Plant transcriptional regulators of abiotic stress

• US 20060008874A1
• Filed: 05/04/2004
• Published: 01/12/2006
• Est. Priority Date: 09/22/1998
• Status: Active Grant

• Current Assignee: Mendel Biotechnology, Inc. (KOCH Industries Incorporated)
• Original Assignee: Mendel Biotechnology, Inc. (KOCH Industries Incorporated)
• Inventors: Reuber, T. Lynne, Libby, Jeffrey M., Gutterson, Neal I., Kumimoto, Roderick W., Ratcliffe, Oliver, Creelman, Robert A.
• Granted Patent: US 8,283,519 B2
• Time in Patent Office: Days
• US Class Current: 435/69.100
• CPC Class Codes: C07K 14/415 from plants
• CPC Class Codes: C12N 15/8261 with agronomic (input) traits, e.g. crop yield
• CPC Class Codes: C12N 15/8273 for drought, cold, salt resistance
• CPC Class Codes: Y02A 40/146 Genetically Modified [GMO] plants, e.g. transgenic plants

Abstract

The invention relates to plant transcription factor polypeptides, polynucleotides that encode them, homologs from a variety of plant species, variants of naturally-occurring sequences, and methods of using the polynucleotides and polypeptides to produce transgenic plants having advantageous properties, including improved cold and other osmotic stress tolerance, as compared to wild-type or reference plants. The invention also pertains to expression systems that may be used to regulate these transcription factor polynucleotides, providing constitutive, transient, inducible and tissue-specific regulation.

Claims

1. A recombinant polynucleotide comprising a nucleotide sequence that hybridizes over its full length to SEQ ID NO:

1 or its complement under stringent conditions that include two wash steps of 6×

SSC at 65°

C., each step being 10-30 minutes in duration.

2. The recombinant polynucleotide of claim 1, wherein the recombinant polynucleotide is operably linked to at least one regulatory element capable of regulating expression of the recombinant polynucleotide when the recombinant polynucleotide is transformed into a plant.

3. The recombinant polynucleotide of claim 1, wherein the recombinant polynucleotide is incorporated into an expression vector.

4. The recombinant polynucleotide of claim 3, wherein the recombinant polynucleotide is incorporated into a cultured host cell.

5. The recombinant polynucleotide of claim 1, wherein the recombinant polynucleotide encodes a polypeptide comprising SEQ ID NO:

2.

6. The recombinant polynucleotide of claim 1, wherein the recombinant polynucleotide comprises SEQ ID NO:

1.

21. A method for producing a transgenic plant having increased tolerance to an abiotic stress, the method steps comprising:

(a) introducing the expression vector according to claim 3 into a plant to produce the transgenic plant; and

(b) identifying an abiotic stress tolerance trait of the transgenic plant.

22. The method of claim 21, the method steps further comprising:

(c) selfing or crossing the abiotic stress tolerant plant with itself or another plant, respectively, to produce seed.

23. The method of claim 21, wherein said abiotic stress tolerance is cold.

24. The method of claim 21, wherein said abiotic stress tolerance is an osmotic stress.

25. The method of claim 24, wherein said osmotic stress is selected from the group consisting of heat, desiccation, drought, freezing, and high salt.

7. A transgenic plant having increased abiotic stress tolerance, wherein said transgenic plant comprises a recombinant polynucleotide comprising a nucleotide sequence that encodes a member of the G867 clade of transcription factor polypeptides;

wherein the member of the G867 clade of transcription factor polypeptides comprises an AP2 domain, a DML motif, and a B3 domain, in order from N-terminal to C-terminal.

8. The transgenic plant of claim 7, wherein the AP2 domain, the DML motif, and the B3 domain are at least 81%, 76%, and 78% identical to the AP2 domain, the DML motif, and the B3 domain of SEQ ID NO:

2, respectively.

