Isolated polynucleotides and polypeptides, and methods of using same for increasing nitrogen use efficiency, yield, growth rate, vigor, biomass, oil content, and/or abiotic stress tolerance

Patent metadata

Field	Value
Publication number	US-10457954-B2
Application number	US-201113819777-A
Country	US
Kind code	B2
Filing date	Aug 23, 2011
Priority date	Aug 30, 2010
Publication date	Oct 29, 2019
Grant date	Oct 29, 2019

Abstract

Provided are isolated polynucleotides and nucleic acid constructs which comprise a nucleic acid sequence at least 80% identical to a nucleic acid sequence selected form the group consisting of SEQ ID NOs: 277, 1-276, 278-469 and 785-2397; and isolated polypeptides which comprise an amino acid sequence at least 80% homologous to an amino acid sequence selected from the group consisting of SEQ ID NOs: 482, 470-481, 483-784 and 2398-3818. Also provided are transgenic cells and plants expressing same and methods of using same for increasing nitrogen use efficiency, yield, biomass, growth rate, vigor, oil content, fiber yield, fiber quality, and/or abiotic stress tolerance of a plant.

First claim

What is claimed is:

1. A method of increasing harvest index, leaf blade area, plot coverage, and/or rosette area of a plant, comprising:
(a) over-expressing within the plant an exogenous polynucleotide comprising a nucleic acid sequence encoding the polypeptide selected from the group consisting of SEQ ID NOs: 656, 3402, 3403, 3404 and 3405, wherein said exogenous polynucleotide is operably linked to a heterologous constitutive promoter, and
(b) selecting the plant resulting from step (a), said plant overexpressing said exogenous polynucleotide, under nitrogen-limiting conditions which comprise between 0.75-3 millimolar (m M) nitrogen, for an increased harvest index, leaf blade area, plot coverage, and/or rosette area as compared to a non-transformed plant which is grown under the same growth conditions,
(c) isolating the plant or a regenerable portion of said plant selected according to step (b) having said increased harvest index, leaf blade area, plot coverage, and/or rosette area under said nitrogen-limiting conditions so as to obtain an isolated plant or a regenerable portion of said selected plant, and
(d) planting said isolated plant obtained in step (c) or regenerating a plant from said regenerable portion obtained in step (c) to thereby obtain a plant characterized by said increased harvest index, leaf blade area, plot coverage, and/or rosette area under said nitrogen-limiting conditions as compared to said non-transformed plant which is grown under the same growth conditions,
thereby increasing the harvest index, leaf blade area, plot coverage, and/or rosette area of the plant.

2. The method of claim 1 , wherein said exogenous polynucleotide comprises the nucleic acid sequence selected from the group consisting of SEQ ID NOs: 443, 187, 1935, 1936, 1937, 1938, or a codon optimized sequence thereof.

3. The method of claim 1 , wherein said exogenous polynucleotide comprises the nucleic acid sequence selected from the group consisting of SEQ ID NOs: 443, 187, 1935-1938.

4. A method of increasing harvest index, leaf blade area, plot coverage, and/or rosette area of a plant, comprising:
(a) over-expressing within the plant an exogenous polynucleotide comprising a nucleic acid sequence encoding the polypeptide sequence set forth by SEQ ID NO: 656 or an orthologue polypeptide having at least 98% sequence identity to SEQ ID NO: 656, wherein said exogenous polynucleotide is operably linked to a heterologous constitutive promoter, and
(b) selecting the plant resulting from step (a), said plant overexpressing said exogenous polynucleotide, under nitrogen-limiting conditions which comprise between 0.75-3 millimolar (m M) nitrogen, for an increased harvest index, leaf blade area, plot coverage, and/or rosette area as compared to a non-transformed plant which is grown under the same growth conditions,
(c) isolating the plant or a regenerable portion of said plant selected according to step (b) having said increased harvest index, leaf blade area, plot coverage, and/or rosette area under said nitrogen-limiting conditions so as to obtain an isolated plant or a regenerable portion of said selected plant, and
(d) planting said isolated plant obtained in step (c) or regenerating a plant from said regenerable portion obtained in step (c) to thereby obtain a plant characterized by said increased harvest index, leaf blade area, plot coverage, and/or rosette area under said nitrogen-limiting conditions as compared to said non-transformed plant which is grown under the same growth conditions,
thereby increasing the harvest index, leaf blade area, plot coverage, and/or rosette area of the plant.

