Grass fungal endophytes and uses thereof

• US 8,975,489 B2
• Filed: 11/29/2011
• Issued: 03/10/2015
• Est. Priority Date: 12/02/2010
• Status: Active Grant

• Current Assignee: Noble Research Institute, LLC
• Original Assignee: The Samuel Roberts Noble Foundation
• Inventors: Craven, Kelly
• Primary Examiner(s): Fox, David T
• Assistant Examiner(s): Visone, Lee A
• Application Number:
  US13/306,841
• Publication Number:
  US 20120144533A1
• Time in Patent Office: 1,197 Days
• Field of Search: None
• US Class Current: 800/320
• CPC Class Codes: A01N 63/30 Microbial fungi; Substances produced thereby or obtained therefrom
• CPC Class Codes: A23K 10/10 obtained by microbiological or biochemical processes using chemicals or microorganisms for ensilaging of green fodder A23K30/15
• CPC Class Codes: A23K 10/18 of live microorganisms
• CPC Class Codes: C10L 1/02 essentially based on components consisting of carbon, hydrogen, and oxygen only
• CPC Class Codes: C10L 1/023 for spark ignition
• CPC Class Codes: C12N 1/145 Fungal isolates
• CPC Class Codes: C12P 7/10 substrate containing cellulosic material
• CPC Class Codes: C12R 2001/645 Fungi ; Processes using fungi
• CPC Class Codes: Y02E 50/10 Biofuels, e.g. bio-diesel
• CPC Class Codes: Y02E 50/30 Fuel from waste, e.g. synthetic alcohol or diesel

Abstract

The invention provides isolated fungal endophytes and synthetic combinations thereof with host grass plants. Methods for inoculating grass plant with the endophytes, for propagating the grass-endophyte combinations, and for producing feeds and biofuels from grass-endophyte combinations are also provided.

Claims

1. A synthetic combination of a grass plant and an Acremonium spp. fungal endophyte, wherein the fungal endophyte comprises a rDNA sequence comprising SEQ ID NO:

82, and wherein the fungal endophyte colonizes a root tissue of the grass plant.

2. The synthetic combination of claim 1, wherein the grass plant is an agronomically elite grass plant.

3. The synthetic combination of claim 1, wherein the grass plant is a transgenic grass plant.

4. The synthetic combination of claim 3, wherein the transgenic grass plant comprises a transgene that confers herbicide tolerance, drought resistance, insect resistance, fungus resistance, virus resistance, bacteria resistance, male sterility, cold tolerance, salt tolerance, increased yield, enhanced nutrient use efficiency, increased fermentable carbohydrate content, or reduced lignin content.

5. The synthetic combination of claim 4, wherein the transgenic grass plant comprises a transgene that confers reduced lignin content.

6. The synthetic combination of claim 1, wherein the grass plant is wheat, durum wheat, tall wheatgrass, western wheatgrass, maize, rice, sorghum, meadow fescue, tall fescue, cereal rye, Russian wild rye, oats, bermudagrass, Kentucky bluegrass, big bluestem, little bluestem, Miscanthus sp., Miscanthus×

giganteus, blue grama, black grama, side-oat grama, johnsongrass, buffalograss, creeping bentgrass, switchgrass, or sugarcane.

7. The synthetic combination of claim 6, wherein the grass plant is a switchgrass plant.

8. The synthetic combination of claim 7, wherein the switchgrass plant is an agronomically elite switchgrass plant.

9. The synthetic combination of claim 1, wherein the fungal endophyte colonizes a stem tissue of the grass plant.

10. The synthetic combination of claim 1, wherein the grass plant displays increased biomass or vigor relative to a host grass plant of the same genotype that lacks the endophyte, when grown under the same conditions.

11. The synthetic combination of claim 1, wherein the host grass plant is artificially inoculated with the endophyte.

12. The synthetic combination of claim 1, wherein the endophyte protects the host grass plant from biotic or abiotic stresses.

