# US 9,872,502 B2 · Brachiaria-urochloa endophytes

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- Number: US 9,872,502 B2
- Document ID: US9872502B2
- Status: Granted
- Kind: B2
- Application number: 14/127,786
- Filed: 2012-06-01
- Published: 2014-07-17
- Granted: 2018-01-23
- Earliest claimed: 2011-06-20
- Assignees: Agriculture Victoria Services PTY LTD
- Applicants: German Carlos Spangenberg; Kathryn Michaela Guthridge
- Inventors: German Carlos Spangenberg; Kathryn Michaela Guthridge
- Web page: https://patentgrep.com/patent/US9872502B2

## CPC

- A01H 15/00: Fungi; Lichens (fungal microorganisms C12N1/14)
- A01N 43/90: having two or more relevant hetero rings, condensed among themselves or with a common carbocyclic ring system
- A01N 63/30: Microbial fungi; Substances produced thereby or obtained therefrom
- A01N 65/00: Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof (containing compounds of determined constitution A01N27/00 - A01N59/00)
- A01N 65/44: Poaceae or Gramineae [Grass family], e.g. bamboo, lemon grass or citronella grass
- C07K 14/37: from fungi
- C12N 1/145: Fungi isolates
- C12N 9/0008: {acting on the aldehyde or oxo group of donors (1.2)}
- C12P 17/182: {Heterocyclic compounds containing nitrogen atoms as the only ring heteroatoms in the condensed system (alloxazine or isoalloxazine, e.g. riboflavine C12P25/00)}
- C12Y 102/01009: Glyceraldehyde-3-phosphate dehydrogenase (NADP+) (1.2.1.9)
- C12R 2001/645: Fungi {; Processes using fungi}
- A01H: NEW PLANTS OR {NON-TRANSGENIC} PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01N: PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF (preservation of food or foodstuff A23); BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES (preparations for medical, dental or toiletry purposes which kill or prevent the growth or proliferation of unwanted organisms A61K); PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12Y: ENZYMES
- C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS

## Abstract

The present invention relates to fungi of Acremonium spp, wherein said fungi are purified or isolated from plants of the Brachiaria-Urochloa complex and wherein, when said fungi are inoculated into a plant, said plant has improved resistance to diseases and/or pests relative to an uninocualated control plant. The present invention also relates to plants inoculated with such fungi, products produced by the fungi and related genes, proteins and methods.

## Claims

### Claim 1

A method of increasing resistance to pests and/or diseases in a plant, said method comprising the step of inoculating said plant with a fungi of Acremonium spp., wherein, when said fungus is inoculated into the plant, said plant has improved resistance to diseases and/or pests relative to an uninoculated control plant and wherein the internal transcribed spacer (ITS) of said fungi has 99% or greater sequence identity with the internal transcribed spacer of Acremonium 9.2A or Acremonium 3.3A.

### Claim 2 (depends on 1)

The method according to claim 1, wherein said increased resistance to pests and/or diseases is selected from the group consisting of insecticidal activity, insect repellent activity and antifungal activity.

### Claim 3 (depends on 1)

The method according to claim 1, wherein said fungus is purified or isolated from a plant of the Brachiaria-Urochloa species complex.

### Claim 4 (depends on 3)

The method according to claim 3, wherein said plant of the Brachiaria-Urochloa complex is selected from the group consisting of Brachiaria brizantha, Brachiaria decumbens, Brachiaria humidicola and Urochloa mosambicensis.

### Claim 5 (depends on 3)

The method according to claim 3, wherein said fungus is isolated by a method including providing a plurality of samples of fungi;
subjecting said fungi to genetic analysis;
subjecting said fungi to metabolic analysis; and
selecting fungi having a desired genetic and metabolic profile.

### Claim 6 (depends on 5)

The method according to claim 5, wherein said method further includes the step of assessing geographic origin of the fungi and selecting fungi having a desired genetic and metabolic profile and a desired geographic origin.

### Claim 7 (depends on 5)

The method according to claim 5, wherein said genetic analysis includes detecting the presence or absence of polymorphic markers.

### Claim 8 (depends on 3)

The method according to claim 3, wherein said plant of the Brachiaria-Urochloa complex is selected from the group consisting of Brachiaria brizantha, Brachiaria decumbens, Brachiaria humidicola, Brachiaria stolonifera, Brachiaria ruziziensis, Urochloa brizantha, Urochloa decumbens, Urochloa humidicola, Urochloa mosambicensis, Brachiaria marlothii, Brachiaria nigropedata, Urochloa dictyoneura, Urochloa oligotricha, Urochloa panicoides, Brachiaria obtusiflora, Brachiaria serrifolia, Urochloa advena, Urochloa arrecta, Urochloa brachyura, Urochloa eminii, Urochloa mollis, Urochloa xantholeuca, Urochloa oligotricha, Urochloa panicoides, Urochloa plantaginea, Urochloa platynota and Urochloa xantholeuca.

### Claim 9 (depends on 1)

The method according to claim 1, wherein said fungus is selected from the group consisting of Acremonium 9.2A, Acremonium 3.3A and Acremonium 5.1B.

### Claim 10 (depends on 1)

The method of claim 1, wherein the inoculated plant is a grass species.

### Claim 11 (depends on 10)

The method of claim 10, wherein the inoculated plant is a forage, turf or bioenergy grass.

### Claim 12 (depends on 11)

The method of claim 11, wherein the inoculated plant is a member of the Brachiaria-Urochloa species complex.

### Claim 13 (depends on 11)

The method of claim 11, wherein the inoculated plant is a member of the genera Lolium or Festuca.

### Claim 14

A method of increasing resistance to pests and/or diseases in a plant, said method comprising the step of inoculating said plant with a fungi of Acremonium spp., wherein, when said fungus is inoculated into the plant, said plant has improved resistance to diseases and/or pests relative to an uninoculated control plant and wherein the internal transcribed spacer (ITS) of said fungi has 99% or greater sequence identity with the internal transcribed spacer of Acremonium 3.3A.