[0397] A variety of colorants can be used, including organic chromophores classified as nitroso; nitro; azo, including monoazo, diazo, and polyazo; acridine, anthraquinone, azazine, diphenylmethane, indoleamine, indophenol, methine, oxazine, phthalocyanine, thiazine, thiazole, triarylmethane, and xanthracene. Other additives may include micronutrients such as salts of iron, manganese, boron, copper, cobalt, molybdenum, and zinc.

[0398] Polymers or other dust control agents can be applied to leave the treatment on the surface of the plant components.

[0399] In some specific embodiments, in addition to microbial cells or spores, the coating may also comprise an adhesive layer. The adhesive should be non-toxic, biodegradable, and adhesive. Examples of such materials include, but are not limited to, polyvinyl acetate; polyvinyl acetate copolymers; polyvinyl alcohol; polyvinyl alcohol copolymers; cellulose, such as methylcellulose, hydroxymethylcellulose, and hydroxymethylpropylcellulose; dextrin; alginate; sugar; molasses; polyvinylpyrrolidone; polysaccharides; proteins; fats; oils; gum arabic; gelatin; syrups; and starch. Further examples can be found, for example, in U.S. Patent No. 7,213,367, which is incorporated herein by reference.

[0400] Plant component treatment formulations may also contain various additives, such as adhesives, dispersants, surfactants, nutrients, and buffering agents. Other common plant component treatment additives include, but are not limited to, coating agents, wetting agents, buffers, and polysaccharides. At least one agriculturally acceptable carrier, such as water, solid, or dry powder, may be added to the plant component treatment formulation. Dry powders may be derived from a variety of materials, such as calcium carbonate, gypsum, vermiculite, talc, humus, activated carbon, and various phosphorus compounds.

[0401] In some embodiments, the plant component coating composition may comprise at least one filler, which is an organic or inorganic, natural or synthetic component, wherein the active components are combined to facilitate their application to the plant component. In various respects, the filler is an inert solid, such as clay, natural or synthetic silicate, silica, resin, wax, solid fertilizer (e.g., ammonium salt), natural soil minerals (such as kaolin, clay, talc, lime, quartz, palygorskite, montmorillonite, bentonite, or diatomaceous earth) or synthetic minerals (such as silica, alumina, or silicates, especially aluminum silicate or magnesium silicate).

[0402] In some embodiments, the plant component treatment formulation may also contain one or more of the following ingredients: other insecticides, including compounds that act only below ground level; fungicides, such as captan, thiram, metalaxyl, fludioxonil, oxadixyl, and isomers of each of these materials; herbicides, including compounds selected from glyphosate, carbamates, thiocarbamates, acetamides, triazines, dinitroaniline, glyceryl ethers, pyridazinones, uracil, phenoxy compounds, urea, and benzoic acid; and herbicide safeners. Examples of such compounds include benzoxazine, diphenylmethyl derivatives, N,N-diallyl dichloroacetamide, various dihaloacetyl, oxazolyl and thiazolyl compounds, acetone, naphthalenecarboxylic anhydride compounds, and oxime derivatives; chemical fertilizers; biofertilizers; and biocontrol agents, such as other naturally occurring or recombinant bacteria and fungi derived from Rhizobium, Bacillus, Pseudomonas, Serratia, Trichoderma, Glomus, Gliocladium, and mycorrhizal fungi. These components may be added as a separate layer on a plant component or alternatively as part of the plant component coating composition disclosed herein.

[0403] In some embodiments, the formulations used in this disclosure for treating plant components may be in the following forms: suspensions; emulsions; slurries of particles in an aqueous medium (e.g., water); wettable powders; wettable particles (dry and flowable); and dried particles. If formulated as a suspension or slurry, the concentration of the active ingredient in the formulation may be from about 0.5% by weight to about 99% by weight (w / w), or from 5% by weight to 40% by weight, or otherwise formulated by those skilled in the art.

[0404] As mentioned above, other conventional inactive or inert ingredients may be incorporated into the formulation. Such inert ingredients include, but are not limited to: conventional binders; dispersants, such as methylcellulose, which acts as a combined dispersant / binder, for example, in plant component treatments; polyvinyl alcohol; lecithin; polymeric dispersants (e.g., polyvinylpyrrolidone / vinyl acetate); thickeners (e.g., clay thickeners used to increase viscosity and reduce sedimentation of particulate suspensions); emulsion stabilizers; surfactants; antifreeze compounds (e.g., urea); dyes; colorants, etc. Further inert ingredients that may be used in this disclosure can be found in McCutcheon's, Volume 1, “Emulsifiers and Detergents,” MC Publishing Company, GlenRock, NJ, USA, 1996, which is incorporated herein by reference.

[0405] The plant component coating formulations of this disclosure can be applied to plant components by a variety of methods, including but not limited to: mixing in a container (e.g., bottle or bag), mechanical application, tumbling, spraying, and immersion. A variety of active or inert materials can be used to contact the plant components with the microbial composition according to this disclosure.

[0406] In some embodiments, the amount of microorganisms or agricultural composition used to treat the plant component will vary depending on the type of plant component and the type of active ingredient, but the treatment will involve contacting the plant component with an agriculturally effective amount of the composition of the present invention.

[0407] As discussed above, an effective amount means an amount of the composition of the present invention sufficient to affect beneficial or desired results. An effective amount may be applied in one or more applications.

[0408] In some implementations, in addition to the coating layer, the plant components may be treated with one or more of the following: other insecticides, including fungicides and herbicides; herbicide-safe agents; fertilizers and / or biocontrol agents. These components may be added as a separate layer or alternatively added to the coating layer.

[0409] In some embodiments, the plant component coating formulations of this disclosure can be applied to the plant components using a variety of techniques and machines, such as fluidized bed technology, roller mill methods, roller electrostatic plant component processors, and drum coaters. Other methods, such as fountain beds, can also be useful. The plant components can be pre-sized before coating. After coating, the plant components are typically dried and then transferred to a sizing machine for further sizing. Such procedures are known in the art.

[0410] In some embodiments, the plant components treated with microorganisms may also be coated with an external film to protect the coating. Such coatings are known in the art and can be applied using fluidized bed and cylindrical film coating techniques.

[0411] In other embodiments of this disclosure, the composition according to this disclosure can be introduced onto a plant component by using a solid matrix. For example, a certain amount of the composition of the present invention can be mixed with a solid matrix material, and then the plant component can be placed in contact with the solid matrix material for a period of time to allow the composition to be introduced into the plant component. The plant component can then optionally be separated from the solid matrix material and stored or used, or the mixture of the solid matrix material and the plant component can be stored or grown directly. Solid matrix materials that can be used in this disclosure include polyacrylamide, starch, clay, silica, alumina, soil, sand, polyurea, polyacrylate, or any other material capable of absorbing or adsorbing the composition of the present invention for a period of time and releasing the composition into or onto the plant component. It is useful to ensure that the composition of the present invention and the solid matrix material are compatible with each other. For example, the solid matrix material should be selected such that it can release the composition at a reasonable rate (e.g., over a period of time of minutes, hours, or days).

[0412] In some embodiments, this disclosure teaches that individual microorganisms or microbial aggregates or microbial communities developed according to the disclosed methods (including any single microorganism or combination of microorganisms disclosed in Table 1 or Table 1A of this specification, or any microorganisms or combinations thereof from Table 1 or Table 1A and combinations thereof with one or more microorganisms from Table 2) can be combined with any plant biostimulant.

