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CN-121991981-A - Potato StCYP481 gene, promoter, vector, recombinant bacterium and application thereof

CN121991981ACN 121991981 ACN121991981 ACN 121991981ACN-121991981-A

Abstract

The invention discloses a potato StCYP481 gene, a promoter, a vector, recombinant bacteria and application thereof, and relates to the technical field of plant genetic engineering. The promoter and nucleotide sequences are shown in SEQ ID NO.1 and SEQ ID NO. 2. The invention also provides a plant expression recombinant vector, which comprises a StCYP481 promoter, proStCYP481-GUS and 35S constructed by a gene sequence, a genetically engineered host cell, and an application of the gene StCYP481 in the cultivation of salt stress resistant transgenic plants. StCYP481 can effectively enhance the salt tolerance of transgenic plants by maintaining photosynthesis capacity, promoting biosynthesis of osmotic adjusting substances and reducing active oxygen accumulation, and lays a theoretical foundation for subsequent promotion of salt stress resistance of potatoes.

Inventors

  • CHEN YING
  • ZHUANG MINGYANG
  • ZHU XIAOBIAO
  • LV ZHAOYAN
  • Hou Hualan
  • PENG YAXUAN
  • SONG JIAFENG
  • Sheng Suao
  • YUE FANG
  • Cheng Jielan
  • WANG LING
  • LI CHUNCHAN

Assignees

  • 安徽农业大学

Dates

Publication Date
20260508
Application Date
20260213

Claims (10)

  1. 1. A potato salt stress resistance gene StCYP481 or a promoter proStCYP481, wherein the nucleotide sequence of proStCYP481 is shown as Seq ID No.1, and the CDS sequence of StCYP481 is shown as Seq ID No. 2.
  2. 2. Use of the promoter proStCYP481 according to claim 1 for the identification of soil salt stress and for the cultivation of transgenic plants resistant to salt stress, wherein the use is over-expression of proStCYP481 in plants for improving the plant's ability to identify soil salt stress.
  3. 3. A recombinant vector 1, wherein the recombinant vector 1 comprises the promoter proStCYP481 of claim 1.
  4. 4. The recombinant vector 1 according to claim 3, wherein the recombinant vector 1 is pBI121-proStCYP 481:GUS, and the recombinant vector 1 is constructed by inserting a promoter proStCYP 481:481 sequence into the cleavage site of the vector pBI121 by homologous recombination, wherein the cleavage site is HindIII and BamHI.
  5. 5. Recombinant bacterium 1, characterized in that the recombinant bacterium 1 comprises the recombinant vector 1 according to any one of claims 3 to 4.
  6. 6. The recombinant bacterium 1 according to claim 5, wherein the recombinant bacterium 1 is constructed by introducing the recombinant vector 1 according to any one of claims 3 to 4 into a cell, wherein the cell is Agrobacterium.
  7. 7. Use of the gene StCYP481 according to claim 1 for increasing salt tolerance in a plant, wherein said use is such that overexpression of the StCYP481 gene in the plant increases the salt stress tolerance of the plant.
  8. 8. A recombinant vector 2, wherein the recombinant vector 2 comprises the gene StCYP481 of claim 1.
  9. 9. The recombinant vector 2 according to claim 8, wherein the recombinant vector 2 is 35S, stCYP481-GFP, and the recombinant vector 2 is constructed by inserting the StCYP481 gene sequence into the cleavage site of the vector pRI101 by homologous recombination, wherein the cleavage site is SalI and BamHI.
  10. 10. Recombinant bacterium 2, characterized in that the recombinant bacterium 2 comprises the recombinant vector 2 according to any one of claims 8-9.

