Accelerated directed evolution of microbial consortia for the development of desirable plant phenotypic traits
The disclosure relates to methods for the screening, identification, and/or application of one or more microorganisms of use in imparting one or more beneficial properties to one or more plants.
1. A method for deriving a microbial community, enriched for spore-forming microbes that are associated with imparting a beneficial phenotypic trait to a plant, comprising:
a) growing a plurality of plants in a growth medium, in the presence of a first microbial community;
b) selecting at least one plant from said plurality, following step a), based upon a beneficial phenotypic trait exhibited by the plant relative to other plants of said plurality;
c) acquiring a second microbial community from the at least one plant selected in step b);
d) repeating steps a) to c) in an iterative manner, wherein the second microbial community acquired in step c) is used as an inoculum in step a) of any successive repeat; and
e) acquiring a microbial community enriched for spore-forming microbes that is associated with imparting a beneficial phenotypic trait to a plant,
wherein the first and/or second microbial community is treated with a composition or exposed to a condition that enriches the first and/or second microbial community for selective survival of spore-forming microbes over other microbes.
2. The method according to claim 1 , wherein the composition or condition is deleterious to the survival of microbes that do not form spores.
3. The method according to claim 1 , wherein the first and/or second microbial community is treated with a composition comprising a solvent or sterilant, and wherein the composition is deleterious to the survival of microbes that do not form spores.
4. The method according to claim 1 , wherein the first and/or second microbial community is treated with a composition comprising an organic solvent, and wherein the composition is deleterious to the survival of microbes that do not form spores.
5. The method according to claim 1 , wherein the first and/or second microbial community is treated with composition comprising an alcohol, and wherein the composition is deleterious to the survival of microbes that do not form spores.
6. The method according to claim 1 , wherein the first microbial community of step a) is treated with a composition comprising an alcohol.
7. The method according to claim 1 , wherein the second microbial community of step c) is treated with a composition comprising an alcohol.
8. The method according to claim 1 , wherein the first microbial community of step a) and the second microbial community of step c) are treated with a composition comprising an alcohol.
9. The method according to claim 5 , wherein the composition comprising the alcohol is applied to the growth medium in step a).
10. The method according to claim 5 , wherein the composition comprising the alcohol is applied to the inoculum in step d).
11. The method according to claim 1 , wherein the first and/or second microbial community is treated with a composition comprising ethanol.
12. The method according to claim 1 , wherein the spore-forming microbes are selected from Fungi, Firmicutes, and/or Actinomycetes.
13. The method according to claim 1 , wherein the spore-forming microbes are selected from members of the genus Bacillus.
14. The method according to claim 1 , wherein said first microbial community in step a) is obtained by a microbial capture step, comprising:
growing a plurality of plants in a growth medium, in the presence of a microbial community; and
acquiring said first microbial community from the plurality of plants, or growth medium surrounding the plurality of plants.
15. The method according to claim 1 , wherein said first microbial community in step a) is obtained by a microbial capture step, comprising:
growing a plurality of plants in a growth medium, in the presence of a microbial community; and
acquiring said first microbial community from the plurality of plants, or growth medium surrounding the plurality of plants; and
treating the acquired first microbial community to or exposing the acquired first microbial community to a condition that is selective for the survival of spore-forming microbes and is deleterious to the survival of microbes that do not form spores.
16. The method according to claim 1 , wherein said first microbial community in step a) is obtained by a microbial capture step comprising:
growing a plurality of plants in a growth medium, in the presence of a microbial community, the growth medium is treated with a composition or exposed to a condition that enriches for selective survival of spore-forming microbes over other microbes; and
acquiring said first microbial community from the plurality of plants, or growth medium surrounding the plurality of plants.
17. The method according to claim 1 , wherein said first microbial community in step a) is obtained by a microbial capture step, comprising:
acquiring a first microbial community from a plurality of plants growing in natural soil; and
treating the acquired first microbial community to composition or exposing the acquired first microbial community to a condition that is selective for the survival of spore-forming microbes and is deleterious to the survival of microbes that do not form spores.
18. The method according to claim 1 , further comprising: isolating at least one microbe from the microbial community acquired in step e).
19. The method according to claim 1 , further comprising: isolating at least one microbe from the microbial community acquired in step e) and utilizing a molecular technique to characterize the isolated microbe.
20. The method according to claim 1 , wherein the plurality of plants are members of the Poaceae family.
21. The method according to claim 1 , wherein the plurality of plants are members of a genus selected from the group consisting of: Zea, Sorghum, Panicum, Oryza, Triticum, Hordeum, Secale, Triticale , or Avena.
22. The method according to claim 1 , wherein the plurality of plants are legumes.
23. The method according to claim 1 , wherein the plurality of plants are cotton plants.
24. The method according to claim 1 , wherein the plurality of plants are soybean plants.
25. The method according to claim 1 , wherein the plurality of plants are Zea mays plants.
26. The method according to claim 1 , wherein the second microbial community acquired in step c) is from a root, stem, seed, and/or foliar tissue from the at least one plant.
27. The method according to claim 1 , wherein the second microbial community acquired in step c) is not acquired from a root nodule from the at least one plant.
28. The method according to claim 1 , wherein the second microbial community acquired in step c) is acquired from above-ground tissue from the at least one plant.
29. The method according to claim 1 , wherein the second microbial community acquired in step c) is acquired from below-ground tissue from the at least one plant.
