Using the Developmental GeneBicoidto Identify Species of Forensically Important Blowflies (Diptera: Calliphoridae)

Abstract

Identifying species of insects used to estimate postmortem interval (PMI) is a major subject in forensic entomology. Because forensic insect specimens are morphologically uniform and are obtained at various developmental stages, DNA markers are greatly needed. To develop new autosomal DNA markers to identify species, partial genomic sequences of the bicoid (bcd) genes, containing the homeobox and its flanking sequences, from 12 blowfly species (Aldrichina grahami, Calliphora vicina, Calliphora lata, Triceratopyga calliphoroides, Chrysomya megacephala, Chrysomya pinguis, Phormia regina, Lucilia ampullacea, Lucilia caesar, Lucilia illustris, Hemipyrellia ligurriens and Lucilia sericata; Calliphoridae: Diptera) were determined and analyzed. This study first sequenced the ten blowfly species other than C. vicina and L. sericata. Based on the bcd sequences of these 12 blowfly species, a phylogenetic tree was constructed that discriminates the subfamilies of Calliphoridae (Luciliinae, Chrysomyinae, and Calliphorinae) and most blowfly species. Even partial genomic sequences of about 500 bp can distinguish most blowfly species. The short intron 2 and coding sequences downstream of the bcd homeobox in exon 3 could be utilized to develop DNA markers for forensic applications. These gene sequences are important in the evolution of insect developmental biology and are potentially useful for identifying insect species in forensic science.

Phenotypic polymorphism of Chrysomya albiceps (Wiedemann) (Diptera: Calliphoridae) may lead to species misidentification

“…Indeed, this result corroborates with those of Park et al (2013) that this region of the developmental bcd may be a suitable molecular marker for forensic science and entomology. In addition, the multigene phylogenetic analyses were able to differentiate the C. putoria individuals collected in Manaus, Amazonas, from those collected in Campinas, SP, Brazil.…”

“…The following primers were used: Chrybcd-Intr2 (5 -CCAAATCAATTACCAAAGCCAGGTG-3 ) and Chrybcd-Exo3 (5 -CATTTGGTGTTGATCCACCACTGCC-3 ), adapted from primers described for Chrysomya species analyzed by Park et al (2013) and designed in this study. PCR conditions were the same as those described for the COI and COII regions.…”

“…ITS2, which has been evaluated for species identification (Nelson et al, 2007;Song et al, 2008a), phylogenetic inferences (Song et al, 2008b;Marinho et al, 2011Marinho et al, , 2012, and bcd, which was recently evaluated as a molecular identification marker (Park et al, 2013), showed a low sequence divergence when compared with mtDNA regions (Table A.2). These may be a consequence of the higher mutation rate of mtDNA than nuclear DNA (Harvey et al, 2003b).…”

Molecular Species Identification of Forensically Important Flies in Korea

“…이 중 검정파리과 파리의 염기서열 대부분은 NCBI Genbank에 등록되었으며, 23,24) 집파리과와 쉬파리과의 염기서열도 제출되어 등록을 기다 리고 있다 (Table 1). 그 외에 핵 DNA 표지자를 발굴하기 위해 abd-a, ubx, dfd, scr, bcd, abd-b 항상성상자(homeobox) 염 기서열이 다수 종에 대하여 분석이 되었으며, [34][35][36][37] 이 중 bcd 염 기서열을 이용한 검정파리과 파리에 대한 multiplex genotyping kit의 특허가 승인되었다. COI 다음으로 많이 사용 되는 표지자인 ITS2에 대한 분석은 현재 연구가 진행 중이다.…”

Is aggregated oviposition by the blow flies Lucilia sericata and Phormia regina (Diptera: Calliphoridae) really pheromone‐mediated?

“…Even if the flies do not signal themselves, their salivary secretions may contain enzymes and microorganisms that initiate metabolism of carrion carbohydrates, proteins or lipids (Dethier, ; Telford et al ., ). These metabolic activities facilitate liquefaction and breakdown of the carrion surface for fly feeding, and in the process produce volatile metabolites (Telford et al ., ; Park et al ., ; Zheng et al ., ) that are likely to attract flies. Irrespective of whether the semiochemicals are produced by the flies, their enzymes or microorganisms, they are likely associated with secretions from the salivary gland in the flies’ head.…”