[0413] In some embodiments, this disclosure teaches agricultural compositions comprising one or more commercially available biostimulants, including but not limited to: Diehard TM Diehard TM Fe, Diehard TM Soluble Kelp, Diehard TM Humate SP, Foliar Plus TM Plant Plus TM Accomplish Soil Builder TM Nutri Life, Soil Solution TM Seed Coat TM PercPlus TM Plant Thrust TM , and wait.

[0414] In some embodiments, an additive effect on the plant phenotypic trait of interest is observed when microorganisms or microbial aggregates identified according to the taught methods are combined with active chemical agents. In other embodiments, a synergistic effect on the plant phenotypic trait of interest is observed when microorganisms or microbial aggregates identified according to the taught methods are combined with active chemical agents.

[0415] In some embodiments, when microorganisms or microbial aggregates identified according to the taught methods are combined with fertilizers, an additive effect on the plant phenotypic trait of interest is observed. In other embodiments, when microorganisms or microbial aggregates identified according to the taught methods are combined with fertilizers, a synergistic effect on the plant phenotypic trait of interest is observed.

[0416] In some embodiments, when microorganisms or microbial aggregates identified according to the taught methods are combined with plant growth regulators, an additive effect on the plant phenotypic trait of interest is observed. In some embodiments, a synergistic effect is observed when microorganisms or microbial aggregates identified according to the taught methods are combined with plant growth regulators. In some aspects, the microorganisms of this disclosure are combined with... The study combined and observed synergistic effects on one or more phenotypic traits of interest.

[0417] In some embodiments, an additive effect on the plant phenotypic trait of interest is observed when microorganisms or microbial aggregates identified according to the taught methods are combined with biostimulants. In some embodiments, a synergistic effect is observed when microorganisms or microbial aggregates identified according to the taught methods are combined with biostimulants.

[0418] The synergistic effect obtained through the taught method can be quantified according to the Colby formula (i.e., (E) = X + Y - (X*Y / 100)). See Colby, RS, “Calculating Synergistic and Antagonistic Responses of Herbicide Combinations,” 1967 Weeds, Vol. 15, pp. 20-22, the full text of which is incorporated herein by reference. Therefore, the term “synergistic” refers to the component that, in its presence, increases the expected effect beyond the cumulative amount.

[0419] The isolated microorganisms and aggregates disclosed herein can synergistically enhance the efficacy of agricultural active compounds and agricultural aids.

[0420] In other implementations, synergistic effects were observed when microorganisms or microbial aggregates identified according to the taught methods were combined with fertilizers.

[0421] Furthermore, in some embodiments, this disclosure utilizes synergistic interactions to define microbial aggregates. That is, in some aspects, this disclosure combines certain isolated microbial species that exert synergistic effects into aggregates that confer beneficial traits to plants, or are associated with enhancing beneficial plant traits.

[0422] The agricultural compositions developed according to this disclosure can be formulated with certain adjuvants to enhance the activity of known active agricultural compounds. This has the advantage of reducing the amount of active ingredient in the formulation while maintaining the efficacy of the active compound, thus keeping costs as low as possible and complying with any official regulations. In standalone cases, it is also possible to broaden the scope of action of the active compound, since plants (where treatment with a specific active ingredient without addition is not sufficiently successful) can actually be successfully treated by adding certain adjuvants as well as the disclosed microbial isolates and aggregates. Furthermore, when environmental conditions are unfavorable, the performance of the active substance can be enhanced in standalone cases through suitable formulations.

[0423] Such adjuvants, which can be used in agricultural compositions, are typically adjuvants. Adjuvants are usually in the form of surfactants or salt compounds. Based on their mode of action, they can be broadly classified as regulators, activators, fertilizers, pH buffers, etc. Regulators affect the wetting, adhesion, and spreading properties of formulations. Activators disrupt the waxy surface of plants and increase the penetration of active ingredients into the surface (short-term (minutes) and long-term (hours)). Fertilizers (such as ammonium sulfate, ammonium nitrate, or urea) increase the absorption and solubility of active ingredients and can reduce antagonistic behavior of active ingredients. pH buffers are routinely used to adjust formulations to the optimal pH.

[0424] For further embodiments of the agricultural compositions disclosed herein, see “Chemistry and Technology of Agrochemical Formulations”, edited by D.A. Knowles, copyright 1998, Kluwer Academic Publishers, which is incorporated herein by reference.

[0425] Plant and agricultural benefits

[0426] A wide variety of plants (including those cultivated in agriculture) can benefit from the application of microorganisms (such as those described herein, including single microorganisms, aggregates, and / or compositions produced therefrom, or any of the foregoing). Many different plants, including mosses, lichens, and algae, can be used in the methods of this disclosure. In embodiments, the plants have economic, social, or environmental value. For example, plants may include those used as food crops, fiber crops, oil crops, for forestry, for the pulp and paper industry, as feedstock for biofuel production, and as ornamental plants.

