title: Specific PCR Identification betweenPeucedanum praeruptorumandAngelica decursivaand Identification between Them and Adulterant Using DNA Barcode.
abstract: BACKGROUND
  The traditional Chinese medicine (TCM) Qianhu and Zihuaqianhu are the dried roots ofPeucedanum praeruptorumandAngelica decursiva, respectively. Since the plant sources of Qianhu and Zihuaqianhu are more complex, the chemical compositions ofP. praeruptorumandA. decursivaare significantly different, and many adulterants exist because of the differences in traditional understanding and medication habits. Therefore, the rapid and accurate identification methods are required.
  
  
  OBJECTIVE
  The aim was to study the feasibility of using DNA barcoding to distinguish between Traditional Chinese medicine Qianhu (Peucedanum praeruptorum), Zihuaqianhu (Angelica decursiva), and common adulterants, based on internal transcribed spacer (ITS) sequences, as well as specific PCR identification betweenP. praeruptorumandA. decursiva.
  
  
  METHODS
  The ITS sequences ofP. praeruptorum,A. decursiva, and adulterant were studied, and a phylogenetic tree was constructed. Based on the ITS barcode, the specific PCR primer pairs QH-CP19s/QH-CP19a and ZHQH-CP3s/ZHQH-CP3a were designed forP. praeruptorumandA. decursiva, respectively. The amplification conditions were optimized, and specific PCR products were obtained.
  
  
  RESULTS
  The results showed that the phylogenetic trees constructed using the BI and MP methods were consistent, andP. praeruptorumandA. decursivasequence haplotypes formed their own monophyly. The experimental results showed that in PCR products, the target bands appeared in the genuine drug and not in the adulterant, which suggests the high specificity of the two primer pairs.
  
  
  CONCLUSIONS
  The ITS sequence was ideal DNA barcode to identifyP. praeruptorum,A. decursiva, and adulterant. The specific PCR is a quick and effective method to distinguish betweenP. praeruptorumandA. decursiva.
  
  
  CONCLUSIONS
  Peucedanum praeruptorumandAngelica decursivasequence haplotypes formed their own monophyly.The ITS sequence was ideal DNA barcode to identifyP. praeruptorum,A. decursiva, and adulterant.Specific PCR is a quick and effective method to distinguish betweenP. praeruptorumandA. decursiva.Abbreviations used:TCM: The traditional Chinese medicine,P.: Peucedanum,A.: Angelica, ITS: The internal transcribed spacer, PCR: Polymerase chain reaction, NCBI: National Center for Biotechnology Information, NI: Number of individuals, HN: Haplotype number; GAN: Gen Bank accession numbers,L.: Ligusticum,O.: Ostericum,A.: Angelica,P.: Pimpinella, BI: Bayesian inference, MP: Maximum parsimony, AIC: Akaike Information Criterion, MCMC: Markov Chains Monte Carlo, TBR: Tree bisection-reconnection, LPP: Length of PCR product, PRP: PCR reaction procedure, SNP: Single nucleotide polymorphisms, PP: Posterior probability, BS: Bootstrap.Qun Zhao.
authors.affiliation: College of Biological and Pharmaceutical Engineering, West Anhui University, Lu'an, Anhui Province, China; State Key Laboratory Breeding Base of Daodi Herbs, China Academy of Chinese Medical Sciences, Beijing, China.
authors.forename: Bang-Xing
authors.initials: BX
authors.lastname: Han
authors.affiliation: State Key Laboratory Breeding Base of Daodi Herbs, China Academy of Chinese Medical Sciences, Beijing, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Yuan
authors.affiliation: State Key Laboratory Breeding Base of Daodi Herbs, China Academy of Chinese Medical Sciences, Beijing, China.
authors.forename: Lu-Qi
authors.initials: LQ
authors.lastname: Huang
authors.affiliation: College of Biological and Pharmaceutical Engineering, West Anhui University, Lu'an, Anhui Province, China; State Key Laboratory Breeding Base of Daodi Herbs, China Academy of Chinese Medical Sciences, Beijing, China.
authors.forename: Qun
authors.initials: Q
authors.lastname: Zhao
authors.affiliation: School of life science, Qingdao agricultural university, Qingdao, Shandong Province, China.
authors.forename: Ling-Ling
authors.initials: LL
authors.lastname: Tan
authors.affiliation: College of Biological and Pharmaceutical Engineering, West Anhui University, Lu'an, Anhui Province, China.
authors.forename: Xiang-Wen
authors.initials: XW
authors.lastname: Song
authors.affiliation: College of Biological and Pharmaceutical Engineering, West Anhui University, Lu'an, Anhui Province, China.
authors.forename: Xiao-Mei
authors.initials: XM
authors.lastname: He
authors.affiliation: Department of Pharmacy, Anhui University of Chinese Medicine, Hefei, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Xu
authors.affiliation: College of Biological and Pharmaceutical Engineering, West Anhui University, Lu'an, Anhui Province, China; Department of Pharmacy, Anhui University of Chinese Medicine, Hefei, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Liu
authors.affiliation: Department of Pharmacy, Anhui University of Chinese Medicine, Hefei, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
keywords: Angelica decursiva; DNA barcode; ITS; Peucedanum praeruptorum; specific PCR