小麦胁迫相关蛋白基因TaSAP12-D的耐盐性分析  

Identification and Analysis of Salt-tolerance of Stress Associated Protein Gene(TaSAP12-D)From Wheat

作　　者：王亦学 郝曜山 张欢欢 董艳辉 王晓清 吴慎杰 WANG Yixue;HAO Yaoshan;ZHANG Huanhuan;DONG Yanhui;WANG Xiaoqing;WU Shenjie(College of Life Sciences,Shanxi Agricultural University,Taiyuan,Shanxi 030031)

机构地区：[1]山西农业大学生命科学学院,山西太原030031

出　　处：《核农学报》2023年第1期42-50,共9页Journal of Nuclear Agricultural Sciences

基　　金：山西省重点研发计划项目(201903D221079);山西省农业科学院博士基金(YBSJJ2005)。

摘　　要：胁迫相关蛋白(SAPs)是一类具有A20/AN1锌指结构域的蛋白,在植物中主要参与逆境胁迫响应。为探究小麦胁迫相关蛋白基因TaSAP12-D在耐盐胁迫中的功能,本研究以小麦品种旱选10号为试材,克隆得到TaSAP12-D基因,利用农杆菌瞬时注射烟草叶片进行亚细胞定位,利用实时荧光定量PCR(qRT-PCR)进行不同组织和盐胁迫条件下的表达模式分析,利用蘸花法将TaSAP12-D转化到拟南芥(Arabidopsis thaliana L.)中并进行耐盐性分析。结果表明,TaSAP12-D基因全长519 bp,编码172个氨基酸,预测蛋白分子量为18.41 kDa,等电点为9.21。亚细胞定位显示,TaSAP12-D在烟草细胞核和细胞质中均有表达。qRT-PCR结果显示,TaSAP12-D在小麦萌发期和幼苗期的胚芽、根和叶中均有表达,其中在幼苗期的叶中表达量最高。在盐胁迫条件下,TaSAP12-D的表达量显著上调。在150 mmol·L^(-1) NaCl处理条件下,过表达TaSAP12-D拟南芥的存活率显著提高,表明TaSAP12-D可以增强转基因拟南芥的耐盐性。另外,在转基因拟南芥中盐胁迫相关应答基因AtP5CS1、AtRD29A、AtLEA、AtSOS1、AtNHX1和AtHKT的表达量均显著上调,推测TaSAP12-D通过调控上述盐胁迫响应基因的表达来增强拟南芥的耐盐性。本研究解析了TaSAP12-D基因在应答盐胁迫的分子调控机制,为改良作物耐盐性提供了重要的候选基因。Stress associated proteins(SAPs)are a group of A20/AN1 zinc-finger domain-containing proteins which are mainly involved in response to abiotic stresses in plants.In order to explore the function of TaSAP12-D in response to salt stress,a stress associated protein gene designated TaSAP12-D was cloned from wheat(Trticum aestivum L.)cultivar Hanxuan 10.The TaSAP12-D::GFP fusion protein construct was transferred into tobacco leaves through Agrobacterium tumefaciens mediated transfromation for subcellular localization.Quantitative real-time PCR was performed to analyze the expression patterns consisting of different tissues and leaves with NaCl treatment.To identify and analysis of salt-tolerance of TaSAP12-D,transgenic Arabidopsis plants were generated by floral infiltration.The results indicated that the full-length sequence of TaSAP12-D gene was 519 bp,encoding a 172-amino acid protein.The predicted relative molecular weight of the protein was 18.41 kDa and the isoelectric point was 9.21.Subcellular localization showed that TaSAP12-D was located in the nucleus and cytoplasm in tobacco leaf cells.The expression of TaSAP12-D was detected in different tissues,including plumule,root and leaf at the germination and seedling stages,and the highest expression occurred in the leaf tissues at the seedling stage.Furthermore,the transcript levels of TaSAP12-D were inducible by salt stress.Under treatment of 150 mmol·L^(-1) NaCl,the survival rate of transgenic Arabidopsis plants was significantly higher than that of control.Overexpression of TaSAP12-D in Arabidopsis results in enhanced salt tolerance.In addition,the expression of salt stress-related genes(AtP5CS1,AtRD29A,AtLEA,AtSOS1,AtNHX1 and AtHKT)was significantly upregulated in transgenic Arabidopsis plants.It is speculated that TaSAP12-D can improve tolerance to salt stress by regulating the expression of salt stress-related genes.The results explored the molecular modulation mechanism of TaSAP12-D in response to salt stress,and provided candidate gene for improving salt

关 键 词：小麦 TaSAP12-D 非生物胁迫 耐盐性 

分 类 号：S512.1[农业科学—作物学]