IP Library Granted Patent US 10,492,497
Granted Patent B2
US 10,492,497 · App. 16/024,320 · Granted Dec 3, 2019

Seed-origin endophyte populations, compositions, and methods of use

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 10,492,497
App. No.
16/024,320
Granted
Dec 3, 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 (35)

1. A synthetic combination comprising a purified bacterial population in association with a seed of a cereal agricultural plant, wherein the purified bacterial population 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 synthetic combination, 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, 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. A population comprising at least 1000 synthetic combinations of claim 1 , wherein the synthetic combinations are disposed within a package and are shelf stable.

3. A population comprising at least 1000 cereal agricultural plants produced by planting a population of synthetic combinations of claim 1 , wherein the plants are grown in a field.

4. The synthetic combination of claim 1 , wherein the bacterial endophyte is obtainable from an interior seed compartment.

5. The synthetic combination of claim 1 , wherein the bacterial endophyte is obtainable from an exterior surface of a seed.

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

7. The synthetic combination of claim 1 , wherein the bacterial endophyte colonizes the roots of a seedling germinated from the seed.

8. The synthetic combination of claim 1 , wherein the bacterial endophyte is obtained from rice seed, maize seed, wheat seed, or barley seed.

9. The synthetic combination of claim 1 , wherein the bacterial population includes two or more bacterial endophytes.

10. The synthetic combination of claim 1 , wherein the synthetic combination further comprises a purified fungal population.

11. The synthetic combination of claim 1 , wherein the benefit occurs under conditions of biotic stress.

12. The synthetic combination of claim 11 , 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.

13. The synthetic combination of claim 1 , wherein the benefit is a larger amount of lateral roots and root-hairs as compared to the plant germinated from the reference seed.

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

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

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

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

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

19. The synthetic combination 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.

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

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

22. The synthetic combination 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.

23. The synthetic combination 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.

24. The synthetic combination 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.

25. The synthetic combination 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.

26. The synthetic combination 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.

27. The synthetic combination 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.

28. The synthetic combination 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 synthetic combination 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 synthetic combination 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 synthetic combination 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 synthetic combination 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 synthetic combination 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 synthetic combination 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.

35. The synthetic combination 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.

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; MARTINEZ, LUIS MIGUEL; JOHNSTON, DAVID MORRIS; MILLET, YVES ALAIN; LYFORD, JEFFREY; NAYDICH, ALEXANDER; SADOWSKI, CRAIG
To: SYMBIOTA, INC.
Reel/Frame 046338/0319 →
CHANGE OF NAME Recorded Jul 12, 2018
From: SYMBIOTA, INC.
To: INDIGO AG, INC.
Reel/Frame 046574/0162 →
Continuity (14)
Continuation 15436609 · 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 20180368421A1 · Dec 27, 2018
Cited By (1)
US 12,490,744