CAJ | 학술논문

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 CodonCode 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。
목적：응용ITS2서렬대약재사간여천사간급기혼위품진행DNA조형마감정연구，보장림상용약안전。방법：제취사간、천사간급기혼위품화근연충적 DNA ，경 PCR확증후측서，통과 CodonCode Aligner V3.7.1대측서서렬진행질량분석화병접，기우MEGA5.1중적K2P모형계산사간여천사간급기혼위품적충내、충간유전거리급구건계통취류수。결과：사간적ITS2서렬비대후장도위272 bp，충내최대K2P거리위0，평균GC함량위52.22%。천사간적ITS2서렬비대후서렬장도위268 bp，충내최대K2P거리위0.004，평균GC함량위67.87%。사간、천사간적충내최대K2P거리균소우여혼위품적충간최소K2P거리。기우ITS2서렬구건NJ취류수，사간、천사간능각자취위일지，정현출교호적단계성，능흔호지여기혼위품백급、산관、황화사간，이급근연충덕국연미、북릉연미명현구분개。결론：ITS2서렬재약재사간여천사간급기혼위품화근연충적감정방면구유흔호적감정효과，대보장사간여천사간적용약안전구유중요의의。
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.