Microbial fermentation methods and compositions

Patent metadata

Field	Value
Publication number	US-9845462-B2
Application number	US-201514876431-A
Country	US
Kind code	B2
Filing date	Oct 6, 2015
Priority date	Jun 1, 2012
Publication date	Dec 19, 2017
Grant date	Dec 19, 2017

Abstract

The present invention provides methods for the cultivation of the Methylobacterium genus of bacteria. In particular the method provides methods for the efficient and inexpensive cultivation of these bacteria. Additionally, the invention provides methods for the utilization of these bacterial cultures to improve plant agriculture.

First claim

What is claimed is:

1. A plant that is at least partially coated with a composition comprising a fermentation product that comprises: (i) an exogenous solid substance wherein a mono-culture or co-culture of Methylobacterium is adhered thereto by having been grown on said solid substance as an axenic culture that is essentially free of contaminating microorganisms; and (ii) an agriculturally acceptable adjuvant, an agriculturally acceptable excipient, or a combination thereof.

2. The plant of claim 1 , wherein said plant is selected from the group consisting of a corn, Brassica sp., alfalfa, rice, rye, sorghum, pearl millet, proso millet, foxtail millet, finger millet, sunflower, safflower, soybean, tobacco, potato, peanuts, cotton, sweet potato, cassava, coffee, coconut, pineapple, citrus trees, cocoa, tea, banana, avocado, fig, guava, mango, olive, papaya, cashew, macadamia, almond, sugar beet, sugarcane, oat, barley, tomato, lettuce, green bean, lima bean, pea, cucurbit, ornamental, and conifer plant.

3. The plant of claim 1 , wherein said composition further comprises at least one pesticide, at least one bacteriostatic agent, or a combination thereof.

4. The plant of claim 3 , wherein said pesticide is selected from the group consisting of an insecticide, a fungicide, a nematocide, and a bacteriocide, and wherein said pesticide results in no more than a 50% inhibition of Methylobacterium growth when the composition is applied to a plant or plant part in comparison to a composition lacking the pesticide.

5. The plant of claim 1 , wherein the solid substance comprises a plurality of particles with adherent Methylobacterium and the Methylobacterium titer of said particles is at least about 5×10 8 colony-forming units per gram of particles to at least about 5×10 13 colony-forming units of Methylobacterium per gram of particles.

6. The plant of claim 1 , wherein the density of adherent Methylobacterium on said solid substance is at least about 1 Methylobacterium/ 2 square micrometers of the exogenous solid substance surface area.

7. The plant of claim 1 , wherein said Methylobacterium in said mono-culture or in said co-culture can colonize a plant or plant part.

8. The plant of claim 1 , wherein the Methylobacterium in said mono-culture or in said co-culture are isolates from a plant or plant part.

9. The plant of claim 1 , wherein said solid substance selected from the group consisting of a polysaccharide, a diatomaceous earth, a salt crystal, and combinations thereof.

10. The plant of claim 9 , wherein said polysaccharide is selected from the group of a cellulosic polysaccharide, a chitinous polysaccharide, and a galactan polysaccharide.

11. The plant of claim 1 , wherein said solid substance having a mono-culture or co-culture of Methylobacterium adhered thereto is an agriculturally acceptable excipient.

12. The plant of claim 1 , wherein said composition further comprises one or more microorganisms of pre-determined identity other than Methylobacterium.

13. The plant of claim 12 , wherein said one or more microorganisms of pre-determined identity are Bacillus sp., Pseudomonas sp., Coniothyrium sp., Pantoea sp., Streptomyces sp., or Trichoderma sp.

14. A plant part that is at least partially coated with a composition that comprises a fermentation product comprising: (i) an exogenous solid substance wherein a mono-culture or co-culture of Methylobacterium is adhered thereto by having been grown on said solid substance as an axenic culture that is essentially free of contaminating microorganisms; and (ii) an agriculturally acceptable adjuvant, an agriculturally acceptable excipient, or a combination thereof.

