Search

CN-122012602-A - Application of alfalfa FTa2 gene in cultivation of cold-resistant plants

CN122012602ACN 122012602 ACN122012602 ACN 122012602ACN-122012602-A

Abstract

The application belongs to the technical field of plant genetic engineering, and provides application of alfalfa FTa2 genes in cultivation of cold-resistant plants, wherein the nucleotide sequence of the alfalfa FTa2 genes is SEQ ID NO.1 or SEQ ID NO.3. The application discovers that the alfalfa cold resistance related gene FTa2 negatively regulates the cold resistance of alfalfa, knocks out or knocks down the FTa2 by utilizing a genome editing technology, can obviously improve the cold resistance of plants, and is particularly characterized in that the survival rate of the plants after freezing treatment is improved, and the relative conductivity of leaves is reduced. The application provides important paradigm and gene resource for cultivating cold-resistant plant varieties.

Inventors

  • DONG JIANGLI
  • Ye Qinyi
  • WANG TAO
  • Xue Qixia

Assignees

  • 中国农业大学

Dates

Publication Date
20260512
Application Date
20260402

Claims (10)

  1. 1. The application of the alfalfa FTa2 gene in cultivating cold-resistant plants is characterized in that the nucleotide sequence of the alfalfa FTa2 gene is SEQ ID NO.1 or SEQ ID NO.3.
  2. 2. The use of claim 1, wherein the alfalfa FTa2 gene in the plant is knocked out or knocked down, or the level or activity of a protein encoded by the alfalfa FTa2 gene in the plant is reduced, to achieve one or more effects selected from (A1) - (A3): (A1) Reducing the relative conductivity of the blade; (A2) The survival rate of the plants at low temperature is improved; (A3) Improving the growth state of plants at low temperature.
  3. 3. The use according to claim 1 or 2, wherein the alfalfa FTa2 gene in the plant is knocked out or knocked down or the level or activity of a protein encoded by the alfalfa FTa2 gene in the plant is reduced using a tool selected from one or more of (B1) - (B5): (B1) A tool based on a CRISPR-Cas9 method, preferably, the tool based on the CRISPR-Cas9 method leads mutation to lead the alfalfa FTa2 gene to terminate translation in advance through gene editing, more preferably, targets in the gene editing are SEQ ID NO.5 and SEQ ID NO.6, or SEQ ID NO.13 and SEQ ID NO.14; (B2) Means based on homologous recombination methods; (B3) A ZFN method-based tool; (B4) RNA interference tools of the alfalfa FTa2 gene, preferably siRNA, miRNA, shRNA, dsRNA; (B5) ASO of said alfalfa FTa2 gene; (B6) Inhibitors, preferably antibodies, of the protein encoded by the alfalfa FTa2 gene are used.
  4. 4. A use according to any one of claims 1-3 for one or more of (C1) - (C3): (C1) The cold resistance of plant plants is improved; (C2) Providing plant propagation material with improved cold resistance, preferably seeds, tissue culture material, organs available for propagation; (C3) Plant lines with improved cold resistance are provided.
  5. 5. Use according to any one of claims 1 to 3, wherein the plant is a alfalfa plant.
  6. 6. The use according to claim 5, wherein the plant is alfalfa or medicago tribulus.
  7. 7. The alfalfa FTa2 gene is characterized in that the nucleotide sequence of the alfalfa FTa2 gene is SEQ ID NO.1 or SEQ ID NO.3.
  8. 8. The protein encoded by the alfalfa FTa2 gene of claim 7.
  9. 9. The protein of claim 8, having the amino acid sequence of SEQ ID No.2 or SEQ ID No.4.
  10. 10. Use of any one of the means (D1) - (D3) for detecting the expression level of the alfalfa FTa2 gene or the expression level of a protein encoded by the alfalfa FTa2 gene: (D1) Judging the cold resistance of alfalfa plants; (D2) Predicting the cold resistance of alfalfa propagation materials; (D3) Cultivating cold-resistant alfalfa varieties; the means for detecting the expression level of the alfalfa FTa2 gene or the expression level of the protein encoded by the alfalfa FTa2 gene is one or more of the following: (E1) A primer; (E2) A probe; (E3) A PCR reagent; (E4) A chip; (E5) An antibody; (E6) ELISA reagents; the nucleotide sequence of the FTa2 gene is SEQ ID NO.1 or SEQ ID NO.3, and the nucleotide sequence of the protein coded by the alfalfa FTa2 gene is SEQ ID NO.1 or SEQ ID NO.3.

