title: Host gene response to endosymbiont and pathogen in the cereal weevil Sitophilus oryzae.
abstract: BACKGROUND
  Insects thriving on nutritionally poor habitats have integrated mutualistic intracellular symbiotic bacteria (endosymbionts) in a bacteria-bearing tissue (the bacteriome) that isolates the endosymbionts and protects them against a host systemic immune response. Whilst the metabolic and physiological features of long-term insect associations have been investigated in detail over the past decades, cellular and immune regulations that determine the host response to endosymbionts and pathogens have attracted interest more recently.
  
  
  RESULTS
  To investigate bacteriome cellular specificities and weevil immune responses to bacteria, we have constructed and sequenced 7 cDNA libraries from Sitophilus oryzae whole larvae and bacteriomes. Bioinformatic analysis of 26,886 ESTs led to the generation of 8,941 weevil unigenes. Based on in silico analysis and on the examination of genes involved in the cellular pathways of potential interest to intracellular symbiosis (i.e. cell growth and apoptosis, autophagy, immunity), we have selected and analyzed 29 genes using qRT-PCR, taking into consideration bacteriome specificity and symbiosis impact on the host response to pathogens. We show that the bacteriome tissue accumulates transcripts from genes involved in cellular development and survival, such as the apoptotic inhibitors iap2 and iap3, and endosomal fusion and trafficking, such as Rab7, Hrs, and SNARE. As regards our investigation into immunity, we first strengthen the bacteriome immunomodulation previously reported in S. zeamais. We show that the sarcotoxin, the c-type lysozyme, and the wpgrp2 genes are downregulated in the S. oryzae bacteriome, when compared to aposymbiotic insects and insects challenged with E. coli. Secondly, transcript level comparison between symbiotic and aposymbiotic larvae provides evidence that the immune systemic response to pathogens is decreased in symbiotic insects, as shown by the relatively high expression of wpgrp2, wpgrp3, coleoptericin-B, diptericin, and sarcotoxin genes in aposymbiotic insects.
  
  
  CONCLUSIONS
  Library sequencing significantly increased the number of unigenes, allowing for improved functional and genetic investigations in the cereal weevil S. oryzae. Transcriptomic analyses support selective and local immune gene expression in the bacteriome tissue and uncover cellular pathways that are of potential interest to bacteriocyte survival and homeostasis. Bacterial challenge experiments have revealed that the systemic immune response would be less induced in a symbiotic insect, thus highlighting new perspectives on host immunity in long-term invertebrate co-evolutionary associations.
authors.forename: Aurélien
authors.initials: A
authors.lastname: Vigneron
authors.forename: Delphine
authors.initials: D
authors.lastname: Charif
authors.forename: Carole
authors.initials: C
authors.lastname: Vincent-Monégat
authors.forename: Agnès
authors.initials: A
authors.lastname: Vallier
authors.forename: Frédérick
authors.initials: F
authors.lastname: Gavory
authors.forename: Patrick
authors.initials: P
authors.lastname: Wincker
authors.forename: Abdelaziz
authors.initials: A
authors.lastname: Heddi
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D020563:Gammaproteobacteria / Q000502:physiology*; D005786:Gene Expression Regulation; D015723:Gene Library; D023281:Genomics / Q000379:methods*; D058507:Host Specificity; D054884:Host-Pathogen Interactions; D019476:Insect Proteins / Q000235:genetics*; D007814:Larva / Q000382:microbiology; D064307:Microbiota; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D013559:Symbiosis; D048889:Weevils / Q000196:embryology / Q000235:genetics* / Q000382:microbiology / Q000502:physiology
chemical_list: D001426:Bacterial Proteins; D019476:Insect Proteins