山茶属植物EST-SSR序列分析

EST-SSR Sequence Analysis of Genus Camellia

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.