# US 10,499,656 B2 · Paenibacillus strain, antifungal compounds, and methods for their use

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- Number: US 10,499,656 B2
- Document ID: US10499656B2
- Status: Granted
- Kind: B2
- Application number: 16/190,556
- Filed: 2018-11-14
- Published: 2019-03-07
- Granted: 2019-12-10
- Earliest claimed: 2015-03-26
- Assignees: Bayer CropScience AG
- Applicants: Bayer CropScience LP
- Inventors: Jeremy Beau; Daniel M. Joo; Patrick Schwientek; Colleen S. Taylor; Bjorn A. Traag
- Web page: https://patentgrep.com/patent/US10499656B2

## CPC

- C07K 11/02: cyclic, e.g. valinomycins {; Derivatives thereof}
- A01N 63/25: Paenibacillus
- C07K 11/00: Depsipeptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K 14/195: from bacteria
- C12N 1/205: Bacterial isolates
- C12N 9/90: Isomerases (5.)
- C12N 9/93: {Ligases (6)}
- C12R 2001/01: Bacteria or Actinomycetales {; using bacteria or Actinomycetales}
- 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)
- 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
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS

## Abstract

The present invention relates to a composition comprising a biologically pure culture of a fungicidal Paenibacillus sp. strain comprising a variant fusaricidin synthetase lacking a functional adenylation domain in the third module. The present invention also provides a composition comprising a biologically pure culture of a fungicidal Paenibacillus sp. strain or a cell-free extract thereof comprising at least one Paeniserine and at least one Paeniprolixin. Also provided are isolated compounds and methods of treating a plant to control a plant disease with the disclosed compositions and compounds.

## Claims

### Claim 1

A composition comprising a biologically pure culture of a fungicidal Paenibacillus sp. strain comprising a variant fusaricidin synthetase lacking a functional adenylation domain in the third module (FusA-A3) or a cell-free extract thereof,
wherein the lack of a functional FusA-A3 inhibits synthesis of fusaricidins with a tyrosine or a phenylalanine at amino acid residue (3) compared to synthesis of fusaricidins by a Paenibacillus sp. strain comprising a wild-type fusaricidin synthetase; and
wherein the variant fusaricidin synthetase comprises a deletion in FusA-A3 of amino acid residues that correspond to positions Asp254, Ala255, Ser258, Thr297, Leu318, Ala320, Gly341, Val349 and Cys350 of SEQ ID NO: 1 or Asp254, Ala255, Ser258, Thr297, Leu318, Ala320, Ala341, Val349 and Cys350 of SEQ ID NO: 9.

### Claim 2 (depends on 1)

The composition of claim 1, comprising a cell-free extract of the biologically pure culture of the fungicidal Paenibacillus sp. strain.

### Claim 3 (depends on 2)

The composition of claim 2, further comprising at least one Paeniserine and at least one Paeniprolixin.

### Claim 4 (depends on 3)

The composition of claim 3, wherein the at least one Paeniserine is selected from the group consisting of Paeniserine A1, Paeniserine A2, Paeniserine A3, Paeniserine A4, Paeniserine B1, Paeniserine B2, Paeniserine B3, Paeniserine B4, Paeniserine C1, Paeniserine C2, and Paeniserine C3.

### Claim 5 (depends on 3)

The composition of claim 3, wherein the at least one Paeniprolixin is selected from the group consisting of Paeniprolixin A1, Paeniprolixin A2, Paeniprolixin B Paeniprolixin B2, Paeniprolixin C1, Paeniprolixin D1, Paeniprolixin E1, Paeniprolixin E2, Paeniprolixin F1, Paeniprolixin F2, Paeniprolixin G1, and Paeniprolixin G2.

### Claim 6 (depends on 1)

The composition of claim 1 comprising a fermentation product of the biologically pure culture of the fungicidal Paenibacillus sp. strain.

### Claim 7 (depends on 6)

The composition of claim 6, wherein the fermentation product is a solid formulation.

### Claim 8 (depends on 6)

The composition of claim 6, wherein the fermentation product is a liquid formulation.

### Claim 9 (depends on 8)

The composition of claim 8 comprising at least about 1×104 CFU of the strain/mL of the liquid formulation.

### Claim 10 (depends on 1)

A method of treating a plant to control a disease, wherein the method comprises applying an effective amount of the composition of claim 1 to the plant, to a part of the plant and/or to a locus of the plant.

### Claim 11 (depends on 10)

The method of claim 10, wherein the composition further comprises at least one Paeniserine and at least one Paeniprolixin.

### Claim 12 (depends on 10)

The method of claim 10, wherein the method comprises applying the composition to foliar plant parts.

### Claim 13 (depends on 10)

The method of claim 10, wherein the composition is applied at about 1×1010 to about 1×1012 colony forming units (CFU) of the fungicidal Paenibacillus sp. strain per hectare.

### Claim 14 (depends on 10)

The method of claim 10, wherein the composition is applied at about 0.5 kg to about 5 kg fermentation solids per hectare.

### Claim 15 (depends on 10)

The method of claim 10, wherein the plant disease is caused by a fungus.

### Claim 16 (depends on 15)

The method of claim 15, wherein the plant disease is mildew or a rust disease.

### Claim 17 (depends on 16)

The method of claim 16, wherein the mildew is powdery mildew or downy mildew.

### Claim 18 (depends on 16)

The method of claim 16, wherein the rust disease is selected from the group consisting of wheat leaf rust, leaf rust of barley, leaf rust of rye, brown leaf rust, crown rust, and stem rust.

### Claim 19 (depends on 15)

The method of claim 15, wherein the fungus is selected from the group consisting of Alternaria alternata, Alternaria solani, Botrytis cinerea, Colletotrichum lagenarium, Fusarium culmorum, Phaeosphaeria nodorum, Zymoseptoria tritici, Phytophthora cryptogea, Phytophthora infestans, Pythium ultimum, Magnaporthe oryzae, Thanatephorus cucumeris, Ustilago segetum var. avenae, Uromyces appendiculatus, and Puccinia triticina.