15. The plant part of claim 14 , wherein said plant part is from a plant selected from the group consisting of a corn, Brassica sp., alfalfa, rice, rye, sorghum, pearl millet, proso millet, foxtail millet, finger millet, sunflower, safflower, soybean, tobacco, potato, peanuts, cotton, sweet potato, cassava, coffee, coconut, pineapple, citrus trees, cocoa, tea, banana, avocado, fig, guava, mango, olive, papaya, cashew, macadamia, almond, sugar beet, sugarcane, oat, barley, tomato, lettuce, green bean, lima bean, pea, cucurbit, ornamental, and conifer plant.

16. The plant part of claim 14 , wherein said composition further comprises at least one pesticide, at least one bacteriostatic agent, or a combination thereof.

17. The plant part of claim 16 , wherein said pesticide is selected from the group consisting of an insecticide, a fungicide, a nematocide, and a bacteriocide, and wherein said pesticide results in no more than a 50% inhibition of Methylobacterium growth when the composition is applied to a plant or plant part in comparison to a composition lacking the pesticide.

18. The plant part of claim 14 , wherein the solid substance comprises a plurality of particles with adherent Methylobacterium and wherein the Methylobacterium titer of said particles is at least about 5×10 8 colony-forming units per gram of particles to at least about 5×10 13 colony-forming units of Methylobacterium per gram of particles.

19. The plant part of claim 14 , wherein the density of adherent Methylobacterium on said solid substance is at least about 1 Methylobacterium/ 2 square micrometers of the exogenous solid substance surface area.

20. The plant part of claim 14 , wherein said Methylobacterium in said mono-culture or in said co-culture can colonize a plant or plant part.

21. The plant part of claim 14 , wherein said Methylobacterium in said mono-culture or in said co-culture are isolates from a plant or plant part.

22. The plant part of claim 14 , wherein said solid substance is selected from the group consisting of a polysaccharide, a diatomaceous earth, a salt crystal, and combinations thereof.

23. The plant part of claim 22 , wherein said polysaccharide is selected from the group of a cellulosic polysaccharide, a chitinous polysaccharide, and a galactan polysaccharide.

24. The plant part of claim 14 , wherein said solid substance having a mono-culture or co-culture of Methylobacterium adhered thereto is an agriculturally acceptable excipient.

25. The plant part of claim 14 , wherein said composition further comprises one or more microorganisms of pre-determined identity other than Methylobacterium.

26. The plant part of claim 25 , wherein said one or more microorganisms of pre-determined identity are Bacillus sp., Pseudomonas sp., Coniothyrium sp., Pantoea sp., Streptomyces sp., or Trichoderma sp.

27. The plant part of claim 14 , wherein the plant part is a seed.

28. A processed plant product obtained from the plant part of claim 14 , wherein said processed product is a meal, paste, flour, flake, or feed, and contains a detectable amount of said exogenous solid substance having a mono-culture or co-culture of Methylobacterium adhered thereto by having been grown on said solid substance as an axenic culture that is essentially free of contaminating microorganisms, and wherein said exogenous solid substance having a mono-culture or co-culture of Methylobacterium adhered thereto is detectable in comparison to a processed product of an untreated plant part wherein the exogenous solid substance is not detectable.

Assignees

• Newleaf Symbiotics Inc

Inventors

• Bogosian Gregg

Classifications

• C12N11/02

  Enzymes or microbial cells immobilised on or in an organic carrier · CPC title

• A01H3/00

  Processes for modifying phenotypes {, e.g. symbiosis with bacteria} (A01H4/00 takes precedence) · CPC title

• C12N1/20 Primary

  Bacteria; Culture media therefor · CPC title

• C12N11/14

  Enzymes or microbial cells immobilised on or in an inorganic carrier · CPC title

• A01C1/06

  Coating or dressing seed · CPC title