Protocols for Efficient Rearing and Functional Analysis of the Endoparasitoid Wasp Asobara japonica and its Host Drosophila melanogaster

Hayato Kondo; Takumi Kamiyama; Anastasiia Staroverova; Ryusuke Niwa; Yuko Shimada-Niwa

doi:10.3791/72035

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Method Article

Protocols for Efficient Rearing and Functional Analysis of the Endoparasitoid Wasp Asobara japonica and its Host Drosophila melanogaster

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DOI:

10.3791/72035

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July 24th, 2026

Hayato Kondo¹ , Takumi Kamiyama² , Anastasiia Staroverova³ , Ryusuke Niwa² , Yuko Shimada-Niwa²

¹Degree Programs in Life and Earth Sciences, Graduate School of Science and Technology, University of Tsukuba, ²Life Science Center for Survival Dynamics, Tsukuba Advanced Research Alliance (TARA), University of Tsukuba, ³Doctoral Program in Humanics, School of Integrative and Global Majors, University of Tsukuba

Summary

The goal of this protocol is to facilitate functional analysis of parasitoid-host interactions in Asobara japonica and Drosophila melanogaster through optimized rearing methods, a single-oviposition infection assay, and RNAi-mediated gene knockdown.

Abstract

Parasitism is a biological interaction in which one organism exploits the body or resources of another (the host), resulting in substantial damage or death to the host. Among parasitic animals, parasitoid wasps are one of the most species-rich lineages, accounting for nearly 20% of all insect species. In particular, endoparasitoid wasps oviposit directly into host bodies and deploy a diverse array of venom factors to manipulate host development, immunity, and physiology. In response to wasp attack, hosts attempt to eliminate parasitoid eggs through innate immune mechanisms. However, the molecular and cellular mechanisms by which individual venom components modulate host biology and promote successful parasitism remain poorly understood. Here, we describe a standardized laboratory protocol for rearing the endoparasitoid wasp Asobara japonica and its host Drosophila melanogaster. Both parthenogenetic and sexual strains of A. japonica are available, and the parthenogenetic strain exhibits a high parasitism success rate, allowing stable maintenance of laboratory stocks for genome analysis and parasitism assays. A single-oviposition infection assay and a double-stranded RNA-based gene knockdown method were optimized for functional analysis of venom genes. Together, these protocols provide a practical experimental framework for dissecting the molecular mechanisms underlying parasitoid-host interactions and will facilitate future research in developmental biology, immunology, and physiology.

Introduction

Parasitism is a lifestyle in which one organism exploits the nutritional resources of another and is widespread across taxa. It is estimated that up to half of all known organisms exhibit parasitism in their life cycle¹. Among parasites, parasitoid wasps are a particularly diverse group, accounting for approximately 20% of all insects². They exploit a wide range of arthropod hosts, such as insects, spiders, and mites, and exhibit remarkable diversity in their life histories and parasitic strategies³. These features make parasitoid wasps one of the most evolutionarily and ecologically successful in....