METHOD FOR AFFECTING PHENOTYPIC ACTIVITY OF ENDOPHYTIC FUNGI

• US 20150368607A1
• Filed: 06/19/2015
• Published: 12/24/2015
• Est. Priority Date: 06/19/2014
• Status: Active Grant

• Current Assignee: Arizona Board of Regents (University of Arizona)
• Original Assignee: Arizona Board of Regents (University of Arizona)
• Inventors: Arnold, Anne Elizabeth, Arendt, Kayla, Baltrus, David A.
• Granted Patent: US 10,000,733 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: C12N 1/14 Fungi culture of mushrooms A01G18/00; as new plants A01H15/00; Culture media therefor
• CPC Class Codes: C12N 1/20 Bacteria; Culture media therefor
• CPC Class Codes: C12Q 1/26 involving oxidoreductase
• CPC Class Codes: C12Q 1/34 involving hydrolase
• CPC Class Codes: G01N 2333/37 from fungi
• CPC Class Codes: G01N 2333/942 acting on beta-1, 4-glucosidic bonds, e.g. cellulase
• CPC Class Codes: G01N 33/56961 Plant cells or fungi

Abstract

Described herein are methods for the alteration and/or transfer of fungal functional traits, e.g., phenotypic activity, via the controlled transfer of endohyphal symbionts, e.g., bacteria, among fungal species. Also described are methods for the identification of endohyphal bacterial symbionts as determinants of cellulase and ligninase activity in fungi, and the use of endohyphal bacterial symbionts to alter the activity, including cellulase and ligninase activities, of the fungi. In particular, the fungi described herein are endophytic fungi, that is, fungi which colonize living, and subsequently senescent, plant tissue.

Claims

1. A method for transferring endohyphal bacterium from a first endophytic fungus to a second endophytic fungus comprising the following steps:

preparing a bacterial inoculum of the endohyphal bacterium from the first endophytic fungus, and inoculating the second endophytic fungus with the bacterial inoculum.

2. The method of claim 1 wherein the first and second endophytic fungi are of same species and strain.

3. The method of claim 1 wherein the first and second endophytic fungi are of different strains of same species.

4. The method of claim 1 wherein the first and second endophytic fungi are of different species.

5. The method of claim 1 wherein the first and second endohyphal bacteria are of same species.

6. The method of claim 1 wherein the first and second endohyphal bacteria are of different strains of same species.

7. The method of claim 1 wherein the first and second endohyphal bacteria are of different species.

8. A method for transferring endohyphal bacteria between a first endophytic fungus and a second endophytic fungus comprising the following steps:

preparing a first bacterial inoculum of the endohyphal bacterium from the first fungus,

preparing a second bacterial inoculum of the endohyphal bacterium from the second fungus, and

cross-inoculating the fungi by inoculating the first fungus with the second bacterial inoculum and inoculating the second fungus with the first bacterial inoculum.

9. A method for examining phenotypic changes in an endophytic fungi of the phylum Ascomyota comprising treating the endophytic fungi with an antibiotic to cure the fungi of its endohyphal bacteria, and comparing phenotypic activity of the fungi before and after treatment with the antibiotic.

10. The method of claim 9 further comprising inoculating the cured fungi with endohyphal bacteria to resynthesize the fungi with endohyphal bacteria symbiont, and comparing phenotypic activity of the cured fungi to the resynthesized fungi.

11. A method for altering enzymatic activity of endophytic fungi comprising transferring endohyphal bacteria from a first endophytic fungus to a second endophytic fungus.

12. The method of claim 11 wherein the enzymatic activity is increased.

13. The method of claim 11 wherein the enzymatic activity is decreased.

14. The method of claim 11 wherein the enzymatic activity is lignocellulytic activity.

15. The method of claim 11 wherein the fungus is Pestalotiopsis and the bacterium is Luteibacter, or wherein the fungus is Microdiplodia and the bacterium is Luteibacter.

16. The method of claim 11 wherein the enzymatic activity is degrading plant material.

17. A method for increasing degradation of plant material comprising increasing enzymatic activity of endophytic fungi in the plant material.