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Updated: May 15, 2026
Generation of Monoclonal Cultures from Wolbachia-infected Drosophila melanogaster JW18 Cell Line
Published on: June 27, 2025
Wolbachia from the planthopper Laodelphax striatellus establishes a robust, persistent, streptomycin-resistant
A M Fallon1, G D Baldridge, L A Higgins
1Department of Entomology, University of Minnesota, 1980 Folwell Ave., St. Paul, MN 55108, USA. fallo002@umn.edu
Abstract:
The obligate intracellular bacterium, Wolbachia pipientis (Rickettsiales: Anaplasmataceae), distorts reproduction of its arthropod hosts to facilitate invasion of naïve populations. This property makes Wolbachia an attractive "gene drive" agent with potential applications in the control of insect vector populations. Genetic manipulation of Wolbachia will require in vitro systems for its propagation, genetic modification, amplification, and introduction into target insects. Here we show that Wolbachia from the planthopper, Laodelphax striatellus, establishes a robust infection in clonal C7-10 Aedes albopictus mosquito cells. Infected cells, designated C/wStr, expressed radiolabeled proteins that were enriched in cells grown in the absence of antibiotics that inhibit Wolbachia, relative to cultures grown in medium containing tetracycline and rifampicin. Using mass spectrometry, we verified that tryptic peptides from an upregulated 24 kDa band predominantly represented proteins encoded by the Wolbachia genome, including the outer surface protein, Wsp. We further showed that resistance of Wolbachia to streptomycin is associated with a K42R mutation in Wolbachia ribosomal protein S12, and that the pattern of amino acid substitutions in ribosomal protein S12 shows distinct differences in the closely related genera, Wolbachia and Rickettsia.
Insights
Wolbachia pipientis, a bacterium used for insect control, can now be cultured in Aedes albopictus mosquito cells. This breakthrough enables genetic manipulation of Wolbachia for vector control applications.
Area of Science:
- Microbiology
- Genetics
- Vector Biology
Background:
- Wolbachia pipientis is an obligate intracellular bacterium that manipulates arthropod reproduction.
- Its potential as a gene drive agent for insect vector control is significant.
- Developing in vitro systems is crucial for genetic manipulation and application of Wolbachia.
Purpose of the Study:
- To establish a robust in vitro system for Wolbachia propagation and genetic modification.
- To investigate the characteristics of Wolbachia infection in mosquito cells.
- To identify genetic markers associated with Wolbachia traits like antibiotic resistance.
Main Methods:
- Infection of clonal Aedes albopictus mosquito cells with Wolbachia from Laodelphax striatellus.
- Radiolabeling of proteins and mass spectrometry to identify Wolbachia-encoded proteins.
- Genetic analysis to identify mutations conferring streptomycin resistance.
Main Results:
- Successfully established a stable Wolbachia infection (C/wStr) in Aedes albopictus cells.
- Identified upregulated Wolbachia-specific proteins, including the outer surface protein Wsp.
- Discovered a K42R mutation in ribosomal protein S12 linked to streptomycin resistance in Wolbachia.
Conclusions:
- The established C/wStr cell line provides a valuable tool for in vitro research on Wolbachia.
- Understanding Wolbachia protein expression and genetic mutations is key for its application in vector control.
- Comparative analysis of ribosomal protein S12 mutations highlights evolutionary divergence between Wolbachia and Rickettsia.
