目的 克隆茶树WRKY转录因子（CsWRKYs）家族中的8个成员，并对其进行生物信息学和非生物胁迫下的表达分析。方法 采用实时荧光定量PCR（qRT-PCR）技术从"铁观音"茶树品种中克隆8个WRKY基因，运用生物信息学的方法对8个WRKY基因编码的蛋白进行理化性质分析，同时通过与拟南芥同源基因的比较进行进化树的构建、多序列比对及保守基序分析。采用qRT-PCR方法检测8个WRKY基因在低温、干旱和脱落酸（abscisic acid，ABA）胁迫处理下的表达量。结果 8个WRKY转录因子基因开放阅读框（ORF）长度分别为1 407、2 208、1 302、849、978、879、1 443和810 bp，分别编码468、735、433、282、325、292、480和269个氨基酸，GenBank登录号分别为MG298951、MG298952、MG298955、MG298956、MG298957、MG298959、MG298960和MG298963。系统进化树及序列比对分析显示，8个CsWRKYs可以分成了2个大组；除CsWRKY39缺少锌指结构外，其他CsWRKYs均含有WRKYGQK保守七肽结构域及锌指结构组成的WRKY结构域。非生物胁迫下的表达模式显示，CsWRKYs基因在低温、干旱和ABA胁迫处理下均有表达且表达受到不同程度的诱导；CsWRKY2、CsWRKY21、CsWRKY23、CsWRKY44和CsWRKY65基因在低温处理后表达量上调超过2，显著响应低温胁迫；CsWRKY21、CsWRKY23、CsWRKY39和CsWRKY65在干旱胁迫处理12 h及ABA处理6 h时均上调表达。结论 克隆获得来自不同组的8个茶树WRKY基因，推测与茶树抗逆密切相关。

Objective To clone eight members of WRKY of transcription factor family in Camellia sinensis, and analyze their bioinformatics and expression under abiotic stress. Methods Eight WRKY transcription factor genes were cloned from Tieguanyin cultivar by RT-PCR, and the physicochemical properties of the eight WRKY protein were analyzed by bioinformatics methods. At the same time, the establishment of phylogenetic tree, comparison of multiple sequences, and analysis of conserved motifs were carried out by comparing WRKY of C. sinensis with homologous genes of Arabidopsis thaliana. Quantitative real-time PCR (qRT-PCR) was used to detect the expression of eight WRKY genes under low temperature, drought, and ABA stress treatment. Results The ORF lengths of eight WRKY genes were 1 407, 2 208, 1 302, 849, 978, 879, 1 443, and 810 bp, encoding 468, 735, 433, 282, 325, 292, 480, and 269 amino acids, respectively. GenBank accession numbers were MG298951, MG298952, MG298955, MG298956, MG298957, MG298959, MG298960, and MG298963, respectively. Phylogenetic tree and sequence alignment analysis showed that eight CsWRKYs could be divided into two groups and contained WRKYGQK conserved domain and zinc finger structures, except that CsWRKY39 lacked zinc finger structure. The expression pattern of CsWRKYs was induced under the condition of low temperature, drought, and ABA stress. The expression of CsWRKY2, CsWRKY21, CsWRKY23, CsWRKY44 and CsWRKY65 increased to more than 2 after low temperature treatment with significant response to low temperature stress. The expression of CsWRKY21, CsWRKY23, CsWRKY3,9 and CsWRKY65 was up-regulated under 12 h of drought stress and 6 h of ABA treatment. This result indicated that CsWRKYs might be closely related to stress response in C. sinensis. Conclusion Eight CsWRKY genes from different groups were cloned, and this result indicated that CsWRKYs might be closely related to stress response in C. sinensis.

国家自然科学基金项目（31600555）；福建省"2011协同创新中心"中国乌龙茶产业协同创新中心专项（闽教科[2015]75号）；福建农林大学科技创新专项基金项目（CXZX2017181）