Syifara Chika, Tiara Dwi Meilina, Muhammad Ramdhani Arfan, Asri Febriana & Arnia Sari Mukaromah 2024. Genetic variation of wax apple (Syzygium samarangense (Blume) Merr. & L.M. Perry) cultivar using internal transcribed spacer 2 (ITS2). Floribunda 7(4): 153–165 — Wax apple (Syzygium samarangense (Blume) Merr. & L.M. Perry) is a tropical fruit, has diverse cultivars and classified as a non-climacteric fruit. The study objectives were to identify the genetic variation of the S. samarangense cultivars in the ITS2 DNA barcode region, analyze the phylogenetic relationships among S. samarangense cultivars and determine successfulness of ITS2 barcode to describe genetic variation of the S. samarangense cultivars from Demak Regency, Central Java, Indonesia. Furthermore, the stages in this study consisted of sampling, environmental parameters analysis, DNA isolation, DNA amplification, electrophoresis, DNA sequencing in the First Base Malaysia, analysis of DNA sequencing data, secondary structure prediction of ITS2, and reconstruction a phylogenetic tree using MEGA 11 and ITS2 database. The result shown appearance of few nucleotide variation among S. samarangense cultivars and there were nucleotides variations between S. samarangense and Syzygium aqueum in the ITS2 region. S. samarangense cultivar Citra Wonosari and cultivar Delima Wonosari had the identical secondary structure of ITS2. However, this was slightly different between S. samarangense cultivar Citra Botorejo secondary structure of ITS2 which consisted of only two loops in helix I and cultivar Citra Tempuran second loop in helix I which bigger than all samples. The phylogenetic tree described the closely relationship between S. samarangense Citra cultivars, and had separated S. samarangense cultivar Delima in the different clade. The genetic relationship of S. samarangense cultivars were more influenced by cultivar type rather than cultivated location. Therefore, ITS2 could be utilized for genetic variation analysis of S. samarangense cultivars.
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