[0427] In other implementations, the plants may be economically, socially, or environmentally undesirable, such as weeds. The following is a list of non-limiting examples of plant types to which the methods of this disclosure can be applied: Food crops Grains, for example Corn, rice, wheat, barley, sorghum, millet, oats, rye, black wheat, and buckwheat; Leafy vegetables, for example Cruciferous vegetables, such as cabbage, broccoli, bok choy, and arugula; salad greens, such as spinach, watercress, and lettuce; Fruits and flowering vegetables, such as Avocados, sweet corn, artichokes; melons, such as zucchini, cucumbers, cantaloupes, squash, and pumpkins; nightshade vegetables / fruits, such as tomatoes, eggplants, and peppers; Legumes, such as Peanuts, peanuts, peas, soybeans, kidney beans, lentils, chickpeas, okra; Bulbous and stem vegetables, For example, asparagus, celery, and alliums ( Allium Crops, such as garlic, onions, and leeks; Roots and tubers Vegetables, such as carrots, beets, bamboo shoots, cassava, yams, ginger, Jerusalem artichokes, parsnip, radishes, potatoes, sweet potatoes, taro, turnips, and wasabi; Sugar crops, including sugar beets ( Beta vulgaris ),sugar cane( Saccharum officinarum ); Crop cultivation for the production of non-alcoholic beverages and stimulants, such as coffee, black tea, herbal tea and green tea, cocoa, and tobacco; Fruit crops Fruits such as true berries (e.g., kiwi, grape, currant, gooseberry, guava, feijoa, pomegranate), citrus fruits (e.g., orange, lemon, lime, grapefruit), superior fruits (e.g., banana, cranberry, blueberry), aggregate fruits (blackberry, raspberry, boysonberry), compound fruits (e.g., pineapple, fig), stone fruits (e.g., apricot, peach, cherry, plum), pome fruits (e.g., apple, pear), and other fruits such as strawberries and sunflower seeds; Cooking and medicinal herbs, such as Rosemary, basil, bay leaf, coriander, mint, dill, St. John's wort ( Hypericum ), foxglove, aloe vera, and rosehip; Spices are produced by crop plants, such as Black pepper, cumin, cinnamon, nutmeg, ginger, cloves, saffron, cardamom, nutmeg skin, red chili pepper, masala, star anise; Planting crops that produce nuts, such as Almonds and walnuts, Brazil nuts, cashews, coconuts, chestnuts, macadamia nuts, pistachios, peanuts, pecans; Crop cultivation for the production of beer, wine and other alcoholic beverages, such as grapes and hops; Oilseed crops, such as Soybeans, peanuts, cotton, olives, sunflowers, sesame, lupins, and brassica crops (e.g., canola / rapeseed); and Edible fungi, such as White mushrooms, shiitake mushrooms, and oyster mushrooms; Plants used in rural agriculture Leguminosae: Clover ( Trifolium ) species, genus Alfalfa ( Medicago ) species and the genus Lobelia ( Lotus Species; white clover (T.repens); Red clover (T. pratense); Caucasian clover （T. ambigum); Earth clover (T. subterraneum); Alfalfa (Medicago sativum); Annual alfalfa; Tribulus terrestris; alfalfa zebrina; red clover (Onobrychis viciifolia); Hundred Veins Root (Lotus corniculatus); Great Hundred Veins Root (Lotus pedunculatus); Seed legumes / dried beans, including peas ( Pisum sativum ),kidney bean( Phaseolus vulgaris ),broad bean( Vicia faba ),green beans( Vigna radiata ),cowpea( Vigna unguiculata ), chickpeas ( Cicer arietum Lupinus (Species of the genus Lupinus) Lupinus species Cereals, including corn / maize ( Zea mays ), sorghum ( Sorghum spp .),Millet( Panicum miliaceum, P. sumatrense ), rice ( Oryza sativa indica, Oryza sativa japonica ),wheat( Triticum aestivum ),barley( Hordeum vulgare ),rye( Secale cereale Black wheat () Triticum X Secale ),oat( Avena sativa ); Forage and woodland grasses: temperate grasses, such as ryegrass ( Lolium ) species; Festuca ( ) Festuca ) species; genus *Gnaphalium* ( Agrostis ) species, perennial ryegrass ( Lolium perenne ); hybrid ryegrass ( Lolium hybridum ); annual ryegrass ( Lolium multiflorum ), tall fescue ( Festuca arundinacea ); cowtail grass ( Festuca pratensis ); Festuca rubra fescue ( ); fescue ( Festuca ovina ); Festuca ryegrass (Festuca ryegrass hybrid ( Lolium X Festuca crosses )); duckgrass ( Dactylis glomerata Kentucky bluegrass ( ); Kentucky bluegrass ( Poa pratensis Kentucky bluegrass (); Poa palustris Kentucky bluegrass in woodland (); Poa nemoralis ); Common Kentucky bluegrass ( Poa trivialis Kentucky bluegrass ( ); Kentucky bluegrass ( Poa compresa ); Broccoli ( Bromus ) species; genus *Gnaphalium* ( Phalaris (Tigris genus) Phleum ) species); Limnipotentifolia ( Arrhenatherum elatius ); genus *Agropyron* ( Agropyron ) species; rough-haired oats ( Avena strigosa Millet (); Setaria italic ); Tropical grasses, such as: *Gnaphalium* ( ) Phalaris ) species; genus *Arm-shaped Grass* ( Brachiaria ) species; genus *Lysimachia* ( Eragrostis ) species; millet ( ) Panicum Species; Bahia grass (Paspalum notatum); genus Paspalum (Brachypodium) ) species; and grasses used for biofuel production, such as switchgrass. (Panicum virgatum) and Miscanthus ( Miscanthus ) species; Fiber crops Cotton, kapok, jute, coconut, sisal, flax (Flame genus) Linum ) species), New Zealand flax (New Zealand hemp genus ( Phormium (Species); planted and natural forest species harvested for paper and engineered wood fiber products, such as coniferous and broadleaf forest species; Tree and shrub species used in planting forestry and biofuel crops Pine (Pinus) Pinus ) species); fir (Dalbergia genus ( Pseudotsuga ) species); spruce (genus spruce ( Picea ) species); cypress (Cypress genus ( Cupressus ) species); Acacia tree (Acacia genus ( Acacia ) species); Alder (Alder genus ( Alnus ) species); oak species (Quercus genus ( Quercus ) species); Sequoia (Giant Sequoia) Sequoiadendron ) species); willow (willow genus ( Salix ) species); birch (Betula genus ( Betula ) species); Cedar (Cedar genus ( Cedurus ) species); Ash (Fraxinus genus ( Fraxinus ) species); larch (Larch genus ( Larix ) species); Eucalyptus genus ( Eucalyptus ) species; bamboo (Bambusae tribe ( Bambuseae ) species) and poplar (poplar genus ( Populus (Species).

[0452] Plants cultivated for conversion into energy, biofuels, or industrial products through extraction, biological, physical, or biochemical processes. Oil-producing plants, such as oil palm, jatropha, soybean, cotton, and flaxseed; latex-producing plants, such as rubber tree (…). Hevea brasiliensis ) and Panama rubber trees ( Castilla elasticaPlants used as direct or indirect feedstocks for the preparation of biofuels (i.e., following chemical, physical (e.g., thermal or catalytic), or biochemical (e.g., enzymatic pretreatment), or biological (e.g., microbial fermentation) conversions during the preparation of biofuels, industrial solvents, or chemical products (e.g., ethanol or butanol, propylene glycol, or other fuels or industrial materials), including sugar crops (e.g., sugar beets, sugarcane), starch-producing crops (e.g., C3 and C4 cereals and tuber crops), cellulosic crops such as trees (e.g., pine, eucalyptus), and grasses and gramineous plants such as bamboo, switchgrass, and miscanthus; crops used in energy, biofuel, or industrial chemical production by gasification and / or microbial or catalytic conversion into biofuels or other industrial feedstocks (e.g., solvents or plastics, producing or not producing biochar), such as biomass crops such as coniferous, eucalyptus, tropical or broadleaf trees, grasses and gramineous plants such as bamboo, switchgrass, miscanthus, sugarcane, or cork such as poplar and willow; and biomass crops used for the preparation of biochar; Crops that produce natural products that can be used in the pharmaceutical, agro-nutritional, and cosmeceutical industries. Crops that produce pharmaceutical precursors or compounds or nutritional food and cosmeceutical compounds and materials, such as star anise (shikimic acid), Japanese knotweed (resveratrol), and kiwifruit (soluble fiber, proteolytic enzymes). Flowering, ornamental, and landscaping plants grown for their aesthetic appeal or environmental characteristics Flowers, such as roses, tulips, and chrysanthemums; Ornamental shrubs, such as boxwood, privet, rose, azalea, and ivy. Beautifying plants, such as sycamore, Mexican orange, mouse thorn, euphorbia, and mosses. Mosses, such as peat moss Planting plants for bioremediation In some respects, the microorganisms of this disclosure are applied to hybrid plants to enhance the beneficial traits of said hybrids. In other respects, the microorganisms of this disclosure are applied to genetically modified plants to enhance the beneficial traits of said GM plants. The microorganisms taught herein can be applied to hybrids and GM plants, and thus can maximize the superior genetic and trait techniques of these plants.

[0463] It should be understood that plants may be provided as seeds, seedlings, cuttings, propagules, or any other plant material or tissue form capable of growth. In one embodiment, seeds may be surface-sterilized with materials such as sodium hypochlorite or mercuric chloride to remove microorganisms from contaminated surfaces. In one embodiment, propagules are grown in pure cultures, such as as sterile seedlings in tissue cultures, before being placed in a plant growth medium.

