Abstract

Associations between microbes and animals are ubiquitous and hosts may benefit from harbouring microbial communities through improved resource exploitation or resistance to environmental stress. The pea aphid, Acyrthosiphon pisum, is the host of heritable bacterial symbionts, including the obligate endosymbiont Buchnera aphidicola and several facultative symbionts. While obligate symbionts supply aphids with key nutrients, facultative symbionts influence their hosts in many ways such as protection against natural enemies, heat tolerance, color change and reproduction alteration. The pea aphid also encompasses multiple plant-specialized biotypes, each adapted to one or a few legume species. Facultative symbiont communities differ strongly between biotypes, although bacterial involvement in plant specialization is uncertain. Here, we analyse the diversity of bacterial communities associated with nine biotypes of the pea aphid complex using amplicon pyrosequencing of 16S rRNA genes. Combined clustering and phylogenetic analyses of 16S sequences allowed identifying 21 bacterial OTUs (Operational Taxonomic Unit). More than 98% of the sequencing reads were assigned to known pea aphid symbionts. The presence of Wolbachia was confirmed in A. pisum while Erwinia and Pantoea, two gut associates, were detected in multiple samples. The diversity of bacterial communities harboured by pea aphid biotypes was very low, ranging from 3 to 11 OTUs across samples. Bacterial communities differed more between than within biotypes but this difference did not correlate with the genetic divergence between biotypes. Altogether, these results confirm that the aphid microbiota is dominated by a few heritable symbionts and that plant specialization is an important structuring factor of bacterial communities associated with the pea aphid complex. However, since we examined the microbiota of aphid samples kept a few generations in controlled conditions, it may be that bacterial diversity was underestimated due to the possible loss of environmental or transient taxa.

Highlights

  • Sustained associations between microbes and eukaryotic hosts are widespread

  • The classification from Greengenes database resulted in 21 unique bacterial genera that belonged to 4 phyla, 6 classes, 8 orders and 14 families (S3 Table)

  • We confirmed the presence of Wolbachia, which was recently reported in North American A. pisum by [14], in two pea aphid samples

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Summary

Introduction

Sustained associations between microbes and eukaryotic hosts are widespread. This is true for insects which are engaged in multiple forms of interactions with micro-organisms referred to as symbiosis in its broad sense (i.e. all types of close associations between organisms of different species). The development of Generation Sequencing techniques (NGS) permits to assess without a priori the diversity and structure of insect microbial communities to better understand both the influence of microbes on their host populations and the dynamics of symbiotic associations. It is possible to characterize and analyze the whole microbial community of a given host, these techniques have been mainly restricted so far to insect model species or species of medical importance [5], [6], [7], [8]

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