摘要
目的:应用ITS2序列对药材射干与川射干及其混伪品进行DNA条形码鉴定研究,保障临床用药安全。方法:提取射干、川射干及其混伪品和近缘种的DNA,经PCR扩增后测序,通过Codon Code Aligner V3.7.1对测序序列进行质量分析和拼接,基于MEGA5.1中的K2P模型计算射干与川射干及其混伪品的种内、种间遗传距离及构建系统聚类树。结果:射干的ITS2序列比对后长度为272 bp,种内最大K2P距离为0,平均GC含量为52.22%。川射干的ITS2序列比对后序列长度为268 bp,种内最大K2P距离为0.004,平均GC含量为67.87%。射干、川射干的种内最大K2P距离均小于与混伪品的种间最小K2P距离。基于ITS2序列构建NJ聚类树,射干、川射干能各自聚为一支,呈现出较好的单系性,能很好地与其混伪品白及、山菅、黄花射干,以及近缘种德国鸢尾、北陵鸢尾明显区分开。结论:ITS2序列在药材射干与川射干及其混伪品和近缘种的鉴定方面具有很好的鉴定效果,对保障射干与川射干的用药安全具有重要意义。
This study aimed to identify Belamcandae Rhizoma, Iridis tectori Rhizoma and their adulterants by ITS2 sequence. All the DNA samples of Belamcandae Rhizoma, Iridis tectori Rhizoma and their adulterants were extracted. The ITS2 sequence were succesfully amplified, and purified PCR products were sequenced. All the sequences were assembled using the CondonCode Aligner V3.7.1. The Kimura 2-Parameter （K2P） genetic distances and the Neighbor-Joining （NJ） phylogenetic tree were calculated by using MEGA5.1. Results indicated that the maximum intraspecific genetic distances of Belamcandae Rhizoma was 0, and the average GC content was 52.22%;the maximum intraspecific genetic distances of Iridis tectori Rhizoma was 0.004, and the average GC content was 67.87%. The maximum K2P intraspecific genetic distance of Belamcanda chinensis, Iris tectorum were both lower than the minimum interspecific genetic distance of adulterants. Additionally, the ITS2 sequences in each of these polytypic species were separated into pairs of divergent clusters in the NJ tree. The NJ tree based on ITS2 sequence indicated that Belamcandae Rhizoma, Iridis tectori Rhizoma and their adulterants could be distinguished clearly. It could be concluded that ITS2 barcode can be used to correctly identify Belamcandae Rhizoma, Iridis tectori Rhizoma from their adulterants, and ensure their safety in use.
出处
《世界科学技术-中医药现代化》
北大核心
2014年第11期2355-2360,共6页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
科学技术部重大新药创制国家重大科技专项(2014ZX09304307001-014):中药新药安全性检测技术与标准-中药原料混伪品分子基因鉴定技术与标准
主课题负责人:肖红斌
子课题负责人:陈士林
关键词
射干
川射干
ITS2
混伪品
Belamcandae Rhizoma, Iridis tectori Rhizoma, ITS2, adulterants
