Rice high temperature resistance gene and use in crop breeding resistance to high temperature thereof
The present invention provides a new plant gene-rice high temperature resistance 1 gene (Rice High Temperature Resistance 1, HTR1) and encoded protein thereof. Also disclosed is the use of the high temperature resistance gene, especially for the enhancement of high-temperature resistance of plants in plant variety improvement and cross breeding.
1. A method for obtaining a high temperature resistant line of a plant, wherein the method comprises steps of:
(a) transforming plant cells with a recombinant DNA construct, wherein said recombinant DNA construct comprises a polynucleotide molecule which comprises a promoter operably linked to a nucleotide sequence encoding a rice α2 subunit protein of 26S proteasome having an amino acid sequence, wherein the amino acid at position 99 is mutated from histidine to arginine as compared to the corresponding amino acid position 99 in the amino acid sequence of SEQ ID NO: 4, wherein said rice α2 subunit protein of 26S proteasome is selected from the group consisting of (i) the α2 subunit protein of 26S proteasome of rice having the amino acid sequence as set forth in SEQ ID NO: 2, and (ii) an α2 subunit protein of 26S proteasome of rice having an amino acid sequence which has at least 95% amino acid sequence identity to the amino acid sequence of SEQ ID NO: 2, wherein said transformation is through Agrobacterium tumefaciens mediated transformation, and wherein said transformation results in stable integration of said recombinant DNA construct into the genome of said transformed plant cells;
(b) regenerating transformed plants from said transformed plant cells; and
(c) selecting a transformed plant from step (b), which overexpresses said rice α2 subunit protein of 26S proteasome, and exhibits high temperature resistance when grown under a high temperature stress as compared to a control plant of the same species grown under identical conditions.
2. The method of claim 1 , wherein said rice α2 subunit protein of 26S proteasome has one or more characteristics selected from the group consisting of:
(i) after degradation and denaturation treatment under said high temperature stress, the activity of said rice α2 subunit protein of 26S proteasome is significantly higher than the activity of the polypeptide as shown in SEQ ID NO: 4 subjected to identical degradation and denaturation treatment; and
(ii) under said high temperature stress, the stability of said rice α2 subunit protein of 26S proteasome is significantly higher than the stability of the polypeptide as shown in SEQ ID NO: 4 subjected to said high temperature stress condition.
3. The method of claim 1 , wherein said rice α2 subunit protein of 26S proteasome is set forth in SEQ ID NO: 2.
4. The method of claim 1 , wherein said plant cells are selected from the group consisting of rice, maize, sorghum and wheat.
5. The method of claim 1 , wherein the nucleotide sequence encoding said rice α2 subunit protein of 26S proteasome is selected from the group consisting of:
the nucleotide sequence as set forth in SEQ ID NO: 1, and
the nucleotide sequence as set forth in SEQ ID NO: 29.
6. The method of claim 1 , wherein said promoter is a high temperature stress responsive promoter, and wherein said high temperature stress is from 35° C. to 45° C.
7. The method of claim 1 , wherein said promoter is set forth in SEQ ID NO: 27.
8. The method of claim 1 , wherein the nucleotide sequence encoding said rice α2 subunit protein of 26S proteasome is set forth in SEQ ID NO: 1.