Plant transcriptional regulators of abiotic stress

• US 8,283,519 B2
• Filed: 05/04/2004
• Issued: 10/09/2012
• 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: Creelman, Robert A., Ratcliffe, Oliver, Kumimoto, Roderick W., Gutterson, Neal I., Reuber, T. Lynne, Libby, Jeffrey M., Heard, Jacqueline E., Riechmann, Jose Luis, Pineda, Omaira
• Primary Examiner(s): Ketter, Jim
• Application Number:
  US10/838,616
• Publication Number:
  US 20060008874A1
• Time in Patent Office: 3,080 Days
• Field of Search: None
• US Class Current: 800/262
• 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 transgenic plant that 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 B3 domain and a DML motif, wherein the AP2 domain, the B3 domain and the DML motif are at least 81%, 78% and 76% identical to the AP2 domain and the B3 domain and the DML motif of SEQ ID NO;

2, respectively.

2. The transgenic plant of claim 1, wherein the member of the G867 clade of transcription factor polypeptides is at least 64% identical to SEQ ID NO:

2.

3. The transgenic plant of claim 1, wherein the transgenic plant has greater tolerance to osmotic stress or cold than a control plant.

4. The transgenic plant of claim 3, wherein the osmotic stress is 150 mM NaCl or 9.4% (w/v) sucrose.

5. The transgenic plant of claim 1, 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 clade of transcription factor polypeptides.

8. The transgenic plant 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.

9. The transgenic plant of claim 8, wherein the regulatory element comprises an inducible or tissue specific promoter.

10. The transgenic plant of claim 8, wherein the regulatory element is selected from the group consisting of a SUC2 promoter, an RBCS3 promoter, an ARSK1 promoter, and an RD29A promoter.

11. A transgenic seed produced from the transgenic plant according to claim 1.

26. The method of claim 9, wherein the AP2 domain and the B3 domain are at least 86% and 87% identical to the AP2 domain and the B3 domain of SEQ ID NO:

2, respectively.

27. A transgenic plant of claim 3, wherein the greater osmotic stress tolerance is greater tolerance to salt or sucrose.

6. A transgenic plant, wherein said transgenic plant comprises a recombinant polynucleotide comprising a nucleotide sequence that encodes SEQ ID NO:

2.

7. The transgenic plant of claim 6, wherein said nucleotide sequence comprises SEQ ID NO:

1.

12. A method for producing a transgenic plant having greater tolerance to an osmotic stress or cold than a control plant, the method steps comprising:

(a) introducing an expression vector into a plant to produce the transgenic plant;

wherein the expression vector comprises 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 B3 domain and a DML motif; and

wherein the AP2 domain, the B3 domain and the DML motif are at least 81%, 78% and 76% identical to the AP2 domain, the B3 domain and the DML motif of SEQ ID NO;

2, respectively,

(b) growing the transgenic plant; and

(c) selecting the transgenic plant on the basis of its greater tolerance to osmotic stress or cold than the control plant.

13. The method of claim 12, the method steps further comprising:

(d) selfing or crossing the transgenic plant with itself or another plant, respectively, to produce a transgenic seed.

14. The method of claim 12, wherein said greater osmotic stress tolerance is greater tolerance to salt or sucrose.

15. The method of claim 12, wherein said greater tolerance to an increased osmotic stress is greater tolerance to 150 mM NaCl or 9.4% (w/v) sucrose.

16. The method of claim 12, wherein said member of the G867 clade of transcription factor polypeptides is at least 64% identical to SEQ ID NO:

2.

17. A method of claim 12,

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

18. The method of claim 17, wherein the AP2 domain, the DML motif, and the B3 domain are at least 86%, 90%, and 87% identical to the AP2 domain, the DML motif, and the B3 domain of SEQ ID NO:

2, respectively.

19. The method of claim 17, wherein the transformed plant has a morphology that is substantially similar to the control plant.

20. The method of claim 17, wherein said greater osmotic stress tolerance is greater tolerance to salt or sucrose.

21. The method of claim 17, wherein said greater osmotic stress tolerance is greater tolerance to 150 mM NaCl or 9.4% (w/v) sucrose.

22. The method of claim 17, 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.

23. A transgenic seed produced from the transformed plant produced by the method according to claim 17.

24. The method of claim 17, the method steps further comprising:

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

(e) growing a progeny plant from the transgenic seed;

wherein the progeny plant has greater tolerance to an osmotic stress or cold than the control plant.

25. A transgenic seed produced from the progeny plant produced by the method according to claim 24.

28. A method for producing a transgenic plant, the method comprising:

(a) providing an expression vector comprising a polynucleotide sequence encoding SEQ ID NO;

2; 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;

(c) growing the transgenic plant; and

(d) selecting the transformed plant on the basis of its greater tolerance to an osmotic stress or cold than a control plant.