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CN-121975855-A - Application of blueberry VcDELLA-1 gene in improving high-temperature stress resistance of plants

CN121975855ACN 121975855 ACN121975855 ACN 121975855ACN-121975855-A

Abstract

The application discloses an application of a blueberry VcDELLA-1 gene in improving the high temperature stress resistance of plants, and belongs to the field of plant genetic engineering. Based on blueberry heat stress transcriptome data analysis, only VcDELLA-1 in the DELLA family is found to contain DLPYAD motif, and the gene expression quantity in the salicylic acid treatment group is obviously higher than that in the control group, so that the gene is a blueberry heat-resistant key gene. Under the high temperature and salicylic acid treatment, vcDELLA-1 in the transgenic VcDELLA-1 gene Arabidopsis and blueberry plants is obviously up-regulated for expression, the curling shrinkage degree of the leaves is light, the activity of superoxide dismutase is obviously reduced, the content of peroxidase, malondialdehyde and hydrogen peroxide is obviously increased, and the related genes of heat stress are greatly expressed. VcDELLA-1 gene can raise high-temperature resistance of plant by regulating and controlling heat stress related gene expression so as to provide important gene resource for cultivating new variety of high-temperature resistant plant.

Inventors

  • WU YAQIONG
  • Yu Ruisi
  • ZHANG PANPAN
  • Wang que
  • ZHANG CHUNHONG
  • YANG HAIYAN
  • LI WEILIN

Assignees

  • 江苏省中国科学院植物研究所

Dates

Publication Date
20260505
Application Date
20260327

Claims (10)

  1. 1. The application of the blueberry VcDELLA-1 gene in improving the high temperature stress resistance of plants is characterized by comprising the step of overexpressing the blueberry VcDELLA-1 gene in the plants, wherein the nucleotide sequence of the blueberry VcDELLA-1 gene is shown as SEQ ID NO. 1.
  2. 2. The application according to claim 1, characterized in that said application comprises the steps of: S1, constructing a plant expression vector of VcDELLA-1 genes; S2, transforming a plant expression vector of VcDELLA-1 genes into plant tissues or cells; s3, culturing and screening to obtain transgenic plants with improved high-temperature stress resistance; the plant is blueberry or Arabidopsis thaliana.
  3. 3. The use according to claim 2, wherein the construction method of the plant expression vector of VcDELLA-1 gene is specifically that the expression cassette of blueberry VcDELLA-1 gene is inserted into a vector backbone comprising plant selectable marker genes and/or terminator sequences.
  4. 4. The use according to claim 3, wherein the expression cassette of the blueberry VcDELLA-1 gene is constructed by connecting the blueberry VcDELLA-1 gene with a plant functional promoter to form an expression cassette.
  5. 5. The use according to claim 4, wherein the plant functional promoter is a cauliflower mosaic virus 35S promoter.
  6. 6. The use according to claim 2, wherein said transformation is performed by agrobacterium-mediated genetic transformation.
  7. 7. The use according to claim 1, wherein the improvement of the plant's high temperature stress tolerance is manifested in that the transgenic plant has at least one characteristic selected from the group consisting of: (a) The damage degree of the blade is reduced; (b) The superoxide dismutase activity is improved; (c) The activity of catalase is improved; (d) The activity of peroxidase is improved; (e) The content of malondialdehyde is reduced; (f) Up-regulation of heat stress related gene expression levels.
  8. 8. The use according to claim 7, wherein the heat stress related gene is selected from at least one of the following genes ：McMYB52、CHIP、SIL、HSF16、ERECTA、ZmCIPH、bZIP-2、bZIP-3、HSF A-3、HSP20-1、HSP20-2、HSP20-3、HSF C-1、HSP70、WRKY.
  9. 9. Use of a transgenic plant or part thereof obtained by the use according to any one of claims 1 to 8 for the preparation of a high temperature tolerant plant variety.
  10. 10. Use of a transgenic plant or part thereof obtained by the use of any one of claims 1-8 as a model for studying the mechanism of heat resistant molecules or the pathway of high temperature stress response in plants.

