Gao Suxia, Liu Hongyan, Wang Fei
Institute of Plant Protection, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China.
Zhongguo Zhong Yao Za Zhi. 2010 Mar;35(6):690-3. doi: 10.4268/cjcmm20100606.
To provide molecular evidences for phylogenetic analysis by studying ITS sequences of Rehmannia glutinosa from different areas.
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.
The results showed that the size of the ITS of R. glutionsa 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 sequence 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.
The genetic relationship among R. glutinosa varieties was very close, there was no distinct systematic differentiation.
通过研究不同产地地黄的ITS序列,为系统发育分析提供分子证据。
采用CTAB法从地黄叶片中提取DNA。对PCR扩增产物进行克隆。用MEGA4.0软件进行数据分析。
结果表明,所测地黄ITS长度为613~614 bp,长度变异仅1 bp。ITS1序列长度为224~225 bp,G+C含量在60.4%~63%之间。ITS2序列长度为224~225 bp,G+C含量在57.1%~65.3%之间。5.8S rDNA序列长度为164 bp,在这些物种中非常保守。基于ITS序列数据构建的系统发育树表明,北京2号地黄与其他地黄的亲缘关系较远。野生地黄品种内存在明显的多样性,而栽培地黄品种间无差异。在栽培地黄品种中,河南地黄与其他省份地黄之间无差异。在野生品种中,河南神农山和青天河的地黄与山东栽培地黄的系统关系较近,而河南和山西的野生地黄品种与栽培品种之间无差异。
地黄品种间遗传关系非常密切,无明显的系统分化。