# US 2015/0147754 A1 · Herbicide-resistant sunflower plants with multiple herbicide resistant alleles of ahasl1 and methods of use

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- Number: US 2015/0147754 A1
- Document ID: US20150147754A1
- Status: Published application
- Kind: A1
- Application number: 14/087,041
- Filed: 2013-11-22
- Published: 2015-05-28
- Earliest claimed: 2013-11-22
- Assignees: Nidera S.A.; BASF Agrochemical Products B.V.
- Applicants: Nidera S.A.; BASF Agrochemical Products B.V.
- Inventors: Carlos Sala; Mariano Bulos; Sherry R. Whitt; Brigitte J. Weston; Bijay K. Singh; Adriana M. Echarte
- Web page: https://patentgrep.com/patent/US20150147754A1

## CPC

- C12Q 1/6895: for plants, fungi or algae
- C12Q 2600/156: Polymorphic or mutational markers
- C12Q 2600/172: Haplotypes
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

Herbicide resistant sunflower plants comprising two different herbicide-resistant alleles of the sunflower acetohydroxyacid synthase large subunit 1 (AHASL1) gene are described. Methods for making these sunflower plants and methods for controlling weeds or other undesired vegetation growing in the vicinity of these sunflower plants are disclosed. Such methods involve the use of acetohydroxyacid synthase-inhibiting herbicides. Methods for controlling parasitic weeds growing on sunflower plants are also described. Additionally provided are methods for determining the genotype of sunflower plants for AHASL1 gene.

## Claims

### Claim at position 1

1-27. (canceled)

### Claim 28

A method for genotyping sunflower AHASL1, said method comprising the steps of:
(a) obtaining genomic DNA from a sunflower plant;
(b) using said DNA as a template for a first polymerase chain reaction (PCR) amplification comprising said DNA, polymerase, deoxyribonucleotide triphosphates, a forward AHASL1 primer and a reverse wild-type AHASL1 primer, wherein said reverse wild-type AHASL1 primer anneals to a nucleotide sequence comprising the nucleotide sequence set forth in SEQ ID NO:13, wherein the nucleotide that is at the 3′ end nucleotide of said reverse wild-type AHASL1 primer is the complement of the nucleotide that is at position 1 of the nucleotide sequence set forth in SEQ ID NO:13;
(c) using said DNA as a template for a second PCR amplification comprising said DNA, polymerase, deoxyribonucleotide triphosphates, said forward AHASL1 primer and a mutant reverse AHASL1 primer, wherein said reverse mutant AHASL1 primer anneals to a nucleotide sequence comprising the nucleotide sequence set forth in SEQ ID NO: 14, wherein the nucleotide that is at the 3′ end nucleotide of said reverse mutant AHASL1 primer is the complement of the nucleotide that is at position 1 of the nucleotide sequence set forth in SEQ ID NO:14; and
(d) detecting the products of said first and said second PCR amplifications;
wherein said forward AHASL1 primer comprises a nucleotide sequence that corresponds to a region of the sunflower AHASL1 gene that is 5′ of the (ACC)n region.

### Claim 29 (depends on 28)

The method of claim 28, wherein said reverse wild-type AHASL1 primer comprises the nucleotide sequence set forth in SEQ ID NO:4.

### Claim 30 (depends on 28)

The method of claim 28, wherein said reverse mutant AHASL1 primer comprises the nucleotide sequence set forth in SEQ ID NO:5.

### Claim 31 (depends on 28)

The method of claim 28, wherein said forward AHASL1 primer anneals to a nucleotide sequence comprising the complement of the nucleotide sequence set forth in SEQ ID NO:12.

### Claim 32 (depends on 28)

The method of claim 28, wherein said forward AHASL1 primer comprises the nucleotide sequence set forth in SEQ ID NO:3.

### Claim 33

A kit for genotyping sunflower AHASL1, said kit comprising
(a) a forward AHASL1 primer comprising a nucleotide sequence that corresponds to a region of the sunflower AHASL1 gene that is 5′ of the (ACC)n region;
(b) a reverse wild-type AHASL1 primer, wherein said reverse wild-type AHASL1 primer anneals to a nucleotide sequence comprising the nucleotide sequence set forth in SEQ ID NO:13, wherein the nucleotide that is at the 3′ end nucleotide of said reverse wild-type AHASL1 primer is the complement of the nucleotide that is at position 1 of the nucleotide sequence set forth in SEQ ID NO:13; and
(c) a reverse mutant AHASL1 primer, wherein said reverse mutant AHASL1 primer anneals to a nucleotide sequence comprising the nucleotide sequence set forth in SEQ ID NO:14 wherein the nucleotide that is at the 3′ end nucleotide of said reverse mutant AHASL1 primer is the complement of the nucleotide that is at position 1 of the nucleotide sequence set forth in SEQ ID NO:14.

### Claim 34 (depends on 33)

The kit of claim 33, further comprising a polymerase enzyme capable of catalyzing the PCR amplification of a first fragment of a sunflower AHASL1 gene and a second fragment of a sunflower AHASL1 gene, wherein the first fragment is between said annealing site of said forward AHASL1 primer and said annealing site of said reverse wild-type AHASL1 primer in a sunflower AHASL1 gene and the second fragment is between said annealing site of said forward AHASL1 primer and said annealing site of said reverse mutant AHASL1 primer in a sunflower AHASL1 gene.

### Claim 35 (depends on 33)

The kit of claim 33, wherein said forward AHASL1 primer anneals to a nucleotide sequence comprising the complement of the nucleotide sequence set forth in SEQ ID NO:12.

### Claim 36 (depends on 33)

The kit of claim 33, wherein said forward AHASL1 primer comprises the nucleotide sequence set forth in SEQ ID NO:3.

### Claim 37 (depends on 33)

The kit of claim 33, wherein said reverse wild-type AHASL1 primer comprises the nucleotide sequence set forth in SEQ ID NO:4.

### Claim 38 (depends on 33)

The kit of claim 33, wherein said reverse mutant AHASL1 primer comprises the nucleotide sequence set forth in SEQ ID NO:5.