Rapid Identification of Chinese Medicinal Plants of Arctium by PCR-RFLP
Objective With the deepening of domestic scholars'research on medicinal plants of Arctium in the past decade,a rapid and accurate molecular identification method is needed for the identification of Arctium in China.Therefore,this study aims to establish a rapid identification method for medicinal plants of Arctium in China using polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP).Method By conducting restriction endonuclease map analysis on commonly used i-dentification DNA barcodes of two medicinal plants of Arctium in China,Zhiwu Niubang(A.Lappa L.)and Maotou Niubang(A.tomentosum M.),it is found that Zhiwu Niubang(A.Lappa L.)has a single nucleotide polymorphism(SNP)site in the internal transcribed spacer 1(ITS1)fragment,which happens to be the cleavage site of BsaA Ⅰ enzyme(YAC/GTR),and design primers to amplify the target fragment based on the enzyme cleavage site.In addition,it established the PCR system:pre denaturation at 95℃ for 5 min,cyclic reactions for 40 times(90℃ for 20 s,60℃ for 20 s,72℃ for 20 s),extension at 72℃ for 5 min,and in-sulation at 4℃.The restriction endonuclease BsaA Ⅰ digestion system was established,and agarose gel electrophoresis was made to observe the results.Result The electrophoresis results showed that after BsaA Ⅰ enzyme digestion,Zhiwu Niubang(A.Lappa L.)would produce short bands with lengths of 101 bp and 125 bp,respectively,while Maotou Niubang(A.tomentosum M.)will still have a long band of 226 bp without being digested,successfully distinguished two medicinal plants.This method is simple,fast and accurate,and meets the needs of daily identification of Arctium plants in China.Conclusion This experiment started with the ITS1 sequence of Zhiwu Niubang(A.Lappa L.)and Maotou Niubang(A.tomentosum M.),and used PCR-RFLP technolo-gy to complete the identification study of the two plants for the first time,establishing a PCR-RFLP identification method for Arc-tium plants in China.
万方数据
维普
