Identification of Marker Nucleotides for the Molecular Authentication of Arisaematis Rhizoma Based on the DNA Barcode Sequences

Abstract

ABSTRACT Objectives : Official Arisaematis Rhizoma is described only three species, Arisaema amurnse , Arisaema erubescens , and Arisaema heterophyllum, in national Pharmacopoeia. However, other Arisaema species, Arisaema ringens, Arisaema takesimense and Arisaema serratum, also have been distributed as an inauthentic Arisaematis Rhizoma in the herbal market. To develop a reliable molecular a uthentication method for Arisaematis Rhizoma in species level, we analyzed DNA barcode regions using six Arisaema species.Methods : Thirty-eight samples of six Arisaema plants species (A. amurense, A. amurense f. serratum, A. heterophyllum, A. takesimense, and A. serratum) were collected from different habitate and nucleotide sequences of DNA barcode regions (rDNA-ITS, matK, and rbcL gene) were analyzed after PCR amplification. The species-spec ific sequences and phylogenetic relations were estimated using entire sequence s of three DNA barcodes based on the analysis of ClastalW and UPGMA, respectively.Results : The comparative analysis of DNA barcode sequences were revealed inter-species specific nucleotides to distinguish the medicinal plant of Arisaema Rhizoma in species levels excluding between A. amurense and its subspecies (A. amurense f. serratum) and A. takesimense and A. serratum, respectively. However, we obtained sequence differences enough to discriminate authentic and inaut hentic Arisaematis Rhizoma. Therefore, we suggest that these SNP type molecular genetic markers were an r eliable method avaliable to identify official herbal medicines.Conclusions : These marker nucleotides could be useful to identify the official herbal medicines by providing definitive information that can identify original medicinal pla nt and distinguish from inauthentic adulterants and substitutes.Key words : Arisaema species, Arisaematis Rhizoma, DNA barcode, Marker Nucleotide, Molecular Authentication