北美海蓬子Na^+/H^+逆向运输蛋白基因的克隆及拟南芥转化分析

Cloning and Transgene Analysis of a Novel Na～+/H～+ Antiporter Gene from Salicornia Bigelovii

Na+/H+逆向运输蛋白在植物耐盐性方面起着极为重要的作用。根据在不同植物中功能相同的同源基因保守序列设计引物,通过RT-PCR方法从真盐生植物北美海蓬子(Salicornia Bigelovii Torr.)中克隆出包括有1683bp的完整编码区在内的2591bp(GenBank登陆号为DQ157454)的Na+/H+逆向运输蛋白基因(SbNHX1)cDNA序列。将该基因编码区序列构建到植物表达载体pVKH-35S-pA上,并通过农杆菌介导转化到模式植物拟南芥(Arabidopsis thaliana)中,得到T3代转入单基因种子。200mmol/L的NaCl胁迫培养结果显示,转基因拟南芥在200mmol/L的NaCl胁迫条件下生长状况好于野生型植株,植物表现有一定的耐盐性。

Na^＋/H^＋ antiporters play an important role in plant salt-tolerance. The primers were designed according to the conserved region of the homologous gene in other plants. The 2591bp cDNA in length was cloned by RT-PCR from Salicornia Bigelovii Tort., which includes a completed reading frame of 1 683 bp. The cDNA sequence was named as SbNHX1 and the accession number in GeneBank is DQ157454. The completed open reading frame of the gene was inserted to the plant expression vectors of pVKH-35S-pA, and then was transformed to A rabidopsis thaliana through Agrobacterium. The transgenic Arabidopsis were grown better than wild-type Arabidopsis in the MS medium with 200 mmol/L sodium chloride, or in soil watered with 200 mmol/L sodium chloride.