茶树CsASR基因的克隆及其表达分析  

Cloning and Expression Analysis of Cs ASR Gene in Tea Plant(Camellia sinensis)

作　　者：岳川[1,2] 曹红利[1,2] 郝心愿[2] 郭玉琼[1] 叶乃兴[1] 王新超[2] 杨亚军[2] YUE Chuan;CAO Hongli;HAO Xinyuan;GUO Yuqiong;YE Naixing;WANG Xinchao;YANG Yajun(College of Horticulture, Fujian Agriculture and Forestry University/Key Laboratory of Tea Science in Universities of Fujian Province/Collaborative Innovation Center of Chinese Oolong Tea Industry, Fuzhou 350002, China;Tea Research Institute of Chinese Academy of Agricultural Sciences, National Center for Tea Improvement, Key Laboratory of Tea Plant Biology and Resources Utilization Ministry of Agriculture, Hangzhou 310008, China)

机构地区：[1]福建农林大学园艺学院/茶学福建省高校重点实验室/中国乌龙茶协同创新中心,福建福州350002 [2]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,浙江杭州310008

出　　处：《茶叶科学》2017年第4期399-410,共12页Journal of Tea Science

基　　金：国家自然科学基金(31600555);福建省自然科学基金项目(2017J01616);国家茶叶产业技术体系(CARS-23)

摘　　要：逆境胁迫影响茶树生长发育及茶叶品质,ASR(abscisic acid,stress,ripening-induced)基因在植物抗逆响应中具有重要功能。本研究以龙井43品种茶树为材料,从中克隆了CsASR基因的全长cDNA序列、基因组序列及其启动子序列,分析了该基因的生物信息学特征及在组织间和不同胁迫处理下的表达模式。结果显示,CsASR的cDNA序列全长875 bp,含有546 bp的ORF序列,编码181个氨基酸,蛋白质分子量19.89 kD,理论等电点5.69;CsASR蛋白结构序列中74.5%的序列为无序结构,是一种无序蛋白;CsASR的C-端含有ABA/WDS功能结构域,主要定位于细胞质和细胞核中;茶树CsASR与海枣的ASR相似性最高,为87%,而在进化树中与枣的关系最近。CsASR基因含2个外显子,第1个外显子长363 bp,第2个外显子长183 bp,内含子较大为2 750 bp,内含子中含7种简单重复序列和2种DNA转座子序列。克隆获得起始密码子ATG上游2 554 bp的启动子区序列,该启动子上含有干旱、低温、高温以及ABA等相关的顺式作用元件。CsASR在根中的表达量最低;ABA抑制CsASR的表达,而干旱、NaCl和低温胁迫能够显著上调CsASR的表达。表明CsASR基因可能与茶树抗逆密切相关。Abiotic stress severely affects the growth and development of tea plant and the quality of tea.ASR(abscisic acid,stress,ripening)genes play crucial roles in the plant response to stresses.The full-length cDNA,genomic sequence and the promoter sequence of CsASR gene were cloned from tea cultivar Longjing43in this study,The bioinformatic and the expression analysis of CsASR in tissues under different stress treatments were performed.The results revealed that the full-length cDNA of CsASR was875bp,containing a546bp ORF encoding181amino acid.The predict protein molecular and theoretic isoelectric point of CsASR were19.89kD and5.69.Most regions of the amino acid sequence(74.5%)were predicted as the non-ordered regions,indicating that CsASR is a disordered protein.C-terminal of CsASR contained an ABA/WDS functional domain which was primarily located in both the cytoplasm and the nucleus.The homologous alignment and phylogenetic tree analysis showed that CsASR had the highest similarity(87%)with Phoenix dactylifera,and had the closest genetic relationship with Ziziphus nummularia.The genomic sequence of CsASR gene was comprised by two exons with363bp and183bp in length,respectively,and had a2750bp intron which contained seven simple repeats and two DNA transposons.The promoter sequence of CsASR was2554bp in length and was predicted to contain several stress-responsive elements related to drought,cold,high temperature stresses and ABA-signaling.The expression analysis showed that CsASR had the lowest level in roots,and its expression was repressed by ABA treatment.While the drought,NaCl and cold stresses could significantly up-regulate the expression of CsASR.The results revealed that CsASR might be closely related to stress response in tea plant.

关 键 词：茶树 ASR基因 逆境胁迫 基因克隆 

分 类 号：Q943.2[生物学—植物学] S571.1[农业科学—茶叶生产加工]