论文部分内容阅读
目的评价ITS/ITS2条形码序列在花椒药材鉴定中的准确性与稳定性。方法以多基原药材花椒为研究对象,采用改良的试剂盒法提取药材基因组DNA,PCR扩增ITS序列,利用基于隐马尔可夫模型的HMMer注释方法获得ITS2序列。通过分析花椒药材及其同属混伪品的种内、种间变异,利用中药材DNA条形码鉴定系统和NJ系统聚类树对花椒药材进行鉴定分析。结果花椒药材的ITS序列长度为618~620 bp,ITS2序列长度为224 bp,两种基原物种花椒和青椒的ITS/ITS2序列种内平均K2P遗传距离均远小于其与混伪品的种间K2P遗传距离;中药材DNA条形码鉴定系统和NJ树均可以准确区分花椒药材及其同属混伪品。结论 ITS/ITS2序列作为DNA条形码能稳定、准确鉴别花椒药材。
Objective To evaluate the accuracy and stability of ITS / ITS2 barcode sequences in the identification of Chinese medicinal materials. METHODS: The genomic DNA of medicinal plants was extracted by improved kit method. The ITS sequences were amplified by PCR. The ITS2 sequences were obtained by HMMer annotation method based on Hidden Markov Model. By analyzing the intraspecific and interspecific variation of the medicinal materials of Zanthoxylum bungeanum and its generic counterparts, the Chinese cabbage DNA barcode identification system and the NJ phylogenetic tree were used to identify and analyze Zanthoxylum bungeanum. Results ITS sequence of Zanthoxylum bungeanum was 618-620 bp and ITS2 was 224 bp in length. The average K 2 P genetic distance of ITS / ITS 2 sequences of Zanthoxyli and Zanthoxylum bungeanum was significantly smaller than that of Zanthoxylum bungeanum K2P genetic distance; Chinese herbal medicines DNA barcode identification system and NJ tree can accurately distinguish the pepper and its belong to the same mixed material. Conclusion ITS / ITS2 sequence can be used as a DNA barcode to identify the Chinese medicinal materials.