小麦TaGPX8基因的克隆与表达分析  

Cloning and Expression Analysis of TaGPX8 Gene in Wheat(Triticum aestivum)

作　　者：张华东 李雅倩 董飞燕 丁芳草 刘孟伟 许嘉盛 高春保[1,2] 王晓玲[1] 刘易科[2] 方正武[1] ZHANG Hua-Dong;LI Ya-Qian;DONG Fei-Yan;DING Fang-Cao;LIU Meng-Wei;XU Jia-Sheng;GAO Chun-Bao;WANG Xiao-Ling;LIU Yi-Ke;FANG Zheng-Wu(College of Agronomy/MARA Key Laboratory of Sustainable Crop Production in the Middle Reaches of the Yangtze River(Co-construction by Ministry and Province),Yangtze University,Jingzhou 434025,China;Food Crops Institute/Hubei Provincial Key Laboratory of Germplasm Innovation and Genetic Improvement of Food Crops/Wheat Disease Biology Research Station on Central China,Ministry of Agriculture and Rural Affairs,Hubei Academy of Agricultural Sciences,Wuhan 430064,China;Wuhan University of Bioengineering,Wuhan 430403,China)

机构地区：[1]长江大学农学院/农业农村部长江中游作物绿色高效生产重点实验室(部省共建),荆州434025 [2]湖北省农业科学院粮食作物研究所/粮食作物种质创新与遗传改良湖北省重点实验室/农业农村部华中地区小麦病害生物学科学观测实验站,武汉430064 [3]武汉生物工程学院,武汉430403

出　　处：《农业生物技术学报》2023年第2期232-241,共10页Journal of Agricultural Biotechnology

基　　金：湖北省自然科学基金(2021CFB393);湖北省中央引导地方科技发展专项(2020ZYYD011);国家小麦产业技术体系建设专项(CARS-03)。

摘　　要：谷胱甘肽过氧化物酶(glutathione peroxidase, GPX)作为植物最主要的抗氧化酶之一,在植物逆境胁迫应答中发挥着重要作用。本课题组前期对小麦(Triticum aestivum) GPX基因家族的转录组数据进行分析时发现,TaGPX8(IWGSC数据库登录号:TraesCS4D02G162000.1)基因具有受逆境诱导表达的特性,本研究对TaGPX8进行了克隆,并对其编码蛋白进行了生物信息学分析和亚细胞定位,同时对盐和干旱胁迫下根和叶中TaGPX8的表达进行了分析。基因结构分析显示,TaGPX8全长14 754 bp,包含6个外显子和5个内含子,CDS全长564 bp,编码187个氨基酸残基,编码蛋白相对分子质量是21.330 kD,理论等电点为6.62,亲水性平均值为-0.664;启动子序列分析发现TaGPX8的启动子序列中含有多个参与激素反应、光反应和响应胁迫的顺式作用元件;qPCR结果显示,TaGPX8基因响应干旱胁迫和盐胁迫,干旱胁迫12 h时TaGPX8在叶片中的表达量与对照无显著差异,干旱胁迫24 h时被显著诱导,表达量上调,为对照的12倍,48 h时表达量降低,与对照无显著差异,且干旱胁迫48 h时在根部的表达也有所上升;TaGPX8在盐胁迫12 h叶片中表达受到一定程度抑制,但在处理24 h表达量升高,之后又降低与对照无显著差异,而在根中的表达受到抑制。酵母转录激活活性分析显示TaGPX8基因没有转录自激活活性。亚细胞定位显示TaGPX8定位于细胞核和细胞膜。本研究为深入探究TaGPX8基因的功能提供了参考。Glutathione peroxidase(GPX), as one of the most important antioxidant enzymes in plants, plays an important role in plant stress response. In the analysis of the transcriptome data of the wheat(Triticum aestivum) GPX gene family, it was found that the TaGPX8(IWGSC accession No. TraesCS4D02G162000.1) had the characteristics of being induced by stress. In this study, TaGPX8 was cloned, its protein was analyzed by bioinformatics and subcellular localized, and the expression of TaGPX8 in roots and leaves under salt and drought stress were analyzed.The results of gene structure analysis showed that TaGPX8 had a full length of 14 754 bp, including 6 exons and 5 introns, and the full length of CDS was 564 bp, encoding 187 amino acid residues. The relative molecular mass of the encoded protein was 21.330 kD, its theoretical isoelectric point was 6.62, and its average hydrophilicity was -0.664. Promoter sequence analysis found that the promoter sequence of TaGPX8 contained some cis-acting elements involved in hormone response, light response, and stress response. qPCR results revealed that TaGPX8 gene responded to drought and salt stress. Under drought stress, the expression level of TaGPX8 in leaves at 12 h had no significant difference with the control, but was significantly induced at 24 h, its expression level was up regulated, which was 12 times that of the control, and the expression level returned to no significant difference with the control in 48 h. In addition, the expression in roots also increased at 48 h of drought stress treatment. Under salt stress, the expression of TaGPX8 in leaves was inhibited to a certain extent at 12 h of treatment, but the expression level increased at 24 h of treatment,and then recovered to no significant difference with the control at 48 h. However, the expression in the root was suppressed. The yeast transcription activation experiments verified that TaGPX8 did not have transcriptional activation activity. Subcellular localization showed that the TaGPX8 protein was localized in th

关 键 词：小麦 TaGPX8 生物信息学分析 qPCR 自激活 亚细胞定位 

分 类 号：S512[农业科学—作物学] S188