Description

Application of alfalfa FTa2 gene in cultivation of cold-resistant plants Technical Field The application belongs to the technical field of plant genetic engineering, and particularly provides application of alfalfa FTa2 genes in cultivation of cold-resistant plants. Background Plants are subjected to various biotic and abiotic stresses during the growth and development process, and in recent years, the severe temperature fluctuation caused by the global climate change obviously inhibits the growth of the plants and threatens the productivity of crops. Among them, low temperature stress is a serious natural disaster, which not only affects the growth, development, yield and quality of plants, but also limits the geographical distribution of many plants. The cold-resistant related genes of the plants are deeply excavated, so that the mechanism of the plants for resisting the low-temperature stress is analyzed, the plants are modified by utilizing plant genetic engineering, the tolerance of the plants to the low-temperature stress is further improved, the dependence of grains on the environment is reduced, the grain yield under the low-temperature condition is improved, and the human beings are benefited. Alfalfa is an important feed source as a fourth important cash crop in the world, and the excavation of the key genes of alfalfa for resisting low temperature can provide gene resources for cultivating cold-resistant plant varieties. Disclosure of Invention In one aspect, the application provides application of alfalfa FTa2 genes in cultivation of cold-resistant plants, wherein the nucleotide sequence of the alfalfa FTa2 genes is SEQ ID NO.1 or SEQ ID NO.3. Further, knocking out or knocking down the alfalfa FTa2 gene in the plant or reducing the level or activity of a protein encoded by the alfalfa FTa2 gene in the plant in the application to achieve one or more effects selected from (A1) - (A3): (A1) Reducing the relative conductivity of the blade; (A2) The survival rate of the plants at low temperature is improved; (A3) Improving the growth state of plants at low temperature. Further, the use in the application uses a tool selected from one or more of (B1) - (B5) to knock-out or knock-down the alfalfa FTa2 gene in the plant, or to reduce the level or activity of a protein encoded by the alfalfa FTa2 gene in the plant: (B1) A tool based on a CRISPR-Cas9 method, preferably, the tool based on the CRISPR-Cas9 method leads mutation to lead the alfalfa FTa2 gene to terminate translation in advance through gene editing, more preferably, targets in the gene editing are SEQ ID NO.5 and SEQ ID NO.6, or SEQ ID NO.13 and SEQ ID NO.14; (B2) Means based on homologous recombination methods; (B3) A ZFN method-based tool; (B4) RNA interference tools of the alfalfa FTa2 gene, preferably siRNA, miRNA, shRNA, dsRNA; (B5) ASO of said alfalfa FTa2 gene; (B6) Inhibitors, preferably antibodies, of the protein encoded by the alfalfa FTa2 gene are used. The above-described tools can be routinely designed, obtained, and modified by those skilled in the art using known tools, and carried using vectors known in the art. Further, the application is for one or more of (C1) - (C3) (C1) The cold resistance of plant plants is improved; (C2) Providing plant propagation material with improved cold resistance, preferably seeds, tissue culture material, organs available for propagation; (C3) Plant lines with improved cold resistance are provided. Further, the plant is an alfalfa plant. Further, the plant is alfalfa or medicago tribulus. In another aspect, the present application provides a method of improving cold resistance in a plant, the method comprising knocking out or knocking down an alfalfa FTa2 gene in the plant, the nucleotide sequence of the alfalfa FTa2 gene being SEQ ID No.1 or SEQ ID No.3. In another aspect, the application provides alfalfa FTa2 genes, wherein the nucleotide sequence of the alfalfa FTa2 genes is SEQ ID NO.1 or SEQ ID NO.3. In another aspect, the present application provides a protein encoded by the alfalfa FTa2 gene described above. Further, the amino acid sequence of the protein is SEQ ID NO.2 or SEQ ID NO.4. In another aspect, the present application provides the use of any one of the means (D1) - (D3) for detecting the expression level of the alfalfa FTa2 gene or the expression level of a protein encoded by the alfalfa FTa2 gene: (D1) Judging the cold resistance of alfalfa plants; (D2) Predicting the cold resistance of alfalfa propagation materials; (D3) Cultivating cold-resistant alfalfa varieties; the means for detecting the expression level of the alfalfa FTa2 gene or the expression level of the protein encoded by the alfalfa FTa2 gene is one or more of the following: (E1) A primer; (E2) A probe; (E3) A PCR reagent; (E4) A chip; (E5) An antibody; (E6) ELISA reagents; the nucleotide sequence of the FTa2 gene is SEQ ID NO.1 or SEQ ID NO.3, and the nucleotide sequence of the protein coded by the alfalfa FTa2 ge