[0464] Application method

[0465] Microorganisms can be applied to plants, seedlings, cuttings, propagules, etc., and / or growth media containing said plants, using any suitable technique known in the art.

[0466] However, as an example, isolated microorganisms, aggregates, or compositions containing them and / or compositions derived therefrom can be applied to plants, seedlings, cuttings, propagules, etc. by spraying, coating, sprinkling, or any other method known in the art.

[0467] In another embodiment, the isolated microorganisms, aggregates, or compositions containing them can be applied directly to plant seeds prior to sowing.

[0468] In another embodiment, the isolated microorganisms, aggregates, or compositions containing them can be applied directly to plant seeds in the form of seed coating.

[0469] In one embodiment of this disclosure, the isolated microorganisms, aggregates, or compositions containing them are supplied in the form of granules or fillers applied to plant growth media or soil irrigation.

[0470] In other embodiments, the isolated microorganisms, aggregates, or compositions containing them are supplied in the form of foliar application, such as foliar sprays or liquid compositions. Foliar sprays or liquid applications may be applied to growing plants or growth media, such as soil.

[0471] In some embodiments, the isolated microorganisms, aggregates, or compositions comprising them are supplied in forms selected from: soil irrigation agents, foliar sprays, impregnation treatments, furrow treatments, soil conditioners, granules, broadcast treatments, post-harvest disease control treatments, or seed treatments. In some embodiments, the agricultural composition may be applied alone or in a rotary spraying procedure.

[0472] In some embodiments, the isolated microorganisms, aggregates, or compositions containing them are compatible with the tank. In some embodiments, the agricultural composition is compatible with tanks containing other agricultural products. In some embodiments, the agricultural composition is compatible with equipment used for ground, air, and irrigation application.

[0473] In another embodiment, the isolated microorganisms, aggregates, or compositions containing them may be formulated into granules and applied next to the seeds during planting. Alternatively, the granules may be applied after planting. Or, the granules may be applied before planting.

[0474] In some embodiments, isolated microorganisms, aggregates, or compositions containing them are applied to plants or growth media in the form of topical application and / or irrigation to improve crop growth, yield, and quality. Topical application can be carried out using dry mixtures, powders, or dusting compositions, or it can be a liquid-based formulation.

[0475] In embodiments, the isolated microorganisms, aggregates, or compositions containing them may be formulated as: (1) solutions; (2) wettable powders; (3) spreadable powders; (4) soluble powders; (5) emulsions or suspension concentrates; (6) seed dressings or seed coatings; (7) tablets; (8) water-dispersible granules; (9) water-soluble granules (slow-release or immediate-release); (10) microencapsulated granules or suspensions; (11) as irrigation components; and (12) components of fertilizers, pesticides, and other compatibility modifiers, etc. In some respects, the compositions may be diluted in an aqueous medium prior to conventional spray application. The compositions of this disclosure may be applied to soil, plants, seeds, rhizosphere, root sheath, or other areas where application of the microbial composition will be beneficial. Furthermore, shock treatment may be used as a means of introducing endophytic microorganisms.

[0476] The composition is applied to the leaves of plants in various ways. It can be applied to the leaves as an emulsion or suspension concentrate, a liquid solution, or a foliar spray. Application can be carried out in a laboratory, growth chamber, greenhouse, or field.

[0477] In another embodiment, microorganisms can be inoculated into plants by pruning roots or stems and exposing the plant surface to microorganisms by spraying, dipping, or otherwise applying a liquid microbial suspension, gel, or powder.

[0478] In another embodiment, the microorganisms can be injected directly into the leaf or root tissue, or otherwise inoculated directly into or onto the leaf or root pruning site, or into detached plumules, radicles, or coleoptiles. These inoculated plants can then be further exposed to a growth medium containing additional microorganisms; however, this is not necessary.

[0479] In other embodiments, particularly where the microorganisms are unculturable, the microorganisms can be transferred to the plant by any one or a combination of the following methods: grafting, explant insertion, aspiration, electroporation, injury, root pruning, inducing stomatal opening, or any physical, chemical, or biological treatment that provides an opportunity for the microorganisms to enter the plant cells or intercellular spaces. Many alternative techniques are readily apparent to those skilled in the art.

[0480] In one embodiment, the microorganisms infiltrate parts of the plant (such as roots, stems, leaves, and / or reproductive plant parts (becoming endophytes)), and / or grow on the surface of roots, stems, leaves, and / or reproductive plant parts (becoming epiphytes) and / or grow in the plant rhizosphere. In one embodiment, the microorganisms form a symbiotic relationship with the plant.

[0481] aspect

[0482] Some aspects of this disclosure include, but are not limited to:

[0483] Aspect 1: An isolated bacterial strain selected from the group consisting of: *Bacillus tekirae* (strain ID: BCP-80, SEQ ID NO:1) deposited as NRRL accession number B-67810; *Bacillus methyltrophicus* (strain ID: BCP-60, SEQ ID NO:2) deposited as NRRL accession number B-67812; *Bacillus amyloliquefaciens* (strain ID: BCP-69, SEQ ID NO:3) deposited as NRRL accession number B-67815; *Bacillus amyloliquefaciens* (strain ID: BCP-77A, SEQ ID NO:4) deposited as NRRL accession number B-67947; and *Bacillus amyloliquefaciens* (strain ID: BCP-77B, SEQ ID NO:4) deposited as NRRL accession number B-67949. NO:5); Bacillus alginate (strain ID: BCP-68A, SEQ ID NO:6) deposited under NRRL accession number B-67813; Bacillus alginate (strain ID: BCP-68B, SEQ ID NO:7); Bacillus alginate (strain ID: BCP-68C, SEQ ID NO:8); Bacillus alginate (strain ID: BCP-68D, SEQ ID NO:) deposited under NRRL accession number B-67811; *Discocephalus aureus* / *Arthropoda oligosporus* (strain ID: BCP-93, SEQ ID NO:10) deposited under NRRL accession number B-67878; *Bacillus pumilus* (strain ID: BCP-89B, SEQ ID NO:5) deposited under NRRL accession number B-67878. NO:11); and Bacillus spindleii (strain ID: BCP-91, SEQ ID NO:12) deposited as NRRL accession number B-67871, or isolated strains of Bacillus spindleii with substantially similar morphological and physiological characteristics, substantially similar genetic characteristics, their progeny, mutants or gene-edited, altered or modified variants.

