CAJ | 학술논문

目的：结合ITS基因和COI基因对《四川省中药材标准》收录的凉山虫草和近缘物种及当地市场上出现的混伪品进行分子鉴定，考察其方法的可行性。方法：对收集的28份样品通过DNA提取、PCR扩增，分别获得其ITS序列和COI序列。用Codon Code Aligner V3.7.1，Mega 5.0等软件进行比对分析，构建基于ITS序列和COI序列的样品的NJ树，进行聚类分析。结果：凉山虫草和近缘物种及混伪品寄生真菌的ITS序列种间最小K-2P距离大于各物种种内最大K-2P距离：种内变异小，种间差异较大，从而基于ITS序列的NJ树能将凉山虫草与其他近缘物种及其混伪品很好地归属；凉山虫草和近缘物种及其混伪品寄主昆虫的COI序列种间最小K-2P距离大于各物种种内最大K-2P距离：种内变异小，种间差异较大，通过遗传距离及NJ树的分析能将凉山虫草与其他近缘物种其及其混伪品及其他近缘物种很好地区分。结论：结合ITS序列和COI序列的DNA条形码技术可以很好地鉴定凉山虫草与其近缘物种及其混伪品，并对虫草属的系统学和分类研究提供了技术支持。
목적：결합ITS기인화COI기인대《사천성중약재표준》수록적량산충초화근연물충급당지시장상출현적혼위품진행분자감정，고찰기방법적가행성。방법：대수집적28빈양품통과DNA제취、PCR확증，분별획득기ITS서렬화COI서렬。용Codon Code Aligner V3.7.1，Mega 5.0등연건진행비대분석，구건기우ITS서렬화COI서렬적양품적NJ수，진행취류분석。결과：량산충초화근연물충급혼위품기생진균적ITS서렬충간최소K-2P거리대우각물충충내최대K-2P거리：충내변이소，충간차이교대，종이기우ITS서렬적NJ수능장량산충초여기타근연물충급기혼위품흔호지귀속；량산충초화근연물충급기혼위품기주곤충적COI서렬충간최소K-2P거리대우각물충충내최대K-2P거리：충내변이소，충간차이교대，통과유전거리급NJ수적분석능장량산충초여기타근연물충기급기혼위품급기타근연물충흔호지구분。결론：결합ITS서렬화COI서렬적DNA조형마기술가이흔호지감정량산충초여기근연물충급기혼위품，병대충초속적계통학화분류연구제공료기술지지。
This study was aimed to identify and distinguish Metacordyceps liangshanensis recorded by the Sichuan Province Traditional Chinese Medicine Standard from its adulterants and its relative species by combining ITS and COI barcode sequences in order to study the feasibility of this new method. After extracting DNA of 28 species of Cordyceps samples, DNA were amplified and sequenced. And then, ITS and COI sequences were received. Codon-Code Aligner V3.7.1 and Mega 5.0 were used to analyze the variable site and construct the N-J tree. The results showed that the minimum ITS inter-specific K-2P distance was relatively higher than the maximum intra-specific K-2P distance. The inter-specific sequence divergence between M. liangshanensis and its adulterants exhibited high while intra-specific sequence divergence exhibited low. And COI one was the same case. N-J tree of both ITS and COI indicated that same genus belonged together and each species belonged to relatively independent branch. It was concluded that based on the ITS and COI gene, the technology of DNA barcode can be an excellent identification of M. liangshanensis, its adulterants and its relative species. It provided technical support for the further research on species molecular identification and phylogenetics of Cordyceps .