SEED-ORIGIN ENDOPHYTE POPULATIONS, COMPOSITIONS, AND METHODS OF USE

• US 20150020239A1
• Filed: 06/26/2014
• Published: 01/15/2015
• Est. Priority Date: 06/26/2013
• Status: Active Grant

• Current Assignee: Indigo Ag, Inc.
• Original Assignee: Symbiota, Inc.
• Inventors: von Maltzahn, Geoffrey, Flavell, Richard Bailey, Toledo, Gerardo V., Djonovic, Slavica, Mrquez, Luis Miguel, Johnston, David Morris, Millet, Yves Alain, Lyford, Jeffrey, Naydich, Alexander, Sadowski, Craig
• Granted Patent: US 9,113,636 B2
• Time in Patent Office: Days
• US Class Current: 800/298
• CPC Class Codes: A01C 1/00 Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
• CPC Class Codes: A01C 1/06 Coating or dressing seed
• CPC Class Codes: A01N 25/00 Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application , e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
• CPC Class Codes: A01N 63/20 Bacteria; Substances produced thereby or obtained therefrom
• CPC Class Codes: A01N 63/30 Microbial fungi; Substances produced thereby or obtained therefrom
• CPC Class Codes: G01N 33/5097 involving plant cells immunoassays of plant cells G01N33/56961

Abstract

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 the agricultural plant derived from the seed or seedling. For example, this application provides purified bacterial populations that include novel seed-origin bacterial endophytes, and synthetic combinations of seeds and/or seedlings (e.g., cereal seeds and/or seedlings) with heterologous seed-derived bacterial endophytes.

Claims

1. A synthetic combination comprising a purified bacterial population in association with a seed or seedling of a cereal agricultural plant, wherein the purified bacterial population comprises a seed-origin bacterial endophyte capable of at least one of:

production of an auxin, nitrogen fixation, production of an antimicrobial, production of a siderophore, mineral phosphate solubilization, production of a cellulase, production of a chitinase, production of a xylanase, and acetoin production, or a combination of two or more thereof, wherein the seed-origin bacterial endophyte is present in the synthetic combination in an amount effective to provide a benefit to the seed or seedling or the cereal agricultural plant derived from the seed or seedling.

2. The synthetic combination of claim 1, wherein the synthetic combination is disposed within a package and is shelf stable.

3. The synthetic combination of claim 1, wherein the purified bacterial population is localized on the surface of the seed or seedling.

4. The synthetic combination of claim 1, wherein the seed-origin bacterial endophyte is present at a concentration of at least 1 CFU/seed on the surface of a seed.

5. The synthetic combination of claim 1, wherein the seed-origin bacterial endophyte is obtained or is obtainable from an interior seed compartment.

6. The synthetic combination of claim 1, wherein the seed-origin bacterial endophyte is obtained or obtainable from the interior seed compartment of a different cultivar, variety or crop as compared to the seed or seedling.

7. The synthetic combination of claim 1, wherein the seed-origin bacterial endophyte is obtained or obtainable from an exterior surface of a different cultivar, variety or crop as compared to the seed or seedling.

8. The synthetic combination of claim 1, wherein the seed-origin bacterial endophyte is heterologous to the microbial population within the seed or seedling.

9. The synthetic combination of claim 1, wherein the benefit is heritable by progeny of the agricultural plant.

10. The synthetic combination of claim 1, wherein the seed-origin bacterial endophyte is capable of at least two of the following:

production of an auxin, nitrogen fixation, production of an antimicrobial, production of a siderophore, mineral phosphate solubilization, production of a cellulase, production of a chitinase, production of a xylanase, and acetoin production.

11. The synthetic combination of claim 1, wherein the benefit comprises one or a plurality of benefits selected from the group consisting of increased root biomass, increased root length, increased height, increased shoot length, increased leaf number, increased water use efficiency, increased overall biomass, increase grain yield, increased photosynthesis rate, increased tolerance to drought, increased heat tolerance, increased salt tolerance, increased resistance to nematode stress, increased resistance to a fungal pathogen, increased resistance to a bacterial pathogen, increased resistance to a viral pathogen, a detectable modulation in the level of a metabolite, and a detectable modulation in the proteome relative to a reference plant.

12. The synthetic combination of claim 11, wherein the benefit comprises at least two benefits, and wherein the synthetic combination comprises two or more seed-origin bacterial endophyte.

13. A synthetic combination of a purified bacterial population in association with a seed or seedling of a cereal agricultural plant, wherein the synthetic combination is produced by the step of contacting the seed or seedling with a formulation comprising a purified bacterial population, wherein the purified bacterial population comprises an effective amount of a seed-origin bacterial endophyte capable of conferring a benefit on the contacted seed or seedling or cereal agricultural plant derived from the seed or seedling, the benefit selected from the group consisting of:

increased root biomass, increased root length, increased height, increased shoot length, increased leaf number, increased water use efficiency, increased overall biomass, increase grain yield, increased photosynthesis rate, increased tolerance to drought, increased heat tolerance, increased salt tolerance, increased resistance to nematode stress, increased resistance to a fungal pathogen, increased resistance to a bacterial pathogen, increased resistance to a viral pathogen, a detectable modulation in the level of a metabolite, and a detectable modulation in the proteome relative to a reference plant, under conditions such that the formulation becomes disposed upon an exterior surface of the seed or seedling in a manner effective for the seed-origin bacterial endophyte to provide the benefit to the seed or seedling or to the cereal agricultural plant derived from the seed or seedling, and wherein the seed-origin bacterial endophyte is capable of host colonization and/or replication within a tissue of the cereal agricultural plant.

14. An agricultural formulation comprising a purified bacterial population consisting essentially of a seed-origin bacterial endophyte capable of conferring on a seed, seedling, or agricultural plant a benefit selected from:

increased tolerance to drought, increased heat tolerance, and increased salt tolerance, wherein the seed-origin bacterial endophyte is present in an amount effective to provide the benefit to a seed or seedling to which the formulation is administered or to an agricultural plant derived from the seed or seedling to which the formulation is administered, and an agriculturally acceptable carrier.

15. The agricultural formulation of claim 14, wherein the seed-origin bacterial endophyte is obtained or obtainable from an interior seed compartment.

16. The agricultural formulation of claim 14, wherein the seed-origin bacterial endophyte is obtained or obtainable from an exterior surface of a seed.

17. The agricultural formulation of claim 14, wherein the seed-origin bacterial endophyte is capable of:

production of an auxin, nitrogen fixation, production of an antimicrobial, production of a siderophore, mineral phosphate solubilization, production of a cellulase, production of a chitinase, production of a xylanase, and acetoin production, or a combination of two or more thereof.

18. The agricultural formulation of claim 14, wherein the seed-origin bacterial endophyte is capable of generating a bacterial network in the agricultural plant derived from the seed or seedling or in the seed or seedling to which the formulation is administered, or in the surrounding environment of the plant, seed, or seedling, and wherein the bacterial network is capable of causing a detectable modulation in the level of a metabolite in the seed, seedling, or plant or a detectable modulation in the proteome of the agricultural plant derived from the seed or seedling.

19. The agricultural formulation of claim 14, wherein the purified bacterial population consists essentially of two seed-origin bacterial endophytes.

20. The agricultural formulation of claim 14, wherein the seed-origin bacterial endophyte comprises a 16S nucleic acid sequence at least 97% identical to a 16S nucleic acid sequence of a bacterial endophyte set forth in Table 1.