30. The method according to claim 1 , wherein the at least one plant selected in step b) exhibits a superior beneficial phenotypic trait, relative to other plants of said plurality, selected from the group consisting of: growth rate, height, weight, production of metabolite, production of protein, production of carbohydrate, production of oil, leaf size, stem size, general health, survivability, speed of seed germination, quantity of biomass produced, root growth, leaf growth, shoot growth, grain yield, fiber yield, oil yield, seed yield, stress response, and combinations thereof.
31. The method according to claim 1 , wherein said first microbial community in step a) is obtained from a pre-existing microbial collection, database, library, or depository.
32. An accelerated microbial selection method for deriving a microbial community, enriched for heat-tolerant microbes that are associated with imparting a beneficial phenotypic trait to a plant, comprising:
a) growing a plurality of plants in a growth medium, in the presence of a first microbial community;
b) selecting at least one plant from said plurality, following step a), based upon a beneficial phenotypic trait exhibited by the plant relative to other plants of said plurality;
c) acquiring a second microbial community from the at least one plant selected in step b);
d) repeating steps a) to c) in an iterative manner, wherein the second microbial community acquired in step c) is used as an inoculum in step a) of any successive repeat; and
e) acquiring a microbial community enriched for heat-tolerant microbes that is associated with imparting a beneficial phenotypic trait to a plant,
wherein the first and/or second microbial community is pasteurized for selective survival of heat-tolerent microbes over other microbes.
33. The method according to claim 32 , wherein the pasteurization is selective for the survival of heat-tolerant microbes and is deleterious to the survival of microbes that are not heat tolerant.
34. The method according to claim 32 , wherein the pasteurization occurs for between 10 to 20 allowing for the survival of heat-tolerant microbes, but deleterious to the survival of microbes that are not heat-tolerant.
35. The method according to claim 32 , wherein the first microbial community of step a) is pasteurized.
36. The method according to claim 32 , wherein second microbial community of step c) is pasteurized.
37. The method according to claim 32 , wherein the first microbial community of step a) and the second microbial community of step c) are pasteurized.
38. The method according to claim 32 , wherein the growth medium in step a) is pasteurized.
39. The method according to claim 32 , wherein the inoculum in step d) is pasteurized.
40. The method according to claim 32 , wherein the heat-tolerant microbes are members of the genus Bacillus.
41. The method according to claim 32 , wherein said first microbial community in step a) is obtained by a microbial capture step, comprising:
growing a plurality of plants in a growth medium, in the presence of a microbial community; and
acquiring said first microbial community from the plurality of plants, or growth medium surrounding the plurality of plants.
42. The method according to claim 32 , wherein said first microbial community in step a) is obtained by a microbial capture step, comprising:
growing a plurality of plants in a growth medium, in the presence of a microbial community; and
acquiring said first microbial community from the plurality of plants, or growth medium surrounding the plurality of plants; and
pasteurizing the acquired first microbial community, wherein pasteurization is selective for the survival of heat-tolerant microbes and is deleterious to the survival of microbes that are not heat tolerant.
43. The method according to claim 32 , wherein said first microbial community in step a) is obtained by a microbial capture step, comprising:
growing a plurality of plants in a growth medium, in the presence of a microbial community, wherein the growth medium is pasteurized; and
acquiring said first microbial community from the plurality of plants, or growth medium surrounding the plurality of plants;
wherein the pasteurization ensures the selective survival of heat-tolerant microbes.
44. The method according to claim 32 , wherein said first microbial community in step a) is obtained by a microbial capture step, comprising:
acquiring a first microbial community from a plurality of plants growing in natural soil; and
pasteurizing the acquired first microbial community; wherein pasteurization is selective for the survival of heat-tolerant microbes and is deleterious to the survival of microbes that are not heat tolerant.
45. The method according to claim 32 , further comprising: isolating at least one microbe from the microbial community acquired in step e).
46. The method according to claim 32 , further comprising: isolating at least one microbe from the microbial community acquired in step e) and utilizing a molecular technique to characterize the isolated microbe.
47. The method according to claim 32 , wherein the plurality of plants are members of the Poaceae family.
48. The method according to claim 32 , wherein the plurality of plants are members of a genus selected from the group consisting of: Zea, Sorghum, Panicum, Oryza, Triticum, Hordeum, Secale, Triticale , or Avena.
49. The method according to claim 32 , wherein the plurality of plants are legumes.
50. The method according to claim 32 , wherein the plurality of plants are cotton plants.
51. The method according to claim 32 , wherein the plurality of plants are soybean plants.
52. The method according to claim 32 , wherein the plurality of plants are Zea mays plants.
53. The method according to claim 32 , wherein the second microbial community acquired in step c) is from a root, stem, seed, and/or foliar tissue from the at least one plant.
54. The method according to claim 32 , wherein the second microbial community acquired in step c) is not acquired from a root nodule from the at least one plant.
55. The method according to claim 32 , wherein the second microbial community acquired in step c) is acquired from above-ground tissue from the at least one plant.
56. The method according to claim 32 , wherein the second microbial community acquired in step c) is acquired from below-ground tissue from the at least one plant.
57. The method according to claim 33 , wherein the at least one plant selected in step b) exhibits a superior beneficial phenotypic trait, relative to other plants of said plurality, selected from the group consisting of: growth rate, height, weight, production of metabolite, production of protein, production of carbohydrate, production of oil, leaf size, stem size, general health, survivability, speed of seed germination, quantity of biomass produced, root growth, leaf growth, shoot growth, grain yield, fiber yield, oil yield, seed yield, stress response, and combinations thereof.
58. The method according to claim 32 , wherein said first microbial community in step a) is obtained from a pre-existing microbial collection, database, library, or depository.