Developing molecular marker using Internal Transcribed Spacer (ITS) regions for Zingiber spp. identification

In the order Zingiberales, the pantropic Zingiberaceae is the largest family with over 1200 species and 53 genera with varieties of medicinal and culinary uses. There is a very limited information exist using Internal Transcribed Spacer (ITS) regions as a molecular marker in the identification of Zingiber and Etlingera species. This study aims to develop a molecular marker for Zingiber and Etlingera species identification and differentiation. The Polymerase Chain Reaction (PCR) method was used in the identification of Zingiber and Etlingera species using Internal Transcribed Spacer regions in this study. The leaf samples of Zingiber officinale and Etlingera elatior were collected around Kelantan, Malaysia and Cetyl Trimethyl Ammonium Bromide (CTAB) method was used to isolate genomic DNA. Then the isolated DNA were used for ITS sequences amplification and analysis. The amplified ITS regions of Zingiber officinale and Etlingera elatior were sent for Sanger sequencing and the ITS sequences obtained were used to identify the species through Basic Local Alignment Search Tool. The sequencing results for isolated DNA Zingiber officinale showed 91% match with Zingiber officinale in National Center for Biotechnology Information Genbank (NCBI) database and 99% similar with Etlingera elatior. The molecular phylogenetic trees were constructed and indicated that Zingiber species and Etlingera species were separated into two cluster. Hence, it indicate that ITS can be used as the molecular marker in future for Zingiber and Etlingera species identification.