茶树根系发育相关基因CsRDAs的克隆与功能分析  

Cloning and Function Analysis of Root Development Associated Genes CsRDAs in Tea Plants(Camellia sinensis)

作　　者：陈蓉 刘慧 付萍 赵德刚[1] 黄小贞 CHEN Rong;LIU Hui;FU Ping;ZHAO De-Gang;HUANG Xiao-Zhen(The Key Laboratory of Plant Resources Conservation and Germplasm Innovationin Mountainous Region(Ministry of Education)/College of Life Sciences,Guizhou University,Guiyang 550025,China;College of Tea Sciences,Guizhou University,Guiyang 550025,China)

机构地区：[1]贵州大学生命科学学院/山地植物资源保护与种质创新省部共建教育部重点实验室,贵阳550025 [2]贵州大学茶学院,贵阳550025

出　　处：《农业生物技术学报》2022年第1期63-74,共12页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(31960615)。

摘　　要：茶树(Camellia sinensis)作为国家重要的木本经济作物,其根系发育会直接影响茶叶产量和品质。为了进一步利用茶树优良品种资源,并从分子水平上解析茶树根系的发育过程,本研究从前期构建的黔茶1号(C. sinensis cv. Qiancha 1, QC1)和黔湄601号(C. sinensis cv. QianMei601, QM601)胚根转录组文库中,筛选到了1个根系发育相关基因,命名为CsRDA1 (root development associated gene 1)。利用逆转录(reverse transcription-PCR, RT-PCR)技术从QC1中克隆得到了CsRDA1及其3个同源基因的cDNA序列,分别命名为CsRDA1 (GenBank No. MW451597)、CsRDA2 (GenBank No. MZ516824)、CsRDA3 (GenBank No. MZ516825)和CsRDA4 (GenBank No. MZ516826)。系统发育与基序分析结果表明,茶树CsRDAs成员进化上来源相似,保守基序一致。qPCR结果表明,CsRDAs在不同组织器官中的表达各具特异性,提示其可能参与不同器官的发育调控。本氏烟草(Nicotiana benthamiana)叶片瞬时转染结果表明,CsRDA1和CsRDA2主要定位于细胞核和细胞质中,CsRDA3定位于叶绿体和细胞质中。进一步利用农杆菌(Agrobacterium tumefaciens)介导的愈伤转化法获得过表达CsRDA1的转基因水稻(Oryza sativa)。初步结果发现,转基因植株相比对照,总根数增加。该研究表明CsRDA1可能参与了茶树根系发育的调控,为进一步阐明茶树胚根系发育调控网络提供了参考依据。Tea plants(Camellia sinensis), as an important commercial crop, are one of the popular and economic beverages worldwide. Root formation of tea plants is essential for higher yields and stable quality.To elucidate the programmed root developmental processes, the root development associated genes(CsRDAs)were isolated and cloned by reverse transcription-PCR(RT-PCR), which was based on the root transcriptome library of Guizhou landrace tea cultivars, called C. sinensis cv. Qiancha 1(QC1) and C. sinensis cv.QianMei601(QM601). They were named CsRDA1(GenBank No. MW451597), CsRDA2(GenBank No.MZ516824), CsRDA3(GenBank No. MZ516825) and CsRDA4(GenBank No. MZ516826), respectively.Phylogenetic and the domain analysis showed that the CsRDAs were evolutionally conservative and contained similar motifs. qPCR results showed that the 4 members of CsRDAs could specifically express in different tissues and developmental stages, which showed that their functions may be tissue specific and developmentally controlled. Meanwhile, transient expression of CsRDAs fusion protein in Nicotiana benthamiana leaves revealed that CsRDA1 and CsRDA2 mainly localized in the nucleus and cytoplasm, while CsRDA3 localized in the chloroplast and cytoplasm. Furthermore, the transgenic rice(Oryza sativa) plants which overexpressed CsRDA1 were successfully obtained by Agrobacterium tumefaciens-mediated callus transformation. Preliminary results showed that constitutive expression of CsRDA1 increased the number of roots in rice plants. These data suggested that CsRDA1 may participate in the regulation of the development of tea plant root system. It provides a theoretical reference for further elucidating the regulation network of tea plant radicle development.

关 键 词：茶树 根系发育 转录组 蛋白定位 

分 类 号：S571[农业科学—作物学]