RT Journal Article SR Electronic T1 Insect—symbiont gene expression in the midgut bacteriocytes of a blood-sucking parasite JF bioRxiv FD Cold Spring Harbor Laboratory SP 572495 DO 10.1101/572495 A1 Husnik Filip A1 Hypsa Vaclav A1 Darby Alistair YR 2019 UL http://biorxiv.org/content/early/2019/03/08/572495.abstract AB Background Animals interact with a diverse array of both beneficial and detrimental microorganisms. These interactions sometimes spark obligate symbioses where the host depends on beneficial bacteria for survival and reproduction. In insects, these obligate symbioses in many cases allow feeding on nutritionally unbalanced diets such as plant sap and vertebrate blood. It is, however, still not clear how are these obligate intracellular symbioses maintained at the cellular level for up to several hundred million years. Exact mechanisms driving host-symbiont interactions are only understood for a handful of model species and data on blood-feeding hosts with intracellular bacteria are particularly scarce.Results Here, we analyzed interactions at the symbiotic interfaces of an obligately blood-sucking parasite of sheep, the louse fly Melophagus ovinus. We assembled a reference transcriptome from one male and one female individual and used RNA-Seq with five biological replicates to compare expression in the midgut cells housing bacteria to the rest of the gut (foregut-hindgut). We focused on nutritional and immunity interactions between the insect host and its obligate symbiont Arsenophonus melophagi, and also generated lower-coverage data for three facultative bacterial symbionts (Sodalis melophagi, Bartonella melophagi, and Wolbachia sp.) and one facultative eukaryote Trypanosoma melophagium. We found strong evidence for the importance of zinc in the system likely caused by symbionts using zinc-dependent proteases when acquiring amino acids, and for likely different immunity mechanisms controlling the symbionts than in closely related tsetse flies.Conclusions Our results show that cellular and nutritional interactions between this blood-sucking insect and its symbionts are less intimate than what was previously found in some (but not all) plant-sap sucking insects such as aphids, psyllids, whiteflies, and mealybugs. This finding is likely interconnected to several features observed in symbionts in blood-sucking arthropods, particularly their midgut intracellular localization (as opposed to being localized in truly specialized bacteriocytes), intracytoplasmic presence (as opposed to having an outermost host-derived ‘symbiosomal’ membrane), less severe genome reduction, and relatively recent associations caused by frequent evolutionary losses and replacements.Data deposition Raw RNA-Seq data were made available through the European Nucleotide Archive (ENA) database under the study accession number PRJEB30632. All assemblies and additional large supplementary files are available on FigShare [https://doi.org/10.6084/m9.figshare.6146777.v1]. All commands used for data analyses are available on Github [https://github.com/filip-husnik/melophagus].COGClusters of Orthologous GroupsEAAsEssential Amino AcidsFPKMFragments Per Kilobase MillionGNBPGram-Negative Binding ProteinHGTHorizontal Gene TransferTPMTranscripts Per Kilobase MillionTMMTrimmed Mean of M-values normalization methodNR and NTNon-redundant protein and nucleotide databasesORFOpen Reading FrameRNA-SeqRNA SequencingPGRPPeptidoglycan Recognition ProteinIMDImmunodeficiency signaling pathway