Abstract

Ants are evolutionarily successful species and occupy diverse trophic and habitat niches on the earth. To fulfill dietary requirements, ants have established commensalism with both sap-feeding insects and bacteria. In this study, we used high-throughput sequencing of the bacterial 16S rRNA gene to characterize the bacterial composition and structure of the digestive tracts in three species of Formica ants and Lasius niger (Linnaeus)—species that predominantly feed on honeydew secreted by aphids. We found that bacterial communities displayed species- and colony-level signatures, and that bacterial communities in the infrabuccal pockets and crops were different from those in the midguts and hindguts. Lactobacillus and Wolbachia were dominant in the infrabuccal pockets and crops of workers, whereas Wolbachia was dominant in the midguts, hindguts and brood (larvae, pupae and cocoons). To learn more about the dominant Lactobacillus in ants, we assessed its prevalence in a wide range of aphid-tending ants using diagnostic PCR. We found that Lactobacillus was more prevalent in Formicinae than in Myrmicinae species. We also isolated four strains of lactic acid bacteria (Lactobacillus sanfranciscensis, Lactobacillus lindneri, Weissella cibaria and Fructobacillus sp.) from the infrabuccal pockets and crops of aphid-tending ants using a culture-dependent method. Two predominant lactic acid bacterial isolates, Lactobacillus sanfranciscensis (La2) and Weissella cibaria (La3), exhibited abilities in catabolizing sugars (sucrose, trehalose, melezitose and raffinose) known to be constituents of hemipteran honeydew. These findings contribute to further understanding the association between ants, aphids and bacteria, and provide additional information on the function of lactic acid bacteria in ants.

Highlights

  • Ants are highly evolved social insects that live in organized colonies with labor division

  • Weissella cibaria (La3) can catabolize sucrose, melezitose, and raffinose (Figure 5B and Supplementary Figures 3A–F). These results revealed that lactic acid bacteria facilitate ants in utilizing sugars contained in honeydew. This is the first study to investigate the bacterial communities of ants across different species and several colonies, especially in the context of ant-aphid mutualism

  • We aimed to identify the factors that shape the variations in the bacterial communities and their functions

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Summary

Introduction

Ants are highly evolved social insects that live in organized colonies with labor division. The extraordinary abundance and species richness of ants have facilitated their evolutionary success in nature. Ants have evolved significant differentiation in feeding habits and related foraging behavior to utilize a variety of food resources. One of the most interesting behaviors of ants is the habit of tending sap-feeding insects such as aphids and mealybugs throughout their 115–168 million years history (Currie et al, 1999; Stadler and Dixon, 2005; Moreau et al, 2006). Sap-feeding insects excrete carbohydrate-rich honeydew that serve as a food resource for natural enemies, e.g., ants and hyperparasitoids, and these insects defend the sap-feeders from predators and parasites as a reward (Way, 1963; Hölldobler and Wilson, 1990; van Neerbos et al, 2020)

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