茶树冷诱导基因CsMAPK15的克隆与鉴定  

Cloning and Identification of Cold-induced Gene CsMAPK15 in Tea Plants(Camellia sinensis)

作　　者：薛承进 赵兰馨 赵德刚 黄小贞 XUE Cheng-Jin;ZHAO Lan-Xin;ZHAO De-Gang;HUANG Xiao-Zhen(College of Life Sciences/Institute of Agro-bioengineering/Key Laboratory of Plant Resources Conservation and Germplasm Innovationin in Mountainous Region(Ministry of Education),Guizhou University,Guiyang 550025,China;College of Tea Sciences,Guizhou University,Guiyang 550025,China;GuizhouAcademy ofAgricultural Sciences,Guiyang 550006,China)

机构地区：[1]贵州大学生命科学学院/农业生物工程研究院/山地植物资源保护与种质创新教育部重点实验室,贵阳550025 [2]贵州大学茶学院,贵阳550025 [3]贵州省农业科学研究院,贵阳550006

出　　处：《农业生物技术学报》2023年第2期311-321,共11页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(31960615)。

摘　　要：茶树(Camellia sinensis)是重要的木本经济植物之一,容易遭受低温冻害,严重影响茶叶产量和品质。为了深入探讨茶树低温胁迫响应的分子调控机理,本研究利用前期构建的抗寒性强的’黔茶1号’(Qiancha 1,QC1)和抗寒性弱的’黔湄601’(Qianmei 601, QM601)的低温响应转录组文库,分析了茶树丝裂原活化蛋白激酶(mitogen-activated protein kinase, MAPK)基因家族对冷信号的响应,并从中筛选克隆了1个冷诱导的MAPK基因(转录组文库编号:CSS0027011),命名为CsMAPK15(GenBank No. ON39909)。CsMAPK15基因开放阅读框包含1 701个碱基,编码566个氨基酸,有1个保守的MAPK结构域。系统发育与蛋白保守基序分析结果表明,CsMAPK15蛋白具有3个保守基序,与拟南芥(Arabidopsis thaliana) AtMAPK16(AT5G19010)编码氨基酸相似性为79.17%,与水稻(Oryza sativa) OsMAPK15(LOC_Os11g17080)编码氨基酸相似性为80.51%。组织器官表达模式分析表明,CsMAPK15在茶树的根组织中表达量最高。启动子序列分析发现,CsMAPK15启动子具有多个顺式作用元件,包括胁迫响应元件(stress-responsive element,STRE)和水杨酸(salicylic acid, SA)响应元件(TCA-element)等。qPCR检测进一步证实CsMAPK15能够响应冷信号和SA信号。利用愈伤转化获得持续表达CsMAPK15的转基因水稻,初步分析结果显示,正常生长条件下,与野生型植株相比,转基因植株的生长发育没有显著改变。本研究为进一步阐明CsMAPK15在植物抗寒性调控中的作用提供了研究材料和理论依据。Tea plants(Camellia sinensis), as one of the important woody economic plants, are vulnerable to low temperature and freezing injury, which seriously affecting the yield and quality of tea. In order to further clarify the molecular mechanism on cold response of tea plants, in this study, based on the cold treatment transcriptome library of 2 Guizhou cultivars, Qiancha 1(QC1) and Qianmei 601(QM601), a cold-upregulated differentially expressed mitogen-activated protein kinase(MAPK) gene(transcriptome library number:CSS0027011) was isolated and cloned, and named as CsMAPK15(GenBank No. ON39909). The open reading frame of CsMAPK15 contains 1 701 nucleotides encoding a protein of 566 amino acids. Motif analysis revealed that CsMAPK15 had 3 conserved motifs. Phylogenetic analysis showed that Cs MAPK15 shared high sequence similarity with homologs from other species. It shared 79.17% identity with AtMAPK16(AT5G19010) from Arabidopsis and 80.51 % similarity with OsMAPK15(LOC_Os07g47490) from rice(Oryza sativa). The tissue pattern analysis revealed that expression level of CsMAPK15 was highest in roots. Promoter analysis showed that the CsMAPK15 promoter had several cis-acting elements, including stress responsive element(STRE) and salicylic acid(SA) responsive element(TCA element). qPCR confirmed that CsMAPK15could response to cold signal and SA. Furthermore, the transgenic rice plants which overexpressed CsMAPK15were successfully obtained by callus transformation. The preliminary results showed that there was no significant change in plant growth and development between transgenic plants and wild-type plants. This study provides research materials and theoretical basis for further clarifying the role of CsMAPK15 in plant cold resistance regulation.

关 键 词：茶树 冷胁迫 丝裂原活化蛋白激酶(MAPK) 基因家族 转录组分析 

分 类 号：S1[农业科学—农业基础科学] S6