9. The transgenic plant of claim 7, wherein the abiotic stress is cold.

10. The transgenic plant of claim 7, wherein the abiotic stress is an osmotic stress.

11. The transgenic plant of claim 10, wherein the osmotic stress is selected from the group consisting of heat, drought, desiccation, freezing, and high salt.

12. The transgenic plant of claim 7, wherein:

(a) the AP2 domain binds to a transcription-regulating region comprising the motif CAACA; and

(b) the B3 domain binds to a transcription regulating region comprising the motif CACCTG; and

wherein said binding of the AP2 domain and the B3 domain cooperatively enhances DNA binding affinity of the member of the G867 dade of transcription factor polypeptides.

13. The transgenic plant of claim 7, wherein the member of the G867 clade of transcription factor polypeptides comprises SEQ ID NO:

2.

14. The transgenic plant of claim 7, wherein the recombinant polynucleotide comprises SEQ ID NO:

1.

15. The transgenic plant of claim 7, wherein the member of the G867 lade of transcription factor polypeptides comprises SEQ ID NO:

64.

16. The transgenic plant of claim 7, wherein said the member of the G867 lade of transcription factor polypeptides is selected from the group consisting of SEQ ID NOs:

2, 4, 6, 8, 18, 22, 28, 32, and 36.

17. The transgenic plant of claim 7, wherein the recombinant polynucleotide is operably linked to at least one regulatory element capable of regulating expression of the recombinant polynucleotide when the recombinant polynucleotide is transformed into a plant.

18. The transgenic plant of claim 17, wherein the regulatory element comprises an inducible or tissue specific promoter.

19. The transgenic plant of claim 17, wherein the regulatory element is selected from the group consisting of a SUC2 promoter, an RBCS3 promoter, an ARSK1 promoter, and an RD29A promoter.

20. Seed produced from the transgenic plant according to claim 7.

26. A method for producing a transformed plant with greater tolerance to an abiotic stress than a control plant, the method comprising:

(a) providing an expression vector comprising a polynucleotide sequence encoding a member of the G867 clade of transcription factor polypeptides;

wherein the member of the G867 clade of transcription factor polypeptides comprises an AP2 domain, a DML motif, and a B3 domain, in order from N-terminal to C-terminal; and

wherein the polynucleotide sequence is operably linked to an inducible or tissue specific promoter that controls expression of the polynucleotide sequence;

(b) transforming a target plant with the expression vector to produce the transformed plant; and

(c) growing the transformed plant comprising the expression vector.

27. The method of claim 26, wherein the AP2 domain, the DML motif, and the B3 domain are at least 81%, 76%, and 78% identical to the AP2 domain, the DML motif, and the B3 domain of SEQ ID NO:

2, respectively.

28. The method of claim 26, wherein the transformed plant has a morphology that is substantially similar to the control plant.

29. The method of claim 26, wherein said abiotic stress tolerance is cold.

30. The method of claim 26, wherein said abiotic stress tolerance is an osmotic stress.

31. The method of claim 30, wherein said osmotic stress is selected from the group consisting of heat, desiccation, drought, freezing, and high salt.

32. The method of claim 26, wherein the member of the G867 clade of transcription factor polypeptides comprises a polypeptide sequence selected from the group consisting of SEQ ID NO:

2, 4, 6, 8, 18, 22, 28, 32, 34 and 36.

33. The method of claim 26, wherein the inducible or tissue specific promoter is selected from the group consisting of a SUC2 promoter, an RBCS3 promoter, an ARSK1 promoter, and an RD29A promoter.

34. Seed produced from the transformed plant produced by the method according to claim 26.

35. The method of claim 26, the method steps further comprising:

(d) selfing or crossing the transformed plant with itself or another plant, respectively, to produce seed; and

(e) growing a progeny plant from the seed;

wherein the progeny plant has greater tolerance to an abiotic stress than the control plant.

36. Seed produced from the progeny plant produced by the method according to claim 35.