IP Library Granted Patent US 10,499,653
Granted Patent B2
US 10,499,653 · App. 16/024,031 · Granted Dec 10, 2019

Methods of use of seed-origin endophyte populations

Inventors: Geoffrey von Maltzahn (Boston, MA); Richard Bailey Flavell (Thousand Oaks, CA); Gerardo V. Toledo (Belmont, MA); Slavica Djonovic (Malden, MA); Luis Miguel Marquez (Belmont, MA); David Morris Johnston (Cambridge, MA); Yves Alain Millet (Newtonville, MA); Jeffrey Lyford (Hollis, NH); Alexander Naydich (Cambridge, MA); Craig Sadowski (Somerville, MA)
Assignee: Indigo Ag, Inc.
A01N63/00A01C1/06A01N63/02G01N33/5097A01C1/00
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Quick Facts
Patent No.
US 10,499,653
App. No.
16/024,031
Granted
Dec 10, 2019
Kind
B2
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 (34)

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% identical to the 16S nucleic acid sequence of SEQ ID NO: 595, and wherein the benefit is increased nitrogen use efficiency as compared to the plant germinated from the reference seed.

Assignments (5)
RELEASE OF SECURITY INTEREST Recorded Oct 25, 2023
From: CORTLAND CAPITAL MARKET SERVICES LLC
To: INDIGO AG, INC.; INDIGO AGRICULTURE, INC.
Reel/Frame 065344/0780 →
SECURITY INTEREST Recorded Aug 10, 2023
From: INDIGO AGRICULTURE, INC.; INDIGO AG, INC.
To: CORTLAND CAPITAL MARKET SERVICES LLC, AS AGENT
Reel/Frame 064559/0438 →
SECURITY INTEREST Recorded Sep 29, 2020
From: CORTLAND CAPITAL MARKET SERVICES LLC
To: INDIGO AGRICULTURE, INC.; INDIGO AG, INC.
Reel/Frame 053926/0211 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jul 12, 2018
From: VON MALTZAHN, GEOFFREY; FLAVELL, RICHARD; TOLEDO, GERARDO V.; DJONOVIC, SLAVICA; MARQUEZ, LUIS MIGUEL; JOHNSTON, DAVID MORRIS; MILLET, YVES ALAIN; LYFORD, JEFFREY; NAYDICH, ALEXANDER; SADOWSKI, CRAIG
To: SYMBIOTA, INC.
Reel/Frame 046338/0280 →
CHANGE OF NAME Recorded Jul 12, 2018
From: SYMBIOTA, INC.
To: INDIGO AG, INC.
Reel/Frame 046574/0165 →
Continuity (14)
Continuation 15436592 · Feb 17, 2017
Continuation 15145687 · May 3, 2016
Continuation 15017531 · Feb 5, 2016
Continuation 14704891 · May 5, 2015
Continuation 14316469 · Jun 26, 2014
Provisional Application 61935761 · Feb 4, 2014
Provisional Application 61959854 · Sep 4, 2013
Provisional Application 61959870 · Sep 4, 2013
Provisional Application 61959859 · Sep 4, 2013
Provisional Application 61959861 · Sep 4, 2013
Provisional Application 61959847 · Sep 4, 2013
Provisional Application 61959858 · Sep 4, 2013
Provisional Application 61957255 · Jun 26, 2013
Related Publication 20180368419A1 · Dec 27, 2018
Cited By (1)
US 12,490,744