5. A method at generating a plant having increased tolerance to nitrogen limiting conditions, comprising:
(a) over-expressing within a plant an exogenous polynucleotide comprising a nucleic acid sequence encoding the polypeptide sequence set forth by SEQ ID NO: 656, or an analogue polypeptide having at least 96% sequence identity to the polypeptide sequence set forth by SEQ ID NO:656 wherein said exogenous polynucleotide is operably linked to a heterologous constitutive promoter, wherein said orthologue polypeptide is capable of increasing harvest index, leaf blade area, plot coverage, and/or rosette area of a plant under nitrogen limiting conditions which comprise between 0.75-3 mM nitrogen, and
(b) selecting plant resulting from step (a), said plant overexpressing said exogenous polynucleotide, under said nitrogen-limiting conditions which comprise between 0.75-3 mM nitrogen, for an increased harvest index, leaf blade area, plot coverage, and/or rosette area as compared to a non-transformed plant which is grown under the same growth conditions,
(c) isolating the plant or regenerable portion of said plant selected according to step (b) having said increased harvest index, leaf blade area, plot coverage, and/or rosette area under said nitrogen-limiting conditions so as to obtain an isolated plant or a regenerable portion of said selected plant, and
(d) planting said isolated plant obtained in step (c) or regenerating a plant from said regenerable portion obtained in step (c) to thereby obtain a plant characterized by said increased harvest index, leaf blade area, plot coverage, and/or rosette area under said nitrogen-limiting conditions as compared to said non-transformed plant which is grown under the same growth conditions,
thereby generating the plant having the increased tolerance to nitrogen limiting conditions.

6. The method of claim 1 , wherein said polypeptide sequence is set forth by SEQ ID NO: 656.

7. The method of claim 4 , wherein said polypeptide sequence is set forth by SEQ ID NO: 656.

8. The method of claim 1 , wherein said selecting in step (b) is for said increased harvest index under said nitrogen-limiting conditions.

9. The method of claim 1 , wherein said selecting in step (b) is for said increased leaf blade area under said nitrogen-limiting conditions.

10. The method of claim 1 , wherein said selecting in step (b) is for said increased plot coverage under said nitrogen-limiting conditions.

11. The method of claim 1 , wherein said selecting in step (b) is for said increased rosette area under said nitrogen-limiting conditions.

12. The method of claim 4 , wherein said selecting in step (b) is for said increased harvest index under said nitrogen-limiting conditions.

13. The method of claim 4 , wherein said selecting in step (b) is for said increased leaf blade area under said nitrogen-limiting conditions.

14. The method of claim 4 , wherein said selecting in step (b) is for said increased plot coverage under said nitrogen-limiting conditions.

15. The method of claim 4 , wherein said selecting in step (b) is for said increased rosette area under said nitrogen-limiting conditions.

16. The method of claim 1 , wherein said constitutive promoter is selected from the group consisting of a 35S promoter or an At6669 promoter.

17. The method of claim 16 , wherein the sequence of said 35S promoter is set forth by SEQ ID NO: 3827, 3833 or 3834.

18. The method of claim 16 , wherein the sequence of said At6669 promoter is set forth by SEQ ID NO: 3829 or 3826.

19. The method of claim 4 , wherein said constitutive promoter is selected from the group consisting of a 35S promoter or an At6669 promoter.

20. The method of claim 19 , wherein the sequence of said 35S promoter is set forth by SEQ ID NO: 3827, 3833 or 3834.

21. The method of claim 19 , wherein the sequence of said At6669 promoter is set forth by SEQ ID N

Assignees

• Panik David
• Vinocur Basia Judith
• Karchi Hagai
• Evogene Ltd

Inventors

• Panik David
• Vinocur Basia Judith
• Karchi Hagai

Classifications

• C12N15/8247

  involving modified lipid metabolism, e.g. seed oil composition · CPC title

• C12N15/8271 Primary

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

• C12N15/8261

  with agronomic (input) traits, e.g. crop yield · CPC title

• C07K14/415

  from plants · CPC title

• Y02A40/146

  Genetically Modified [GMO] plants, e.g. transgenic plants · CPC title