[0484] Aspect 2: The bacterial strains isolated according to Aspect 1, wherein the isolated bacterial strains have substantially similar morphological and physiological characteristics to strains selected from the group consisting of: *Bacillus tekirae* (strain ID: BCP-80, SEQ ID NO:1) deposited as NRRL accession number B-67810; *Bacillus methyltrophicus* (strain ID: BCP-60, SEQ ID NO:2) deposited as NRRL accession number B-67812; *Bacillus amyloliquefaciens* (strain ID: BCP-69, SEQ ID NO:3) deposited as NRRL accession number B-67815; *Bacillus amyloliquefaciens* (strain ID: BCP-77A, SEQ ID NO:4) deposited as NRRL accession number B-67947; and *Bacillus amyloliquefaciens* (strain ID: BCP-77B, SEQ ID NO:4) deposited as NRRL accession number B-67949. NO:5); Bacillus alginate (strain ID: BCP-68A, SEQ ID NO:6) deposited under NRRL accession number B-67813; Bacillus alginate (strain ID: BCP-68B, SEQ ID NO:7); Bacillus alginate (strain ID: BCP-68C, SEQ ID NO:8); Bacillus alginate (strain ID: BCP-68D, SEQ ID NO:) deposited under NRRL accession number B-67811; *Discocephalus aureus* / *Arthropoda oligosporus* (strain ID: BCP-93, SEQ ID NO:10) deposited under NRRL accession number B-67878; *Bacillus pumilus* (strain ID: BCP-89B, SEQ ID NO:5) deposited under NRRL accession number B-67878. NO:11); and Bacillus spindleii (strain ID: BCP-91, SEQ ID NO:12) deposited as NRRL accession number B-67871, or isolated strains of Bacillus spindleii with substantially similar morphological and physiological characteristics, substantially similar genetic characteristics, their progeny, mutants or gene-edited, altered or modified variants.

[0485] Aspect 3: The bacterial strains isolated according to Aspect 1, wherein the isolated bacterial strains have genetic characteristics substantially similar to those of strains selected from the group consisting of: *Bacillus tekirae* (strain ID: BCP-80, SEQ ID NO:1) deposited as NRRL accession number B-67810; *Bacillus methyltrophicus* (strain ID: BCP-60, SEQ ID NO:2) deposited as NRRL accession number B-67812; *Bacillus amyloliquefaciens* (strain ID: BCP-69, SEQ ID NO:3) deposited as NRRL accession number B-67815; *Bacillus amyloliquefaciens* (strain ID: BCP-77A, SEQ ID NO:4) deposited as NRRL accession number B-67947; and *Bacillus amyloliquefaciens* (strain ID: BCP-77B, SEQ ID NO:4) deposited as NRRL accession number B-67949. NO:5); Bacillus alginate (strain ID: BCP-68A, SEQ ID NO:6) deposited under NRRL accession number B-67813; Bacillus alginate (strain ID: BCP-68B, SEQ ID NO:7); Bacillus alginate (strain ID: BCP-68C, SEQ ID NO:8); Bacillus alginate (strain ID: BCP-68D, SEQ ID NO:) deposited under NRRL accession number B-67811; *Discocephalus aureus* / *Arthropoda oligosporus* (strain ID: BCP-93, SEQ ID NO:10) deposited under NRRL accession number B-67878; *Bacillus pumilus* (strain ID: BCP-89B, SEQ ID NO:5) deposited under NRRL accession number B-67878. NO:11); and Bacillus spindleii (strain ID: BCP-91, SEQ ID NO:12) deposited as NRRL accession number B-67871, or isolated strains of Bacillus spindleii with substantially similar morphological and physiological characteristics, substantially similar genetic characteristics, their progeny, mutants or gene-edited, altered or modified variants.

[0486] Aspect 4: The bacterial strains isolated according to Aspect 1, wherein the isolated bacterial strains are descendants of strains selected from the group consisting of: *Bacillus tekirae* (strain ID: BCP-80, SEQ ID NO:1) deposited as NRRL accession number B-67810; *Bacillus methyltrophicus* (strain ID: BCP-60, SEQ ID NO:2) deposited as NRRL accession number B-67812; *Bacillus amyloliquefaciens* (strain ID: BCP-69, SEQ ID NO:3) deposited as NRRL accession number B-67815; *Bacillus amyloliquefaciens* (strain ID: BCP-77A, SEQ ID NO:4) deposited as NRRL accession number B-67947; and *Bacillus amyloliquefaciens* (strain ID: BCP-77B, SEQ ID NO:4) deposited as NRRL accession number B-67949. NO:5); Bacillus alginate (strain ID: BCP-68A, SEQ ID NO:6) deposited under NRRL accession number B-67813; Bacillus alginate (strain ID: BCP-68B, SEQ ID NO:7); Bacillus alginate (strain ID: BCP-68C, SEQ ID NO:8); Bacillus alginate (strain ID: BCP-68D, SEQ ID NO:) deposited under NRRL accession number B-67811; *Discocephalus aureus* / *Arthropoda oligosporus* (strain ID: BCP-93, SEQ ID NO:10) deposited under NRRL accession number B-67878; *Bacillus pumilus* (strain ID: BCP-89B, SEQ ID NO:5) deposited under NRRL accession number B-67878. NO:11); and Bacillus spindleii (strain ID: BCP-91, SEQ ID NO:12) deposited as NRRL accession number B-67871, or isolated strains of Bacillus spindleii with substantially similar morphological and physiological characteristics, substantially similar genetic characteristics, their progeny, mutants or gene-edited, altered or modified variants.

[0487] Aspect 5: The bacterial strains isolated according to Aspect 1, wherein the isolated bacterial strains are mutants of strains selected from the group consisting of: *Bacillus tekirae* (strain ID: BCP-80, SEQ ID NO:1) deposited as NRRL accession number B-67810; *Bacillus methyltrophicus* (strain ID: BCP-60, SEQ ID NO:2) deposited as NRRL accession number B-67812; *Bacillus amyloliquefaciens* (strain ID: BCP-69, SEQ ID NO:3) deposited as NRRL accession number B-67815; *Bacillus amyloliquefaciens* (strain ID: BCP-77A, SEQ ID NO:4) deposited as NRRL accession number B-67947; and *Bacillus amyloliquefaciens* (strain ID: BCP-77B, SEQ ID NO:4) deposited as NRRL accession number B-67949. NO:5); Bacillus alginate (strain ID: BCP-68A, SEQ ID NO:6) deposited under NRRL accession number B-67813; Bacillus alginate (strain ID: BCP-68B, SEQ ID NO:7); Bacillus alginate (strain ID: BCP-68C, SEQ ID NO:8); Bacillus alginate (strain ID: BCP-68D, SEQ ID NO:) deposited under NRRL accession number B-67811; Aureobacterium chrysogenum / Oligospora spp. (strain ID: BCP-93, SEQ ID NO:10) deposited under NRRL accession number B-67878; Bacillus pumilus (strain ID: BCP-89B, SEQ ID NO:5) deposited under NRRL accession number B-67878. NO:11); and Bacillus spindleii (strain ID: BCP-91, SEQ ID NO:12) deposited as NRRL accession number B-67871, or isolated strains of Bacillus spindleii with substantially similar morphological and physiological characteristics, substantially similar genetic characteristics, their progeny, mutants or gene-edited, altered or modified variants.

