The Partial Nucleotide Sequences of the Mitochondrial Genes, COI and 16s rRNA, of Fireflies in the Genera Pygoluciola, Trisinuata, and Medeopteryx (Coleoptera: Lampyridae)

Abstract

Fireflies in the genera Pygoluciola, Medeopteryx, and Trisinuata distribute throughout lower northern Thailand. Five morphospecies of these fireflies were collected and nucleotide sequences were analyzed. We sequenced the partial of two mitochondrial genes; Cytochrome c oxidase subunit I (COI) and 16s rRNA, and the phylogenetic trees were constructed and compared with the firefly nucleotide sequences in the NCBI database. The COI gene was amplified using LCO1490 and HCO2198 primers while specific primers of LR-J-13020a and LR-N-13398a were used to analyze amplicons of 16s rRNA. The results showed the average GC-contents in the nucleotide of five firefly morphospecies were found to average 30.28% and 22.15% in (COI) and 16s rRNA, respectively. The variable site in COI nucleotide sequences was 399 of the total 520 nucleotides (76.73%) whereas 91 of the total 418 nucleotides (21.77%) were found in 16s rRNA nucleotide sequences. Construction of the phylogenetic trees of those five firefly morphospecies with reference accession samples indicated five clades when analyzed by COI nucleotide sequences and two clades were revealed in 16s rRNA gene. According to these results, COI nucleotides are one of the most effective methods for differentiating between and within firefly species. The addition of the 16s rRNA gene analysis showed an increase in the precision of firefly identification. Those five firefly morphospecies were separated using both the COI and 16s rRNA nucleotide sequences.