CN-122012598-A - Application of rice OsTIP & lt 2 & gt gene in improving resistance of rice to acetochlor and rice breeding material thereof

Abstract

The invention discloses application of a rice OsTIP & lt 2 & gt gene in improving resistance of rice to acetochlor and a rice breeding material thereof. The resistance of rice to acetochlor can be improved by knocking OsTIP <2 >, the OsTIP <2 > gene has an amino acid sequence shown as SEQ ID NO. 2, and the gene is matched with acetochlor for use, so that the rice can be assisted in light and simplified planting, and the rice has important economic value. The herbicide resistance characteristic of the gene editing rice can be stably inherited through sexual or asexual reproduction, and can be used for constructing resistant rice germplasm resources.

Inventors

• XU HANHONG
• XIAO CHUNXIA
• LIN FEI
• Jiang Dengji
• YAO YIFENG

Assignees

• 华南农业大学

Dates

Publication Date

20260512

Application Date

20260316

Claims (5)

1. 1. The application of the rice OsTIP and 2 genes in improving the resistance of the rice to acetochlor is characterized in that the resistance of the rice to the acetochlor is improved by knocking out OsTIP and 2 genes; the amino acid sequence of OsTIP & lt 2 & gt and 2 & lt 2 & gt gene is shown as SEQ ID NO & lt 2 & gt, or the amino acid sequence of derivative protein which is obtained by substituting and/or deleting and/or adding one or more amino acid residues in the sequence shown as SEQ ID NO & lt 2 & gt and still has the same function.
2. 2. The application of the rice OsTIP and 2 genes in improving the resistance of rice to acetochlor according to claim 1, wherein the nucleotide sequence of the OsTIP and 2 genes is shown as SEQ ID NO. 1.
3. 3. The application of rice OsTIP2 and 2 genes in creating amide herbicide resistant germplasm resources in a sexual reproduction or asexual reproduction mode is characterized in that the OsTIP2 and 2 genes have an amino acid sequence shown as SEQ ID NO. 2 or are amino acid sequences of derivative proteins which are obtained by substituting and/or deleting and/or adding one or more amino acid residues in the sequence shown as SEQ ID NO. 2 and still have the same function.
4. 4. A method for culturing OsTIP & lt 2 & gt and 2 & lt 2 & gt transgenic plants is characterized in that OsTIP & lt 2 & gt gene knockout vectors are constructed, the gene knockout vectors are introduced into target plants, and OsTIP & lt 2 & gt gene knockout rice plants are obtained.
5. 5. A rice breeding material is characterized in that a tonoplast aquaporin gene OsTIP and 2 are knocked out by CRISPR/Cas9 gene editing technology.

Description

Application of rice OsTIP & lt 2 & gt gene in improving resistance of rice to acetochlor and rice breeding material thereof Technical Field The invention relates to the technical field of rice breeding, in particular to application of a rice OsTIP gene 2 in improving resistance of rice to acetochlor and a rice breeding material thereof. Background The light simplified planting forms such as direct seeding of rice are the development direction of rice cultivation, wherein the key of ensuring high and stable yield of rice is prevention and control of weeds. Chemical control is the most dominant weeding mode at present, but paddy field weeds are various in variety and have multiple drug resistance, so that the problems of excessive use of herbicide, environmental pollution, pesticide residue, aggravation of weed resistance and the like are easily caused. The application of the amide herbicide acetochlor as a pre-emergence herbicide can reduce the total consumption of the herbicide and improve the weeding simplicity, but is difficult to be applied to paddy fields because of the disadvantage of the herbicide on the growth of rice. According to the invention, the acetochlor transport protein is inactivated by a gene editing technology, so that herbicide is prevented from reaching a rice target part to act, a resistant rice variety is obtained, and the herbicide is matched with acetochlor for use, thereby helping rice to simplify planting and having important economic value. Disclosure of Invention The invention aims to provide rice OsTIP2, application of 2 genes in improving the resistance of rice to acetochlor and a rice breeding material thereof. In order to achieve the above purpose, the technical scheme adopted by the invention is as follows: the invention provides rice OsTIP, application of 2 genes in improving the resistance of rice to acetochlor, wherein OsTIP2 is knocked out, and 2 genes improve the resistance of rice to acetochlor; the amino acid sequence of OsTIP & lt 2 & gt and 2 & lt 2 & gt gene is shown as SEQ ID NO & lt 2 & gt, or the amino acid sequence of derivative protein which is obtained by substituting and/or deleting and/or adding one or more amino acid residues in the sequence shown as SEQ ID NO & lt 2 & gt and still has the same function. Furthermore, the nucleotide sequence of OsTIP & lt 2 & gt gene is shown as SEQ ID NO. 1. The invention also provides application of rice OsTIP and 2 genes in creating amide herbicide resistant germplasm resources in a sexual reproduction or asexual reproduction mode, wherein the amino acid sequence of the OsTIP and 2 genes is shown as SEQ ID NO.2 or the amino acid sequence of derivative proteins with the same functions obtained by substituting and/or deleting and/or adding one or more amino acid residues in the sequence shown as SEQ ID NO. 2. A method for culturing OsTIP & 2 transgenic plant includes such steps as constructing OsTIP & 2 gene knockout carrier, and introducing said carrier into target plant to obtain OsTIP & 2 gene knockout rice plant. A rice breeding material is prepared by knocking out tonoplast aquaporin gene OsTIP and 2 by CRISPR/Cas9 gene editing technique, or by editing nucleotide sequence of rice OsTIP and 2 gene to inhibit expression and/or activity of protein OsTIP and 2 in plant rice. Compared with the prior art, the invention has the outstanding effects that: The invention clones a tonoplast aquaporin family gene OsTIP < 2 >, 2, and the CRISPR/Cas9 gene editing technology is used for knocking out OsTIP < 2 > in wild rice, and after 2, the growth inhibition effect of acetochlor on the wild rice is remarkably relieved, which shows that the gene participates in the targeting transport path of acetochlor in the rice body, and has the potential of cultivating acetochlor-resistant rice germplasm resources. The herbicide resistance characteristic of the gene editing rice can be stably inherited through sexual or asexual reproduction, and can be used for constructing resistant rice germplasm resources. The application of the rice OsTIP2 and 2 genes in improving the resistance of rice to acetochlor and the rice breeding materials thereof are further described below with reference to the accompanying drawings and specific examples. Drawings FIG. 1 is a graph showing the results of subcellular localization of OsTIP2 protein-GFP in rice protoplasts. Scale=10 nm. FIG. 2 shows the sequence alignment of OsTIP, protein 2 and representative aquaporins. FIG. 3 shows OsTIP, molecular docking analysis of protein 2 and acetochlor. FIG. 4 shows the growth of yeast with heterologous expression OsTIP, 2 proteins under acetochlor stress, wherein FIG. A shows the growth phenotype of yeast under acetochlor stress and FIG. B shows the growth curve of recombinant yeast. FIG. 5 shows OsTIP, 2 mutant plant identification. FIG. 6 shows the tolerance of wild rice and OsTIP2 mutants to acetochlor during seed germination, wherein FIG. A shows the growth of plants