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Novel rice DREB transcription factor OsDREBx and coding gene and application thereof

A technology of transcription factor and rice, applied in application, genetic engineering, plant gene improvement, etc., can solve problems such as limited effect

Inactive Publication Date: 2010-11-17
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the stress resistance of plants is a complex quantitative trait, which is the result of the joint action of multiple regulatory mechanisms, the effect of transforming plants with a single functional gene is often limited.

Method used

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  • Novel rice DREB transcription factor OsDREBx and coding gene and application thereof
  • Novel rice DREB transcription factor OsDREBx and coding gene and application thereof
  • Novel rice DREB transcription factor OsDREBx and coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Cloning of OsDREBx gene

[0026] In the present invention, the full-length sequence of the OsDREBx gene is cloned from the indica rice variety 9311 by the following method.

[0027] The total DNA of the indica rice variety 9311 was extracted using a genomic DNA purification kit (purchased from Cybersun Corporation). Design primer sequence as follows:

[0028] P1: 5’ATGGCACTGG GGTTATTGAA CGC 3’

[0029] P2: 5’TCACAAGC CCTCAAAGAA CTGAG 3

[0030] PCR reaction system:

[0031] 10×buffer 2.5μl

[0032] MgCl 2 (25mM) 1.5μl

[0033] Taq enzyme 0.2μl

[0034] dNTPs(2.5mM) 2μl

[0035] Primer 2μl

[0036] Template 5μl

[0037] Final reaction volume 25μl

[0038] Thermal cycle reaction: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 1 min, annealing temperature is determined by primers, 72°C extension for 2 min, 30 cycles; 72°C extension for 5 min; 4°C to stop the reaction. The amplification reaction products were electrophoresed on a 1% agarose gel, and the PCR amplific...

Embodiment 2

[0039] Example 2 Verification of OsDREB gene in yeast one-hybrid binding experiment

[0040] The constructed yeast pGADT7-Rec2 vector containing the OsDREBx gene, and the constructed pHis-DRE4 vector containing four DRE (TACCGACTA) repeats, which can bind to both DREB1 and DREB2, were co-transformed into the Y187 competent strain and spread on On the triple deficiency medium SD / -His / -Leu / -Trp containing 100Mm 3AT, it grows like figure 2 The colony shown in A; At the same time, in the control, whether the yeast pGADT7-Rec2 vector without the OsDREBx gene and the pHis-DRE4 vector containing the four DRE repeats were co-transformed into the Y187 competent strain, or The constructed yeast pGADT7-Rec2 vector containing the OsDREBx gene was co-transformed into the Y187 competent strain with the pHis blank vector not linked to the four DRE repeats, and spread on the triple deficiency medium SD / -His / - containing 100Mm3AT After Leu / -Trp, no colonies grew.

[0041] In addition, in order t...

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PUM

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Abstract

The invention provides a novel rice DREB transcription factor OsDREBx and a coding gene thereof. The protein has an amino acid sequence shown as SEQ ID No.2. The gene can play an important role in stress resistance of plants, such as drought resistance, high-salinity tolerance, low-temperature resistance and the like, and can be used for the crop transgenic breeding research. The gene can transform crops such as rice, wheat, corn, vegetables and the like, endows the crops with corresponding stress resistance, and has important economic value and application prospect.

Description

Technical field [0001] The invention relates to a new rice DREB transcription factor OsDREBx, a gene encoding it, and the application of the gene in plant drought resistance, salt tolerance, low temperature tolerance and other stress resistance. It belongs to the field of biotechnology. Background technique [0002] Adversity conditions such as drought, low temperature, and high salinity are currently the main non-biological limiting factors that endanger crops, which seriously affect crop growth, yield and quality. In particular, my country's crop production areas are mostly distributed in different latitudes and different altitudes, and various natural disasters (such as drought, low temperature, etc.) occur frequently, often causing large-scale crop production declines. According to statistics, the average annual drought area in my country is about 21.59 million hectares, and 10 billion kilograms of grain is lost every year due to drought. Suitable temperature is one of the ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00
Inventor 徐正君江斌李利华高晓玲徐永菊朱建清邓晓建王平荣黄静
Owner SICHUAN AGRI UNIV