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Published on: August 14, 2017
Maintenance and Evaluation of Wolbachia Male-Killers of Dipterans
Margaret E Schedl1, Ngoc Huan Nguyen1, Robert L Unckless1
1Department of Molecular Biosciences, University of Kansas, Lawrence, KS, USA.
Abstract:
As research on Wolbachia male-killers of dipterans increases over time, several standard methods have emerged for the validation and characterization of these Wolbachia strains. Here, we describe typical steps for maintenance and evaluation of male-killers in the laboratory. Drosophila that host Wolbachia male-killers include both mushroom-feeders and flies reared on standard media, which require different maintenance protocols. In addition, male killing in flies typically occurs during embryonic development, and as such, many experiments require embryo tissue samples. We provide step-by-step instructions for standard methods of isolation, rearing, confirmation of Wolbachia male-killing as the basis of a sex ratio bias, and collection of embryos from these species. Thus, anyone intending to study a male-killer in the lab will have the complete set of protocols necessary to collect and rear them, validate them, and collect embryonic tissue for further study.
Insights
This study details laboratory protocols for rearing and validating Wolbachia male-killers in flies. It provides essential methods for researchers studying these bacteria and their impact on insect sex ratios.
Area of Science:
- Entomology
- Microbiology
- Genetics
Background:
- Research on Wolbachia male-killers in dipterans is expanding.
- Standardized methods are needed for characterizing these bacterial strains.
Purpose of the Study:
- To describe standard laboratory protocols for maintaining and evaluating Wolbachia male-killers.
- To provide a comprehensive guide for researchers studying these organisms.
Main Methods:
- Detailed step-by-step instructions for isolation and rearing of Drosophila species hosting male-killers.
- Protocols for confirming Wolbachia-induced sex ratio bias.
- Methods for collecting embryonic tissue samples.
Main Results:
- Established protocols for diverse Drosophila species, including mushroom-feeders and those on standard media.
- Validated methods for confirming male-killing as the cause of sex ratio distortion.
- Efficient techniques for embryo collection from infected flies.
Conclusions:
- This work offers a complete set of protocols for researchers to collect, rear, validate, and study Wolbachia male-killers.
- The described methods facilitate further research into the mechanisms and implications of Wolbachia-mediated sex determination in insects.
