Wijayasiriwardene T. D. C. M. K.1*, Herath H. M. I. C.2 and Premakumara G. A. S.1
1. Industrial Technology Institute, Colombo, Sri Lanka.
2. Faculty of Graduate Studies, University of Colombo, Sri Lanka.
Abstract
Harankaha is important medicinal plant, which is used in Sri Lankan traditional medicine. However, three plants are reported under the same vernacular name (e.g. Curcuma albiflora, Curcuma zedoaria, and Zingiber zerumbet) and therefore the raw material may be adulterated or substituted with each other. Phylogenetic analysis of gene sequences and combining with complete genomic sequences helps to identify genetic basis of plants. Standard CTAB method with little modifications was used for the extraction and purification. Extracted DNA was amplified using universal primers for matK genes in chloroplast genome rbcL gene in chloroplast genome by PCR (polymerase chain reaction). The rbcL primers amplified about 600 bp, while matK was about 850 bp. Amplified fragments were sequenced and obtained the DNA sequences for the matK and rbcL genes. Sequenced fragments were analyzed and used for DNA barcoding. DNA barcode was submitted to BOLD (Barcoding of Life Database) online database and then uploaded to the GenBank through the BOLD system (Accession No KF 521885). The distance of interspecific were 0.0010 in rbcL and 0.05 in matK, which is less than or equal to 0.05 matK sequences were selected for identification of C. albiflora, C. zedoaria and Z. zerumbet. C. albiflora appeared as a different group as per the Neighbor-Joining method and therefore, it can be identified as a new group. The matK gene of C. albiflora and other species showed, totally 212 variable sites. However, rbcL gene of C. albiflora and other species showed only 2 variable sites. Therefore, matK gene is suggested to identify C. albiflora from other two species claimed as Harankaha plants.
Key words : Curcuma albiflora, DNA barcoding, MaturaseK