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Plant stress-tolerance-associated protein TaDREB3A and coding gene and application thereof

A stress tolerance and gene technology, applied in the direction of plant genetic improvement, botanical equipment and methods, application, etc., can solve problems such as comprehensive improvement of stress resistance of difficult plants, and achieve the effect of improving drought resistance.

Inactive Publication Date: 2011-11-09
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the stress tolerance of plants is a complex trait regulated by multiple genes, it is difficult to achieve comprehensive improvement of plant stress resistance by introducing a single functional protein gene

Method used

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  • Plant stress-tolerance-associated protein TaDREB3A and coding gene and application thereof
  • Plant stress-tolerance-associated protein TaDREB3A and coding gene and application thereof
  • Plant stress-tolerance-associated protein TaDREB3A and coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1. Cloning of TaDREB3A

[0045] 1. Isolation of mRNA

[0046] The three-leaf stage seedlings of trifoliate wheat (National Germplasm Resource Bank, No. ZM242) grown in hydroponics for about 10 days were subjected to drought treatment for 2 hours, quick-frozen with liquid nitrogen, and stored at -80°C for later use. mRNA isolation was performed using the Quikprep Micro mRNA Purification Kit (Pharmacia).

[0047] 2. Construction and titer determination of cDNA library

[0048] 1. Construction of cDNA library

[0049] Using Timesaver TM cDNA Synthesis Kit (Pharmacia) synthesized the mRNA obtained in step 1 into cDNA double-stranded, and added EcoRI / NotI adaptor; ZAP Express was used Predigested Gigapack IIIGold Cloning Kit (Stratagene) was used to construct the cDNA library, and a total of 500ul library solution was obtained.

[0050] 2. Determination of titer

[0051] (1) Take 1ul of library solution and dilute it 1000 times with SM Buffer;

[0052] (2)...

Embodiment 2

[0128] Example 2. Analysis of the expression characteristics of TaDREB3A by real-time fluorescence quantitative PCR

[0129] 1. Carry out various coercive treatments

[0130] The wheat seedlings with a seedling age of 10 days were subjected to the following treatments

[0131] (1) Drought treatment: take out the water on the roots of the hydroponic wheat seedlings, place them on dry filter paper, and take out after 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, and 24 hours of drought culture. The materials were quick-frozen in liquid nitrogen and stored at -80°C for later use.

[0132] (2) Salting treatment: The wheat seedlings were placed in 2% NaCl and Na 2 SO 4 composed of sodium salt solution (NaCl and Na 2 SO 4 The mass percentage of 3:2), the materials were taken out after 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, and 24 hours of light culture, and were quick-frozen with liquid nitrogen and stored at -80°C for later use.

[0133] (3) Absci...

Embodiment 3

[0152] Example 3. Activation properties of TaDREB3A

[0153] The main rationale for demonstrating the activation properties of transcription factors using the yeast one-hybrid system is as follows image 3 As shown, the DRE cis-acting element and the mutant DRE cis-acting element were constructed to the upstream of the basic promoter Pmin (minimal promoter) of pHISi-1 vector and pLacZi vector, respectively, and the downstream of the Pmin promoter was connected to reporter genes (His3, LacZ and URA3). When the expression vector YEP-GAP (without activation function) linked with the target gene encoding the transcription factor is transformed into yeast cells linked with the DRE cis-acting element and the mutant DRE cis-acting element, if the mutant DRE is linked The reporter gene in yeast cells with a cis-acting element cannot be expressed, but the reporter gene in yeast cells with a specific DRE cis-acting element can be expressed, indicating that the transcription factor can ...

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Abstract

The invention discloses a plant stress-tolerance-associated protein TaDREB3A and a coding gene and application thereof. The protein provided by the invention is one of the following proteins: (a) a protein consisting of an amino acid sequence shown as a sequence 1 in a sequence list, and (b) a protein which is associated with plant stress tolerance and is derived from the sequence 1 by substituting and / or deleting and / or adding one or several amino acid residues for the amino acid sequence shown in the sequence 1. The TaDREB3A provided by the invention can express under the induction of drought, high salt, high temperature, low temperature, pathogenic bacteria, ABA (Abscisic Acid), ethylene, JA (Jasmonic Acid) and SA (Salicylic Acid), and can specifically regulate and control transcription expression of the gene containing a DRE / CRT (Dehydration-Responsive Element / C-Repeat) cis-element (core sequence: CCGAC); therefore, the drought resistance, the salt tolerance, the high temperature resistance and the resistance to the powdery mildew pathogen of the plant are improved. The TaDREB3A provided by the invention provides a base for artificially controlling the expression of genes related to the stress tolerance and the stress resistance, and is about to play an important role in cultivation of plants with enhanced stress tolerance and enhanced stress resistance.

Description

technical field [0001] The invention relates to a plant stress tolerance-related protein TaDREB3A, its coding gene and application. Background technique [0002] Adversity stresses such as drought, high salinity and low temperature are obstacles to the growth and development of wheat. Therefore, understanding the response and signal transduction mechanism of wheat to stress conditions and improving the stress resistance of wheat varieties have become one of the important tasks of wheat genetic research and wheat variety improvement. Under adversity stress, plants will produce a series of responses, accompanied by many physiological, biochemical and developmental changes. Clarifying the response mechanism of plants to stress will provide scientific evidence for the research and application of stress-resistant genetic engineering. At present, the research on plant stress resistance has gradually penetrated into the cellular and molecular levels, combined with genetics and ge...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21A01H5/00
CPCY02A40/135
Inventor 马有志徐兆师李连城陈明李文滨
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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