大豆GmCLC-d1/d2基因参与盐胁迫适应过程的生理功能  

Physiological function of soybean GmCLC-d1/d2gene involved in salt stress adaptation

作　　者：张露 黄巧玥 程聪 刘询 於丙军[1] ZHANG Lu;HUANG Qiaoyue;CHENG Cong;LIU Xun;YU Bingjun(Lab of Plant Stress Biology,College of Life Sciences,Nanjing Agricultural University,Nanjing 210095,China;College of Life Sciences,Xinjang Agricultural University,Urumqi 830052,China)

机构地区：[1]南京农业大学生命科学学院植物逆境生物学实验室,江苏南京210095 [2]新疆农业大学生命科学学院,新疆乌鲁木齐830052

出　　处：《南京农业大学学报》2023年第6期1084-1095,共12页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金项目(31671604);江苏省农业科技自主创新项目[CX(22)2003]。

摘　　要：[目的]本文旨在研究大豆GmCLC-d1/d2基因及其启动子对盐胁迫的响应,并探索其参与植物盐胁迫适应过程的生理机制。[方法]从大豆品种‘Lee68’中克隆了GmCLC-d1/d2基因,分别构建其植株RNA干扰(RNAi)和过表达载体,通过发根农杆菌介导转化获得RNAi和过表达大豆发根组合植株,利用根癌农杆菌介导转化获得过表达拟南芥,分析和比较盐胁迫下植株的形态和生理指标变化。[结果]与转空载大豆发根组合植株相比,盐处理下RNAi-GmCLC-d1/d2植株鲜重、叶片相对含水量(RWC)和叶绿素含量,根和叶过氧化氢酶(CAT)、过氧化物酶(POD)和超氧化物歧化酶(SOD)活性明显下降,根、叶相对电解质渗漏率(REL)和丙二醛(MDA)含量以及根、茎、叶Cl^(-)含量明显上升,NO_(3)^(-)含量显著下降,导致根、茎和叶,特别是地上部Cl^(-)/NO_(3)^(-)值显著上升;而发根过表达大豆植株OE-GmCLC-d1/d2的上述指标表现相反,耐盐性有所增强。盐胁迫下过表达GmCLC-d1/d2拟南芥种子发芽率、幼苗根长以及3周龄植株鲜重、叶片RWC及叶绿素含量等均明显优于野生型,地上部REL和MDA含量显著降低,CAT、POD和SOD活性明显增强;根和地上部Cl^(-)含量明显降低,NO_(3)^(-)含量明显上升,导致根和地上部,特别是后者Cl^(-)/NO_(3)^(-)值显著下降。[结论]GmCLC-d1/d2可通过增强盐胁迫下NO_(3)^(-)在发根过表达大豆组合植株或过表达拟南芥化植株根部的吸收以及向地上部的转运和分配,并在一定程度上抑制Cl^(-)的吸收和转运,以维持植株特别是地上部较低的Cl^(-)/NO_(3)^(-)值,从而增强耐盐性,其中GmCLC-d2基因的效应更明显。[Objectives]This paper aimed to study the responses of soybean GmCLC-d1/d2 genes and their promoters to NaCl stress,and to explore their physiological functions in salt stress adaptation in plants.[Methods]GmCLC-d1/d2 genes were cloned from the soybean‘Lee68’cultivar,their plant RNA interference(RNAi)and overexpression vectors were constructed for obtaining RNAi-and overexpressed(OE)-GmCLC-d1/d2 soybean hairy root composite plants mediated by Agrobacterium rhizogenes and OE-GmCLC-d1/d2 Arabidopsis thaliana mediated by A.tumefaciens,respectively.The changes in plant morphological and physiological indexes under NaCl treatment were analyzed and compared.[Results]Compared with the empty vector-transformed soybean hairy root composite plants under NaCl stress,the plant fresh weight,leaf relative water content(RWC)and chlorophyll content,activities of catalase(CAT),peroxidase(POD)and superoxide dismutase(SOD)in roots and leaves of RNAi-GmCLC-d1/d2 soybean plants obviously decreased,the relative electrolytic leakage(REL)and malondialdehyde(MDA)content in roots and leaves,and Cl^(-)content in roots,stems and leaves apparently increased,but NO_(3)^(-) content significantly decreased,which resulted in Cl^(-)/NO_(3)^(-) ratios in roots,stems and leaves,particularly raised significantly in the shoots.However,the above indexes were opposite in OE-GmCLC-d1/d2 soybean hairy root composite plants,with the enhanced salt tolerance.The seed germination rate,seedling root length,and 3-week-old plant fresh weight,leaf RWC and chlorophyll content of OE-GmCLC-d1/d2 A.thaliana were clearly better than those of wild type(WT)under salt stress,and additionally,REL and MDA content in shoots significantly decreased,the activities of CAT,POD and SOD were improved,while Cl^(-)contents in roots and shoots decreased,and NO_(3)^(-) content obviously increased,which resulted in a significant drop of Cl^(-)/NO_(3)^(-) ratio in roots and shoots,especially in the latter.[Conclusions]GmCLC-d1/d2 genes could facilitate NO_(3)^(-) uptake in the roo

关 键 词：GmCLC-d1/d2基因 大豆发根过表达 过表达拟南芥 Cl^(-)/NO_(3)^(-)值 耐盐性 

分 类 号：Q945.78[生物学—植物学] S565.1[农业科学—作物学]