CAJ | 학술논문

目的：药材天麻由于隶属于单子叶植物兰科天麻属，天麻中含有大量多糖、蛋白质以及大量次生代谢物质，使其提取的DNA质量不高，影响后续的PCR扩增。且ITS2通用引物在单子叶植物部分物种通用性很差。本研究通过对DNA提取过程、PCR扩增过程进行优化，设计出一对用于鉴别天麻药材的ITS2引物，为中药材天麻的鉴定提供一种新方法。方法：以通用ITS2引物扩增出的两条序列为研究对象，经Primer Premier 5.0设计出3对特异性引物，通过对22份样品进行研究，筛选出一条高特异性的ITS2引物。结果：在54益的退火温度下，TM2F-2R对天麻的鉴定效率高达90.9%。结论：TM2F-2R可作为天麻ITS2序列的特异性引物，为天麻的分子鉴定提供了一套准确、稳定的鉴定方法。
목적：약재천마유우대속우단자협식물란과천마속，천마중함유대량다당、단백질이급대량차생대사물질，사기제취적DNA질량불고，영향후속적PCR확증。차ITS2통용인물재단자협식물부분물충통용성흔차。본연구통과대DNA제취과정、PCR확증과정진행우화，설계출일대용우감별천마약재적ITS2인물，위중약재천마적감정제공일충신방법。방법：이통용ITS2인물확증출적량조서렬위연구대상，경Primer Premier 5.0설계출3대특이성인물，통과대22빈양품진행연구，사선출일조고특이성적ITS2인물。결과：재54익적퇴화온도하，TM2F-2R대천마적감정효솔고체90.9%。결론：TM2F-2R가작위천마ITS2서렬적특이성인물，위천마적분자감정제공료일투준학、은정적감정방법。
This study aimed to design a pair of primers for amplifying internal transcribed spacer 2 region which was used to identify Gastrodia elata through optimizing of DNA extraction and PCR amplification process. Two sequences are used for research object by amplification of universal primers. Design three pairs of specific primers by Primer Premier 5.0 and select the highest specificity through the study of 22 samples. The results showed that identification efficiency of the primer named TM2F-2R is as high as 90.9% when Annealing Temperature is equal to 54 degrees Celsius. Therefore, TM2F-2R can be used as primers ITS2 sequences of G. elata, this article provides a set of accu-rate and stable identification methods for G. elata in the molecular identification.