The morphology, geographical distribution, and evolutionary relationships of butterflies belonging to the Pieridae family exhibit significant variation. The current study analyzed the nucleotide substitution pattern and evolutionary relationships of specific Gonepteryx and related Lepidoptera sequences. The 16-nucleotide gene sequence of mitochondrial Cytochrome Oxidase subunit-1 (COI) was retrieved from the NCBI GenBank database. A significant bias toward transitions was revealed by the substitution analysis, with C→T/U exhibiting the highest relative substitution rate (34.88). A significant AT bias was also evident in the nucleotide composition, with A and T/U together comprising about 71.01% of the sequences. A number of strongly supported molecular affinities were identified using the Maximum Likelihood tree. While the G. cleopatra-associated group showed 99% support, the two Gonepteryx farinosa sequences formed a well-supported clade with 100% bootstrap support.

The G. aspasia and G. mahaguru sequences clustered with 65% bootstrap support, but their larger grouping with G. rhamni was 100% supported. One Lepidoptera sp. sequence demonstrated a tight relationship with the G. cleopatra group with an 88% bootstrap value, whereas two Lepidoptera sp. sequences constituted a well-supported group with 90% bootstrap support. On the other hand, certain G. rhamni sequences had bootstrap values of 23% and 29%, indicating weakly supported associations.