Molecular authentication and quantitative analysis of Sarcandra glabra and adulterated chloranthus products using SNP markers

Mitochondrial DNA A DNA Mapp Seq Anal. 2016 Sep;27(5):3618-25. doi: 10.3109/19401736.2015.1079826. Epub 2015 Sep 10.

Abstract

Background: Sarcandra glabra (Thunb.) Nakai is one of the most popular and valuable plant species in the oriental medicinal herb market. Chloranthus (Chloranthaceae) species are the most widely used adulterants, but they are known to have hepatotoxicity effects and different medicinal values.

Objective: The aim of this study is to develop a robust and accurate DNA marker for the qualitative and quantitative analyses of their products.

Materials and methods: Four single nucleotide polymorphism (SNP) sites specific to Sarcandra glabra, Chloranthus spicatus, Chloranthus serratus and Chloranthus henryi were exploited from the trnL-F region in chloroplast DNA, which have a higher copy number in the products than the nuclear DNA. Based on the SNP sites, specific primers were designed to identify the products of Sarcandra glabra, Chloranthus spicatus, Chloranthus serratus and Chloranthus henryi in mixed solutions via multiplexed PCR. The primers were also used to quantitatively analyse the ratio of chloroplast DNA in the mixed products using real-time PCR.

Results: The established multiplexed-PCR and real-time PCR methods were determined to be effective for the authentication and relative quantitative assessments of the products of Sarcandra glabra, its adulterants, and their mixtures.

Conclusion: We therefore present an effective method for monitoring the quality of these products.

Keywords: Chloranthus; Sarcandra glabra; TrnL-F; multiplexed allele-specific PCR; real-time PCR SNP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA, Chloroplast / genetics
  • Genes, Chloroplast
  • Molecular Typing
  • Multiplex Polymerase Chain Reaction
  • Plants, Medicinal / genetics
  • Polymorphism, Single Nucleotide*
  • Sequence Analysis, DNA
  • Sequence Homology, Nucleic Acid
  • Tracheophyta / genetics*

Substances

  • DNA, Chloroplast