GUARD CELL-SPECIFIC EXPRESSION OF TRANSGENES IN COTTON

• US 20140215655A1
• Filed: 08/09/2012
• Published: 07/31/2014
• Est. Priority Date: 08/12/2011
• Status: Abandoned Application

• Current Assignee: Bayer Cropscience NV (Bayer AG)
• Original Assignee: Bayer Cropscience NV (Bayer AG)
• Inventors: Pien, Stphane, Den Boer, Bart
• Application Number:
  US14/238,669
• Publication Number:
  US 20140215655A1
• Time in Patent Office: Days
• US Class Current: 800/287
• CPC Class Codes: C07K 14/415 from plants
• CPC Class Codes: C12N 15/8225 Leaf-specific, e.g. including petioles, stomata
• CPC Class Codes: C12N 15/8273 for drought, cold, salt resistance

Abstract

In one aspect, the application discloses a cotton plant cell comprising (a) a chimeric gene comprising a first nucleic acid sequence comprising at least 700 consecutive nucleotides of SEQ ID NO: 1 or a nucleic acid sequence having at least 80% sequence identity thereto any of which has stomata-preferential promoter activity; (b) a second nucleic acid sequence encoding an expression product of interest; and (c) a transcription termination and polyadenylation sequence. In addition, the present application discloses a cotton plant, a method of expressing a transgene in cotton under stress conditions, a method of producing a cotton plant, a method of detecting the expression of a transgene under stress conditions and a method for modulating the resistance of a cotton plant to stress as characterized in the claims.

Claims

1. A cotton plant cell comprising a chimeric gene comprising

(a) a first nucleic acid sequence comprising at least 700 consecutive nucleotides of SEQ ID NO:

1 or a nucleic acid sequence having at least 80% sequence identity thereto any of which has stomata preferential promoter activity;

(b) a second nucleic acid sequence encoding an expression product of interest; and

optionally

(c) a transcription termination and polyadenylation sequence.

2. The cotton plant cell of claim 1, wherein said expression product of interest is

(i) a protein or peptide which is optionally involved in mediating stomatal aperture or

(ii) an RNA molecule capable of modulating the expression of a gene comprised in said cotton plant, wherein said gene comprised in said cotton plant is optionally involved in mediating stomatal aperture.

3. The cotton plant cell of claim 2, wherein said protein or said gene optionally involved in mediating stomatal aperture is selected from a carbonic anhydrase, an S-type channel, OST1, HT1, a protein involved in mediating ABA responsiveness.

4. The cotton plant cell of claim 2 or 3, wherein said RNA molecule comprises a first and second RNA region whereinsaid first RNA region comprises a nucleotide sequence of at least 19 consecutive nucleotides having at least about 94% sequence identity to the nucleotide sequence of said gene comprised in said cotton plant;

2. said second RNA region comprises a nucleotide sequence complementary to said 19 consecutive nucleotides of said first RNA region; and

3. said first and second RNA region are capable of base-pairing to form a double stranded RNA molecule between at least said 19 consecutive nucleotides of said first and second region.

5. The cotton plant cell of any one of claims 1 to 4, wherein said first nucleic acid sequence comprises the nucleotide sequence of SEQ ID NO:

1.

6. The cotton plant cell of any one of claims 1 to 5, wherein said first nucleic acid sequence consists of SEQ ID NO:

1.

7. A transgenic cotton plant or seed thereof or a cotton plant part comprising

(a) a chimeric gene comprising

a. a first nucleic acid sequence comprising at least 700 consecutive nucleotides of SEQ ID NO:

1 or a nucleic acid sequence having at least 80% sequence identity thereto any of which has stomata preferential promoter activity;

b. a second nucleic acid sequence encoding an expression product of interest; and

c. a transcription termination and polyadenylation sequence;

or

(b) the cotton plant cell according to any one of claims 1 to 6.

8. A method of effecting stomata-preferential or stomata-specific expression of a product of interest in cotton comprising:

(a1) introducing a chimeric gene comprising a first nucleic acid sequence comprising at least 700 consecutive nucleotides of SEQ ID NO;

1 or a nucleic acid sequence having at least 80% sequence identity thereto any of which confers stomata-preferential or stomata-specific expression of said chimeric gene, a second nucleic acid sequence encoding an expression product of interest, and a transcription termination and polyadenylation sequence into a cotton plant and growing the plant;

OR

(a2) growing the cotton plant of claim 7 or growing a plant from the seed of claim 7.

9. A method of producing a cotton plant or of increasing the yield of a cotton plant comprising:

introducing or introgressing a chimeric gene comprising a first nucleic acid sequence comprising at least 700 consecutive nucleotides of SEQ ID NO;

1 or a nucleic acid sequence having at least 80% sequence identity thereto any of which has stomata preferential promoter activity, a second nucleic acid sequence encoding an expression product of interest, and a transcription termination and polyadenylation sequence;

OR

growing the plant of claim 7 or growing a plant from the seed of claim 7.

10. A method of detecting the expression of a transgene comprising

(a) providing the cotton plant cell of any one of claims 1 to 6 or the plant of claim 7, wherein said expression product of interest is the transgene;

and

(b) detecting the expression of the transgene.

12. The method of any one of claims 8 to 10, wherein said chimeric gene is the chimeric gene described in any one of claims 1 to 6.

13. Use of

(a) the cotton plant or seed of claim 7;

(b) a chimeric gene comprising

a. a first nucleic acid sequence comprising at least 700 consecutive nucleotides of SEQ ID NO;

1 or a nucleic acid sequence having at least 80% sequence identity thereto any of which has stomata preferential promoter activity;

b. a second nucleic acid sequence encoding an expression product of interest; and

optionally

c. a transcription termination and polyadenylation sequence;

or

(c) a nucleic acid sequence comprising at least 700 consecutive nucleotides of SEQ ID NO;

1 or a nucleic acid sequence having at least 80% sequence identity thereto any of which has stomata preferential promoter activity;

for stomata-preferential expression of a transgene in cotton, for modulating the water use efficiency of a cotton plant or for increasing cotton yield.

14. The use for stomata-preferential expression of claim 13, wherein said chimeric gene is the chimeric gene described in any one of claims 1 to 6.

11. A method for modulating the water use efficiency of a cotton plant comprising

introducing into a cotton plant a chimeric gene comprising

a. a first nucleic acid sequence comprising at least 700 consecutive nucleotides of SEQ ID NO:

1 or a nucleic acid sequence having at least 80% sequence identity thereto any of which has stomata preferential promoter activity;

b. a second nucleic acid sequence encoding an expression product of interest which is involved in mediating stomatal aperture; and

c. a transcription termination and polyadenylation sequence;

growing said cotton plant.