Discovery of MicroRNAs of the Stable Fly (Diptera: Muscidae) by High-Throughput Sequencing

Abstract

The stable fly, Stomoxys calcitrans (L.), is a serious ectoparasite affecting animal production and health of both animals and humans. Stable fly control relies largely on chemical insecticides; however, the development of insecticide resistance as well as environmental considerations requires continued discovery research to develop novel control technologies. MicroRNAs (miRNAs) are a class of short noncoding RNAs that have been shown to be important regulators of gene expression across a wide variety of organisms, and may provide an innovative approach with regard to development of safer more targeted control technologies. The current study reports discovery ad initial comparative analysis of 88 presumptive miRNA sequences from the stable fly, obtained using high-throughput sequencing of small RNAs. The majority of stable fly miRNAs were 22-23 nt in length. Many miRNAs were arthropod specific, and several mature miRNA sequences showed greater sequence identity to miRNAs from other blood-feeding dipterans such as mosquitoes rather than to Drosophilids. This initial step in characterizing the stable fly microRNAome provides a basis for further analyses of life stage-specific and tissue-specific expression to elucidate their functional roles in stable fly biology.

Identification and characterization of microRNAs in the screwworm flies Cochliomyia hominivorax and Cochliomyia macellaria (Diptera: Calliphoridae)

“…Furthermore, the miRNA repertoire of the screwworm flies and other schizophoran flies ( Fig. 1D) are comparable and quantitatively similar, including the housefly, M. domestica (138 miRNAs; Scott et al, 2014), the stable fly, S. calcitrans (88 miRNAs; Tuckow et al, 2013), the Oriental fruit fly, B. dorsalis (123 miRNAs; Huang et al, 2014) and the sheep blowfly, L. cuprina (118 miRNAs predicted in this study; see Experimental Procedures and Table S2). A total of 69 miRNAs (39%) described for other species of Schizophora could not be identified in C. hominivorax and C. macellaria (Fig.…”

“…B). This is a conserved miRNA characteristic found in many insect species, including other flies (Stark et al ., ; Skalsky et al ., ; Tuckow et al ., ; Huang et al ., ).…”

“…domestica (138 miRNAs; Scott et al ., ), the stable fly, S . calcitrans (88 miRNAs; Tuckow et al ., ), the Oriental fruit fly, B . dorsalis (123 miRNAs; Huang et al ., ) and the sheep blowfly, L .…”

“…melanogaster by other authors (Shcherbata et al ., ; Yang et al ., ), who suggested that bantam‐3p is required for stem cell maintenance in adult flies, the prevention of apoptosis during cell proliferation and the regulation of germline stem cell fate. The miR‐8‐3p is one of the most abundant miRNAs in the small RNA transcriptomes of the screwworm flies (Table S1), as for other insects (Skalsky et al ., ; Cristino et al ., ; Tuckow et al ., ; Huang et al ., ). It has been reported that miR‐8‐3p participates in a large number of biological process, including development (Belles et al ., ), egg and ovarian formation (Lucas et al ., ), neuromotor coordination (Karres et al ., ; Rubio et al ., ) and maintenance of the innate immune system homeostasis (Choi & Hyun, ).…”

“…transcriptomes of the screwworm flies (Table S1), as for other insects (Skalsky et al, 2010;Cristino et al, 2011;Tuckow et al, 2013;Huang et al, 2014). It has been reported that miR-8-3p participates in a large number of biological process, including development (Belles et al, 2012), egg and ovarian formation (Lucas et al, 2015), neuromotor coordination (Karres et al, 2007;Rubio et al, 2013) and maintenance of the innate immune system homeostasis (Choi & Hyun, 2012).…”