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The Function Characterization of Salt-tolerant Correlative Gene TaRSP and TA-pSRP in Wheat

Author: ZhangZuo
Tutor: HuangZhanJing；GeRongChao
School: Hebei Normal
Course: Genetics
Keywords: Wheat TaRSP gene The TA-pSRP gene Salt tolerance Arabidopsis GFP
CLC: S512.1
Type: Master's thesis
Year: 2008
Downloads: 47
Quote: 0
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Abstract

Currently, soil salinization is a global problem, restricting agricultural production The salt damage seriously affect plant growth and development, resulting in crop production. Improve crop salt tolerance has become the urgent need to address the key issues of modern plant breeding work, which GM provides more possibilities for artificial improvement of crop. The field of genetic engineering by changing the expression levels of the gene of the plant itself or the introduction of exogenous salt tolerance gene to the artificial improvement of the crop. Plant salt tolerance gene cloning, identification, and use them, has become a hot research. This paper is to study the two genes is related to salt tolerance. We speculate TaRSP (Triticum asetivum Root Specific Protein) gene according to the part of the online information and TA-pSRP (Triticum asetivum putative syntaxin-related protein) gene in wheat Tolerance Gene. On the basis of the work of previous studies, we identified its salt-tolerant features, in order for wheat salt tolerance to carry out some useful exploration. The gene TaRSP containing a complete open reading frame of 381bp, encoding 126 amino acids. The bioinformatics projections indicate that the conserved domain of the the TaRSP gene containing a HPS_like of. Quantitative PCR results show that, the gene TaRSP gene expression in the roots, and the roots of wheat in the salt stress TaRSP gene expression appears enhanced. In turn can initially speculated that the gene is related to salt tolerance gene. To the TaRSP protein subcellular localization, the TaRSP gene and GFP gene construct fusion expression vector, transformed Arabidopsis screened homozygous transgenic plants. Fluorescence root cells of Arabidopsis thaliana homozygous transgenic plants confocal observation, the results show that TaRSP homozygous transgenic plants are mainly located in the cell membrane and the nucleus, the fluorescence of the control plants transferred to the GFP gene is distributed in the cytoplasm and nucleus and the parts of the cell membrane, suggesting that the gene product TaRSP mainly located in the cell membrane and the nucleus. In order to further determine the TaRSP salt tolerance gene function, we constructed plant expression binary vector, transformed Arabidopsis plants by dip flower of salt tolerance in the obtained single copy transgenic plants under salt stress, respectively, the transgenic seedling root growth as well as the complete plant salt tolerance. were detected. The results show that in the presence of 200 mM NaCl MS medium, transgenic Arabidopsis plant growth status and survival rate was significantly better than the control plants, and the TaRSP gene overexpression can significantly enhance the normal growth of the transgenic plants under salt stress seedling root, thus improving overall plant salt tolerance. Therefore, speculate TaRSP gene plays a very important role in plant salt tolerance. In order to determine role of TaRSP abiotic stress signaling pathway, we have known some of the genes in the Arabidopsis stress signaling pathways quantitative PCR analysis to determine its location in the signal path. In transgenic Arabidopsis over expression TaRSP gene, ADH gene expression was significantly increased, RD29B KIN2, FRY1 SAD1 gene expression is also up-regulated, but did not change significantly reduce the rest of the gene expression in varying degrees. So we may speculate TaRSP gene upstream of the ADH gene. 2. TA-pSRP gene contains a complete open reading frame of 909bp, encoding 302 amino acids. The bioinformatics prediction that TA-pSRP gene containing a SynN conservative domain and a t_SNARE of domain. NLS prediction on the TA-pSRP gene encoding protein found mainly localized in the Golgi. Quantitative PCR results showed that by NaCl, PEG treatment, the wheat roots the TA-pSRP gene expression is upregulated. In turn can initially speculated that the gene is related to salt tolerance gene. In order to further determine the salt tolerance of the TA-pSRP gene function, we constructed a plant expression binary vector, by dip flower Law into Arabidopsis plants, homozygote plants of salt tolerance to salt stress conditions, on seedling root growth and whole plant salt tolerance of transgenic plants were detected. The results showed that the transgenic Arabidopsis plant growth status and survival rate was significantly better than the control plants, containing 200 mM NaCl MS medium, and the TA-pSRP gene overexpression can significantly enhanced the normal growth of the transgenic plants under salt stress seedling root, and to improve overall plant salt tolerance. Therefore, it is speculated that the TA-pSRP gene plays a very important role in plant salt tolerance. In order to determine the role of of TA-pSRP abiotic stress signaling pathways, we have known some of the genes in the Arabidopsis stress signaling pathways quantitative PCR analysis to determine its location in the signal path. ADH in overexpression the TA-pSRP transgenic Arabidopsis, cor15a, FRY1 and SAD1 gene expression significantly increased, RD29B, KIN2 gene expression is also up-regulated, but did not change significantly, reduce the rest of the gene expression in varying degrees. Therefore, we hypothesized that the TA-pSRP gene may COR15a, ADH, FRY1, and SAD1 gene upstream by the the phospholipase pathway and CDPK way of participation in regulation of osmotic stress.

The Function Characterization of Salt-tolerant Correlative Gene TaRSP and TA-pSRP in Wheat

Abstract

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