ANALISIS FILOGENETIK KUPU-KUPU E N D E M I K S U L A W E S ICethosia Myrina BERDASARKAN GEN CYTOCHROME OXIDASE I (COI)

Cethosia myrina is a butterfly species belonging to the family Nymphalidae and is endemic to Sulawesi, Indonesia. The phylogenetic
relationships of this species were investigated through molecular phylogenetic analysis. Phylogenetics is a method used to examine the
evolutionary relationships among living organisms. This study aimed to describe the phylogenetic relationships of C. myrina based on the
mitochondrial Cytochrome Oxidase Subunit I (COI) gene. Samples were collected using an exploratory survey method. Genomic DNA
was isolated using the GS 100gSYNC™ DNA Extraction Kit. DNA amplification was performed using COI primers (LCO1490 forward
and HCO2198 reverse) through the Polymerase Chain Reaction (PCR) technique. Amplified DNA fragments were visualized using 1%
agarose gel electrophoresis, a UV transilluminator, and a gel documentation system. Subsequent bioinformatics analyses were conducted
using Gene Studio, DnaSP, BLAST, DNASTAR, and MESQUITE software. Phylogenetic reconstruction was performed using the
Neighbor-Joining (NJ) and Maximum Likelihood (ML) methods in MEGA version 11, applying the Kimura 2-parameter model with
10,000 bootstrap replicates. The results demonstrated that both the Neighbor-Joining and Maximum Likelihood methods produced
phylogenetic trees that clearly illustrated the evolutionary relationships among the analyzed samples. Genetic variation analysis revealed
the presence of two haplotypes, with a haplotype diversity (Hd) of 0.603 ± 0.165 and a nucleotide diversity (?) of 0.00053 ± 0.00013.
Phylogenetic reconstruction formed a single monophyletic clade of C. myrina supported by a 99% bootstrap value. A genetic distance of
0.00% was observed among populations from Central Sulawesi (CMST1.1, CMST1.2, and CMST1.3), Central Sulawesi (EU275514.1),
and South Sulawesi (HM998338.1), indicating an extremely close genetic relationship among these populations.