Methods of use of seed-origin endophyte populations

• US 9,288,995 B2
• Filed: 05/05/2015
• Issued: 03/22/2016
• 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, Marquez, Luis Miguel, Johnston, David Morris, Millet, Yves Alain, Lyford, Jeffrey, Naydich, Alexander, Sadowski, Craig
• Primary Examiner(s): IBRAHIM, MEDINA AHMED
• Application Number:
  US14/704,891
• Publication Number:
  US 20150320050A1
• Time in Patent Office: 322 Days
• Field of Search: None
• US Class Current: 1/1
• 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 method for increasing tolerance to drought of a cereal agricultural plant, said method comprising:

(a) contacting seed of said plant with a composition comprising a purified bacterial population that comprises a seed bacterial endophyte that is heterologous to the seed and is capable of producing an auxin or acetoin and is capable of increasing the root length of a plant or seedling grown from the seed as compared to a reference plant or seedling grown under the same conditions, said bacterial endophyte has a 16S nucleic acid sequence that is at least 97% identical to a 16S nucleic acid sequence selected from the group consisting of SEQ ID NO;

521, 533, 560, 710, 856, and 1447;

wherein the seed bacterial endophyte is present in the composition in an amount effective to increase the level of at least one protein associated with tolerance to drought within the plant or a seedling grown from the seed, as compared to a reference plant or seedling grown under the same conditions, wherein the protein is a pectinesterase I,

(b) and growing said seed to produce plants colonized with the endophyte bacteria, wherein the plants have increased tolerance to drought stress as compared to a reference plant grown under the same conditions.

3. The method of claim 1, wherein the seed bacterial endophyte is obtainable from an interior seed compartment.

4. The method of claim 1, wherein the seed bacterial endophyte is obtainable from an exterior surface of a seed.

5. The method of claim 1, wherein the seed bacterial endophyte is obtainable from a different cultivar, variety or crop as compared to the seed.

6. The method of claim 1, wherein the seed bacterial endophyte colonizes the roots of the cereal agricultural plant.

7. The method of claim 1, wherein the bacterial population includes two or more bacterial endophytes.

8. The method of claim 1, wherein the bacterial endophyte is obtained from a rice seed.

9. The method of claim 1, wherein the bacterial endophyte is obtained from a maize seed.

10. The method of claim 1, wherein the bacterial endophyte is obtained from a wheat seed.

11. The method of claim 1, wherein the bacterial endophyte is obtained from a barley seed.

2. A method for increasing tolerance to oxidative stress of a cereal agricultural plant, said method comprising

(a) contacting seed of said plant with a composition comprising a purified bacterial population that comprises a seed bacterial endophyte that is heterologous to the seed and is capable of producing an auxin or acetoin and is capable of increasing the root length of a plant or seedling grown from the seed as compared to a reference plant or seedling grown under the same conditions, said bacterial endophyte has a 16S nucleic acid sequence that is at least 97% identical to a 16S nucleic acid sequence selected from the group consisting of SEQ ID NO:

521, 533, 560, 710, 856, and 1447;

wherein the seed bacterial endophyte is present in the composition in an amount effective to increase the level of at least one protein associated with tolerance to oxidative stress within the plant or seedling grown from the seed, as compared to a reference plant or a seedling grown under the same conditions, wherein the protein is a glutathione S-transferase or a peroxidase,

(b) and growing said seed to produce plants colonized with the endophyte bacteria, wherein the plants have increased tolerance to oxidative stress as compared to a reference plant grown under the same conditions.

12. The method of claim 2, wherein the seed bacterial endophyte is obtainable from an interior seed compartment.

13. The method of claim 2, wherein the seed bacterial endophyte is obtainable from an exterior surface of a seed.

14. The method of claim 2, wherein the seed bacterial endophyte is obtainable from a different cultivar, variety or crop as compared to the seed.

15. The method of claim 2, wherein the seed bacterial endophyte colonizes the roots of the cereal agricultural plant.

16. The method of claim 2, wherein the bacterial population includes two or more bacterial endophytes.

17. The method of claim 2, wherein the bacterial endophyte is obtained from a rice seed.

18. The method of claim 2, wherein the bacterial endophyte is obtained from a maize seed.

19. The method of claim 2, wherein the bacterial endophyte is obtained from a wheat seed.

20. The method of claim 2, wherein the bacterial endophyte is obtained from a barley seed.