基于ITS2條形碼的桔梗藥材遺傳多樣性研究

　　[摘要]目的：利用ITS2條形碼探討不同產(chǎn)地桔梗的遺傳多樣性。方法：提取桔梗藥材DNA， PCR擴(kuò)增基于進(jìn)化速率較快的DNA內(nèi)部轉(zhuǎn)錄間隔區(qū)2（ITS2）序列并測序，采用Clustal比對分析不同產(chǎn)地桔梗ITS2序列，應(yīng)用MEGA 5.05軟件計算種內(nèi)Kimura 2-parameter（K2-P）遺傳距離，以桔�？泣h參屬植物黨參的ITS2序列為外展值，運(yùn)用鄰接法（neighbor-joining method）構(gòu)建桔梗系統(tǒng)進(jìn)化樹。結(jié)果：試驗所用全部樣本的K2-P遺傳距離為0～0.930，相同地區(qū)來源的樣本K2-P遺傳距離分布于0～0.178，不同地區(qū)來源的樣本K2-P遺傳距離為0.735～0.930;分析結(jié)果表明，桔梗種內(nèi)遺傳變異程度巨大，與其地理位置顯著相關(guān)。結(jié)論：ITS2條形碼適用于桔梗種內(nèi)遺傳多樣性研究，為進(jìn)一步拓展DNA條形碼技術(shù)的應(yīng)用提供了參考。
　　[關(guān)鍵詞]桔梗;ITS2條形碼;遺傳多樣性
　　Study on genetic polymorphism of Platycodon grandiflorum
　　based on barcoding of ITS2
　　WU Bo1，2， LI Yong-bo^/1， RAO Jiang-bo^/1， ZENG Jin-xiang1，2， ZHU Ji-xiao1，2， FANG Xiang-xiang^/1，
　　LIU Fu-qing^/1， LI Hong-ze^/1， HAN Feng-yu^/1， ZHONG Guo-yue1，2*
　　（1. Research Center of Natural Resourses of Chinese Medicinal Materials and Ethnic Medicine， Jiangxi University of Chinese
　　Medicine， Nanchang 330004， China; 2. Chinese Medicine Germplasm Resource Engineering Technology Research Center，
　　Jiangxi University of Traditional  Chinese Medicine， Nanchang 330004， China;
　　3. Inner Mongolia Mengji Pharmaceutical Technology Co.， Ltd.， Chifeng 024000， China;
　　4. Inner Mongolia Tianqi Han & Mongolia Pharmaceutical Co.， Ltd.， Chifeng 024000， China）
　　[Abstract]Objective： ITS2 of DNA barcoding was used to study genetic polymorphism of Platycodon grandiflorum. Method： Total genomic DNA was isolated from P. grandiflorum. PCR was used to amplified the region of internal transcribed spacer 2 （ITS2）， and PCR products were sequenced. The sequences of ITS2 were analyzed and compared by Clustal. The intraspecies genetic distance was calculated based on Kimura 2-parameter model by using MEGA 5.05. The ITS2 sequence of Codonopsis pilosula was used as the outreach value for plants of the genus， and the phylogenic tree used constructed by Neighbor-Joining（NJ） method. Result： The K2-P′s genetic distance of all samples were ranged from 0 to 0.930. The K2-P′s genetic distance of samples at the same area were ranged from 0 to 0.178. The K2-P′s genetic distance of samples at different areas were ranged from 0.735 to 0.930. The analytical result showed that the degree of genetic variation were heavy in intraspecies of P. grandiflorum and significantly correlated with geographical location. Conclusion： The DNA barcoding of ITS2 can applied to study the intraspecific genetic diversity， it provides a reference for further development of DNA barcoding technology applications.

相關(guān)熱詞搜索：桔梗 條形碼 多樣性 藥材 遺傳