保康柴胡5S-rDNA间隔序列分析及道地性研究

Preliminary Attempt to Identify Geoherbalism of Bupleurum by Sequence Divergence of 5S-rDNA Spacer Region

目的探讨保康柴胡道地性形成的基因基础。方法提取6个柴胡样品的总DNA,进行5S-rDNA间隔序列的PCR扩增和测序,用Mega3.0软件进行序列分析,采用RP-HPLC法测定柴胡药材中柴胡皂苷a、d的含量。结果柴胡样品的5S-rDNA间隔序列具有相似的碱基序列,长度为270-321bp,G＋C%含量为30.7-36.5%,各样品柴胡皂苷a、d的含量有所不同。结论种间5S-rDNA间隔序列差异大于种内,道地药材之间的遗传距离较小,6种柴胡品质的差异可能与碱基序列有关。

Objective To study the formation of geoherbalism of baokang Bupleurum on the molecular level. Methods Genomic DNAs from Bupleurum, the sequences of 5S-rDNA spacer region were amplified and sequenced, and then analysed with Mega3.0. Determination of Saikosaponin a, d in Bupleurum by RP-HPLC. Results The sequences of 5S-rDNA spacer region were similar, the length of them between 270 bp and 321bp, the G ＋ C content is about 30.7 -36.5%. The saikosaponin a, d in Bupleurum were different. Conclusion The genuine distance between species was larger than that within species, the genetic distance among genuine crude drugs was smaller. The different of Saikosaponin a,d in Bupleurum may be resulted from base sequences variation.