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CN-121991969-A - Mulberry heat shock transcription factor MaHSF1, coding gene and application thereof

CN121991969ACN 121991969 ACN121991969 ACN 121991969ACN-121991969-A

Abstract

The application discloses a mulberry heat shock transcription factor MaHSF, a coding gene and application thereof, and aims to solve the technical problem of deficient high-temperature resistant gene resources and germplasm resources of the current mulberry. The application is based on heat shock transcription factor MaHSF which is screened from mulberry HSF gene family, is obviously up-regulated under high temperature stress and can improve the high temperature tolerance of yeast by heterologous expression, the over-expression of the heat shock transcription factor can inhibit the increase of the MDA content of high Wen Zhongbing dialdehyde, promote the increase of the SOD, POD and CAT activities of catalase and the free proline content of osmotic regulating substances, further improve the high temperature tolerance of arabidopsis, and the expression of the promoter has developmental stage specificity and heat stress induction characteristics, thereby indicating the accuracy of a regulating mechanism. The application verifies MaHSF a function and action mechanism of the coding gene thereof for resisting high temperature stress injury for the first time, and provides a solid theoretical basis for the mulberry production to cope with high temperature stress injury and select and breed new varieties with high temperature resistance.

Inventors

  • TIAN YUE
  • HE MEIYU
  • ZHANG KAIXIN
  • LIU LI
  • CAO XU
  • Zhai Zeyang

Assignees

  • 江苏科技大学

Dates

Publication Date
20260508
Application Date
20260212

Claims (10)

  1. 1. The mulberry heat shock transcription factor MaHSF gene is characterized in that the nucleotide sequence of the mulberry heat shock transcription factor MaHSF1 gene is shown in SEQ ID NO. 1.
  2. 2. An expressed protein of the mulberry heat shock transcription factor MaHSF gene as claimed in claim 1, wherein the amino acid sequence is shown in SEQ ID No. 2.
  3. 3. A promoter of mulberry heat shock transcription factor MaHSF gene as claimed in claim 1, wherein the sequence is shown in SEQ ID No. 3.
  4. 4. Use of a mulberry heat shock transcription factor MaHSF gene as claimed in claim 1 for enhancing tolerance of a plant to high temperature stress.
  5. 5. The use of claim 4, wherein the enhancing the tolerance of the plant to high temperature stress is inhibiting the increase in MDA in the plant under high temperature stress, increasing SOD, POD, CAT activity in the plant, and increasing the accumulation of free proline in the plant.
  6. 6. The use according to claim 5, comprising the steps of: (1) Constructing an overexpression vector containing the mulberry heat shock transcription factor MaHSF gene, wherein the overexpression vector is an arabidopsis thaliana stable genetic overexpression vector containing MaHSF1 specific sequence, and is named as pCAMBIA1300-MaHSF1, and primer sequences for amplifying the MaHSF1 specific sequence are SEQ ID NO.4 and SEQ ID NO.5: 1300-MaHSF1-F:aggacaggtacccggggatcATGAAGTCTCAGTTTGTGCG; 1300-MaHSF1-R:atcgtctttgtagtcgtcgatCTTGGGGCTAGAGGCCAAA; Wherein, the lowercase letter is a carrier homology arm; (2) Transforming the over-expression vector constructed in the step (1) into a plant cell; (3) And (3) treating the plant cells obtained in the step (2) for 24 hours at the temperature of 42 ℃ to improve the high-temperature tolerance of the plants.
  7. 7. Use of a promoter of the mulberry heat shock transcription factor MaHSF gene as defined in claim 3 in plant tissue expression or high temperature response expression.
  8. 8. The use according to claim 7, wherein, The expression in plant tissue means that the basic activity is extremely low in the early stage of vegetative growth, and the expression is regulated and controlled in the development stage; The expression in high Wen Xiangying means that the heat stress specificity induces the promoter of the mulberry heat shock transcription factor MaHSF gene to be obviously expressed in vascular tissues and reproductive organs, and has the characteristic of heat response.
  9. 9. The use according to claim 8, comprising the steps of: (1) Constructing an overexpression vector containing a promoter of the mulberry heat shock transcription factor MaHSF gene, wherein the overexpression vector is an arabidopsis thaliana stable genetic overexpression vector containing a MaHSF gene promoter specific sequence, and is named as pBI121-ProMaHSF1, and primer sequences for amplifying the MaHSF gene promoter specific sequence are SEQ ID NO.6 and SEQ ID NO.7: pBI121-ProMaHSF1-F:tatgaccatgattacgccaagctAAGTGTCTTTAAGAGGAGGC; pBI121-ProMaHSF1-R:ggactgaccacccggggatcTTTCACTACAACTCAATCTTGATTT; Wherein, the lowercase letter is a carrier homology arm; (2) Transforming the over-expression vector constructed in the step (1) into a plant cell; (3) The plant cells obtained in the step (2) have high specificity and dynamic expression patterns under the normal development state and depend on the development stage; Or alternatively, the first and second heat exchangers may be, (4) And (3) respectively treating the plant cells obtained in the step (2) for 3h and 6h in a 42 ℃ environment, wherein the expression mode has obvious high temperature stress induction characteristics.
  10. 10. The use according to claim 6 or 9, wherein the plant cells used are all cells of arabidopsis thaliana.

