Methods of use of seed-origin endophyte populations

Patent metadata

Field	Value
Publication number	US-10499653-B2
Application number	US-201816024031-A
Country	US
Kind code	B2
Filing date	Jun 29, 2018
Priority date	Jun 26, 2013
Publication date	Dec 10, 2019
Grant date	Dec 10, 2019

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.

First claim

What is claimed is:

1. A method of treating seeds of a cereal agricultural plant, comprising mechanically or manually inoculating a plurality of cereal agricultural plant seeds with an agricultural formulation comprising an agriculturally acceptable carrier and a purified bacterial population that comprises a seed bacterial endophyte of the genus Enterobacter that is heterologous to the seed, and has a 16S nucleic acid sequence that is at least 99% identical to a 16S nucleic acid sequence selected from the group consisting of SEQ ID NO: 595, 545, and 548, and is capable of producing an auxin or acetoin, wherein the endophyte is heterologously disposed to the seed in an amount effective to colonize a plant germinated from the inoculated seed, wherein the plant germinated from the inoculated seed has a benefit as compared to a plant germinated from a reference seed sowed under the same conditions, and wherein the benefit is selected from the group consisting of: a larger amount of lateral roots and root-hairs, increased shoot length, increased seedling weight, increased shoot biomass, increased nitrogen use efficiency, and increased root biomass.

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

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

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

5. The method of claim 1 , wherein the bacterial endophyte colonizes the roots of a seedling germinated from the inoculated seed.

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

7. The method of claim 1 , wherein the bacterial endophyte is obtained from rice seed, maize seed, wheat seed, or barley seed.

8. The method of claim 1 , wherein the agricultural formulation further comprises a purified fungal population.

9. The method of claim 1 , wherein the benefit occurs under conditions of biotic stress.

10. The method of claim 9 , 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.

11. The method of claim 1 , wherein the benefit is a larger amount of lateral roots as compared to the plant germinated from the reference seed.

12. The method of claim 1 , wherein the benefit is a larger amount of root-hairs as compared to the plant germinated from the reference seed.

13. The method of claim 1 , wherein the benefit is increased shoot length as compared to the plant germinated from the reference seed.

14. The method of claim 1 , wherein the benefit is increased seedling weight as compared to the plant germinated from the reference seed.

15. The method of claim 1 , wherein the benefit is increased shoot biomass as compared to the plant germinated from the reference seed.

16. The method of claim 1 , wherein the benefit is increased nitrogen use efficiency as compared to the plant germinated from the reference seed.

17. The method of claim 1 , wherein the benefit is increased root biomass as compared to the plant germinated from the reference seed.

18. The method of claim 1 , wherein the seed bacterial endophyte of the genus Enterobacter has a 16S nucleic acid sequence that is 100% identical to the 16S nucleic acid sequence selected from the group consisting of SEQ ID NO: 595, 545 and 548.

19. The method of claim 1 , wherein the effective amount is at least 10 3 CFU/seed on the surface of the seed.

20. The method of claim 1 , wherein the cereal agricultural plant is selected from the group consisting of: maize, rice, barley, wheat, and oats.

21. The method of claim 1 , wherein the seed bacterial endophyte of the genus Enterobacter has a 16S nucleic acid sequence that is at least 99% identical to the 16S nucleic acid sequence of SEQ ID NO: 595.

22. The method of claim 1 , wherein the seed bacterial endophyte of the genus Enterobacter has a 16S nucleic acid sequence that is at least 99% identical to the 16S nucleic acid of SEQ ID NO: 545.

23. The method of claim 1 , wherein the seed bacterial endophyte of the genus Enterobacter has a 16S nucleic acid sequence that is at least 99% identical to the 16S nucleic acid sequence of SEQ ID NO: 548.

24. The method of claim 1 , wherein the seed bacterial endophyte of the genus Enterobacter has a 16S nucleic acid sequence that is 100% identical to the 16S nucleic acid sequence of SEQ ID NO: 595.

25. The method of claim 1 , wherein the seed bacterial endophyte of the genus Enterobacter has a 16S nucleic acid sequence that is 100% identical to the 16S nucleic acid of SEQ ID NO: 545.

26. The method of claim 1 , wherein the seed bacterial endophyte of the genus Enterobacter has a 16S nucleic acid sequence that is 100% identical to the 16S nucleic acid sequence of SEQ ID NO: 548.

27. The method of claim 1 , wherein the seed bacterial endophyte of the genus Enterobacter has a 16S nucleic acid sequence that is 100% identical to the 16S nucleic acid sequence of SEQ ID NO: 595, and wherein the benefit is increased root biomass as compared to the plant germinated from the reference seed.

28. The method of claim 1 , wherein the seed bacterial endophyte of the genus Enterobacter has a 16S nucleic acid sequence that is 100% identical to the 16S nucleic acid sequence of SEQ ID NO: 595, and wherein the benefit is increased shoot length as compared to the plant germinated from the reference seed.

29. The method of claim 1 , wherein the seed bacterial endophyte of the genus Enterobacter has a 16S nucleic acid sequence that is 100% identical to the 16S nucleic acid of SEQ ID NO: 545, and wherein the benefit is increased shoot length as compared to the plant germinated from the reference seed.

30. The method of claim 1 , wherein the seed bacterial endophyte of the genus Enterobacter has a 16S nucleic acid sequence that is 100% identical to the 16S nucleic acid sequence of SEQ ID NO: 548, and wherein the benefit is increased shoot length as compared to the plant germinated from the reference seed.

31. The method of claim 1 , wherein the seed bacterial endophyte of the genus Enterobacter has a 16S nucleic acid sequence that is 100% identical to the 16S nucleic acid sequence of SEQ ID NO: 595, and wherein the benefit is increased seedling weight as compared to the plant germinated from the reference seed.

32. The method of claim 1 , wherein the seed bacterial endophyte of the genus Enterobacter has a 16S nucleic acid sequence that is 100% identical to the 16S nucleic acid of SEQ ID NO: 545, and wherein the benefit is increased seedling weight as compared to the plant germinated from the reference seed.

33. The method of claim 1 , wherein the seed bacterial endophyte of the genus Enterobacter has a 16S nucleic acid sequence that is 100% identical to the 16S nucleic acid sequence of SEQ ID NO: 548, and wherein the benefit is increased seedling weight as compared to the plant germinated from the reference seed.

34. The method of claim 1 , wherein the seed bacterial endophyte of the genus Enterobacter has a 16S nucleic acid sequence that is 100%

Assignees

• Indigo Ag 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

Classifications

• G01N33/5097

  involving plant cells (immunoassays of plant cells G01N33/56961) · CPC title

• A01N25/00

  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 title

• A01C1/06 Primary

  Coating or dressing seed · CPC title

• A01C1/00 Primary

  Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting · CPC title

• A01N63/00 Primary

  Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates (containing compounds of determined constitution A01N27/00 - A01N59/00; unicellular algae A01N65/03) · CPC title