[0488] Aspect 6: The bacterial strains isolated according to Aspect 1, wherein the isolated bacterial strains are gene-edited, altered, or modified variants of strains selected from the group consisting of: *Bacillus tekirae* (strain ID: BCP-80, SEQ ID NO:1) deposited as NRRL accession number B-67810; *Bacillus methyltrophicus* (strain ID: BCP-60, SEQ ID NO:2) deposited as NRRL accession number B-67812; *Bacillus amyloliquefaciens* (strain ID: BCP-69, SEQ ID NO:3) deposited as NRRL accession number B-67815; *Bacillus amyloliquefaciens* (strain ID: BCP-77A, SEQ ID NO:4) deposited as NRRL accession number B-67947; and *Bacillus amyloliquefaciens* (strain ID: BCP-77B, SEQ ID NO:4) deposited as NRRL accession number B-67949. NO:5); Bacillus alginate (strain ID: BCP-68A, SEQ ID NO:6) deposited under NRRL accession number B-67813; Bacillus alginate (strain ID: BCP-68B, SEQ ID NO:7); Bacillus alginate (strain ID: BCP-68C, SEQ ID NO:8); Bacillus alginate (strain ID: BCP-68D, SEQ ID NO:) deposited under NRRL accession number B-67811; *Discocephalus aureus* / *Arthropoda oligosporus* (strain ID: BCP-93, SEQ ID NO:10) deposited under NRRL accession number B-67878; *Bacillus pumilus* (strain ID: BCP-89B, SEQ ID NO:5) deposited under NRRL accession number B-67878. NO:11); and Bacillus spindleii (strain ID: BCP-91, SEQ ID NO:12) deposited as NRRL accession number B-67871, or isolated strains of Bacillus spindleii with substantially similar morphological and physiological characteristics, substantially similar genetic characteristics, their progeny, mutants or gene-edited, altered or modified variants.

[0489] Aspect 7: *Bacillus tekirae* (strain ID: BCP-80, SEQ ID NO:1) deposited under NRRL accession number B-67810; *Bacillus methyltrophicus* (strain ID: BCP-60, SEQ ID NO:2) deposited under NRRL accession number B-67812; *Bacillus amyloliquefaciens* (strain ID: BCP-69, SEQ ID NO:3) deposited under NRRL accession number B-67815; *Bacillus amyloliquefaciens* (strain ID: BCP-77A, SEQ ID NO:4) deposited under NRRL accession number B-67947; *Bacillus amyloliquefaciens* (strain ID: BCP-77B, SEQ ID NO:5) deposited under NRRL accession number B-67949; *Bacillus alginate* (strain ID: BCP-68A, SEQ ID NO:5) deposited under NRRL accession number B-67813. NO:6); Bacillus alginate (strain ID: BCP-68B, SEQ ID NO:7); Bacillus alginate (strain ID: BCP-68C, SEQ ID NO:8); Bacillus alginate deposited under NRRL accession number B-67811 (strain ID: BCP-68D, SEQ ID NO:7); Bacillus aureus / Oligospora spp. deposited under NRRL accession number 67879 (strain ID: BCP-93, SEQ ID NO:10); Bacillus pumilus deposited under NRRL accession number B-67878 (strain ID: BCP-89B, SEQ ID NO:11); and Bacillus fusiformis deposited under NRRL accession number B-67871 (strain ID: BCP-91, SEQ ID NO:7); NO:12) or isolated bacterial strains, their offspring, mutants or gene-edited, altered or modified variants that have substantially similar morphological and physiological characteristics and substantially similar genetic characteristics.

[0490] Aspect 8: An isolated bacterial strain comprising a polynucleotide sequence sharing at least 97% sequence identity with any of SEQ ID NO:1-12.

[0491] Aspect 9: A substantially pure culture of the strain of bacteria isolated according to any one of aspects 1 to 8.

[0492] Aspect 10: Cell-free or inactivated preparations of isolated bacterial strains according to any one of Aspects 1 to 8.

[0493] Aspect 11: Metabolites produced by the bacterial strains isolated according to any one of aspects 1 to 8.

[0494] Aspect 12: An agricultural composition comprising: an isolated strain of microorganisms according to any one of aspects 1 to 8; and an agriculturally acceptable vector.

[0495] Aspect 13: The agricultural composition according to aspect 12, wherein the isolated bacterial strain is present in the composition at a concentration of 1×10^2 to 1×10^12 bacterial cells / gram.

[0496] Aspect 14: The agricultural composition according to aspect 12 or 13, wherein the agricultural composition is formulated as a seed coating, foliar spray, soil irrigation agent, impregnation treatment agent, furrow treatment agent, soil conditioner, granules, broadcast treatment agent, or post-harvest disease control treatment agent.

[0497] Aspect 15: A method for conferring at least one beneficial trait on a plant species, the method comprising: applying an isolated strain of microorganism according to any one of aspects 1 to 8 to the plant species or the growth medium in which the plant species is present.

[0498] Aspect 16: A method for conferring at least one beneficial trait on a plant species, the method comprising: applying an agricultural composition according to any one of aspects 12 to 14 to the plant species or a growth medium in which the plant species is situated.

[0499] Aspect 17: A microbial aggregate comprising at least two microorganisms selected from the group consisting of: A) *Bacillus tekirae*, *Bacillus methyltrophicus*, *Bacillus amyloliquefaciens*, *Bacillus alginate*, *Discocele aureus* / *Oligospora spp.*, *Bacillus pumilus*, and *Bacillus fusiformis*; and B) *Arthrobacter cylindrica*, *Arthrobacter mysore*, *Arthrobacter nicotine*, *Arthrobacter trophoblastus*, *Bacillus megaterium*, *Bacillus subtilis*, *Bacillus thuringiensis*, and *Bacillus thuringiensis*. Bacillus aureus, Bacillus belye, Brugia frostbite, Chlorpyrifos spirochetes, Sacchariformis spp., Bacillus fusiformis, Gyeonggi-do Masseum, Niastrimesteris, Neosphingosine monosporus, Amylolytic Bacillus, Polymyxin-degrading Bacillus, Polymyxin-degrading Bacillus, Pseudomonas fluorescens, Pseudomonas jinjuense, Pseudomonas oryzae, Pseudomonas putidae, Aquatic Laenella, and Perfreeze-expanding Bacillus; at least one of these microorganisms is selected from group A).

[0500] Aspect 18: Microbial aggregates according to aspect 17, wherein at least one of the microorganisms is characterized as having morphological and physiological characteristics substantially similar to those of microorganisms selected from group A).

[0501] Aspect 19: Microbial aggregates according to aspect 17, wherein at least one of the microorganisms is characterized as having genetic characteristics substantially similar to those of microorganisms selected from group A).

[0502] Aspect 20: Microbial aggregates according to aspect 17, wherein at least one of the microorganisms is characterized as a descendant of microorganisms selected from group A).

[0503] Aspect 21: The microbial aggregate according to aspect 17, wherein at least one of the microorganisms is characterized as a mutant of a microorganism selected from group A).

[0504] Aspect 22: Microbial aggregates according to aspect 17, wherein at least one of the microorganisms is characterized as a genetically edited, altered or modified variant of a microorganism selected from group A).

[0505] Aspect 23: A substantially pure culture of microbial aggregates according to any one of aspects 17 to 22.

[0506] Aspect 24: Cell-free or inactivated preparations of microbial aggregates according to any one of Aspects 17 to 22.

[0507] Aspect 25: Metabolites produced by microbial aggregates according to any one of aspects 17 to 22.

[0508] Aspect 26: An agricultural composition comprising: a microbial aggregate according to any one of aspects 17 to 22; and an agriculturally acceptable carrier.

[0509] Aspect 27: The agricultural composition according to aspect 26, wherein the microbial aggregate is present in the composition at a concentration of 1×10^2 to 1×10^12 bacterial cells / gram.

