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Value of FINS Depends on Choice of DNA Locus as Shown in Baihuasheshecao

白花蛇舌草的FINS方法鑑定

摘要


Forensically informative nucleotide sequencing (FINS) is a molecular approach to identify biological materials based on DNA sequences. It focuses on the clustering of samples with reference species through an analysis of the phylogenetic relationship among the species. The choice of loci is critical to the successful use of FINS for identification. In this study, we investigated the influence of six loci (ITS, matK, rbcL, rps16, trnH-psbA and trnL-trnF) on the application of FINS for the identification of traditional Chinese medicine as demonstrated in Baihuasheshecao, a traditional anti-tumor herb derived from Hedyotis diffusa willd. A total of 36 plant samples of Hedyotis species and eight samples of Baihuasheshecao commodities were collected from Hong Kong, Macao, Taiwan, mainland China and the USA. Internal transcribed spacers (ITS) sequences displayed the highest rate of formation of single species cluster (89%), followed by rps16 (78%), matK (74%), trnL-trnF (72%), trnH-psbA (67%) and rbcL (61%). All the loci could discriminate H. diffusa from the common substitute species H. corymbosa, H. pinifolia and H. tenelliflora. FINS identification based on ITS, trnH-psbA and trnL-trnF revealed that five Baihuasheshecao commodities were genuine herb derived from H. diffusa, while the other three commodities were substitutes derived from H. corymbosa. MatK, rbcL and rps16 could not be amplified from all commodity samples and were thus inapplicable for FINS identification of dried the samples. In conclusion, the success of FINS identification depends on the DNA loci as they show different abilities to differentiate genuine species from substitutes, to discriminate closely related species and to generate quality sequences. In this study, the eligibility of DNA locus for FINS identification of Baihuasheshecao commodities, in descending order, is ITS > trnL-trnF > trnH-psbA > rps16 > matK > rbcL.

關鍵字

白花蛇舌草 FINS 耳草屬 分子鑑定

並列摘要


Forensically informative nucleotide sequencing (FINS) is a molecular approach to identify biological materials based on DNA sequences. It focuses on the clustering of samples with reference species through an analysis of the phylogenetic relationship among the species. The choice of loci is critical to the successful use of FINS for identification. In this study, we investigated the influence of six loci (ITS, matK, rbcL, rps16, trnH-psbA and trnL-trnF) on the application of FINS for the identification of traditional Chinese medicine as demonstrated in Baihuasheshecao, a traditional anti-tumor herb derived from Hedyotis diffusa willd. A total of 36 plant samples of Hedyotis species and eight samples of Baihuasheshecao commodities were collected from Hong Kong, Macao, Taiwan, mainland China and the USA. Internal transcribed spacers (ITS) sequences displayed the highest rate of formation of single species cluster (89%), followed by rps16 (78%), matK (74%), trnL-trnF (72%), trnH-psbA (67%) and rbcL (61%). All the loci could discriminate H. diffusa from the common substitute species H. corymbosa, H. pinifolia and H. tenelliflora. FINS identification based on ITS, trnH-psbA and trnL-trnF revealed that five Baihuasheshecao commodities were genuine herb derived from H. diffusa, while the other three commodities were substitutes derived from H. corymbosa. MatK, rbcL and rps16 could not be amplified from all commodity samples and were thus inapplicable for FINS identification of dried the samples. In conclusion, the success of FINS identification depends on the DNA loci as they show different abilities to differentiate genuine species from substitutes, to discriminate closely related species and to generate quality sequences. In this study, the eligibility of DNA locus for FINS identification of Baihuasheshecao commodities, in descending order, is ITS > trnL-trnF > trnH-psbA > rps16 > matK > rbcL.

參考文獻


Li, M., Zhang, Y. B., But, P. P. H. and Shaw, P. C. 2011. Forensically informative nucleotide sequencing (FINS) for the authentication of Chinese medicinal materials. Chin. Med. 6: 42.
Chan, W. H., Ling, K. H., Shaw, P. C., Chiu, S. W. and But, P. P. H. 2012. Application of FINS and multiplex PCR for detecting genuine abalone products. Food Control 23: 137-142.
Chapela, M. J., Sotelo, C. G., Calo-Mata, P., Pérez-Martín, R. I., Rehbein, H., Hold, G. L., Quinteiro, J., Rey-Méndez, M., Rosa, C. and Santos, A. T. 2002. Identification of Cephalopod species (Ommastrephidae and Loliginidae) in seafood products by forensically informative nucleotide sequencing (FINS). J. Food Sci. 67: 1672-1676.
Guha, S. and Kashyap, V. K. 2006. Molecular identification of lizard by RAPD & FINS of mitochondrial 16s rRNA Gene. Leg. Med. 8: 5-10.
Lau, D. T. W., Shaw, P. C., Wang, J. and But, P. P. H. 2001. Authentication of medicinal Dendrobium species by the internal transcribed spacer of ribosomal DNA. Planta Med. 67: 456-460.

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