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Genome and developmental transcriptome of the forensically important blowfly, Phormia Regina

Lin, Sheng-Hao1; Bellantuono, Anthony1; Wells, Jeffrey1; DeGennaro, Matthew1

Published Jul 08, 2025; Updated Oct 27, 2025 on Dryad. https://doi.org/10.5061/dryad.w6m905r04

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Abstract

Forensic entomologists often rely on the morphological traits of insects, such as length and weight, to estimate the time of death. Blowflies like Phormia regina are particularly significant in North American forensic investigations. However, age estimation of older P. regina maggots becomes challenging due to limited morphological changes during the lengthy L3 larval phase of development. To address this gap, we used transcriptomic profiling of blowfly maggots to generate molecular markers that specify their age. We first characterized maggot weight, behavior, and mRNA in 10-hour increments during development. At 27.5°C, the weight of the maggots increased from when first recorded at 70 through 100 hours and then remained stable from 110 hours to pupation. The behavioral transition between the feeding and the post-feeding wandering stage usually took place between 100 and 120 hours.  Second, we built a chromosomal-scale Phormia regina genome annotated with long mRNA reads to provide a reliable database to uncover transcriptomic signatures during larval development. We applied differential gene expression analysis (DEGs), weighted gene co-expression network analysis (WGCNA), and the generalized linear model (GLM) to identify nine candidate genes that all three statistical analyses indicated are useful for delineating the age of otherwise indeterminate L3 maggots. In turn, these genes could be used to design a quantitative PCR protocol for more accurate estimates of the time of death.