RT Journal Article SR Electronic T1 Genetic and behavioral analyses suggest that larval and adult stages of Lucilia cuprina employ different sensory systems to detect rotten beef JF bioRxiv FD Cold Spring Harbor Laboratory SP 2024.12.20.629795 DO 10.1101/2024.12.20.629795 A1 Wulff, Juan P. A1 Laminack, Rachel K. A1 Scott, Maxwell J. YR 2025 UL http://biorxiv.org/content/early/2025/01/09/2024.12.20.629795.abstract AB Background The blowfly Lucilia cuprina is a destructive parasite of sheep that causes flystrike or myiasis. Larvae consume the animal’s living flesh, producing large wounds that can lead to death. Growing resistance to conventional control methods has prompted the analysis of alternative strategies.Methods An RNA-Seq analysis was used to identify sensory receptors and other genes relevant to the physiology of L. cuprina larvae. Adult females and larvae of the same species carrying a loss-of-function mutation for the L. cuprina odorant coreceptor gene (LcupOrco) were obtained by gene editing. Their response to fresh and rotten meat at different temperatures was evaluated.Results The RNA-Seq analysis of whole larvae at different stages and third instar head and gut tissues, suggested that odorant (OR), gustatory, ionotropic and pickpocket receptors may not play a central role in the L. cuprina larval sensory signaling and digestive systems. Rather, ATP-binding cassettes (ABCs) were highly enriched in head and gut RNA, and odorant-binding proteins (OBPs) only in the head. To confirm that ORs are not essential for larval detection of rotten beef, diet-choice assays were performed including larvae and adults homozygous for a null mutation in LcupOrco. While the attraction of adult females to rotten beef was fully disrupted, LcupOrco mutant larvae showed no change in diet preference.Conclusions The expression pattern of the ABC and OBP gene families suggests a central role in the sensory system of the L. cuprina larva for these receptors. Behavioral assays showed that ORs are essential for the adult female response to rotten beef, but not for larval behavior. These findings are consistent with high levels of expression of LcupOrco in the adult female antenna but very low expression in larvae.Competing Interest StatementThe authors have declared no competing interest.ABCATP-binding cassette transporterAKHAdipokinetic hormoneAMTammonium transporterBHBenjamini-HochbergBLASTbasic local alignment search toolCcysteineCDScoding sequenceCD36/SNMPCD36 and sensory neuron membrane proteinsCSPchemosensory proteinCocoldCYPcytochrome P450DEdifferential expressedDGEdifferential gene expressionDH3diuretic hormone class 2DNAdeoxyribonucleic acidDMDSdimethyl disulfideDMTSdimethyl trisulfideEexonEHeclosion hormoneFBfat bodyFCFold-ChangeFrfreshGgutggramgDNAgenomic DNAGRgustatory receptorHheadhhourHohothPahectopascalIintronIPMintegrated pest managementIRionotropic receptoriTOLinteractive Tree of LifeJHjuvenile hormoneKBkilobaseKClpotassium chlorideLlarvaLSleft sideMLMaximum likelihoodML/NPC2Myeloid lipid-recognition (ML) and Niemann-Pick C2 disease proteinsmRNAmessenger RNAMTmalpighian tubulesNCBInational center for biotechnology informationNCnon-choiceNFnuclease freeNJNeighbor-JoiningNPFlong neuropeptide FOBPodorant-binding proteinORodorant receptorOrcoodorant receptor coreceptorPCAprincipal components analysisPEpaired endPPKpickpocket receptorRHrelative humidityRHBGammonium transporter Rhesus type BRNARibonucleic AcidRNA-SeqRNA-sequencingRorottenRSright sideRTroom temperatureSDstandard deviationSEsecretions and excretionsSITsterile insect techniquesNPFshort neuropeptide FSRB1Scavenger Receptor Class B Type ITMtransmembraneTPMtranscripts per millionTRPTransient receptor potentialUTRuntranslatedWLwhole larvawtwild-type