报春PF sHSP21.4基因转化拟南芥的获得及耐热鉴定

根据灰岩皱叶报春PF sHSP 21.4热激蛋白基因序列,采用PCR法从差减文库中克隆得到该基因,并构建pCAMBIA 130 1-PF sHSP21.4植物表达载体,通过根癌农杆菌介导的花器浸染法转化拟南芥,经潮霉素筛选、PCR和RT-PCR法鉴定后,对转基因植株和野生型对照进行高温胁迫,分析其耐热性差异.结果显示:...

Transformation of Primula PF sHSP 21.4 Gene and Heat Stress Tolerance Identification of Transgenic Arabidopsis thaliana

According to the sequence of PF sHSP 21.4 gene from Primula forrestii, PF sHSP 21.4 gene was cloned from a subtractive library by PCR, and the plant expression vector pCAMBIA1301 which harbored PF sHSP 21.4 gene was established, wild type Arabidopsis plants were transformed by Agrobacterium tumefaciens-mediated Floral Dip method, after hygromycin screening, PCR and RT-PCR detetion indicated PF sHSP 21.4 gene had been integrated into the genome. The performance of the transformed plants and the wild type under heat stress was analyzed. The results showed that, after 2 hours’ stress of 43℃, 44℃, the survival rate of transgenic plants was much higher than that of wild type. What’s more, the increase of relative conductance rate in the leaves of transgenic plants was much smaller than that in the leaves of wild-type, and the reduction of relative quantum yield of PSⅡ of transformed in Arabidopsis plants was lesser than that of wild control obviously, for the content of soluble protein, that in the transformed plants increased slight while the content in the wild type was decreased with the degree of stress. Consequently, the experimental results displayed that the PF sHSP 21.4 gene has a great value in improving the heat stress resistance in plants.