title: Development of RNAi in a Drosophila endoparasitoid wasp and demonstration of its efficiency in impairing venom protein production.
abstract: Endoparasitoid wasps are essential regulators of insect pests in ecosystems as well as important biological control auxiliaries. Traits important for parasitism success, such as the injection of venom proteins at oviposition, have thus been mainly studied. However, identification of the key genes involved among the large number of genes identified was still prevented by the lack of functional approaches. Here, we report the development of RNA interference (RNAi) in Leptopilina boulardi, a figitid endoparasitoid that performs its entire development inside the Drosophila host. Having set up conditions for in vitro development of parasitoid late larval stages or pupae, we first targeted the cinnabar gene by microinjecting double-stranded RNA (dsRNA), leading to its silencing and production of red-eyed individuals. We then demonstrated that expression of the gene encoding LbGAP, a virulence factor found in a high amount in L. boulardi venom, could be specifically and almost completely silenced. Finally, a time-course analysis revealed that LbGAP silencing lasted during the entire lifetime of L. boulardi. This is the first report of the efficient silencing of venom protein-encoding genes in parasitoid wasps. Overall, RNAi opens the way for a large-scale functional analysis of parasitoid venom factors as well as other traits involved in parasitism success and more largely in the biology of these ecologically important organisms.
authors.affiliation: INRA, ISA, UMR 1355, Evolution et Spécificité des Interactions Multitrophiques (ESIM), 06903 Sophia Antipolis, France; Université Nice Sophia Antipolis, ISA, 06903 Sophia Antipolis, France; CNRS, ISA, UMR 7254, 06903 Sophia Antipolis, France. Electronic address: colinet@sophia.inra.fr.
authors.forename: Dominique
authors.initials: D
authors.lastname: Colinet
authors.affiliation: INRA, ISA, UMR 1355, Evolution et Spécificité des Interactions Multitrophiques (ESIM), 06903 Sophia Antipolis, France; Université Nice Sophia Antipolis, ISA, 06903 Sophia Antipolis, France; CNRS, ISA, UMR 7254, 06903 Sophia Antipolis, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Kremmer
authors.affiliation: INRA, ISA, UMR 1355, Evolution et Spécificité des Interactions Multitrophiques (ESIM), 06903 Sophia Antipolis, France; Université Nice Sophia Antipolis, ISA, 06903 Sophia Antipolis, France; CNRS, ISA, UMR 7254, 06903 Sophia Antipolis, France.
authors.forename: Séverine
authors.initials: S
authors.lastname: Lemauf
authors.affiliation: INRA, ISA, UMR 1355, Evolution et Spécificité des Interactions Multitrophiques (ESIM), 06903 Sophia Antipolis, France; Université Nice Sophia Antipolis, ISA, 06903 Sophia Antipolis, France; CNRS, ISA, UMR 7254, 06903 Sophia Antipolis, France.
authors.forename: Christian
authors.initials: C
authors.lastname: Rebuf
authors.affiliation: INRA, ISA, UMR 1355, Evolution et Spécificité des Interactions Multitrophiques (ESIM), 06903 Sophia Antipolis, France; Université Nice Sophia Antipolis, ISA, 06903 Sophia Antipolis, France; CNRS, ISA, UMR 7254, 06903 Sophia Antipolis, France.
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Gatti
authors.affiliation: INRA, ISA, UMR 1355, Evolution et Spécificité des Interactions Multitrophiques (ESIM), 06903 Sophia Antipolis, France; Université Nice Sophia Antipolis, ISA, 06903 Sophia Antipolis, France; CNRS, ISA, UMR 7254, 06903 Sophia Antipolis, France.
authors.forename: Marylène
authors.initials: M
authors.lastname: Poirié
mesh_terms: D000818:Animals; D004331:Drosophila melanogaster / Q000469:parasitology*; D005260:Female; D005786:Gene Expression Regulation; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D007814:Larva / Q000378:metabolism / Q000469:parasitology / Q000502:physiology; D008297:Male; D008845:Microinjections; D016133:Polymerase Chain Reaction; D011679:Pupa / Q000378:metabolism / Q000469:parasitology / Q000502:physiology; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000378:metabolism; D037521:Virulence Factors; D014862:Wasp Venoms / Q000235:genetics* / Q000378:metabolism; D014863:Wasps / Q000235:genetics / Q000254:growth & development / Q000502:physiology*
chemical_list: D019476:Insect Proteins; D012330:RNA, Double-Stranded; D037521:Virulence Factors; D014862:Wasp Venoms
keywords: Drosophila; Endoparasitoid; Leptopilina; Parasitism success; RNA interference; Venom