[0510] Aspect 28: The agricultural composition according to aspect 26 or 27, wherein the agricultural composition is formulated as a seed coating, foliar spray, soil irrigation agent, impregnation treatment agent, furrow treatment agent, soil conditioner, granules, broadcast treatment agent, or post-harvest disease control treatment agent.

[0511] Aspect 29: A method for conferring at least one beneficial trait on a plant species, the method comprising: applying a microbial aggregate according to any one of aspects 17 to 22 to the plant species or the growth medium in which the plant species is situated.

[0512] Aspect 30: A method for conferring at least one beneficial trait on a plant species, the method comprising: applying an agricultural composition according to any one of aspects 26 or 27 to the plant species or a growth medium in which the plant species is situated.

[0513] Aspect 31: A microbial aggregate comprising at least two isolated bacterial strains selected from the group consisting of: *Bacillus tekirae* (strain ID: BCP-80, SEQ ID NO:1) deposited as NRRL accession number B-67810; *Bacillus methyltrophicus* (strain ID: BCP-60, SEQ ID NO:2) deposited as NRRL accession number B-67812; *Bacillus amyloliquefaciens* (strain ID: BCP-69, SEQ ID NO:3) deposited as NRRL accession number B-67815; *Bacillus amyloliquefaciens* (strain ID: BCP-77A, SEQ ID NO:4) deposited as NRRL accession number B-67947; and *Bacillus amyloliquefaciens* (strain ID: BCP-77B, SEQ ID NO:4) deposited as NRRL accession number B-67949. NO:5); Bacillus alginate (strain ID: BCP-68A, SEQ ID NO:6) deposited under NRRL accession number B-67813; Bacillus alginate (strain ID: BCP-68B, SEQ ID NO:7); Bacillus alginate (strain ID: BCP-68C, SEQ ID NO:8); Bacillus alginate (strain ID: BCP-68D, SEQ ID NO:) deposited under NRRL accession number B-67811; *Discocephalus aureus* / *Arthropoda oligosporus* (strain ID: BCP-93, SEQ ID NO:10) deposited under NRRL accession number B-67878; *Bacillus pumilus* (strain ID: BCP-89B, SEQ ID NO:5) deposited under NRRL accession number B-67878. NO:11); and *Bacillus fusiformis* (strain ID: BCP-91, SEQ ID NO:12) deposited as NRRL accession number B-67871, or isolated bacterial strains, their progeny, mutants, or gene-edited, altered, or modified variants, and combinations thereof, having substantially similar morphological and physiological characteristics and substantially similar genetic characteristics; and *Arthrobacter cylindrica* (SEQ ID NO:23) deposited as NRRL accession number B-67183; *Arthrobacter cylindrica* (SEQ ID NO:22), *Arthrobacter mysore* (SEQ ID NO:24) deposited as NRRL accession number B-67184; *Arthrobacter nicotinate* (SEQ ID NO:25), *Arthrobacter trophoblast* (SEQ ID NO:25) deposited as NRRL accession number B-67289; NO:26), Bacillus megaterium (SEQ ID NO:27), deposited as NRRL accession number B-67370, and Bacillus megaterium (SEQ ID NO:26).NO:28), Bacillus megaterium (SEQ ID NO:29), Bacillus subtilis (SEQ ID NO:30), Bacillus subtilis (SEQ ID NO:31), Bacillus subtilis (SEQ ID NO:32), Bacillus thuringiensis (SEQ ID NO:33), Bacillus belye (SEQ ID NO:34) deposited under NRRL accession number B-50614, Bacillus frostridae (SEQ ID NO:35) deposited under NRRL accession number B-67360, Cyclopyralid (SEQ ID NO:36) deposited under NRRL accession number B-67236, Cyclopyralid (SEQ ID NO:37) deposited under NRRL accession number B-67197, Sacchariformis (SEQ ID NO:38) deposited under NRRL accession number B-67171, and Sacchariformis (SEQ ID NO:38) deposited under NRRL accession number B-67946. NO:39), *Bacillus fusiformis* (SEQ ID NO:40), *Bacillus nigra* (SEQ ID NO:41) deposited under NRRL accession number B-67198, *Bacillus nigra* (SEQ ID NO:43) deposited under NRRL accession number B-67235, *Bacillus nigra* (SEQ ID NO:44) deposited under NRRL accession number B-67199, *Bacillus nigra* (SEQ ID NO:42), *Neosphingosine monocytogenes* (SEQ ID NO:45) deposited under NRRL accession number B-67945, *Bacillus amyloliquefaciens* (SEQ ID NO:46), *Bacillus polymyxa* (SEQ ID NO:47) deposited under NRRL accession number B-67204, *Bacillus polymyxa* (SEQ ID NO:48), *Pseudomonas fluorescens* (SEQ ID NO:49), *Pseudomonas fluorescens* (SEQ ID NO:50), *Pseudomonas fluorescens* (SEQ ID NO:39), *Pseudomonas fluorescens* (SEQ ID NO:50), *Pseudomonas fluorescens* (SEQ ID NO:49), *Pseudomonas fluorescens* (SEQ ID NO:50), *Pseudomonas fluorescens* (SEQ ID NO:49), *Pseudomonas fluorescens* (SEQ ID NO:42 ... NO:51), *Pseudomonas fluorescens* (SEQ ID NO:52), *Pseudomonas fluorescens* (SEQ ID NO:53), *Pseudomonas jinjuensis* deposited under NRRL accession number B-67207 (SEQ ID NO:54), *Pseudomonas kimoides* deposited under NRRL accession number B-67225 (SEQ ID NO:55), *Pseudomonas kimoides* (SEQ ID NO:56), *Pseudomonas kimoides* (SEQ ID NO:57), *Pseudomonas kimoides* (SEQ ID NO:58), *Pseudomonas putida* (SEQ ID NO:59), *Pseudomonas putida* (SEQ ID NO:60), *Pseudomonas putida* (SEQ ID NO:61), *Pseudomonas putida* (SEQ ID NO:62), *Pseudomonas putida* (SEQ ID NO:62), *Pseudomonas putida* (SEQ ID NO:51), *Pseudomonas putida* (SEQ ID NO:62), *Pseudomonas putida* (SEQ ID NO:53), *Pseudomonas jinjuensis* deposited under NRRL accession number B-67207 (SEQ ID NO:54), *Pseudomonas kimoides* (SEQ ID NO:55), *Pseudomonas putida* (SEQ ID NO:56), *Pseudomonas putida* (SEQ ID NO:57), *Pseudomonas putida* (SEQ ID NO:58), *Pseudomonas putida* (SEQ ID NO:59), *Pseudomonas putida* (SEQ ID NO:60), *Pseudomonas putida* (SEQ ID NO:61), *Pseudomonas putida* (SEQ ID NO:62), *PseuNO:63), *Pseudomonas putida* (SEQ ID NO:64), *Pseudomonas putida* (SEQ ID NO:65), *Pseudomonas putida* (SEQ ID NO:66), *Laenia aquaticis* (SEQ ID NO:67), *Bacillus expanderus* permanently frozen as deposited under NRRL accession number B-67301 (SEQ ID NO:68) and *Bacillus expanderus* permanently frozen as deposited under NRRL accession number B-67302 (SEQ ID NO:69), and combinations thereof, or isolated bacterial strains having substantially similar morphological and physiological characteristics, substantially similar genetic characteristics, their progeny, mutants, or gene-edited, altered, or modified variants; wherein at least one isolated bacterial strain is selected from group A).

