报春PF sHSP 17-1基因转化拟南芥及耐热鉴定

Transformation of Primula PF sHSP 17-1 Gene and Heat Stress Tolerance Identification of Transgenic Arabidopsis thaliana

根据灰岩皱叶报春PF sHSP 17-1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pCAMBIA1301-PF sHSP 17-1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,在40℃、42℃、44℃处理下,转基因拟南芥的相对电导率和丙二醛含量升高的幅度均显著低于野生型对照,而脯氨酸含量的积累明显高于对照,可溶性蛋白的含量在热胁迫下也比对照稳定,仅有轻微下降。试验证明,PF sHSP 17-1基因对植物的耐热性有重要作用。

According to the sequence of PF sHSP 17-1 gene from P.forrestii,PF sHSP 17-1 gene was cloned from a subtractive library by PCR,and the plant expression vector pCAMBIA1301 which harbored PF sHSP 17-1 gene was established,wild type Arabidopsis plants were transformed by Agrobacterium tumefaciens-mediated Floral Dip method,after hygromycin screening,PCR and RT-PCR detection indicated PF sHSP 17-1 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 40℃,42℃,44℃,the increase of relative conductance rate and malondianldehyde（MDA）in transgenic plants was much smaller than that in the wild-type,while the proline contents inclined more dramatically,and the content of soluble protein in the transformed plants demonstrated greater stability and resistance compared with that in the wild control.Consequently,the experimental results displayed that the PFsHSP17-1 gene has a great value in improving the heat stress resistance in plants.