Abstract

BackgroundWolbachia is a bacterial endosymbiont that naturally infects a wide range of insect species, and causes drastic changes to host biology. Stable infections of Wolbachia in mosquitoes can inhibit infection with medically important pathogens such as dengue virus and malaria-causing Plasmodium parasites. However, some native Wolbachia strains can enhance infection with certain pathogens, as is the case for the mosquito Aedes fluviatilis, where infection with Plasmodium gallinaceum is enhanced by the native wFlu Wolbachia strain. To better understand the biological interactions between mosquitoes and native Wolbachia infections, and to investigate the process of pathogen enhancement, we used RNA-Seq to generate the transcriptome of Ae. fluviatilis with and without Wolbachia infection.ResultsIn total, we generated 22,280,160 Illumina paired-end reads from Wolbachia-infected and uninfected mosquitoes, and used these to make a de novo transcriptome assembly, resulting in 58,013 contigs with a median sequence length of 443 bp and an N50 of 2454 bp. Contigs were annotated through local alignments using BlastX, and associated with both gene ontology and KEGG orthology terms. Through baySeq, we identified 159 contigs that were significantly upregulated due to Wolbachia infection, and 98 that were downregulated. Critically, we saw no changes to Toll or IMD immune gene transcription, but did see evidence that wFlu infection altered the expression of several bacterial recognition genes, and immune-related genes that could influence Plasmodium infection. wFlu infection also had a widespread effect on a number of host physiological processes including protein, carbohydrate and lipid metabolism, and oxidative stress. We then compared our data set with transcriptomic data for other Wolbachia infections in Aedes aegypti, and identified a core set of 15 gene groups associated with Wolbachia infection in mosquitoes.ConclusionsWhile the scale of transcriptional changes associated with wFlu infection might be small, the scope is rather large, which confirms that native Wolbachia infections maintain intricate molecular relationships with their mosquito hosts even after lengthy periods of co-evolution. We have also identified several potential means through which wFlu infection might influence Plasmodium infection in Ae. fluviatilis, and these genes should form the basis of future investigation into the enhancement of Plasmodium by Wolbachia.

Highlights

  • Wolbachia is a bacterial endosymbiont that naturally infects a wide range of insect species, and causes drastic changes to host biology

  • RNA sequencing and de novo transcriptome assembly We generated a total of 22,280,160 Illumina paired-end reads across 6 Ae. fluviatilis libraries - 3 with Wolbachia infection, and 3 where the native infection had been cleared by treatment with tetracycline (Tet)

  • Our data indicated that Wolbachia-infected Aedes fluviatilis (wFlu) infection led to the differential expression of 257 genes, and while the scale of these changes was not as extreme as what is sometimes seen with Wolbachia transinfections in Ae. aegypti [10, 11, 14], the effect was still broad in scope and encompassed a wide range of biological processes, many of which are held in common with Wolbachia infections in other mosquitoes

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Summary

Introduction

Wolbachia is a bacterial endosymbiont that naturally infects a wide range of insect species, and causes drastic changes to host biology. Wolbachia induce a wide range of physiological manipulations in different insect hosts, with manipulation of reproductive biology promoting maternal transmission and bacterial propagation [2, 3]. It is through this ability to alter host biology that Wolbachia have gained interest as a form of biological control for the mosquito-transmitted pathogens that are responsible for diseases such as malaria, dengue fever, chikungunya and Zika fever, which represent a serious threat to human health across the globe [4, 5]. CI-causing strains can be used to suppress mosquito populations that are uninfected by Wolbachia through the release of infected males, similar to the sterile insect technique, or to control Wolbachiainfected populations by releasing mosquitoes infected with a different strain, as this crashes the population [8]

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