Genetic Transformation Of TaPrx Gene In Wheat And Identification Of Its Drought And Salt Tolerance

Wheat can produce and accumulate different types of reactive oxygen species(ROS) which was harmful and even could make plants to death when under drought, salinity, high temperature and other abiotic stress environments. Peroxidase(Prx), however, can protect plants’ from suffering of stree environment and increase the plants’ resistance to abiotic stress by removing ROS. In this study, we cloned a TaPrx gene from wheat cultivar Apogee73S2 and built two related expression vectors in the mean while. Common wheat cultivar “Kenong 199” was used for genetic transformation which is mediated by genegun and two transgenic plants were obtained after biolaphos screening and PCR detection. After the homozygous plants were gotten, we identified the drought and salt tolerance of all transgenic plants. Meanwhile we used tobacco and onion to ensure the subcellular localization of TaPrx. Main results were shown as belows:1. Construction of relative expression vectors: TaPrx gene was cloned into two separate vector p BIN-eGFP which was a subcellular localization vector and PC186 which was genegun transgenic vector, and we obtained the subcellular localization vector pBIN-Prx-eGFP and the genegun transgenic vector PC186-Bar-Hpt-Prx.2. Transgenic of TaPrx using genegun method: 2082 calli induced from immature embryos(cultivar Kenong 199) were obtained by tissue culture. 174 regenerating plants were obtained after screening all of the plants generated by genegun transformation using biolaphos, among them 2 postive plants were detected by PCR. RNAs of transgenic acceptor Kenong199 and homozygous transgenic plants were extracted for qRT-PCR testing. It was found that the expression level of TaPrx in transgenic plants was ?? times higher than in Kenong199.3. Functional verification of draught and salinity resistance of transgenic plants: It was found that transgenic plants showed significant resistance to drought and salinity compared with Kenong199.4. Subcellular localization analysis of TaPrx: Tobacco leaves and onion epidemic cells were used expectively to make sure the subcellular localization of TaPrx and we found that TaPrx was localized in cell nucleus.