茶树钙调素基因CsCaMs的克隆及其低温胁迫下的表达分析  

Molecular Cloning and Expression of CaM Family Gene Cs CaMs Under Cold Stress in Tea Plant[Camellia sinensis(L.) O. Kuntze]

作　　者：黄玉婷[1] 钱文俊[1,2] 王玉春[1,2] 曹红利[1] 王璐[1] 郝心愿[1] 王新超[1] 杨亚军[1] 

机构地区：[1]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 [2]西北农林科技大学园艺学院,杨凌712100

出　　处：《植物遗传资源学报》2016年第5期906-912,共7页Journal of Plant Genetic Resources

基　　金：国家自然科学基金(31170650);国家茶叶产业技术体系(CARS-23);浙江省农业新品种选育重大专项(2012C2905-3);中国农科院科技创新工程(CAAS-ASTIP-2014-TRICAAS)

摘　　要：钙调素(CaM)是植物钙离子信号通道的主要参与者,参与低温胁迫下多种植物的抗寒生理作用。本研究根据钙调素基因相关表达序列标签(EST)序列,借助RACE-PCR技术,获得CsCaM1和CsCaM2两条cDNA全长序列,GenBank登录号分别为KT238971和KT238972,长度分别为693 bp和841 bp,均包含450 bp的完整开放阅读框(ORF),编码149个氨基酸,两条氨基酸序列仅一个氨基酸有差异,且均含有4个植物CaM家族的共同特征手型结构EF(EF-hand)。采用实时荧光定量PCR(qRT-PCR)分析CsCaMs在茶树低温胁迫下各种处理中的表达模式。结果表明,CsCaMs无组织表达特异性,低温胁迫处理和CaCl2均能诱导CsCaMs的表达,而钙调素拮抗剂W7与钙离子通道抑制剂LaCl3则会抑制其表达。本研究结果对阐明茶树抗寒性的分子机理有一定理论意义,为茶树的抗寒性育种提供参考。Calmodulin（CaM） is a major participant in calcium signaling pathway, involved in multiple physiological processes of plant under cold stress. Using RACE-PCR and RT-PCR, two full-length cDNAs of CsCaM were cloned from tea plant based on the available EST sequences. CsCaM1 had 693 bp in length, and CsCaM2 had 841 bp in length. Both of them contained a 450 bp open reading frame（ORF） encoding 149 amino acid residues. Cs- CaMs had four EF-hands, which were CaM gene families＇ characteristic. CsCaM1 and CsCaM2 were submitted to GenBank with accession number of KT238971 and KT238972, respectively. The expression profiles of CsCaMs under cold treatment were investigated using quantitative real-time PCR. Results indicated that the expressions of CsCaMs were induced gradually with the extension of cold treatment. Under CaCl2 treatment, the expressions of CsCaMs were up-regulated. However,the expressions of CsCaMs were down-regulated under W7 and LaCl3 treatments. Above results demonstrated that CsCaMs might play an important role in cold tolerance of tea plant.

关 键 词：茶树 钙调素 低温胁迫 表达分析 

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