Agricultural endophyte-plant compositions, and methods of use
This application relates to methods and materials for providing a benefit to a seed or seedling of an agricultural plant (e.g., an agricultural grass plant or a Glycine plant), or the agricultural plant derived from the seed or seedling. For example, this application provides purified bacterial populations that include novel seed bacterial endophytes, and synthetic combinations of seeds and/or seedlings (e.g., cereal or soy seeds and/or cereal or soy seedlings) with heterologous seed-derived bacterial endophytes.
1. A method for preparing an agricultural seed preparation, comprising:
a) manually or mechanically inoculating a plurality of Glycine agricultural plant seeds with a formulation comprising a purified bacterial population at a concentration of at least 10 4 CFU/ml in a liquid formulation or at least 10 4 CFU/gm in a non-liquid formulation, the bacterial population comprising a Ochrobactrum bacterial endophyte comprising a 16S nucleic acid sequence comprising SEQ ID NO: 710 and exhibiting at least one of production of an auxin or production of acetoin, wherein the inoculated seeds and/or the Glycine agricultural plant grown therefrom comprises an increased production of a polypeptide of Aspartate aminotransferase glyoxysomal isozyme AAT1 precursor, wherein the Ochrobactrum bacterial endophyte is present in the formulation in an amount capable of providing a benefit to the inoculated Glycine plant seeds or to the Glycine agricultural plants grown from the inoculated Glycine plant seeds compared to reference Glycine seeds or reference Glycine agricultural plants, wherein the formulation further comprises a stabilizer, a fungicide, a preservative, or a combination thereof; and
b) packaging the inoculated seeds in a container.
2. The method of claim 1 , wherein the bacterial endophyte is amplified within a tissue of the Glycine agricultural plant.
3. The method of claim 1 , wherein the inoculated seeds are packaged in the container such that they are substantially stable at about 25 degrees C. for at least 7 days.
4. The method of claim 1 , wherein the inoculating in (a) comprises spraying, immersing, coating, encapsulating, injecting or dusting the seeds with the formulation.
5. The method of claim 1 , wherein the purified bacterial population comprises a plurality of seed bacterial endophyte entities.
6. The method of claim 1 , wherein the inoculating results in a concentration of at least 10 4 CFU of the bacterial endophyte on the surface of the seed.
7. The method of claim 1 , wherein the bacterial population is obtained from an interior seed compartment and/or wherein the bacterial population comprises a seed bacterial endophyte.
8. The method of claim 1 , wherein the bacterial population is obtained from a plant species other than the seeds to which the formulation is contacted.
9. The method of claim 1 , wherein the bacterial population is obtained from a plant species other than the seeds to which the formulation is contacted, and wherein the bacterial population is propagated in culture.
10. The method of claim 1 , wherein the bacterial population is obtained from a plant cultivar or a seed thereof different from the cultivar of the seeds to which the formulation is contacted.
11. The method of claim 1 , wherein the bacterial population is obtained from a surface sterilized seed.
12. The method of claim 1 , wherein the benefit is increased tolerance to drought.
13. A method of making a seed treatment, the method comprising:
(a) inoculating an isolated cultured Ochrobactrum bacterial endophyte cell or spore comprising a 16S nucleic acid sequence comprising SEQ ID NO: 710 into a liquid medium;
(b) culturing the Ochrobactrum bacterial endophyte cell or spore in the medium at a temperature of about 4 to about 37 degrees C. until the cell or spore density in the medium is at least 10 4 CFU/mL; and
(c) collecting the vegetative cells or spores under conditions suitable to produce a seed treatment.
14. A synthetic combination comprising a purified bacterial population in association with a Glycine agricultural plant seed, wherein the purified bacterial population is heterologously disposed on the seed surface and comprises a Ochrobactrum bacterial endophyte comprising a 16S rDNA sequence comprising SEQ ID NO: 710 and capable of producing an auxin or an acetoin, and wherein the bacterial endophyte is present in the synthetic combination in an amount effective to provide a benefit to the seed or the Glycine agricultural plant produced from the synthetic combination compared to a reference plant produced from a reference seed.
15. The synthetic combination of claim 14 , wherein the purified bacterial population comprises a plurality of bacterial endophyte entities.
16. An article of manufacture, comprising: packaging material and a plurality of the synthetic combination of claim 14 .
17. The synthetic combination of claim 14 , further comprising at least one of an adhesion agent, a pesticide, and a fertilizer.
18. The method of claim 12 , wherein the increased tolerance to drought occurs under conditions of biotic stress, optionally wherein the biotic stress is selected from the group consisting of nematode stress, insect herbivory stress, fungal pathogen stress, bacterial pathogen stress, and viral pathogen stress.
19. The synthetic combination of claim 14 , further comprising a purified fungal population.
20. The synthetic combination of claim 14 , wherein the bacterial endophyte colonizes roots of the Glycine agricultural plant derived from the synthetic combination.
21. The synthetic combination of claim 14 , wherein the Glycine seed is a Glycine max seed.
22. The article of manufacture of claim 16 , wherein (a) the plurality of the synthetic combination are packaged in the packaging material such that they are substantially stable at about 25 degrees C. for at least 7 days; (b) the packaging material is selected from a bag, box, bin, envelope, carton, or container; (c) the packaging material comprises a desiccant; and/or (d) the packaging material comprises an anti-fungal agent.