宁夏农林科技2026，Vol.67Issue(5)：9-16,8.DOI:10.3969/j.issn.1002-204x.2026.05.002
肉苁蓉饮片基原植物PCR-RFLP鉴别方法研究
Research on PCR-RFLP Identification Method for Original Plant of Cistanche Herbs Slices
摘要
Abstract
[Objective]To establish a polymerase chain reaction restriction fragment length polymorphism(PCR-RFLP)identification method for Cistanche deserticola and Cistanche tubulosa,the original plants of Cistanche herbs slices.[Methods]The ITS sequences of C.deserticola and C.tubulosa were compared and analyzed with SanpGene software to screen for heterogeneous restriction endonuclease sites,and PCR amplification products of their ITS sequences were indentified using restriction endonucleases.The DNA extraction method for C.deserticola and C.tubulosa slices,annealing temperature,cycle number,and amplification efficiency of different enzymes for PCR amplification,as well as the enzyme digestion reaction time and substrate amount were optimized,and the PCR-RFLP identification method was established.On this basis,the stability and accuracy of this method were investigated.[Results]The suitable extraction method for DNA from herbs slices was the reagent kit method,and the suitable annealing temperature and cycle number for PCR amplification were 47℃and 35,respectively.The suitable substrate amount for the NcoI restriction enzyme digestion reaction was 7.5 μL,and the suitable temperature and reaction time was 37℃and 60 min,respectively.On the above conditions,the PCR amplification product of C.deserticola was digested into two bands,which were located between 100 bp-250 bp and 500 bp-750 bp,respectively.However,the PCR amplification product of C.tubulosa was not digested and displayed one band,and only a distinct band at 750 bp was observed.[Conclusions]The study established a PCR-RFLP identification method for the original plants of the medicinal herb C.deserticola and C.tubulosa.This method was faster and more accurate than microscopic and component identification,and could provide technological reference for the identification of Cistanche herbs slices in the market.关键词
肉苁蓉饮片/基原植物鉴定/肉苁蓉/管花肉苁蓉/PCR-RFLP/ITS序列Key words
Cistanche herbs slices/Identification of original plants/Cistanche deserticola/Cistanche tubulosa/PCR-RFLP/ITS sequence分类
医药卫生引用本文复制引用
施文韬,李鹏飞,崔佳惠..肉苁蓉饮片基原植物PCR-RFLP鉴别方法研究[J].宁夏农林科技,2026,67(5):9-16,8.基金项目
宁夏回族自治区药品监督管理局中药质量控制重点试验室科技项目"肉苁蓉饮片基原植物种类分子鉴定技术研究". （）
