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CN-121087063-B - Cold-adaptation regulatory gene RAF3/RAF6 and application thereof

CN121087063BCN 121087063 BCN121087063 BCN 121087063BCN-121087063-B

Abstract

The invention provides a cold adaptation regulating gene RAF3/RAF6 and application thereof, belonging to the technical field of genetic engineering. The invention provides a cold adaptation regulatory gene RAF3/RAF6, wherein the genome sequence of the RAF3 is shown as SEQ ID No.1, and the genome sequence of the RAF6 is shown as SEQ ID No. 2. The invention also provides the coded protein of the RAF3/RAF6, and after the expression level of the RAF3/RAF6 is reduced or the activity or the content of the protein is reduced, the cold tolerance and photosynthetic efficiency of plants can be improved, namely, under the cold condition, the RAF3 and/or RAF6 reduce photosynthesis, simultaneously the expression of cold response genes is enhanced, the growth of the plants is finally inhibited, and the adaptation of the plants to cold stress is promoted.

Inventors

  • JIN HONGLEI
  • WANG HONGBIN
  • DUAN SUJUAN
  • LIN SHUANG
  • HONG LIU
  • PAN JIAYI
  • CHEN XIYAN

Assignees

  • 广州中医药大学(广州中医药研究院)

Dates

Publication Date
20260512
Application Date
20250916

Claims (4)

  1. 1. The application of the cold adaptation regulating gene in regulating and controlling the cold tolerance of plants is characterized in that the regulation is to inhibit the expression quantity of the cold adaptation regulating gene and improve the cold tolerance of the plants, wherein the cold adaptation regulating gene is RAF3 or RAF6, the genome sequence of the RAF3 is shown as SEQ ID No.1, and the genome sequence of the RAF6 is shown as SEQ ID No. 2; The plant is Arabidopsis thaliana.
  2. 2. The use according to claim 1, wherein the CDS sequence of RAF3 is shown in SEQ ID No.3 and the CDS sequence of RAF6 is shown in SEQ ID No. 4.
  3. 3. The application of a reagent for inhibiting the expression of a cold adaptation regulating gene in improving the cold tolerance of plants is characterized in that the cold adaptation regulating gene is RAF3 or RAF6, wherein the genome sequence of the RAF3 is shown as SEQ ID No.1, and the genome sequence of the RAF6 is shown as SEQ ID No. 2; The plant is Arabidopsis thaliana.
  4. 4. A breeding method of cold-tolerant plants is characterized by comprising the steps of inhibiting the expression level of cold-adaptive regulation genes in target plants, wherein the cold-adaptive regulation genes are RAF3 or RAF6, the genome sequence of the RAF3 is shown as SEQ ID No.1, and the genome sequence of the RAF6 is shown as SEQ ID No. 2; The target plant is arabidopsis thaliana.

Description

Cold-adaptation regulatory gene RAF3/RAF6 and application thereof Technical Field The invention belongs to the technical field of genetic engineering, and particularly relates to a cold adaptation regulating gene RAF3/RAF6 and application thereof. Background The low temperature environment is one of the important factors limiting agricultural production, and the growth and development of plants can be severely inhibited under the low temperature environment. The plant itself has a complete coping mechanism when facing low temperature environment, and cold stress can seriously affect the plant as chloroplast which is sensitive in plant cells and plays a role of a signal sensor. Taking rice (Oryzasativa) as an example, one key point in a rice breeding program is to improve crop productivity by optimizing the composition and plant architecture of grain yield. However, rice growth is often severely affected by various biotic or abiotic stresses. Frost, chill and low temperature cold damage are major weather disasters that occur frequently in late spring and early autumn. Worldwide, loss of agriculture and forestry production caused by frequent occurrence of low temperature disasters is in the hundreds of billions of dollars. Therefore, research on the cold resistance related genes of plants has very important significance for improving the tolerance of the plants to low-temperature stress. In recent years, with the development of molecular biology, many developments have been made in the study of the theory of rice cold resistance, including that some key cold resistance related genes or QTLs are cloned successively. By utilizing the key genes, the tolerance of plants to low temperature can be effectively improved. However, in the current genetic engineering operation, most of the gene engineering operation utilizes a constitutive promoter to drive the expression of the key genes, and the obtained transgenic plant can show stronger low-temperature tolerance, but the constitutive promoter often brings about unnecessary expression, such as plant dwarf, development delay and material energy waste, which is not beneficial to potential practical application and popularization. Disclosure of Invention The invention provides a cold adaptation regulating gene RAF3/RAF6 and application thereof, wherein the gene RAF3/RAF6 negatively regulates the cold tolerance of plants, and can increase the cold tolerance and photosynthetic efficiency of plants by inhibiting the expression of the gene RAF3/RAF6 in target plants or reducing the content and/or activity of RAF protein in target plants. The invention provides a cold adaptation regulating gene, which comprises RAF3 or RAF6, wherein the genome sequence of the RAF3 is shown as SEQ ID No.1, and the genome sequence of the RAF6 is shown as SEQ ID No. 2. In a preferred mode of the invention, the CDS sequence of the RAF3 is shown as SEQ ID No.3, and the CDS sequence of the RAF6 is shown as SEQ ID No. 4. The invention also provides the protein encoded by the cold adaptation control gene. In a preferred mode of the invention, the amino acid sequence of the protein encoded by the RAF3 gene is shown as SEQ ID No.5, and the amino acid sequence of the protein encoded by the RAF6 gene is shown as SEQ ID No. 6. The invention also provides application of the cold adaptation regulating gene or the protein in regulating cold tolerance of plants. In a preferred mode of the invention, the regulation comprises inhibiting the expression level of the cold-adaptation regulatory gene or reducing the content and/or activity of the protein, and improving the cold tolerance of the plant. The invention also provides an application of the agent for inhibiting the expression of the cold adaptation regulating gene or the agent for reducing the content and/or activity of the protein in improving the cold tolerance of plants. The invention also provides a breeding method of the cold-tolerant plant, which comprises the steps of inhibiting the expression quantity of the cold-adaptive regulation gene in the target plant or reducing the content and/or activity of the protein. In a preferred form of the invention, the plant of interest comprises a dicot. The invention also provides a biological material for inhibiting the cold adaptation regulation gene expression. The invention has the beneficial effects that the mutant is screened from an arabidopsis mutant library by utilizing a forward genetics method, and the cold-adaptation regulatory gene RAF3/RAF6 is obtained by cloning through molecular identification, wherein the genome sequence of the RAF3 is shown as SEQ ID No.1, the genome sequence of the RAF6 is shown as SEQ ID No.2, and the cold-adaptation regulatory gene RAF3/RAF6 belongs to MAPKKK (MAP kinase kinase kinase) kinase members through comparison. The embodiment of the invention proves that the cold adaptation regulating gene is downwards regulated to express in the plant, so that the cold tolerance a