Protein related to heat resistance property and coding gene and application thereof

A technology that encodes genes and proteins, used in applications, genetic engineering, plant genetic improvement, etc.

Inactive Publication Date: 2009-11-25
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In wheat, there are only reports about the expression of TaMBF1a gene being induced by pathogen inf...

Method used

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  • Protein related to heat resistance property and coding gene and application thereof
  • Protein related to heat resistance property and coding gene and application thereof
  • Protein related to heat resistance property and coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Cloning of embodiment 1, TaMBF1c gene

[0039] (1) Electronic cloning of wheat TaMBF1c gene sequence

[0040] Take Affymetrix The sequence with the probe number Ta.12225.1.S1_x_at in the wheat Genome array was used as the basic sequence, and ORF finder analysis was performed on the NCBI (www.ncbi.nlm.nih.gov) website, and it was found that the sequence already contained a complete ORF sequence. This sequence was used as the basis for designing gene-specific primers.

[0041] (2) Cloning of cDNA and untranslated region sequence of wheat TaMBF1c gene

[0042] Primers were designed according to the sequence obtained by electronic cloning, and the primer pair used was TaMBF1c-1: 5'-AAGACAGAGGCGAGGAGAAG-3' and TaMBF 1c-2: 5'-ACCACAAAGCAACCCGAGA-3'.

[0043]The experimental material is a common wheat variety TAM107 (Triticum aestivum L), that is, 5 minutes, then rinsed with distilled water 3 times, and germinated at room temperature for 24 hours. The seeds with consisten...

Embodiment 2

[0048] Example 2, Induced expression pattern of TaMBF1c gene under high temperature stress

[0049] Wheat seedlings grown under normal conditions for ten days as described above were subjected to the following heat stress treatment: (1) directly treated at 40°C for 0.25, 0.5, 1, 2, 12 and 24 hours, respectively named as 0.25h, 0.5h, 1h, 2h, 12h and 24h; (2) 34℃ heat exercise for 3 hours and 40℃ high temperature treatment for 0.25, 0.5, 1 and 2 hours, respectively named 0.25sh, 0.5sh, 1sh and 2sh; (3) After 3 hours of heat exercise at 34°C, the growth resumed under normal conditions for 0, 1, 2 and 12 hours, which were named 3s, 1sc, 2sc and 12sc, respectively, and the material without any high temperature treatment was named ck. The treated leaves were taken, and RNA was extracted for Northern hybridization. Use the primer pair TaMBF1c-1: 5'-AAGACAGAGGCGAGGAGAAG-3' and TaMBF1c-3: 5'-CGGAGGCATTCTTGTTCGTC-3' to amplify the fragment of the TaMBF1c gene and recover it, and use th...

Embodiment 3

[0051] Example 3, Induced expression pattern of TaMBF1c gene under other stresses

[0052] With 20% PEG-6000, 250mM NaCl, oxidative stress (100μM H 2 o 2 ) treatment, and plant hormones ACC (200 μM), ABA (100 μM), and MeJA (100 μM) were used to treat wheat seedlings respectively, and wheat leaves with different treatment times were taken to extract RNA for Northern hybridization. The probes and labeling methods used were the same as above. The specific methods of coercion treatment are as follows:

[0053] High-salt treatment: Add NaCl solution to the culture flask to a final concentration of 250mM.

[0054] PEG treatment: add PEG-6000 solution to the culture flask to a final concentration of 20% (mass percentage).

[0055] ABA treatment: ABA (analytical grade, purchased from Sigma-aldrich) was dissolved in DMSO to make a 10 mM stock solution, and then added to the culture flask to a final concentration of 100 uM.

[0056] ACC treatment: Dissolve ACC in water to make 10mM ...

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Abstract

The invention discloses a protein related to heat resistance property and coding gene and application thereof. The name of the protein related to plant heat resistance property provided by the invention is TaMBFlc (Triticum aestivum Multiprotein Binding Factorlc), the protein is originated from the common wheat (Triticum aestivum L.), is the protein of the following 1) or 2): 1) the protein composed of amino acid sequence shown by the sequence 2 in the sequence table; 2) the protein derived from the 1) related to the heat resistance property with amino-acid residue sequence of the sequence in the sequence table by the substitution and/or deletion and/or adding of one or several amino-acid residues. The protein TaMBFlc related to heat resistance property and coding gene provided by the invention has important theory and actual meanings for cultivating new species such as crops, forests and grasses with improved stress resistance, can be used for the cultivating and the identifying of stress resistance plant species required by the farming and animal husbandry and ecologic environment harnessing.

Description

technical field [0001] The present invention relates to a protein related to stress resistance and its coding gene and application, in particular to a protein related to heat resistance and its coding gene TaMBF1c and application. Background technique [0002] As the global population increases, the demand for food is increasing. As a major food crop in the world, the high temperature stress in the late growth period of wheat seriously affects its yield and quality, and has a great impact on people's lives. Therefore, we urgently need to study the mechanism of wheat heat resistance and develop heat resistance related genes and apply them to wheat breeding to produce heat-tolerant varieties. [0003] Multiprotein bridging factor 1 (MBF1), as a transcriptional co-activator, can activate the expression of downstream genes that depend on this transcription factor by linking TATA-Box Binding Protein (TBP) and a certain transcription factor. Although the MBF1 family genes were f...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/00C12N15/11C12N15/81
Inventor 孙其信秦丹丹彭惠茹倪中福姚颖垠梁荣奇解超杰刘志勇杜金昆
Owner CHINA AGRI UNIV
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