CYCLIN-DEPENDENT KINASE INHIBITORS AS PLANT GROWTH REGULATORS

• US 20060253930A1
• Filed: 07/11/2006
• Published: 11/09/2006
• Est. Priority Date: 06/08/1998
• Status: Active Grant

• Current Assignee: Her Majesty The Queen in Right of Canada as represented by the Minister of Agriculture and Agri-Food, National Research Council Canada, University Of Saskatchewan
• Original Assignee: Her Majesty The Queen In Right of Canada As Represented By The Minister of National Defence, National Research Council Canada, University Of Saskatchewan
• Inventors: Wang, Hong, Crosby, William L., Fowke, Larry C.
• Granted Patent: US 7,732,663 B2
• Time in Patent Office: Days
• US Class Current: 800/285
• 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: Y02A 40/146 Genetically Modified [GMO] plants, e.g. transgenic plants

Abstract

Methods for using cyclin-dependent kinase (CDK) inhibitor genes, or anti-sense constructs complementary to such genes, to modify the growth and development of plant cells and organs are disclosed. Also provided are methods of modifying the development of plant cells and plants by transforming plant cells with nucleic acids encoding cyclin-dependent kinase inhibitor polypeptides, or anti-sense constructs complementary to such nucleic acids, to produce transformed plant cells, and then culturing the plant cells or regenerating a plant under conditions wherein the cyclin-dependent kinase inhibitor, or the anti-sense construct, is expressed. A variety of CDK inhibitor genes, and corresponding anti-sense constructs, are disclosed for use in a variety of plants. The nucleic acid encoding the cyclin-dependent kinase inhibitor may be operably linked to a tissue-specific promoter. Other provided aspects are modified transgenic plants and plant tissues. Also provided are methods of identifying nucleic acids that encode cyclin-dependent kinase inhibitors that are active in plants to modify the development of the plant.

Claims

1. A method of modifying development of a plant by down-regulating ICK1 polypeptide expression, wherein the plant comprises an ICK1 nucleic acid, comprising:

transforming a plant cell with;

(1) an anti-sense coding sequence complementary to a region of the nucleic acid encoding the cyclin-dependent kinase inhibitor polypeptide, or

(2) an enzymatic RNA encoding sequence capable of catalyzing cleavage of RNA transcribed from the nucleic acid encoding the cyclin-dependent kinase inhibitor polypeptide, to produce a transformed plant cell; and

growing the transformed plant cell or progeny of the transformed plant cell to produce a transformed plant under conditions wherein the anti-sense nucleic acid or enzymatic RNA is produced in a proliferative tissue of the transformed plant to inhibit expression of the ICK1 nucleic acid in the proliferative tissue of the transformed plant, thereby modifying development of the plant,

wherein the ICK1 nucleic acid comprises the nucleic acid sequence set forth in SEQ ID NO;

1 or SEQ ID NO;

3, or a nucleic acid sequence having at least 95% sequence identity with the nucleic acid sequence set forth in SEQ ID NO;

1 or SEQ ID NO;

3.

2. The method of claim 1, wherein the ICK1 polypeptide comprises the amino acid sequence set for in SEQ ID NO:

10.

3. The method of claim 1, wherein the plant is a member of the Cruciferae family.

4. The method of claim 1, wherein the plant is a member of the Brassica genus.

5. The method of claim 1, wherein the anti-sense coding sequence is complimentary to a region encoding the amino acid sequence selected from the group consisting of positions 1-72, 1-108, 1-152, 1-191, 1-162, 3-72, 73-108, 73-191, 109-153, 130-191, 153-162, 163-175, 163-191, and 176-191 of the amino acid sequence set forth in SEQ ID NO:

2, or the corresponding nucleic acid positions as set forth in SEQ ID NO;

3, or a nucleic acid sequence having at least 95% sequence identity therewith.

6. The method of claims 1, wherein the anti-sense coding sequence or the enzymatic RNA encoding sequence is operably linked to a constitutive promoter.

7. The method of claim 1, wherein the anti-sense coding sequence or the enzymatic RNA encoding sequence is operably linked to a tissue-specific promoter.

8. The method of claim 7, wherein the tissue-specific promoter is the AP3 promoter.

9. The method of claim 7, wherein the tissue-specific promoter mediates expression of the anti-sense coding sequence or the enzymatic RNA encoding sequence in petal or stamen primordia.

10. The method of claim 1, wherein modifying development of the plant makes the plant male sterile.

11. The method of claim 1, wherein the development of the tissue in the plant is modified so that petals on the transformed plant are altered or absent.

12. The method of claim 1, wherein the enzymatic RNA encoding sequence comprises a ribozyme or a hammerhead RNA structure.

13. A method for down-regulating expression of ICK1 in a plant cell, comprising:

expressing in the plant cell an anti-sense oligonucleotide, anti-sense RNA molecule, or anti-sense DNA molecule complementary to a region of a ICK1 nucleic acid;

or

expressing in the plant cell an enzymatic RNA molecule capable of catalyzing cleavage of ICK1 RNA,

wherein the cyclin-dependent kinase inhibitor is encoded by a nucleic acid molecule as set forth in SEQ ID NO;

1 or SEQ ID NO;

3, or a nucleic acid sequence having at least 95% sequence identity with the nucleic acid sequence set forth in SEQ ID NO;

1 or SEQ ID NO;

3.