Screening of DNA Barcoding Sequences for Identification of Multiple Species of Aquilaria L / 中国药学杂志

ABSTRACT

OBJECTIVE: To screen out efficient sequences from DNA barcoding sequences in order to rapidly identify multiple species of Aquilaria. METHODS: Five species of Aquilaria were collected from eight countries as test materials. The ITS2 sequence in nuclear genome as well as matK, rbcL and trnH-psbA sequences from chloroplast genome were analyzed and evaluated. Phylogenetic trees based on different sequence combinations were built to evaluate the identification efficiency of Aquilaria species. RESULTS: All four kinds of sequences exhibited relatively high success rate of PCR amplification and sequencing. matK had the most variation sites and SNPs while trnH-psbA had the fewest. Barcoding gap test demonstrated that the identification effect of sequences from high to low was matK>ITS2>rbcL>trnH-psbA. According to the homogeneity test and average node supporting rate, phylogenetic trees based on the combination of matK+ITS2+rbcL could clearly divide different species into separated clusters with high node supporting rate. The matK showed the best identification effect when combined with other sequences as shown by the BBA method of Taxon DNA. CONCLUSION: When using DNA barcoding technique to identify Aquilaria species, matK has the highest identification rate among single barcoding candidate sequences. And the combination of matK+ITS2+rbcL can precisely distinguish multiple Aquilaria species.