13. The synthetic combination of claim 12 wherein the abiotic stress is selected from the group consisting of:

water deficiency, nutrient deficiency, heat stress, salt toxicity, aluminum toxicity, heavy metal toxicity, and freezing temperatures.

14. The synthetic combination of claim 12 wherein the biotic stress is selected from the group consisting of:

insect infestation, rust infection, nematode infestation, and herbivore grazing.

15. The synthetic combination of claim 1, wherein the combination is achieved by introduction of the endophyte to the host grass by a method selected from the group consisting of:

inoculation, infection, grafting, and combinations thereof.

16. The synthetic combination of claim 1, wherein the host plant is a forage grass host plant.

17. A commodity product comprising a synthetic combination of a harvested grass plant or part thereof and an Acremonium spp. fungal endophyte comprising a rDNA sequence comprising SEQ ID NO:

82, and wherein the fungal endophyte colonizes a root tissue of the grass plant.

18. The commodity product of claim 17, wherein the product is a biofuel feedstock or an animal feed.

19. A seed comprising a synthetic combination of an Acremonium spp. fungal endophyte and a grass plant embryo, wherein the fungal endophyte comprises a rDNA sequence comprising SEQ ID NO:

82, and wherein the fungal endophyte colonizes a root tissue of the grass plant.

20. A grass seed comprising a seed-coat comprising an Acremonium spp. fungal endophyte comprising a rDNA sequence comprising SEQ ID NO:

82, and wherein the fungal endophyte colonizes a root tissue of the grass plant.

21. A method for propagating a grass plant-fungal endophyte combination, comprising:

a) obtaining a synthetic combination of a grass plant and an Acremonium spp. fungal endophyte comprising a rDNA sequence comprising SEQ ID NO;

82; and

b) vegetatively reproducing the grass plant tissue, wherein the fungal endophyte colonizes a root tissue of the grass plant.

22. A method for cultivating a grass plant comprising:

contacting the host grass plant with an Acremonium spp. fungal endophyte, such that the endophyte colonizes the plant, wherein the fungal endophyte comprises a rDNA sequence comprising SEQ ID NO;

82, and wherein the fungal endophyte colonizes a root tissue of the grass plant.

23. The method of claim 22, wherein the grass plant has enhanced root growth, more tillers, enhanced total biomass, or enhanced seed yield relative to a host grass plant of the same genotype that lacks the endophyte, when grown under the same conditions.

24. The method of claim 22, wherein the grass plant displays tolerance to stress as relative to a host grass plant of the same genotype that lacks the endophyte, when grown under the same conditions.

25. The method of claim 24, wherein said stress is selected from the group consisting of a biotic stress, a pest stress, an insect stress, an abiotic stress, and a water deficit stress.

26. The method of claim 24, wherein the stress is biotic stress caused by at least one organism selected from the group consisting of a mammalian herbivore, a microbial pathogen, and an insect.

27. The method of claim 24, wherein the stress is abiotic stress selected from the group consisting of:

water deficiency, nutrient deficiency, heat stress, salt toxicity, aluminum toxicity, heavy metal toxicity, and freezing temperatures.

28. The method of claim 22, wherein colonization of the grass is achieved by introduction of the fungal endophyte to the grass by a method selected from the group consisting of inoculation, infection, grafting, and combinations thereof.

29. A method for increasing the biomass of a grass plant comprising:

contacting the grass plant with an Acremonium spp. fungal endophyte comprising a rDNA sequence comprising SEQ ID NO;

82, wherein the fungal endophyte colonizes a root tissue of the grass plant, and wherein the plant exhibits increased biomass relative to a grass plant of the same genotype that lacks the endophyte, when grown under the same conditions.

30. A method for producing a biofuel comprising:

obtaining a grass plant comprising an Acremonium spp. fungal endophyte comprising a rDNA sequence comprising SEQ ID NO;

82, wherein the fungal endophyte colonizes a root tissue of the grass plant; and

producing biofuel therefrom.

31. The method of claim 30, wherein the biofuel is ethanol.