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. 2020 Feb;29(1):1-8.
doi: 10.1111/imb.12604. Epub 2019 Jul 3.

Wolbachia transinfections in Culex quinquefasciatus generate cytoplasmic incompatibility

T H Ant  1   2 , C Herd  1   2 , F Louis  3 , A B Failloux  3 , S P Sinkins  1   2
Affiliations

Wolbachia transinfections in Culex quinquefasciatus generate cytoplasmic incompatibility

T H Ant et al. Insect Mol Biol. 2020 Feb.

Abstract

Culex quinquefasciatus is an important mosquito vector of a number of viral and protozoan pathogens of humans and animals, and naturally carries the endosymbiont Wolbachia pipientis, strain wPip. Wolbachia are used in two distinct vector control strategies: firstly, population suppression caused by mating incompatibilities between mass-released transinfected males and wild females; and secondly, the spread of pathogen transmission-blocking strains through populations. Using embryonic microinjection, two novel Wolbachia transinfections were generated in C. quinquefasciatus using strains native to the mosquito Aedes albopictus: a wAlbB single infection, and a wPip plus wAlbA superinfection. The wAlbB infection showed full bidirectional cytoplasmic incompatibility (CI) with wild-type C. quinquefasciatus in reciprocal crosses. The wPipwAlbA superinfection showed complete unidirectional CI, and therefore population invasion potential. Whereas the wAlbB strain showed comparatively low overall densities, similar to the native wPip, the wPipwAlbA superinfection reached over 400-fold higher densities in the salivary glands compared to the native wPip, suggesting it may be a candidate for pathogen transmission blocking.

Keywords: Culex quinquefasciatus; Wolbachia; cytoplasmic incompatibility; incompatible insect technique; population replacement; transinfection.

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Figures

Figure 1
Figure 1
Percentage egg hatching rates from individual egg rafts resulting from crosses between the wild‐type Wolbachia wPip, the wAlbB and the Wolbachia ‐ve (PelU, antibiotic‐treated) lines. Boxplots show median values and interquartile ranges. Dots show hatching rates from individual egg rafts.
Figure 2
Figure 2
Percentage egg hatching rates from individual egg rafts resulting from crosses between the wild‐type wPip, the wPipwAlbA and the Wolbachia‐ve (PelU, antibiotic‐treated) lines. Boxplots show median values and interquartile ranges. Dots show hatching rates from individual egg rafts.
Figure 3
Figure 3
Average egg number per egg raft from Wolbachia‐transinfected, wPip and PelU lines over the first gonotrophic cycle. Eggs from the rafts of 12–15 females were counted. Error bars show standard deviation.
Figure 4
Figure 4
Wolbachia densities in ovary (O), salivary glands (SG) and whole female carcasses (WC) for the wPipwAlbA, wAlbB and wPip lines. Bar charts show mean densities and error bars show standard deviation. Each bar summarizes data from five biological repeats, each with either three whole female carcasses, or the dissected tissues from five females. wsp, Wolbachia surface protein.
Figure 5
Figure 5
Expression of immune genes in the wAlbB and wPipwAlbA lines normalized initially to the 18S ribosomal RNA house‐keeping gene and then to expression in the wPip line. Error bars show 95% confidence intervals from five biological replicates, each containing cDNA from a pool of three females. Def1, Defensin1; LRIM1, leucine‐rich repeat immune protein 1; Rel1, a homologue of Drosophila dorsal; Rel2, an NF‐κB transcriptional factor.
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