DNA barcoding for authentication of the endangered plants in genus Aquilaria

Worathat Thitikornpong, Chanida Palanuvej, Nijsiri Ruangrungsi

Abstract


            Agarwood is a type of wood that is used in traditional Thai medicines and has been applied in several recipes; in particular, this wood is considered as a costly resinous non-wood product. However, the morphological characteristics of the plant in the genus Aquilaria are quite similar, and so the authentication steps need a botanical expert for the purposes of identification and then also require an efficient tool for species-level discrimination. DNA barcoding is an approved molecular technique which uses a short sequence as a barcoding region specific for identified species. In this study, we established the DNA barcoding sequences from six candidate of barcoding loci (ITS, matK, rbcL, rpoC1, psbA-trnH intergenic spacer, and ycf1) from three species of Aquilaria, and the outgroup (Enkleia siamensis (Kurz) Nervling). The phylogenetic tree of each locus was reconstructed and the genetic distances were also determined using a maximum likelihood method. Our results showed that all samples in each locus were successfully amplified and sequenced with universal primers. The ITS was only performed with suitable markers for Aquilaria species identification, which were considered from ML phylogenetic tree reconstruction and the optimum length of genetic distance. Other cpDNA barcoding regions; matK, rbcL, rpoC1, ycf1and trnH-psbA, showed lower discrimination power, so these loci were deemed inappropriate for species identification. Notwithstanding, each barcoding locus can be suitable for genus discrimination in the Thymelaeaceae family. From our findings, DNA barcoding is able to be used as an efficient and reliable tool in Aquilaria species authentication, which can be applied for quality control in the agarwood in the global market.


Full Text:

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