中文摘要：

目的探索一种适用于柴胡药材于根DNA提取的方法，为实现以分子标记方法辨别柴胡药材奠定基础。方法比较研究了分别用CTAB法、SDS法和高盐低pH法等3种方法提取柴胡样品基因组DNA。柴胡药材干根经过PVP以及TNE缓冲液进行不同的预处理，以CTAB法抽提DNA。以EcoRⅠ／MseⅠ双酶切琼脂糖凝胶电泳及RAPD扩增检测提取DNA的质量。采用NTSYS-pc软件计算Jaccard遗传相似系数，以非加权配对算术平均数法（UPGMA）建立聚类图。结果常规DNA提取方法提取药材干根DNA溶液经4℃放置后，黏稠并褐变，严重影响酶切和RAPD扩增。样品预处理优化条件是研磨时加入3％PVP，TNE缓冲液于0℃浸提2次，每次30min。以该优化的预处理方法处理6个柴胡药材干根样品，提取DNA条带清晰，酶切完全，RAPD扩增图谱清晰稳定，共获得有效扩增条带28条，其中多态性条带为20条，占71．43％。聚类分析表明，6个栽培品中，原产地为山西灵丘外地引种（LQWY）和甘肃陇西（LX）、山西灵丘（LQ）和山西方山（FSH）的亲缘关系较近，陕西商洛（SHL）和其他5个栽培品的亲缘关系较远。结论柴胡药材干根经过预处理后，可采用常规的DNA提取方法抽提DNA，所提DNA适合于酶切、RAPD等分子标记分析。

英文摘要：

Objective To study the genomic DNA extraction from the dried roots of Bupleurum chinense. Methods Genomic DNA was extracted from the dried roots of B. chinense by means of three methods of routine CTAB, SDS, and low pH extraction medium with high salt. Based on CTAB method, the pretreatment of samples was modified by adding PVP to grind and TNE buffer to pre-extract. The DNA samples were analyzed by restriction EcoR I/Mse I double enzyme digestion, agarose gel electrophoresis, and RAPD amplification. The Jaccard coefficient was worked out by NTSYS-pc software, and a cluster dendrogram of six samples was established based on UPGMA. Results The DNA could be extracted, but not be completely restricted and effectively amplified, the solution of the DNA became ropy and brown by stored up at 4 ℃, which was extracted from the dried roots by routine DNA extraction methods. After the samples of the dried roots were pretreated by PVP and TNE buffer, the DNA extracted by CTAB method could be completely restricted and effectively amplified. The pretreatment process was that samples of the dried roots were ground with 3 % PVP and then extracted with TNE buffer twice at 0 ℃, each 30 min. The DNA extracted with the modified method from the six dried roots samples could be digested completely by restriction endonuclease. Moreover, the fingerprints were clear and stable by RAPD with three selected primers. A total of 28 bands were amplified, among which 20 bands were polymorphic, accounting for 71.43%. The cluster analysis indicated that there was closer genetic relationship between LQWY and LX, LQ and FSH cultivars of B. chinense, respectively. The relationship of cultivar SHL was far from these of other five cultivars. Conclusion The DNA extracted by routine DNA extraction methods could be completely restricted and effectively amplified after the samples of the dried roots are pretreated by PVP and TNE buffer.