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DOI: 10.1055/s-0028-1084977
An assessment of nrITS sequence for use in barcoding traditional Chinese medicinal herbs
DNA barcoding has been applied to systematic and phylogenetic studies on animals [1] and limited numbers of plant taxa [2,3]. In this study, we attempted to use the nuclear ribosomal internal transcribed spacer (nrITS) as a barcoding marker candidate for classification of traditional Chinese medicinal herbs (TCM) which are lacking of adequate methods for morphological and chemical identification.
The ITS region of DNA extracted from 49 TCM samples was successfully amplified by PCR with primers T3F (5'-GATCAATGCAGACCCATGAAT-3') and T3R (5'-TGATGTGCCTACAACTTGGAG-3'), and then the PCR products were directly sequenced with primer pairs T3F/T3R and 5.8S1/5.8S2 (5'-GCAACTTGCGTTCAAAGACTC-3', 5'-GATGAAGAACGTAGCGAAATG-3'). 152 polymorphic loci have been obtained by Clustal alignment of the 49 unique sequences. Also, we observed average K2P (Kimura two parameter) distances of 0.15 and 0.067 within genus and species. This result suggested that nrITS sequence can be a universal DNA barcode for use in barcoding TCM herbs.
References:1. Hebert, P.D. et al., (2003) Proc Biol Sci. 270 Suppl1: p. S96–9.
2. Ward, R.D. et al., (2005) Philos Trans R Soc Lond B Biol Sci. 360: 1847–57.
3. Kress, W.J. et al., (2005) Proc Natl Acad Sci U S A. 102: 8369–74.