地黄资源遗传多样性分析

Genetic diversity of Rehmannia glutinosa germplasms

目的：研究不同地区栽培地黄及野生地黄品种间ITS序列差异，为地黄的系统发育和分子鉴定提供依据。方法：用CTAB法提取地黄叶片中总DNA，对PCR产物进行克隆测序，采用MEGA4．0进行系统进化分析。结果：各供试地黄的ITS区序列总长度为613～614bp，长度变异仅1bp，其中ITS1长度为224～225bp，G＋C占60．4％一63％；ITS2长度为224—225bp，G＋C占57．1％～65．3％；5．8S长度高度保守，均为164bp。系统发育分析表明：北京2号地黄与其他地黄资源差异性较大，亲缘关系较远。野生地黄居群内差异较小，居群间没有差异。河南栽培地黄与山西、山东省栽培地黄问没有差异；野生地黄资源中神农山和青天河一带地黄与山东省栽培地黄亲缘关系最近，而河南栽培地黄和山西省栽培地黄与野生地黄没有差异。结论：不同地区地黄资源种问亲缘关系较近，系统分化不明显。

Objective ： To provide molecular evidences for phylogenetic analysis by studying ITS sequences of Rehmannia glutinosa from different areas. Method ： The DNAs were extracted from leaves of R. glutinosa by means of CTAB method. The products of PCR amplification were cloned . The data were analyzed by MEGA4.0 software. Result ： The results showed that the size of the ITS of R. glutiousa tested was from 613 to 614 bp and the length variation was only 1 bp. The sequence of ITS1 was 224-225 bp, and G ＋ C content varied from 60.4% to 63%. The sequence of ITS2 was 224-225 bp and G ＋ C content varied from 57.1% to 65.3%. The se- quence of 5.8S rDNA was 164 bp, it＇s very conservative in these species. Phylogram tree based on ITS sequence data indicated that the kinship between Bejing No. 2 R. glutinosa and the others were far. There was obvious diversity within wild R. glutinosa varieties, while there was no different among cultivated R. glutinosa varieties. In cultivated R. glutinosa varieties, there was no diversity between R. glutinosa varieties from Henan and those from others provinces. In wild varieties, R. glutinosa from Shengnongshan and Qingtianhe of Henan province showed a closer systematic relationship with cultivated R. glutinosa from Shandong province, while there was no difference between wild R. glutinosa varieties and cultivated varieties from Henan and Shanxi provinces. Conclusion： The genetic rela- tionship among R. glutinosa varieties was very close, there was no distinct systematic differentiation.