基于ITS全序列分析的重楼常见混伪品鉴定研究

Molecular identification of Paridis Rhizoma and its adulterants by ITS sequence analysis

目的：利用ITS（internal transcr ibed spacer）全序列对重楼常见混伪品进行分子鉴定。方法：对研究材料进行PCR扩增并双向测序，所得序列用ClustalW软件进行多重排定。序列变异位点采用PAUP4．0b10进行统计分析，并构建最大简约法（MP）树。结果：各样本的ITS序列长度为622～639bp，其中ITS1序列有74个变异位点，多于ITS2区和5．8s区。系统发育树上显示不同来源的样本根据亲缘关系的远近各聚一支，在序列比对图中能找到重楼混伪品的鉴别位点。结论：ITS序列能够将正品重楼与其常见混伪品样品进行快速区分，该方法可为鉴定中药重楼提供科学依据与方法参考。

Objection： To identity Paridis Rhizoma and its common adulterants by ITS sequence analysis. Methods： First, the materials were analyzed by PCR and bidirectional sequencing, and the sequence was multiply scheduled by software Clustal W. Second, the variable sites of the sequence were statistically analyzed by PAUP 4.0b10, and MP tree was constructed. Results： The total lengths of the ITS sequence were 622 - 639 bp in different samples. The ITS1 sequence had 74 variable sites, more than the ITS2 and 5.8s area. The samples with close genetic relationship were gathered together on the MP tress and could be distinguished from its adulterants. Conclusion： ITS1 sequence analysis can distinguish the genuine Paridis Rhizoma and its adulterants. But it is necessary to combine other gene sequence analysis in order to get accurate and rapid identification result.