Luo Xiaoli, Xiao Juanli, Wang Zhian, Zhang Anhong, Tian Yingchuan, Wu Jiahe
Institute of Cotton Research, Shanxi Academy of Agricultural Science, Yuncheng 044000, China.
Sheng Wu Gong Cheng Xue Bao. 2008 Aug;24(8):1464-9.
The open reading frame of Spinacia oleracea Betaine Aldehyde Dehydrogenase (SoBADH) was retrieved from Spinacia oleracea and inserted into the Agrobacterium tumefaciens binary vector pBin438, which was driven by CaMV35S promoter, and produced the new binary vector pBSB. A. tumefaciens LBA4404 carrying this plasmid was used in genetic transformation of plants. Forty-five primary transgenic plants were detected by PCR and verified by the Southern blotting from 65 regenerated plants, of which 27 transgenic plants had only one copy of T-DNA. The Northern blotting and Western blotting analysis indicated that the SoBADH gene had been transcribed mRNA and expression protein in the transgenic cotton lines. The testing of SoBADH activity of transgenic plant leaves showed that the enzyme activity was much higher than that of the non-transgenic cotton. The growth of transgenic plants was well under the salinity and freezing stress, whereas the non-transgenic plant grew poorly and even died. Challenging with salinity, the height and fresh weight of transgenic plants was higher compared with those of non-transgenic plants. Under the freezing stress, the relative conductivity of leaf electrolyte leakage of the transgenic cotton lines was lower than that of non-transgenic plants. These results demonstrated that the SoBADH gene could over express in the exogenous plants, and could be used in genetic engineering for cotton stress resistance.
从菠菜中获取菠菜甜菜碱醛脱氢酶(SoBADH)的开放阅读框,并将其插入由花椰菜花叶病毒35S启动子驱动的根癌农杆菌双元载体pBin438中,构建了新的双元载体pBSB。携带该质粒的根癌农杆菌LBA4404用于植物的遗传转化。通过PCR从65株再生植株中检测到45株初级转基因植株,并通过Southern杂交进行了验证,其中27株转基因植株仅含有一个T-DNA拷贝。Northern杂交和Western杂交分析表明,SoBADH基因在转基因棉花品系中已转录成mRNA并表达了蛋白。转基因植株叶片的SoBADH活性检测表明,其酶活性远高于非转基因棉花。在盐胁迫和冻害胁迫下,转基因植株生长良好,而非转基因植株生长不良甚至死亡。在盐胁迫下,转基因植株的株高和鲜重均高于非转基因植株。在冻害胁迫下,转基因棉花品系叶片电解质渗漏的相对电导率低于非转基因植株。这些结果表明,SoBADH基因能够在外源植物中过量表达,可用于棉花抗逆性的基因工程。