Identification of molecular marker for Genus Etlingera based on Internal Transcribed Spacer (ITS) region

This research aimed to identify the specific molecular marker for ginger (Zingiberaceae) as there is very limited molecular study on ginger especially for genus Etlingera. Three selected species from genus Etlingera was selected which are E. elatior, E. megalocheilos and E. littoralis. These three species were chosen from the twelve Etlingera species found in Peninsular of Malaysia. The DNA from these three species were isolated by using Cetyl Trimethylammonium Bromide (CTAB) extraction method and the results from the extraction were then used to conduct Polymerase Chain Reaction (PCR) with the Internal Transcribed Spacer region as the target sequence region. The Internal Transcribed Spacer specific primer was chosen and amplified via PCR. Results from two of the species were sent for sequencing and the sequences obtained were used for species identification through Basic Local Alignment Search Tool (BLAST). The results show these two species correctly belong to genus Etlingera (Zingiberaceae) by comparing with the closest match from Nucleotide BLAST. Three phylogenetic trees were constructed by using MEGA7 Software to show the phylogeny relationship among selected species in Zingiberaceae. Lastly specific primers for two species of Etlingera were designed by using Primer3 Plus. In a nutshell, the designed specific primers for E. elatior and E. megalocheilos identification were developed.