Description

Application of blueberry VcDELLA-1 gene in improving high-temperature stress resistance of plants Technical Field The invention belongs to the technical field of plant genetic engineering, and particularly relates to application of a blueberry VcDELLA-1 gene in improving high-temperature stress resistance of plants. Background Plants are often exposed to high temperature stress during growth and development. Global climate warming results in extremely high Wen Tianqi frequency, high temperature stress has become one of the major abiotic stress factors limiting crop yield and quality. High temperature stress can cause a series of physiological and biochemical changes such as active oxygen accumulation, protein denaturation and inactivation, membrane system damage and the like in plants, and even cause death of the plants when serious. Therefore, the method for excavating and identifying the heat-resistant key genes of the plants and analyzing the heat-resistant molecular mechanisms thereof has great significance for cultivating new varieties of heat-resistant crops by genetic engineering means. The DELLA protein is a key negative regulator in Gibberellin (GA) signaling pathway, belonging to a subfamily of GRAS transcription factor families. DELLA proteins are named for their highly N-terminal conserved DELLA domains, which are the core regions of GA signaling and transduction. Research shows that the DELLA protein not only participates in regulating the growth and development processes of plants, such as seed germination, stem elongation, flowering time, fruit development and the like, but also plays an important role in the response of the plants to various abiotic stresses. For example, DELLA proteins (GAI, RGA, RGL, RGL2, RGL 3) in arabidopsis are involved in regulating the response processes to low temperature, drought, and salt stresses, and overexpression of specific DELLA family members can enhance plant tolerance to adversity. However, there is relatively little research on the function of DELLA proteins in plant heat tolerance, especially in woody plants, which has not been systematically reported. Blueberry (Vaccinium spp.) is a small berry fruit tree with high economic value, and the fruit is rich in anthocyanin, vitamins and various bioactive components, so that the blueberry is favored by consumers. However, blueberries are sensitive to high-temperature stress, and the high temperature in summer often causes leaf burn, photosynthesis degradation and fruit quality degradation, so that sustainable development of the blueberry industry is severely restricted. At present, the excavation and function verification research on blueberry heat-resistant gene resources is still in a starting stage, and the gene resources which can be used for heat-resistant breeding are relatively deficient. Whether the DELLA family genes exist in blueberries or not and the action mechanism of the DELLA family genes in the heat-resistant regulation of the blueberries are not reported to date. Therefore, the DELLA family genes are cloned from blueberries, the functions of the DELLA family genes in improving the high-temperature stress resistance of plants are explored, and not only can plant heat-resistant gene resources be enriched, but also theoretical basis and gene resources are provided for improving the heat resistance of blueberries and other crops through genetic engineering means. Aiming at the requirements, vcDELLA-1 genes are cloned from blueberries for the first time, and the functions of the VcDELLA-1 genes in improving the heat resistance of plants are verified. Disclosure of Invention Aiming at the problems of lack of plant high temperature resistant gene resources, unknown functions of blueberry DELLA family genes, lack of technical means for effectively improving heat resistance of blueberries and the like in the prior art, the invention provides application of a blueberry VcDELLA-1 gene in improving high temperature stress resistance of plants. In order to solve the technical problems, the invention adopts the following technical scheme: The application of the blueberry VcDELLA-1 gene in improving the high temperature stress resistance of plants comprises the step of over-expressing the blueberry VcDELLA-1 gene in the plants, wherein the nucleotide sequence of the blueberry VcDELLA-1 gene is shown as SEQ ID NO. 1. In some embodiments, the application comprises the steps of: S1, constructing a plant expression vector of VcDELLA-1 genes; S2, transforming a plant expression vector of VcDELLA-1 genes into plant tissues or cells; s3, culturing and screening to obtain transgenic plants with improved high-temperature stress resistance; the plant is blueberry or Arabidopsis thaliana. In some embodiments, the method for constructing the plant expression vector of VcDELLA-1 gene specifically comprises inserting the expression cassette of blueberry VcDELLA-1 gene into a vector backbone comprising a plant selectable