Thaumatin-like protein related to wheat leaf rust resistance and encoding gene and application thereof

A sweet-like protein, resistance-related technology, applied to sweet-like protein related to wheat leaf rust resistance and its encoding gene and application field, can solve the problem of the small number of defense genes and limit the research and resistance of wheat leaf rust resistance mechanism Resource reserve utilization, complex structure, etc.

Inactive Publication Date: 2015-08-26
HEBEI AGRICULTURAL UNIV.
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

[0004]Because the wheat genome is relatively large (1.6×109 bp) and complex in structure, the number of defense genes directly cloned from it is small

Method used

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  • Thaumatin-like protein related to wheat leaf rust resistance and encoding gene and application thereof
  • Thaumatin-like protein related to wheat leaf rust resistance and encoding gene and application thereof
  • Thaumatin-like protein related to wheat leaf rust resistance and encoding gene and application thereof

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Embodiment Construction

[0044] 25 μl PCR reaction system: 100 ng template cDNA or gDNA, 10 μmol / L primer 1 μL, 10 mmol / L dNTP 0.5 μL, 1U Taq DNA polymerase 0.3 μL, 10×PCR buffer 2.5 μL, add sterilized ddH 2 0 to 25 μL.

[0045] The PCR reaction program was: pre-denaturation at 94°C for 1 min; denaturation at 94°C for 30 s, annealing at 62°C for 1 min, extension at 72°C for 2 min, 35 cycles; extension at 72°C for 10 min.

[0046] The PCR amplification product was recovered by gel and connected with PGM-T vector and transformed into Escherichia coli DH5α, positive clones were identified by colony PCR amplification supplemented by blue-white screening, and the single colony shaker of positive clones was sent to Shanghai Sangon Company for sequencing.

[0047] Using the online software of bioinformatics in DNAMAN and NCBI database to analyze TcLr19PR5 Genes are analyzed. Composition analysis, physical and chemical properties analysis, open reading frame search and translation of nucleic acid and am...

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Abstract

The invention discloses a thaumatin-like protein gene TcLr19PR5 related to wheat leaf rust resistance and a gene encoded through the thaumatin-like protein gene TcLr19PR5. Through RT-PCR and the electronic cloning technology, a 516bp thaumatin-like protein gene is cloned, and 171 amino acid residues are coded; and real-time fluorescence quantification PCR analysis shows that at different time points after puccinia triticina is inoculated, the expression quantity of the gene in non-affine combinations is higher than that of the gene in affine combinations, it is indicated that expressions are induced through the wheat puccinia triticina, the expression amount of stems is higher than the expressions of the leaves and the roots, and meanwhile it is clear that the expressions are induced by adversity signal molecules of abscisic acid (ABA) and salicylic acid (SA). According to the thaumatin-like protein gene and the gene, basic data are provided for in-depth exploration of wheat and puccinia triticina interaction molecular mechanisms, and the theory basis is provided for cultivating leaf-rust-resistance wheat varieties.

Description

technical field [0001] The invention relates to a sweet-like protein related to wheat leaf rust resistance, its encoding gene and application. Background technique [0002] wheat( Triticum aestivum Linn.), as an important food crop, occupies an important position in the food industry and agricultural production (Cao Yaping, 2008). Wheat leaf rust is one of the most important diseases affecting wheat yield and quality (Bolton et al., 2008). In recent years, plant genetic engineering technology has developed rapidly, and the molecular breeding approach centered on the cloning and transformation of disease resistance genes and various resistance-related genes has become a new direction of breeding. [0003] Disease process-related proteins refer to a class of proteins encoded by host plants and induced in pathological or pathologically related environments (Van et al., 1999). According to the classification system based on plant origin, electrophoretic mobility (molecular w...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/56C12N9/24
Inventor 王海燕刘大群高琳
Owner HEBEI AGRICULTURAL UNIV.
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