[0514] Aspect 32: The microbial aggregate according to aspect 32, wherein the microbial aggregate comprises Bacillus alginate (strain ID: BCP-68A, SEQ ID NO:6), Bacillus alginate (strain ID: BCP-68B, SEQ ID NO:7), Bacillus alginate (strain ID: BCP-68C, SEQ ID NO:8) deposited as NRRL accession number NRLLB-67813 and Bacillus alginate (strain ID: BCP-68D, SEQ ID NO:9) deposited as NRRL accession number B-67811.

[0515] Aspect 33: The microbial aggregate according to aspect 32, wherein the microbial aggregate comprises Bacillus amyloliquefaciens (strain ID: BCP-77A, SEQ ID NO:4) deposited as NRRL accession number B-67947 and Bacillus amyloliquefaciens (strain ID: 77-B, SEQ ID NO:5) deposited as NRRL accession number B-67949.

[0516] Aspect 34: The microbial aggregate according to aspect 32, wherein the microbial aggregate comprises Bacillus pumilus (strain ID: BCP-89B, SEQ ID NO:11) deposited as NRRL accession number B-67878 and Bacillus belesi (SEQ ID NO:34) deposited as NRRL accession number B-50614.

[0517] Aspect 35: The microbial aggregate according to aspect 32, wherein at least one of the isolated bacterial strains is characterized as having morphological and physiological characteristics substantially similar to those of the isolated bacterial strains selected from group A).

[0518] Aspect 36: The microbial aggregate according to aspect 32, wherein at least one of the isolated bacterial strains is characterized as having genetic characteristics substantially similar to those of the isolated bacterial strains selected from group A).

[0519] Aspect 37: The microbial aggregate according to aspect 32, wherein at least one of the isolated bacterial strains is characterized as a descendant of an isolated bacterial strain selected from group A).

[0520] Aspect 38: The microbial aggregate according to aspect 32, wherein at least one of the isolated bacterial strains is characterized as a mutant of an isolated bacterial strain selected from group A).

[0521] Aspect 39: The microbial aggregate according to aspect 32, wherein at least one of the isolated bacterial strains is characterized as a genetically edited, altered or modified variant selected from the isolated bacterial strains of group A).

[0522] Aspect 40: A substantially pure culture of microbial aggregates according to any one of aspects 32 to 39.

[0523] Aspect 41: Cell-free or inactivated preparations of microbial aggregates according to any one of Aspects 32 to 39.

[0524] Aspect 42: Metabolites produced by microbial aggregates according to any one of aspects 32 to 39.

[0525] Aspect 43: An agricultural composition comprising: a microbial aggregate according to any one of aspects 32 to 39; and an agriculturally acceptable carrier.

[0526] Aspect 44: The agricultural composition according to aspect 43, wherein the microbial aggregate is present in the composition at a concentration of 1×10^2 to 1×10^12 bacterial cells / gram.

[0527] Aspect 45: The agricultural composition according to aspect 43 or 44, wherein the agricultural composition is formulated as a seed coating, foliar spray, soil irrigation agent, impregnation treatment agent, furrow treatment agent, soil conditioner, granules, broadcast treatment agent, or post-harvest disease control treatment agent.

[0528] Aspect 46: A method for conferring at least one beneficial trait on a plant species, the method comprising: applying a microbial aggregate according to any one of aspects 32 to 39 to the plant species or the growth medium in which the plant species is situ...

Claims

1. A synthetic composition comprising: a. Microorganisms, exudates derived therefrom, or culture broths derived therefrom, wherein said microorganisms are NRRL Registry No. B-67815; and b. At least one heterologous composition selected from the group consisting of: plant components, formulation components, agricultural compositions, and any combination and / or multiple thereof; The microorganisms are at least 10% of the liquid preparation 2 At least 10 CFU / mL or in non-liquid formulations 2 A concentration of CFU / gram is present.

2. The synthetic composition according to claim 1, wherein the plant component is a seed.

3. The synthetic composition according to claim 2, wherein the seed contains a genetically modified organism.

4. A product comprising the synthetic composition according to claim 1, wherein the synthetic composition is substantially enclosed within an object selected from the group consisting of: tubes, bottles, wide-mouth bottles, ampoules, packaging, utensils, bags, boxes, storage boxes, envelopes, cartons, containers, silos, shipping containers, carriages, and crates.

5. The synthetic composition according to claim 1, wherein the plant component is obtained from fruits, vegetables or row crops.

6. The synthetic composition according to claim 1, wherein the plant component is obtained from plants selected from the group consisting of: corn, soybean, wheat, cotton, cucumber, tomato, pepper, potato, strawberry, orange, lemon, lime, apple, green beans, zucchini, pea, lettuce, broccoli, celery, cauliflower, sorghum, and canola.

7. The synthetic composition according to claim 1, wherein the agricultural composition comprises a growth medium.

8. The synthetic composition according to claim 7, wherein the growth medium comprises soil.

9. A method for cultivating improved plants, the method comprising placing a plurality of synthetic compositions according to claim 1 in the soil in a regular pattern having substantially equal spacing between each of the synthetic compositions.

10. A method for regulating an agronomically important trait in a plant or a component thereof obtained from or derived from the synthetic composition of claim 1, the method comprising placing the synthetic composition in a growth medium; wherein the plant or a component thereof obtained from or derived from the synthetic composition exhibits an improved phenotype compared to a plant or a component thereof not containing the synthetic composition of claim 1.

11. The method of claim 10, wherein the agronomically important trait is selected from the group consisting of: yield, growth, plant health, seedling biomass, normalized difference vegetation index, nitrogen use efficiency, greenness retention, leaf area, nutrient use efficiency, biofilm formation, and rhizosphere competition.

12. The method of claim 10, wherein the microorganism, exudate therefrom, or culture broth therefrom is associated with the plant component by an indirect method selected from the group consisting of: furrow application, soil irrigation application, and lateral application.

13. The method of claim 10, wherein the microorganism, exudate, or culture broth derived therefrom is associated with the plant component by coating the plant component with a liquid formulation of the microorganism, its exudate, or its culture broth.

14. The method of claim 10, wherein the microorganism, exudate, or culture broth derived therefrom is associated with the plant component by coating the plant component with a substantially non-liquid formulation of the microorganism, its exudate, or its culture broth.

15. The method of claim 10, wherein the plant component is a seed, leaf, root, and / or whole plant.

16. The method of claim 10, wherein the plant or a component thereof is a harvested product.

17. The method of claim 11, wherein the harvested product is fruit, vegetable, seed and / or fiber.

18. The method of claim 10, wherein the agronomically important trait is yield.

19. A method for regulating an agronomically important trait in a harvested product, the method comprising introducing a synthetic composition according to claim 1 into the harvested product and / or the plant from which the harvested product is obtained, wherein the agronomically important trait is selected from the group consisting of: increased harvest yield, increased product quality, increased product size, and improved product health.

20. The method of claim 19, wherein the harvested product is fruit.

21. The method of claim 19, wherein the harvested product is vegetables.

22. The method of claim 19, wherein the harvested product is a seed.

23. The method of claim 19, wherein the harvested product is fiber.

24. The synthetic composition of claim 1, further comprising soil, wherein the microorganism is heterologously treated into the soil.

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