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Cloning of Salinity Stress-related Gene in Triticale and Its Function Verification

Author: WangBaiYu
Tutor: CaoLianPu；KongGuangChao
School: Shihezi University
Course: Crop Genetics and Breeding
Keywords: Triticale CA gene Salinity Clone
CLC: S512.4
Type: Master's thesis
Year: 2010
Downloads: 95
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Abstract


OBJECTIVE: land salinization is seriously affected agricultural ecological environment and sustainable development is an important factor limiting agricultural production. Salinization of arable land planted crops showed a substantial reduction or even death. Currently, cultivate excellent salinity crop varieties is one way to solve this problem. Therefore, the application of molecular biology and genetic engineering to cultivate the new Saline crop varieties has become an important part of modern agricultural research. Methods: The study mainly study materials hexaploid triticale varieties, triticale cDNA-AFLP technique, and as a basis for screening to salt stress gene fragments amplified using RACE technology to carbonic anhydrase enzyme full-length cDNA of the gene and protein expression by semi-quantitative RT-PCR and enzyme activity analysis techniques, analysis, and use of Agrobacterium-mediated transformation of Arabidopsis the gene's function. Results: 1. Using cDNA-AFLP method using 64 pairs of primers screened six closely related to salt tolerance gene fragment from the the small rye salt stress with different salinity performance before and after the materials, such as phosphorylation of serine decarboxylation The enzyme, carbonic anhydrase, receptor-like kinase, optical systems II32kDa protein, light system I hydrophobic protein (PSI), the stress response protein differential cDNA fragment using cDNA-AFLP technique for the first time in triticale breeding. 2 to No. 42, carbonic anhydrase cDNA fragment was amplified by RACE technique to the entire length of the the Triticale carbonic anhydrase (CA) cDNA, its sequence length is 1114bp, gene BANK Registry Number: GU145385, wherein the 5 'non- The coding region of 24bp, and the open reading frame of 778bp, 3 'non-coding region of 310bp, encoding 266 amino acids. The triticale the CA cDNA full-length sequence homology in the NCBI, compared with barley β-CA gene (accession number: L36959.1) 92% homology. Clustal x software of its protein structure prediction, the the CA gene encoding the protein has a zinc atom binding residues typical β-CA gene family, initially conclude that the gene for the the the triticale β-CA. 3. Use of activity analysis and semi-quantitative RT-PCR technology, the of triticale CA gene in the expression of the protein and molecular levels. The results showed that the CA activity in the seedling of triticale stress response to salt stress. CA activity of the response to salt stress there are also significant differences in the salt tolerance of different varieties of triticale, triticale CA is a salt stress response upregulated genes. Initially established system of CA gene Agrobacterium-mediated transformation of Arabidopsis smooth T0 generation plants, laid the foundation for the later further study its function. Conclusion: This study established the of hexaploid Triticale Salt cloning of cDNA-AFLP technique, Triticale carbonic anhydrase (CA) length cDNA was cloned into, and analysis by the expression of the enzyme protein and RNA levels showed that the gene for a salt stress response genes, candidate genes and theoretical basis of this study for the molecular breeding of triticale salinity.

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CLC: > Agricultural Sciences > Crop > Cereal crops > Wheat > Triticale
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