Abstract
Drosophila melanogaster has been proven again to be an exceptional experimental tool for studying the efficacy of entomopathogenic nematodes (EPN) as biocontrol agents. Aiming to examine the molecular basis of the insect immune response to EPN infection with or without its mutualistic bacteria, we present a set of established routine techniques for generating axenic culture of infective juveniles of Heterorhabditis bacteriophora and Steinernema carpocapsae nematodes from their non-axenic culture, followed by a reliable and adaptable infection method of D. melanogaster larvae with the EPN stock. Finally, the sensitivity of quantitative real-time PCR method enables us to measure the immune gene transcript levels to explore the dynamics of this complex host-parasite interaction through various timescales and challenge conditions.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Lacey LA, Grzywacz D, Shapiro-Ilan DI, Frutos R, Brownbridge M, Goettel MS (2015) Insect pathogens as biological control agents: Back to the future. J Invertebr Pathol 132:1–41
Patrnogic J, Heryanto C, Eleftherianos I (2018) Transcriptional up-regulation of the TGF-beta intracellular signaling transducer mad of Drosophila larvae in response to parasitic nematode infection. Innate Immun 24:349–356
Yadav S, Gupta S, Eleftherianos I (2018) Differential regulation of immune signaling and survival response in Drosophila melanogaster larvae upon Steinernema carpocapsae nematode infection. Insects 9:E17
Hallem EA, Rengarajan M, Ciche TA, Sternberg PW (2007) Nematodes, bacteria, and flies: a tripartite model for nematode parasitism. Curr Biol 17:898–904
Taylor SC, Nadeau K, Abbasi M, Lachance C, Nguyen M, Fenrich J (2019) The ultimate qPCR experiment: producing publication quality, reproducible data the first time. Trends Biotechnol 37(7):761–774
Yang Y, Wen Y, Cai YN et al (2015) Serine proteases of parasitic helminths. Korean J Parasitol 53:1–11
Yadav S, Daugherty S, Shetty AC, Eleftherianos I (2017) RNAseq analysis of the Drosophila response to the entomopathogenic nematode Steinernema. G3 (Bethesda) 7:1955–1967
White GF (1927) A method for obtaining infective nematode larvae from cultures. Science 66:302–303
Yadav S, Shokal U, Forst F, Eleftherianos I (2015) An improved method for generating axenic entomopathogenic nematodes. BMC Res Notes 8:461
Chomczynski P, Sacchi N (1982) Single-step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159
Matta BP, Bitner-Mathé BC (2011) Getting real with real-time qPCR: a case study of reference gene selection for morphological variation in Drosophila melanogaster wings. Dev Genes Evol 221:49–57
KJ L, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods 25:402–408
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Heryanto, C., Kenney, E., Eleftherianos, I. (2020). A Workflow for Infection of Drosophila with Entomopathogenic Nematodes to Monitor Immune Gene Transcriptional Activity. In: Sandrelli, F., Tettamanti, G. (eds) Immunity in Insects. Springer Protocols Handbooks. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0259-1_13
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0259-1_13
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0258-4
Online ISBN: 978-1-0716-0259-1
eBook Packages: Springer Protocols