Identification Of Baphicacanthus Cusia Rhizoma Et Radix,Dioscorea Opposita Rhizoma And Its Adulterants Using The DNA Barcoding Technique

Objective:Now the adulterants of the Baphicacanthus cusia rhizoma et radix and Dioscorea opposita rhizoma are full of the market, and the authentic and its adulterants appearance are similar, the general scientific research personnel is difficult to distinguish.In this study, I compared Baphicacanthus cusia rhizoma et radix and Dioscorea opposita rhizoma with its adulterants by DNA barcoding candidate sequences:rbcL, psbA-trnH, matK and nuclear gene ITS2sequence, in orser to establish the DNA barcoding method for identification of Baphicacanthus cusia rhizoma et radix, Dioscorea opposita rhizoma and its adulterants method, provide reference for the DNA barcoding technology in traditional Chinese medicine authentication system to.Methods:This study chose60Baphicacanthus cusia rhizoma et radix samples form12producing area,5Baphicacanthus cusia rhizoma et radix and its adulterants species from4genus which are form2families inclued7samples,7Dioscorea opposita rhizoma and its adulterants samples from3genus which are form3families inclued13samples. And then the experiment selected4andidate sequence and amplified then in all df the nuclear genome DNA sequences of ITS2, psbA-trnH, rbcL chloroplast genome DNA and matK four DNA barcoding sequence, the sequence and Genbank registered sequence were analyzed, the characteristics of each species, sequence between genetic distance and according to their genetic distance cluster analysis tree construction.Results:From the extraction of DNA, PCR amplification and electrophoresis conditions of the view, Baphicacanthus cusia rhizoma et radix and its adulterants four sequences were effective.From the genetic distances, phylogenetic tree construction and investigation results of the view,60Baphicacanthus cusia rhizoma et radix samples form12producing area have less differences,.There are only four variable sites in ITS2sequence, forming three haplotypes; and there are many differences between Baphicacanthus cusia rhizoma et radix and its adulterants。The ITS2sequence were the best sequence for the identification of Baphicacanthus cusia rhizoma et radix and its adulterants.From the extraction of DNA PCR amplification and electrophoresis conditions of the view, ITS2sequence are poor of PCR amplification and electrophoresis conditions,and the other sequences are effective.From the genetic distances, phylogenetic tree construction and investigation results of the view, single sequence is not suitable for the identification of Dioscorea opposita rhizomaand its adulterants. We need to use multiple combinations of sequences their identification, in which psbA-trnH+rbcL+matK composite sequence work best.Conclusion:DNA barcoding can be used for the identification of traditional Chinese medicine.And ITS2sequence can be used for identification of Baphicacanthus cusia rhizoma et radix and its adulterants,psbA-trnH+rbcL+matK sequence can be used for identification of Dioscorea opposita rhizoma and its adulterants.