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CN-121975846-A - Application of OsMTA gene in regulation and control of drought stress response of plants

CN121975846ACN 121975846 ACN121975846 ACN 121975846ACN-121975846-A

Abstract

The invention relates to the field of biotechnology, in particular to application of OsMTA gene in regulating and controlling drought stress response of plants. The invention carries out CRISPR gene editing on OsMTA genes to obtain transgenic rice. Compared with the acceptor rice ZH11, after drought treatment is carried out on two different strains of the transgenic rice OsMTA gene knockout mutant, the phenotype is observed and the survival rate after recovery treatment is counted, so that the OsMTA mutant presents a drought tolerance phenotype compared with a wild type, and OsMTA3 plays an important role in the process of responding to rice drought stress.

Inventors

  • WANG LEI
  • CHEN LINGLING
  • XU HANG
  • ZUO YI

Assignees

  • 中国科学院植物研究所

Dates

Publication Date
20260505
Application Date
20260224

Claims (10)

  1. The application of the OsMTA3 gene in regulating and controlling plant drought stress response is characterized in that the nucleotide sequence coded by the OsMTA gene is shown as SEQ ID NO. 1.
  2. 2. The use according to claim 1, wherein the OsMTA gene encodes a protein having the amino acid sequence shown in SEQ ID NO. 3.
  3. 3. The use according to claim 1, wherein the regulation is by knocking out OsMTA gene or reducing the expression level of OsMTA gene to increase drought tolerance in plants.
  4. 4. The use of claim 3, wherein the knockout is a knockout of OsMTA gene using CRISPR/Cas 9 gene editing techniques.
  5. 5. The use according to claim 4, wherein the nucleotide sequence of the target site for the OsMTA gene knockout is shown in SEQ ID NO. 6.
  6. 6. The use according to claim 3, wherein the plant is a dicotyledonous plant or a monocotyledonous plant.
  7. 7. A method of growing a transgenic plant with increased drought tolerance comprising the steps of: (1) Designing a CRISPR/Cas9 editing vector according to the target site of OsMTA gene of claim 1; (2) And infecting the CRISPR/Cas9 editing vector into a plant through agrobacterium, detecting according to a designed primer, and screening to obtain a T0 generation positive plant with the function-defective mutation, wherein the T0 generation positive plant shows improved drought tolerance.
  8. 8. The method of claim 7, wherein the CRISPR/Cas9 editing vector of step (1) has the target site shown in SEQ ID No. 6.
  9. 9. The method of claim 7, wherein the nucleotide sequence of the primer in step (2) is shown in SEQ ID NO. 7-8.
  10. 10. The method of claim 7, wherein the plant of step (2) is a dicotyledonous plant or a monocotyledonous plant.

Description

Application of OsMTA gene in regulation and control of drought stress response of plants Technical Field The invention belongs to the technical field of biology, and particularly relates to application of OsMTA gene in regulation and control of drought stress response of plants. Background As a staple food for nearly half of the world population, rice is one of the most critical crops to maintain food safety. Drought stress finally causes serious yield reduction and quality reduction by interfering physiological metabolism of rice and inhibiting growth and development, and is one of main abiotic stress in global rice production. When the rice is subjected to drought stress, the rice can be damaged disadvantageously (1) the drought causes the plants to have difficult water absorption, the cell turgor pressure is reduced, the leaf wilting and curling are caused, and the whole rice dies when serious. (2) Photosynthesis is inhibited, the absorption of CO 2 is reduced by closing pores, the synthesis of chlorophyll is hindered, the photosynthetic efficiency is reduced, and the accumulation of organic matters is reduced. (3) Metabolic disorders, impaired protein synthesis, accumulation of harmful substances (e.g., active oxygen), and impaired cell membrane structure. (4) Substance transport is hindered and moisture shortages affect nutrient transport, resulting in maldistribution of substances required for growth. Therefore, the breeding of drought-resistant varieties is one of important measures for drought. The RNA m 6 A modification related to the research belongs to RNA methylation modification, and specifically refers to a very conservative chemical modification type formed by replacing hydrogen atom (H) at N 6 position of RNA with methyl (CH 3). The rice drought stress control factor OsMTA in the report belongs to RNA methylase and is responsible for m 6 A modification of hydrogen atom (H) to be converted into methyl (CH 3). The study finds that after carrying out CRISPR gene editing knockout on OsMTA genes of rice, compared with flowers 11 (ZH 11) in acceptor rice, two different strains of osmta knockout mutants after drought stress treatment show drought tolerance phenotypes, and proves that OsMTA genes play an important role in mediating RNA m 6 A modification and then responding to rice drought stress, and have an important significance in improving rice drought tolerance through OsMTA 3. Disclosure of Invention Based on the defects in the prior art, the invention aims to provide an application of OsMTA gene in regulating and controlling plant drought stress response. After OsMTA gene in the plant is knocked out, osMTA protein expression level is greatly reduced, and rice drought tolerance can be improved. In order to achieve the above object, the present invention provides the following technical solutions: the invention provides an application of OsMTA gene in regulating plant drought stress response, wherein the nucleotide sequence coded by OsMTA gene is shown as SEQ ID NO. 1. Preferably, the amino acid sequence of the protein encoded by the OsMTA gene is shown as SEQ ID NO. 3. Preferably, the regulation is by knocking out OsMTA gene or reducing the expression level of OsMTA gene to improve drought tolerance of plants. After the gene in the plant is knocked out, the expression quantity is greatly reduced, and the drought tolerance of the plant can be improved. Preferably, the knockout is a knockout of OsMTA genes using CRISPR/Cas 9 gene editing techniques. Preferably, the nucleotide sequence of the target site of the OsMTA gene is shown as SEQ ID NO. 6. Preferably, the plant is a dicotyledonous plant or a monocotyledonous plant, more preferably rice. The invention also provides a method for cultivating transgenic plants with improved drought tolerance, comprising the following steps: (1) Designing a CRISPR/Cas9 editing vector according to a target site of OsMTA genes; (2) And infecting the CRISPR/Cas9 editing vector into a plant through agrobacterium, detecting according to a designed primer, and screening to obtain a T0 generation positive plant with the function-defective mutation, wherein the T0 generation positive plant shows improved drought tolerance. Preferably, the target site of the CRISPR/Cas9 editing vector in the step (1) is shown as SEQ ID NO. 6. Preferably, the nucleotide sequence of the primer in the step (2) is shown as SEQ ID NO. 7-8. Preferably, the plant of step (2) is a dicotyledonous plant or a monocotyledonous plant, more preferably rice. The invention has the following technical effects and advantages: Experiments prove that OsMTA genes in wild rice are knocked down, and drought-tolerant transgenic rice can be obtained. OsMTA3 gene expression was reduced in transgenic rice compared to recipient rice. Therefore, osMTA genes are related to rice response to drought stress, and a theoretical basis is laid for cultivating transgenic plants for positively regulating and controlling dr