Description

Potato StCYP481 gene, promoter, vector, recombinant bacterium and application thereof Technical Field The invention relates to the technical field of plant genetic engineering, in particular to a potato StCYP481 gene, a promoter, a vector, recombinant bacteria and application thereof. Background Potato (Solanum tuberosum) is an annual herb whose tubers are edible. Potatoes are widely planted worldwide because of their high carbohydrate content, dietary fiber and various micronutrients, and have become the fourth largest food crop following rice, corn and wheat. However, as a staple food crop susceptible to abiotic stress, potato plants are increasingly exposed to a threat of climate change, with significant reductions in yield and quality often occurring under adverse environmental conditions. Soil salinization, which is one of the main abiotic stresses, can severely inhibit plant growth and development, resulting in great loss of crop production. Salt stress can significantly inhibit potato growth and reduce yield, thereby threatening agricultural production and global food safety. Therefore, screening of potato salt stress resistance genes has become a key research direction for breeding improvement thereof. Disclosure of Invention The technical problem to be solved by the invention is how to provide a potato StCYP481 gene, a promoter, a vector, recombinant bacteria and application thereof. The invention solves the technical problems by the following technical means: the first aspect of the invention provides a potato salt stress resistance gene StCYP481 and a promoter proStCYP thereof, wherein the nucleotide sequence of proStCYP481 is shown as Seq ID NO.1, and the CDS sequence of the StCYP481 is shown as Seq ID NO. 2. The invention discovers a potato gene StCYP481 and a promoter proStCYP481 thereof for the first time and application thereof in cultivating and breeding transgenic plants resistant to salt stress. The potato promoter proStCYP comprises the steps of (1) extracting potato genome DNA, (2) designing specific amplification primers proStCYP481-F and proStCYP481-R, and (3) carrying out PCR amplification by taking the potato genome DNA as a template to obtain a promoter proStCYP481 sequence, wherein the sequences of the specific amplification primers proStCYP481-F and proStCYP481-R are respectively shown as Seq ID NO.3 and Seq ID NO. 4. The potato gene StCYP481 cloning step comprises the steps of (1) extracting potato total RNA and reversely transcribing the potato total RNA into cDNA, (2) designing specific amplification primers StCYP481-F and StCYP481-R, (3) carrying out PCR amplification by taking the cDNA as a template to obtain a target gene StCYP481, wherein the sequences of the specific amplification primers StCYP481-F and StCYP481-R are respectively shown as Seq ID NO.5 and Seq ID NO. 6. The second aspect of the invention provides application of the promoter proStCYP481 in identifying soil salt stress and cultivating a salt stress resistant transgenic plant. The application is that over-expression proStCYP481 in plants can improve the capability of plants to recognize soil salt stress. The application comprises the following specific steps: (1) Cloning potato promoter proStCYP481,481; (2) Constructing a potato promoter proStCYP481 over-expression vector; (3) The potato promoter proStCYP481 over-expression vector is used for transforming a target plant, and the transgenic plant with improved soil salt stress identification capability is obtained through identification. In a third aspect, the present invention provides a recombinant vector 1, wherein the recombinant vector 1 comprises the above-described promoter proStCYP481. The recombinant vector 1 is pBI121-proStCYP 481:481, GUS. The construction method of the recombinant vector 1 comprises the step of inserting a promoter proStCYP481 sequence into the cleavage site of a vector pBI121 by using a homologous recombination method. The cleavage sites are HindIII and BamHI. The fourth aspect of the invention provides a recombinant bacterium 1, which comprises the recombinant vector pBI121-proStCYP 481:481, GUS. The construction method of the recombinant strain is as follows, namely, introducing GUS into a strain body, wherein the recombinant vector pBI121-proStCYP481 is constructed by the potato salt stress response promoter proStCYP. The thallus is agrobacterium. In a fifth aspect, the present invention provides the use of the gene StCYP481 described above for increasing salt tolerance in plants. The application is that the overexpression of the StCYP481 gene in the plant can improve the salt stress resistance of the plant. The application comprises the following specific steps: (1) Cloning potato gene StCYP481; (2) Constructing a potato gene StCYP481 overexpression vector; (3) The potato gene StCYP481 over-expression vector is used for transforming a target plant, and the transgenic plant with improved salt stress resistance is obtained throu