Transcriptional factor relevant to resistant adversity from Arabidopsis thaliana, coded gene, and application

A transcription factor and stress tolerance technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problem of no ABRE consensus sequence, etc., achieve the effect of improving tolerance and broad application prospects

Inactive Publication Date: 2007-10-10
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Interestingly, the DRE in the promoter region of the RD22 gene is also affected by ABA, but it does not have a typical ABRE consensus sequence. Analysis shows that there is a DNA sequence containing both MYC and MYB recognition sites in the gene promoter, A total of 67bp, which may be the region induced by ABA and dehydration reaction

Method used

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  • Transcriptional factor relevant to resistant adversity from Arabidopsis thaliana, coded gene, and application
  • Transcriptional factor relevant to resistant adversity from Arabidopsis thaliana, coded gene, and application
  • Transcriptional factor relevant to resistant adversity from Arabidopsis thaliana, coded gene, and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1. Screening of Arabidopsis thaliana R2R3 MYB-type transcription factor coding gene DRM related to stress tolerance and acquisition of its cDNA

[0047] After the seeds of Arabidopsis ecotype Columbia (Col-0) were vernalized at 4°C for 3-5 days, they were treated with 70% alcohol for 2-3 minutes on an ultra-clean bench, and then washed once with sterile water (1-2 minutes). ), then the seeds were treated with 15% sodium hypochlorite for 15 minutes, washed 5-6 times with sterile water, fully oscillated and mixed each time, and finally the seeds were suspended in 0.1% agar, and evenly spread on the culture medium containing MS medium Seal the Petri dish with a parafilm, place it at 23°C, and grow in the culture room with 24h continuous light.

[0048] After two weeks of growth with the above-mentioned Arabidopsis materials, the seedlings were taken, ground with liquid nitrogen, and the total RNA of the leaves was extracted with an RNA extraction kit (QIAGEN) accor...

Embodiment 2

[0049] Example 2, the expression characteristics of the R2R3 MYB transcription factor encoding gene DRM related to stress tolerance in Arabidopsis under abiotic stress

[0050] 1. Detection of the expression characteristics of the R2R3 MYB-type transcription factor coding gene DRM in Arabidopsis thaliana under abiotic stress

[0051] The Arabidopsis ecotype Columbia was subjected to drought, NaCl, and ABA treatments respectively to analyze the expression of the Arabidopsis DRM obtained in Example 1 under abiotic stress. The specific method was: plant the seeds of Arabidopsis Columbia in pots After growing for 2 weeks, the seedlings were subjected to the following stress treatments:

[0052] Drought treatment: Arabidopsis thaliana seedlings were taken out from the soil to absorb the water on the roots, placed on dry filter paper, and then cultured under light conditions for 0 hour (control), 1 hour, 3 hours, 6 hours and 12 hours respectively. sampling.

[0053] Salt treatment...

Embodiment 3

[0065] Example 3, Functional identification of DRM-encoded protein

[0066] Detect the impact of overexpressing DRM on plants with the following transgenic test, and the specific process includes the following steps:

[0067] 1. Construction of DRM gene overexpression vector using 35S strong promoter

[0068] The recombinant vector pGEM-T-DRM carrying the DRM gene obtained in step 1 was double-digested with restriction endonucleases BamH I and Sma I, and the 858bp DRM gene fragment was recovered and purified. The cut vector pJIT163 (http: / / www.pgreen.ac.uk) was ligated to obtain a recombinant vector containing DRM and GFP coding regions, and then the recombinant vector was double-digested with restriction endonucleases Kpn I and Xho I , and then combined with the plant expression vector pBinPlus (Van Engelen, F.A., Molthoff, J.W., Conner, A.J., Nap, J.P., Pereira, A. and Stiekema, W.J.1995. based on pBIN19.Transgenic Res.4, 288-290) ligation, the ligation product was transfo...

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Abstract

This invention discloses stress-tolerant R2R3 MYB transcription factor, its coding gene and application. The stress-tolerant R2R3 MYB transcription factor is derived from Arabidoesis thaliana, and can be applied in culturing stress-tolerant plants that can resist abscisic acid, high salt and draught. The transcription factor is one of the following amino acid residue sequences: (1) SEQ ID NO.1; (2) protein by substituting, deleting or adding 1-10 amino acid residues of SEQ ID NO.1, which can regulate the stress tolerance of plants. The protein and its coding gene have important meaning to research on plant stress-tolerant mechanism, and method for improving draught, salt and stress tolerance of plants. The protein and its coding gene have potential application in stress-tolerant genetic engineering of plants.

Description

technical field [0001] The present invention relates to transcription factors related to adversity stress in plants and their coding genes and applications, in particular to an R2R3 MYB type transcription factor derived from Arabidopsis thaliana and related to abscisic acid resistance, drought and high-salt resistance and other stress tolerance. The coding gene thereof and its application in cultivating plants with improved resistance to abscisic acid, drought and high salinity. Background technique [0002] At present, drought is one of the important factors affecting plant growth and development and reducing crop yield, and the plant hormone abscisic acid (ABA) is a stress signal that plays a very important role in regulating the entire growth and development of plants. When plants are stressed by drought, etc., ABA can serve as an initial endogenous signal, causing plants to close stomata or change the expression of certain genes, etc., causing a series of adaptive respon...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/63C12N5/10C12N15/82
Inventor 王道文丁振华陈锋秦焕菊刘昕
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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