DNA fingerprint analysis of stemona in Thailand

Abstract

In Thailand many species of Stemona were reported and biological activities such as insecticide, insect repellency and anti-tumor were different. Thus, accurate identification of Stemona is needed in order to ensure their efficacies. The identification based on morphological characters of each species alone is difficult because the morphology of Stemona is similar and they are often sold as crude drug which lose their original feature. The purpose of this study was to analyze DNA fingerprint of six Stemona in Thailand; S. tuberosa Lour., S. collinsae Craib, S. phyllantha Gagnep., S. burkillii Prain, S. aphylla craib and Stemona sp., and developed PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) as a genetic marker in order to use as a convenient tool for identification. The nucleotide sequences of three DNA regions; matK, trnH-psbA and ITS1, were exploited to identify these six Stemona species in Thailand. A result of the comparison of partial matK could be classified Stemona into two groups, S. tuberosa group and S. collinsae group, concerning with their morphology and chemical composition. The nucleotide sequences of trnH-psbA and ITS1 could be used to discriminate six Stemona spcies in Thailand. On the basis of difference among the partial matK gene and ITS1 region, the PCR-RFLP analysis was performed. The restriction patterns showed distinct and polymorphic fingerprints among Stemona spp. and were able to apply for crude drug identification. These results exhibited that the obtained nucleotide sequences could be used to identify Stemona in Thailand. PCR-RFLP genetic marker developed here could be used as a convenient tool for authentication of Stemona and also be applied to their commercial crude drugs