DNA barcoding expands reference libraries through the discovery of novel muscid species in Quetta, Balochistan, Pakistan

Hina Ali Ahmed, Sana Ahmed, Afroz Rais, Rabail Urooj, Razia Kanwal, Bibi Malaela, Nazeer Ahmed, Shahjahan Shabbir Ahmed Rana, Hafiz Muhammad Ali, Abdul Qadir

Abstract


Muscid flies are biological vectors of several disease pathogens and are medically and forensically significant insects. The study explored the diversity of dipterans using DNA barcoding and generated DNA barcodes (22 BINs; 4 unique, 18 non-unique) of 283 specimens, belonging to 6 genera and 7 species across 3 sub-families. The mitochondrial gene cytochrome C oxidase subunit-I (COI; 658bp) was accessed to examine the genetic diversity among 7 different muscid species. Coenosia attenuatta was identified as most common, representing 83% of collected specimens. According to barcode probe, largest conspecific divergence was 1.43%, whereas divergence from nearest neighbour species ranged between 7.41 to 13.55%. The barcoding of fly species showed a divergence of <2% compared to sequences available in Barcode of Life Data (BOLD) Systems and a significant difference was observed between maximum intra-specific and minimum nearest neighbour distances in all seven species. The Neighbour-Joining cluster analysis demonstrated that all species formed a monophyletic cluster, supported by high bootstrap values. The data identified 7 species matching to Barcode Index Number (BIN), 5 being novel (BOLD IDs: ADZ8375, AEA1946, ADZ9134, AEA2865, AEA0284) and reported first time in Pakistan. Moreover, these results revealed the significance of COI gene sequencing in exploring the diversification of insects and contributed a novel information regarding DNA barcode of a highly diverse and under-studied order of insects.

Keywords


DNA barcoding, Muscidae, Species identification, Reference library expansion, Biodiversity assessment

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References


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DOI: https://doi.org/10.33804/pp.010.02.6117

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