中药材连翘道地性的分子生物学探讨

A Preliminary Study on the Molecular and Chemical Characteristics of Authenticity (Daodixing)of a Traditional Chinese Medicinal Plant-Forsythia suspense Vahl

运用随机扩增多态DNA(RAPD)方法研究了来自山西黎城、长治、平顺、壶关、屯留及安泽6个地区11个连翘地方栽培品系的亲缘关系,并用HPLC方法测定连翘主要指标成分———连翘甙含量的变异情况.RAPD分析结果表明,连翘栽培的地方品系间存在明显的DNA水平上的多样性变异,运用Philip3.6软件包以及Wagner算法,可分为A、B、C3个不同分支.连翘甙含量分析结果显示,2个遗传关系相近分支B和C的连翘甙含量差异不显著(t=0.44,p=0.67;平均连翘甙含量分别为0.53mg/g和0.67mg/g),这2个类群与第3个类群遗传差异大,连翘甙含量差异也很大(8.7mg/g).这种DNA分子多样性差异与化学指标成分含量差异的相关性说明连翘的遗传基础可能对化学指标成分的形成和积累有显著的影响.研究结果显示运用RAPD分析手段进行药材道地性研究具有可行性.研究结果不仅对于连翘的道地性研究和中药材GAP(GoodAgriculturalPractice)生产有一定的指导意义,而且对于其他道地中药材的研究也具有一定参考价值.

To address the authenticity (daodixing, a concept used to judge the quality of Traditional Chinese Medicines (TCM) mainly based on the growing environments as well as the varieties of medicinal plants) of a Traditional Chinese Medicinal plant, Forsythia suspensa, the genetic relationship of 11 cultivars from sixdifferent regions of Shanxi province (Licheng,Changzhi, Tunliu, Huguan, Pingshun, Anze) was analyzed using the randomly amplified polymorphic DNAs (RAPD) technique. The contents of forsythin, an indicative component of Forsythia suspensa, in these cultivars were analyzed using high performance liquid chromatography(HPLC). The RAPD analysis showed that there was a significant genetic diversity among the 11 cultivars. These cultivars were arranged into three genetic branches (A, B and C) using the Philip3.6 software and Wagner′s method. The branch A containing the only cultivar from Huguan was genetically distant from the other two branches, B and C, which were relatively close. The HPLC analysis showed that there was also a great variation in the contents of forsythin among these cultivars. Most dramatic variations were found between the branchA (8.7 mg/g) and the branchB or the branchC. However no significant variations were found within the three branches, nor between the branchB (the mean 0.53mg/g)and the branchC (the mean 0.67mg/g). These results suggest that the formation and accumulation of forsythin could be affected substantially by the genetic basis of Forsythia suspense. It is particularly true in this case since these cultivars were collected from relatively less diverse environments. This work has shown some promise that the authenticity of Traditional Chinese Medicinal plants can be addressed using RAPD and HPLC techniques.