山茶属植物EST-SSR序列分析  

EST-SSR Sequence Analysis of Genus Camellia

作　　者：董存丽 寇国艳 罗春梅 张燕 杨海艳[2] 范树国[2] 陈迁进 于建功 绍胜富 将昌杰 罗燕英 邱璐[2] Cunli Dong;Guoyan Kou;Chunmei Luo;Yan Zhang;Haiyan Yang;Shuguo Fan;Qianjin Chen;Jiangong Yu;Shengfu Shao;Changjie Jiang;Yangying Luo;Lu Qiu(Chuxiong Administration,Ailao Mountain National Nature Reserve of Yunnan,Chuxiong 675000,China;School of Resources,Environment&Chemistry,Chuxiong Normal University,Chuxiong,Yunnan Province 675000;Department of Modern Landscape Engineering,Chuxiong Technician College,Chuxiong,Yunnan 675000,China;Institute of Cash Crops,Chuxiong Academy of Agricultural Sciences,Chuxiong 675000,China;Forestry Seedling Station of Jinhua City,Jinhua 321000,China;Jinhua International Camellia Species Park,Jinhua 321000,China;Golden Flower Tea Park,Guangxi Province,Nanning 530000,China)

机构地区：[1]云南哀牢山国家级自然保护区楚雄管护局,中国云南楚雄675000 [2]楚雄师范学院资源环境与化学学院,中国云南楚雄675000 [3]楚雄技师学院现代园林工程系,中国云南楚雄675000 [4]楚雄州农业科学院经济作物研究所,中国云南楚雄675000 [5]浙江省金华市林业种苗站,浙江金华321000 [6]浙江省金华市国际茶花物种园,浙江金华321000 [7]广西金花茶公园,中国南宁530000

出　　处：《楚雄师范学院学报》2021年第3期39-50,共12页Journal of Chuxiong Normal University

基　　金：National natural science foundation of China(No.31460238)。

摘　　要：研究通过SSR、EST-SSR分子标记技术对179种山茶属植物进行亲缘关系分析。采用改良的CTAB法提取茶花基因组DNA,用1%的琼脂糖凝胶电泳检测DNA纯度,用2%的琼脂糖凝胶电泳检测PCR扩增产物。扩增产物测序结果显示,EST-SSR序列片段大小为194 bp,序列最长的是崇左金花茶,长度为210 bp,序列最短为凹脉金花茶,长度为152 bp。在NCBI上寻找同源系列,用Cluster软件进行系列比对,发现不同山茶属植物EST-SSR系列差异较大,其中缺失位点为1752个,突变位点813个。用MEGA6.0软件画系统树,对茶花进行亲缘关系分析,39种茶花聚为红山茶组、金花茶组、糙果茶组、连蕊茶组四组,结果与张宏达形态学分类结果基本一致。结论:EST-SSR分子标记用于茶花系统分类及物种鉴定效果较好。179 plants of Camellia genus were analyzed using SSR,EST-SSR molecular markers technology to detect their genetic relationship.DNAs of the 179 Camellia plants were extracted by improved CTAB,then tested for the quality of DNAs by way of 1 percent of agarose gel electrophoresis,and tested for PCR amplification by way of 2 percent of agarose gel electrophoresis.The results of amplification products show,the EST-SSR sequence fragment size is 194bp,the longest being Chongzhu(C.theopsis)of 210bp and shortest being Aomai(C.impressinervis)of 152bp.Referring to the homologous series in NCBI and compared using Cluster X software,great differences were found in the EST-SSRs of different camellia plants,with 1752 deletion loci and 813 mutation sites.Phylogenetic trees were constructed using MEGA 6.0 to analyze their genetic relationship,39 camellia plants were classified into four groups,i.e.Sect.Camellia,Sect.Chrysantha,Sect.Furfuracea and Sect.Theopsis.The results are basically in consistency with Zhang Hongda morphological classification.This indicates that EST-SSR molecular marker is effective in phylogenetic classification and identification of Camellia.

关 键 词：山茶属 SSR EST-SSR 系统树 亲缘关系 

分 类 号：Q949.758.4[生物学—植物学]