Means and methods for the reduction of photorespiration in crops

Patent metadata

Field	Value
Publication number	US-10017778-B2
Application number	US-201414777963-A
Country	US
Kind code	B2
Filing date	Mar 21, 2014
Priority date	Mar 21, 2013
Publication date	Jul 10, 2018
Grant date	Jul 10, 2018

Abstract

The present invention relates to the field of plant molecular biology and concerns methods for enhancing the abiotic stress tolerance in plants by modulating the expression of the glyoxylate reductase gene. The present invention also provides chimeric constructs useful in the methods in the invention. In addition, the invention provides transgenic plants having an enhanced abiotic stress resistance, in particular an enhanced tolerance to high light conditions and an improved CO 2 fixation.

First claim

The invention claimed is:

1. A plant comprising plant cells comprising a chimeric gene, said chimeric gene comprising the following elements:
i) a plant-expressible promoter,
ii) a DNA region which when transcribed yields a glyoxylate reductase inhibitory RNA molecule; and optionally
iii) a 3′ end region involved in transcription termination and polyadenylation, wherein said plant has suppressed photorespiration and improved CO2 fixation or wherein said plant has improved tolerance to high light stress conditions, when compared to a control plant.

2. The plant according to claim 1 , which is oilseed rape, cotton, corn, rice, wheat, vegetable plants, sugarcane, sugar beets, or soybean.

3. Seeds or propagating material of a plant according to claim 1 , comprising a chimeric gene a plant expressible promoter, a DNA region which when transcribed yields a glyoxylate reductase inhibitory RNA molecule, and optionally a 3′ end region involved in transcription termination and polyadenylation.

4. A method to produce a plant with suppressed photorespiration and improved CO2 fixation or to produce a plant with increased tolerance to high light stress conditions comprising the following steps
a) providing transgenic plant cells with a chimeric gene to create transgenic plant cells, said chimeric gene comprising the following operably linked DNA fragments
1) a plant-expressible promoter;
2) a DNA region, which when transcribed yields a glyoxylate reductase inhibitory RNA molecule; and optionally
3) a 3′ end region involved in transcription termination and polyadenylation.

5. The method of claim 4 , further comprising the further steps of:
b) regenerating a population of transgenic plant lines from said transgenic plant cell; and optionally
c) identifying a plant with suppressed photorespiration and improved CO2 fixation, optionally wherein said selection occurs by growing the population of transgenic plant lines under high light stress conditions.

6. The method of claim 4 , wherein said inhibitory glyoxylate reductase RNA molecule comprises at least 19 nucleotides having at least 90% identity to the nucleotide sequence of a glyoxylate reductase gene present in said plant cells.

7. The method of claim 4 , wherein said inhibitory glyoxylate reductase RNA molecule comprises at least 19 nucleotides having at least 90% identity to the complement of the nucleotide sequence of a glyoxylate reductase gene present in said plant cells.

8. The method of claim 4 , wherein said inhibitory glyoxylate reductase RNA molecule comprises a sense region comprising a nucleotide sequence of at least 19 nucleotides having at least 90% identity to the nucleotide sequence of the glyoxylate reductase gene present in said plant cells and an antisense region comprising a nucleotide sequence of at least 19 nucleotides having at least 90% identity to the complement of the nucleotide sequence of the glyoxylate reductase gene present in said plant cells, wherein said sense and antisense region are capable of forming a double stranded RNA region comprising said at least 19 nucleotides.

9. A method to produce a plant with suppressed photorespiration and improved CO2 fixation or with improved tolerance to stress conditions, comprising the following steps
a) subjecting a plant cell line or a plant to mutagenesis;
b) identifying those plant cells or plants that have a mutation in an endogenous glyoxylate reductase gene resulting in a reduction of the glyoxylate reductase activity;
c) optionally subjecting the identified plant cells or plants to abiotic stress conditions;
d) identifying plant cells or plants with suppressed photorespiration and improved CO2 fixation or with increased tolerance to high light stress conditions.

10. A method according to claim 4 , wherein the plant is oilseed rape, cotton, corn, rice, wheat, vegetable plants, sugarcane, sugar beets, or soybean.

11. The plant according to claim 1 , wherein said DNA region which when transcribed yields a glyoxylate reductase inhibitory RNA molecule comprises:
a. a nucleotide sequence of at least 19 nucleotides having at least 90% identity to a nucleotide sequence encoding a protein comprising the amino acid sequence of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101 or 103;
b. a nucleotide sequence of at least 19 nucleotides having at least 90% identity to the complement of a nucleotide sequence encoding a protein comprising the amino acid sequence of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101 or 103;
c. a nucleotide sequence of at least 19 nucleotides having at least 90% identity to the nucleotide sequence of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100 or 102; or
d. a nucleotide sequence of at least 19 nucleotides having at least 90% identity to the complement of a nucleotide sequence of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100 or 102.

12. The method according to claim 4 , wherein said DNA region which when transcribed yields a glyoxylate reductase inhibitory RNA molecule comprises:
a. a nucleotide sequence of at least 19 nucleotides having at least 90% identity to a nucleotide sequence encoding a protein comprising the amino acid sequence of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101 or 103;
b. a nucleotide sequence of at least 19 nucleotides having at least 90% identity to the complement of a nucleotide sequence encoding a protein comprising the amino acid sequence of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101 or 103;
c. a nucleotide sequence of at least 19 nucleotides having at least 90% identity to the nucleotide sequence of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100 or 102; or
d. a nucleotide sequence of at least 19 nucleotides having at least 90% identity to the complement of a nucleotide sequence of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100 or 102.

Assignees

• Vib Vzw
• Univ Gent

Inventors

• Denecker Jordi
• Hoeberichts Frank
• Muhlenbock Per
• Van Breusegem Frank
• Van Der Kelen Katrien

Classifications

• C12N9/0006

  acting on CH-OH groups as donors (1.1) · CPC title

• C12N15/8271

  for stress resistance, e.g. heavy metal resistance · CPC title

• C12N15/8218

  Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS] · CPC title

• C12N15/01

  Preparation of mutants without inserting foreign genetic material therein; Screening processes therefor · CPC title

• C12N15/8269 Primary

  Photosynthesis · CPC title