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作 者 ：马静1；曲春浦2；许志茹1,2*；杨传平2；刘关君2

期 刊 ：植物研究 2015年 35卷 2期 页码：208-217

关键词：铜伴侣蛋白基因；铜锌超氧化物歧化酶基因；双价表达载体；耐盐性；

Keywords:PsATX, PsSOD, dual-gene expression vector, salt tolerance,

摘 要 ：为验证西伯利亚蓼铜伴侣蛋白基因（PsATX）和铜锌超氧化物歧化酶基因（PsSOD）共转化的烟草是否具有耐盐功能，分别构建了植物单价表达载体pROKⅡ-PsATX、pROKⅡ-PsSOD和双价表达载体pROKⅡ-PsATX-PsSOD，通过农杆菌介导遗传转化烟草，PCR验证转基因植株后测定了盐胁迫条件下转基因烟草的丙二醛（MDA）、活性氧、脯氨酸含量及超氧化物歧化酶（SOD）活性，以此鉴定转基因植株的耐盐性。结果表明：（1）PsATX和PsSOD基因已成功整合到烟草基因组中；（2）在100 mmol·L^-1 NaCl处理条件下，转双价基因植株的MDA、活性氧、脯氨酸含量均比野生型和转单价基因植株低，而SOD活性则高于这两类植株。证明PsATX和PsSOD基因共转化的烟草植株比转单价基因植株具有更强的耐NaCl能力。本研究将为研究PsATX和PsSOD基因的抗逆机制奠定分子基础。

Abstract:We analyzed the salt tolerance of tobacco plants which expressed both PsATX and PsSOD genes of Polygonum sibiricum Laxm., constructed univalent expression vector pROKⅡ-PsATX, pROKⅡ-PsSOD and dual-gene expression vector pROKⅡ-PsATX-PsSOD respectively, and then transferred them into tobacco mediated by Agrobacterium tumefaciens. The transgenic tobacco were identified by PCR method. We measured the content of malondialdehyde(MDA), reactive oxygen, proline(Pro), and the activity of superoxide dismutase(SOD) in the transgenic tobacco under salt stress condition to identify the salt tolerance of these plants. The transgenic tobacco containing PsATX, PsSOD and PsATX+PsSOD genes respectively were obtained successfully. The integration of the foreign PsATX and PsSOD genes were confirmed by molecular identification. Under the treatment of 100 mmol·L^-1 NaCl solution, the content of MDA, reactive oxygen and proline in the co-transformed plants was lower than that of wild type and univalent transgenic plants, but the SOD activity were higher than them. By the salt test, the transgenic tobacco contained PsATX+PsSOD genes were more tolerant to salt stress than the transgenic tobacco contained PsATX or PsSOD genes.

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