Regulatory genes for plant differentiation and growth, and use of the same
View Patent ↗An objective of the present invention is to provide gibberellin-binding proteins, genes encoding such proteins, and applications therefor. The proteins of the present invention function as cytoplasmic receptors for gibberellin and mediate gibberellin response in plants. Plants overexpressing such genes exhibited gibberellin-hypersensitive phenotypes, such as increased plant height. In contrast, plants in which the genes were mutated exhibited gibberellin-insensitive phenotypes, and thus became dwarfed. Accordingly, plant differentiation and growth can be regulated by introducing the gibberellin-binding genes of the present invention or by suppressing the expression of same.
1. A method for binding a protein with gibberellin, which comprises the step of contacting the protein with gibberellin, wherein the protein is selected from the group consisting of:
(i) a protein comprising the amino acid sequence of SEQ ID NO: 2,
(ii) a protein comprising an amino acid sequence having 95% or higher identity to SEQ ID NO: 2 and has the activity of binding to gibberellin, and
(iii) a protein having the activity of binding to gibberellin and is encoded by a nucleic acid that hybridizes to the complement of SEQ ID NO:1 at 65° C. in a solution containing 5×SSC, 7% (W/V) SDS, 100 μg/ml denatured salmon sperm DNA, 5×Denhardt's solution, followed by washing in 0.1×SSC at 65° C.
2. A method for detecting gibberellin binding, which comprises the steps of contacting a protein with gibberellin and detecting the binding between the protein and gibberellin, wherein the protein is selected from the group consisting of:
(i) a protein comprising the amino acid sequence of SEQ ID NO: 2,
(ii) a protein comprising an amino acid sequence having 95% or higher identity to SEQ ID NO: 2 and has the activity of binding to gibberellin, and
(iii) a protein having the activity of binding to gibberellin and is encoded by a nucleic acid that hybridizes to the complement of SEQ ID NO:1 at 65° C. in a solution containing 5×SSC, 7% (W/V) SDS, 100 μg/ml denatured salmon sperm DNA, 5×Denhardt's solution, followed by washing in 0.1×SSC at 65° C.
3. A method for assaying a compound that regulates the interaction between gibberellin and a protein, which comprises the steps of:
(a) contacting a test compound, gibberellin, and the protein; and
(b) detecting the binding between gibberellin and the protein, wherein the protein is selected from the group consisting of:
(i) a protein comprising the amino acid sequence of SEQ ID NO: 2,
(ii) a protein comprising an amino acid sequence having 95% or higher identity to SEQ ID NO: 2 and has the activity of binding to gibberellin, and
(iii) a protein having the activity of binding to gibberellin and is encoded by a nucleic acid that hybridizes to the complement of SEQ ID NO:1 at 65° C. in a solution containing 5×SSC, 7% (W/V) SDS, 100 μg/ml denatured salmon sperm DNA, 5×Denhardt's solution, followed by washing in 0.1×SSC at 65° C.
4. A method for selecting a compound that inhibits the interaction between gibberellin and a protein, which comprises the steps of:
(a) contacting a test compound, gibberellin, and the protein;
(b) detecting the binding between gibberellin and the protein; and
(c) selecting a compound that inhibits the binding, wherein the protein is selected from the group consisting of:
(i) a protein comprising the amino acid sequence of SEQ ID NO: 2,
(ii) a protein comprising an amino acid sequence having 95% or higher identity to SEQ ID NO: 2 and has the activity of binding to gibberellin, and
(iii) a protein having the activity of binding to gibberellin and is encoded by a nucleic acid that hybridizes to the complement of SEQ ID NO:1 at 65° C. in a solution containing 5×SSC, 7% (W/V) SDS, 100 μg/ml denatured salmon sperm DNA, 5×Denhardt's solution, followed by washing in 0.1×SSC at 65° C.
5. A method for binding a first protein with a DELLA protein, which comprises the step of contacting the proteins, wherein the first protein is selected from the group consisting of:
(i) a protein comprising the amino acid sequence of SEQ ID NO: 2,
(ii) a protein comprising an amino acid sequence having 95% or higher identity to SEQ ID NO: 2 and has the activity of binding to the DELLA protein, and
(iii) a protein having the activity of binding to the DELLA protein and is encoded by a nucleic acid that hybridizes to the complement of SEQ ID NO:1 at 65° C. in a solution containing 5×SSC, 7% (W/V) SDS, 100 μg/ml denatured salmon sperm DNA, 5×Denhardt's solution, followed by washing in 0.1×SSC at 65° C.
6. An isolated and purified complex comprising gibberellin and a protein selected from the group consisting of:
(i) a protein comprising the amino acid sequence of SEQ ID NO: 2,
(ii) a protein comprising an amino acid sequence having 95% or higher identity to SEQ ID NO: 2 and has the activity of binding to gibberellin, and
(iii) a protein having the activity of binding to gibberellin and is encoded by a nucleic acid that hybridizes to the complement of SEQ ID NO:1 at 65° C. in a solution containing 5×SSC, 7% (W/V) SDS, 100 μg/ml denatured salmon sperm DNA, 5×Denhardt's solution, followed by washing in 0.1×SSC at 65° C.
7. The complex of claim 6 , which further comprises a DELLA protein.
8. An isolated and purified complex comprising a DELLA protein and a protein which selected from the group consisting of:
(i) a protein comprising the amino acid sequence of SEQ ID NO: 2,
(ii) a protein comprising an amino acid sequence having 95% or higher identity to SEQ ID NO: 2 and has the activity of binding to the DELLA protein, or
(iii) a protein having the activity of binding to the DELLA protein and is encoded by a nucleic acid that hybridizes to the complement of SEQ ID NO:1 at 65° C. in a solution containing 5×SSC, 7% (W/V) SDS, 100 μg/ml denatured salmon sperm DNA, 5×Denhardt's solution, followed by washing in 0.1×SSC at 65° C.