First Insights into Genetic Diversity of Indonesian Typhonium flagelliforme Using Transcriptome-Based EST- SSR Markers

Typhonium flagelliforme , an Araceae species recognized for its anticancer metabolites distributed throughout its roots, tubers, stems, and leaves, remains underexplored at the molecular level. In this study, we developed expressed sequence tag–simple sequence repeat (EST-SSR) markers from the T. flagelliforme transcriptome and evaluated their utility in assessing the genetic diversity of Indonesian germplasm as well as their transferability to other Araceae species. Transcriptome assembly yielded 41,392 unigenes, of which 9,868 were successfully annotated. Mining of 18,225 unigenes identified 24,404 potential EST-SSRs, with an overall frequency of one SSR per 4.36 kb and di-nucleotide repeats (45.83%) being the most abundant motif. From these, 38 primer pairs were randomly selected, and 32 produced clear amplicons, of which 23 (60.5%) were polymorphic across 14 accessions. A total of 15 alleles were detected, with an average of 2.87 alleles per locus. The mean observed and expected heterozygosity were 0.238 and 0.620, respectively, while the polymorphism information content (PIC) ranged from 0.416 to 0.580, with an average of 0.515, indicating a moderate level of polymorphism. Genetic structure analysis and principal coordinate analysis (PCoA) consistently grouped the accessions into three distinct genetic clusters, underscoring the effectiveness of the developed markers in resolving population structure. To our knowledge, this represents the first genetic diversity analysis of Indonesian T. flagelliforme germplasm using EST-SSR markers. The newly developed resources provide a valuable foundation for molecular characterization, germplasm conservation, and breeding applications in T. flagelliforme and potentially related Araceae species.