Description

Mulberry heat shock transcription factor MaHSF1, coding gene and application thereof Technical Field The invention belongs to the technical fields of molecular biology and genetic engineering, and particularly relates to a mulberry heat shock transcription factor MaHSF1, a coding gene and application thereof. Background With global warming, high temperature stress has become one of the main abiotic stress factors that restrict plant growth and development, and reduce crop yield and quality. Plants as sessile organisms cannot evade high temperature environments by moving, and thus complex stress control networks have evolved to cope with high temperature stress, where heat shock factors (Heat Shock Factors, HSFs) are a family of core transcription factors that mediate plant heat resistance. HSFs regulate the expression of Heat Shock Proteins (HSPs) and other stress related genes by identifying and combining cis-acting elements of a downstream heat response gene promoter region, thereby enhancing the tolerance of plants to high temperatures. Plants HSFs can be classified into A, B, C, and class a HSFs is more than a transcriptional activator, which plays a key role in heat resistance control, with relatively little research on the HSFA6b subclass, according to structural features. At present, the HSF genes are cloned from various plants such as arabidopsis thaliana, rice, tea trees and the like, but the functions of the HSF genes of different species have species specificity, and the regulation mechanism of most HSF genes is not yet fully elucidated. The mulberry is an important perennial woody economic plant, is not only a main feed source of silkworms, but also has wide application in the fields of medicine, feed, functional food, ecological protection and the like. The mulberry has wide distribution range, forms certain environmental adaptability in long-term evolution, but the high-temperature stress still can lead to leaf wilting, photosynthesis efficiency reduction and growth resistance, and seriously affects the yield and ecological value. Currently, identification and functional studies on mulberry HSF genes are limited. Therefore, the genes with heat-resistant function in the mulberry are excavated, new gene resources and technical approaches are provided for the genetic improvement of heat resistance of plants, and the method has important theoretical significance and application value. Disclosure of Invention The invention aims at solving the problems in the prior art, and aims at providing a mulberry heat shock transcription factor MaHSF gene and providing application of the mulberry heat shock transcription factor MaHSF gene. In order to solve the technical problems, the technical scheme adopted by the invention is as follows: A mulberry heat shock transcription factor MaHSF gene has a nucleotide sequence shown in SEQ ID NO. 1. The invention also provides an expression protein of the mulberry heat shock transcription factor MaHSF gene, and the amino acid sequence of the expression protein is shown as SEQ ID NO. 2. The invention also provides a promoter of the mulberry heat shock transcription factor MaHSF gene, the sequence of which is shown as SEQ ID NO. 3. The invention also provides application of the mulberry heat shock transcription factor MaHSF gene in enhancing the tolerance of plants to high temperature stress. A further improvement is that the enhancement of the tolerance of the plant to high temperature stress means that the increase of MDA of the plant is inhibited under high temperature stress, the activity of SOD, POD, CAT in the plant is improved, and the accumulation of free proline in the plant is improved. The method specifically comprises the following steps: (1) Constructing an over-expression vector containing the mulberry heat shock transcription factor MaHSF gene: The overexpression vector is an arabidopsis stable genetic overexpression vector containing MaHSF specific sequences, is named as pCAMBIA1300-MaHSF1, and the primer sequences for amplifying the MaHSF specific sequences are SEQ ID NO.4 and SEQ ID NO.5: 1300-MaHSF1-F:aggacaggtacccggggatcATGAAGTCTCAGTTTGTGCG; 1300-MaHSF1-R:atcgtctttgtagtcgtcgatCTTGGGGCTAGAGGCCAAA; Wherein, the lowercase letter is a carrier homology arm; (2) Transforming the over-expression vector constructed in the step (1) into a plant cell; (3) And (3) treating the plant cells obtained in the step (2) for 24 hours at the temperature of 42 ℃ to improve the high-temperature tolerance of the plants. The invention also provides an application of the promoter of the mulberry heat shock transcription factor MaHSF gene in plant tissue expression or high-temperature response expression. The further improvement is that the basic activity is extremely low in early vegetative growth, the expression is regulated in the development stage, and the expression in high Wen Xiangying is that the heat stress specificity induces the promoter of the mulberry heat shock