Application of arabidopsis gene ROI1 in aspect of high temperature stress resistance of plants

A transgenic plant, high temperature resistance technology, applied in the field of genetic engineering, can solve problems such as imperfection and unsystematic understanding

Inactive Publication Date: 2010-07-21
HENAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, people's understanding of the whole process is still not systematic and perfect, and there are still many blind spots in the middle

Method used

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  • Application of arabidopsis gene ROI1 in aspect of high temperature stress resistance of plants
  • Application of arabidopsis gene ROI1 in aspect of high temperature stress resistance of plants
  • Application of arabidopsis gene ROI1 in aspect of high temperature stress resistance of plants

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Experimental program
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Effect test

Embodiment 1

[0023] Screening of mutants related to high temperature stress and oxidation and cloning of mutant genes

[0024] like figure 1 shown in H 2 O 2 Under the treatment conditions, the EMS mutagenesis Arabidopsis mutant library was screened by a far-infrared imager, and two mutants related to ROS stress with significantly increased leaf temperature were obtained. We named the two mutants roi1-1 and roi1-1 respectively. roi1-2 (reactive oxygen species insensitive). After 2 hours of high temperature stress treatment at 45°C, it was found by far-infrared analysis that the leaf temperature of roi1-1 and roi1-2 was significantly lower than that of the wild type, and high temperature stress caused wilting and death of roi1-1 and roi1-2 leaves. 2 mutants are against H 2 O 2 Arabidopsis mutants that are insensitive to high temperature stress. Map-based cloning analysis showed that roi1-1 and roi1-2 were mutants produced by mutations at different sites in the At4g33950 gene.

Embodiment 2

[0026] In vivo expression and localization analysis of ROI1 gene

[0027]The promoter region sequence and CDS sequence of ROI1 isolated from the Arabidopsis genome were amplified by PCR, cloned to obtain the GUS reporter vector of pCAMBIA1381 and the GFP cell localization vector of pEGAD, and transformed Arabidopsis plants to obtain transgenic positive plants. GUS histochemical staining and fluorescence localization analysis showed that ROI1 was mainly distributed in the nuclei of vascular and guard cells, which may be related to the response of this gene to control leaf water loss and regulation of stomata. It indicated that the role of ROI1 may be related to the resistance to high temperature stress regulated by stomatal movement.

Embodiment 3

[0029] Functional analysis of ROI1 gene

[0030] like figure 2 As shown, the full-length CDS coding sequence of the isolated ROI1 was amplified by PCR, and the pCAMBIA1205-ROI1 overexpression vector was constructed, and the transgenic positive plants were transformed and screened by Agrobacterium infection.

[0031] The T-DNA insertion mutant SALK_008068(E)(O) for ROI1 was requested from the Arabidopsis Stock Center (NASC).

[0032] Phenotypic analysis of overexpression plants ROI1-OE showed that ROI1 gene overexpression plants (ROI1-OE) could resist high temperature treatment at 45 °C for 24 h, while roi1-1 mutants and T-DNA insertion mutants of ROI1 were treated at 45 °C At 24 hours, the plants had completely withered. Therefore, the expression of ROI1 gene can improve the high temperature stress resistance of plants.

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Abstract

The invention discloses application of an arabidopsis gene ROI1 in the aspect of high temperature stress resistance of plants. An arabidopsis mutant roi1 sensitive to high temperature and oxidation stress is obtained through screening; and the ROI1 gene is obtained by separating the arabidopsis mutant roi1. Experiments show that the arabidopsis ROI1 gene is induced to express by high temperature stress, regulates generation of H2O2 in plant bodies, is combined with heat shock transcription factors HSF3, regulates the activity of the HSF3 and downstream signal molecules in a mode of phosphorylation regulation, and regulates and improves high temperature stress resisting reaction of the plants. Functional analysis and identification of the ROI1 provide basis of theory and method for obtaining new varieties of high temperature stress resisting plants.

Description

technical field [0001] The present invention relates to an application of Arabidopsis gene ROI1, in particular to the application of the gene ROI1 in plant resistance to high temperature stress, and at the same time, it also relates to the application of the gene ROI1 in cultivating high temperature stress resistant transgenic plant varieties, which belongs to genetic engineering field. Background technique [0002] The change of environmental temperature is one of the environmental stress factors that plants must often face. Excessive temperature will seriously affect the growth and development of plants and normal physiological and biochemical processes, which is an important unfavorable factor that limits agricultural production. High temperature stress causes a large amount of reduction in agricultural production. During the long-term evolution of plants, a set of signal transduction regulation mechanism for resisting high temperature stress has been formed, and the reg...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00
Inventor 宋纯鹏安国勇薛小波赵孝亮王棚涛宋玉伟刘浩
Owner HENAN UNIVERSITY
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