CAJ | 학술논문

　　目的：应用ITS2条形码鉴定芡实及其近缘种药材。方法：提取20份芡实及其近缘种药材样品基因组DNA，通过PCR扩增ITS2序列并双向测序，测序后用Sequin 12.3提交至GenBank；从GenBank上下载所有芡实及其近缘种30种102条 ITS2序列；对提交与下载的122条序列，应用MEGA5.1软件进行序列比对，计算种内和种间距离，采用相似性搜索法、最近距离法进行鉴定分析，并构建NJ系统进化树直观反映鉴定结果。结果：芡实药材ITS2序列长度均为214 bp，为1个单倍型，与其近缘种之间遗传距离较远，NJ树结果显示芡实及其近缘种药材可明显区分开，表现出良好的单系性。结论：ITS2序列作为DNA条形码能稳定、准确地鉴别芡实药材，为保障临床安全用药提供了新的技术手段，为拓宽DNA条形码技术在中药鉴定中的应用进行了新的探索。
　　목적：응용ITS2조형마감정검실급기근연충약재。방법：제취20빈검실급기근연충약재양품기인조DNA，통과PCR확증ITS2서렬병쌍향측서，측서후용Sequin 12.3제교지GenBank；종GenBank상하재소유검실급기근연충30충102조 ITS2서렬；대제교여하재적122조서렬，응용MEGA5.1연건진행서렬비대，계산충내화충간거리，채용상사성수색법、최근거리법진행감정분석，병구건NJ계통진화수직관반영감정결과。결과：검실약재ITS2서렬장도균위214 bp，위1개단배형，여기근연충지간유전거리교원，NJ수결과현시검실급기근연충약재가명현구분개，표현출량호적단계성。결론：ITS2서렬작위DNA조형마능은정、준학지감별검실약재，위보장림상안전용약제공료신적기술수단，위탁관DNA조형마기술재중약감정중적응용진행료신적탐색。
Objective: To identify Euryales Semen and its closely related species using the ITS2 barcode. Method:The total genomic DNAs were extracted from twenty samples of Euryales Semen and its closely related species. The ITS2 regions of the samples were amplified and bidirectional sequenced. Obtained sequences were submitted to the GenBank with Sequin 12.3. ITS2 sequences of 102 samples belonging to thirty species were downloaded from GenBank. The 122 ITS2 sequences were aligned and the genetic distances were analyzed with MEGA 5.1. Identification analyses were performed using BLAST1 and nearest distance methods, and were presented intuitively by constructing neighbor-joining (NJ) tree. Result: The length of ITS2 region of Euryales Semen was 214 bp, which was only one haplotype. There was significant divergence of the ITS2 regions among the samples. The NJ tree showed that Euryales Semen could be obviously differed from its closely related species, which had good 408 monophyly. Conclusion: ITS2 regions as a DNA barcode can stably and accurately distinguish Euryales Semen from its closely related species and also provide a new technique to ensure clinical safety in utilization of tradi-tional Chinese medicines. The new exploration could broaden the application of DNA barcoding technology in identification of Traditional Chinese Medicine.