茶树硫酸盐转运蛋白基因CsSULTR3.1的克隆及其对硫和硒的响应分析

园艺学报 2018 45 2 321 332 Acta Horticulturae Sinica doi 10 16420 j issn 0513 353x 2017 0792 http www ahs ac cn 321 收稿日期 收稿日期 2017 11 14 修回日期 修回日期 2017 12 28 基金项目 基金项目 中央级科研院所基本科研业务费专项 1610212016001 国家现代农业产业技术体系建设专项资金项目 CARS 19 中国 农业科学院科技创新工程项目 CAAS ASTIP 2017 TRICAAS 通信作者 Author for correspondence E mail zengjm 茶树硫酸盐转运蛋白基因 CsSULTR3 1 的克隆 及其对硫和硒的响应分析 张晶晶 钱文俊 郝心愿 王 璐 丁长庆 姚利娜 王新超 曾建明 中国农业科学院茶叶研究所 国家茶树改良中心 农业部茶树生物学与资源利用重点实验室 杭州 310008 摘 要 摘 要 从茶树 Camellia sinensis 根系中克隆获得 1 条长度为 1 999 bp 的核酸序列 该序列包 含 1 980 bp 的开放阅读框 ORF 编码 659 个氨基酸 BlastX 同源性比对显示 该基因编码的氨基酸序 列与葡萄 VvSULTR3 1 相似度最高 86 将其命名为 CsSULTR3 1 GenBank 登录号 KY963793 进 一步分析显示 该基因编码的蛋白分子量为 72 39 kD 理论等电点为 7 61 属于非分泌性蛋白 包含 10 个跨膜结构域 以 GFP 作为标记进行亚细胞定位发现 该蛋白定位于细胞质中 qRT PCR 分析显示 CsSULTR3 1 具有组织表达特异性 在茶树茎中表达量最高 CsSULTR3 1 在茶树根系中的表达受 Na2SO4 和 Na2SeO4诱导 60 mg L 1 Na2SO4处理 12 h 后逐渐上调表达 在 48 h 时达到最大值 而 240 mg L 1 Na2SO4处理 12 h 内显著下调表达 随后显著上调并维持相对稳定状态 在不同浓度 Na2SeO4处理条件下 表达量均呈现先降低后升高再降低的趋势 且均在 48 h 时达到最高 关键词 关键词 茶树 硫酸盐转运蛋白 CsSULTR3 1 硫 硒 表达分析 中图分类号 中图分类号 S 571 1 文献标志码 文献标志码 A 文章编号 文章编号 0513 353X 2018 02 0321 12 Cloning and Expression Analysis of CsSULTR3 1 Implicated in Sulfate and Selenate Treatments in Tea Plant Camellia sinensis ZHANG Jingjing QIAN Wenjun HAO Xinyuan WANG Lu DING Changqing YAO Lina WANG Xinchao and ZENG Jianming Tea Research Institute of the Chinese Academy of Agricultural Sciences National Center for Tea Improvement Key Laboratory of Tea Biology and Resources Utilization Ministry of Agriculture Hangzhou 310008 China Abstract In this study a nucleotide sequence in length of 1 999 bp was cloned from tea plant Camellia sinensis termed as CsSULTR3 1 GenBank accession No KY963793 Bioinformatics analysis showed that the sequence contained 1 980 bp Open Reading Frame ORF and encoded 659 amino acid residues BlastX and phylogenetic analysis indicated that CsSULTR3 1 shared the highest identity 86 with SULTR3 1 in Vitis vinifera In addition CsSULTR3 1 was speculated as a non secretory protein with 10 transmembrane domains 72 39 kD molecular weight and 7 61 theoretical isoelectric point Subcellular localization using GFP assays validated that CsSULTR3 1 was located in Zhang Jingjing Qian Wenjun Hao Xinyuan Wang Lu Ding Changqing Yao Lina Wang Xinchao Zeng Jianming Cloning and expression analysis of CsSULTR3 1 implicated in sulfate and selenate treatments in tea plant Camellia sinensis 322 Acta Horticulturae Sinica 2018 45 2 321 332 cytoplasm Results of real time PCR analysis showed that CsSULTR3 1 could be detected in all tested tissues of tea plant and was highly expressed in stems Furthermore CsSULTR3 1 could be induced by the treatments of Na2SO4 and Na2SeO4 respectively Under 60 mg L 1 sulfate condition the expression of CsSULTR3 1 gradually raised after 12 h and reached maximum at 48 h However a significant expression decrease was detected within 12 h under 240 mg L 1 sulfate treatment and an expression stimulation was observed after 24 h treatment In comparison the expression of CsSULTR3 1 showed same change tendency either under high or low selenate treatment condition briefly decreased firstly then increased with a peak at 48 h and followed by another decrease Keywords tea plant Camellia sinensis sulfate transporter CsSULTR3 1 sulfate selenate expression analysis 硫作为半胱氨酸 甲硫氨酸 辅酶 铁氧还蛋白 生物素和硫胺素的组成部分 是植物生长发 育所必需的大量元素 Kramer 1 is a chloroplast localized sulfate transporter in Arabidopsis thaliana Plant Journal for Cell 3 mediates re distribution of sulfur from source to sink organs in Arabidopsis Plant Physiology 131 4 1511 1517 Zhu Yong guan Pilon Smits E A H Zhao Fang jie Williams P N Meharg A A 2009 Selenium in higher plants understanding mechanisms for biofortification and phytoremediation Trends in Plant Science 14 8 436 442 Zuber H Davidian J C Aubert G Aime D Belghazi M Lugan R Heintz D Wirtz M Hell R Tho