Abstract

Some specialist insects feed on plants rich in secondary compounds, which pose a major selective pressure on both the phytophagous and the gut microbiota. However, microbial communities of toxic plant feeders are still poorly characterized. Here, we show the bacterial communities of the gut of two specialized Lepidoptera, Hyles euphorbiae and Brithys crini, which exclusively feed on latex-rich Euphorbia sp. and alkaloid-rich Pancratium maritimum, respectively. A metagenomic analysis based on high-throughput sequencing of the 16S rRNA gene revealed that the gut microbiota of both insects is dominated by the phylum Firmicutes, and especially by the common gut inhabitant Enterococcus sp. Staphylococcus sp. are also found in H. euphorbiae though to a lesser extent. By scanning electron microscopy, we found a dense ring-shaped bacterial biofilm in the hindgut of H. euphorbiae, and identified the most prominent bacterium in the biofilm as Enterococcus casseliflavus through molecular techniques. Interestingly, this species has previously been reported to contribute to the immobilization of latex-like molecules in the larvae of Spodoptera litura, a highly polyphagous lepidopteran. The E. casseliflavus strain was isolated from the gut and its ability to tolerate natural latex was tested under laboratory conditions. This fact, along with the identification of less frequent bacterial species able to degrade alkaloids and/or latex, suggest a putative role of bacterial communities in the tolerance of specialized insects to their toxic diet.

Highlights

  • Plants have biochemical and molecular mechanisms to defend themselves from insects attack

  • The overall taxonomic profiles from the two species exhibited a clear difference: whereas B. crini was characterized by the overwhelming presence of Enterococcus sp., (OTU 1, accounting for 94–99% of reads), H. euphorbiae harbored a more heterogeneous community

  • A species belonging to the Enterobacteriaceae family (OTU 2) and Staphylococcus sp. (OTU 4) were very common in H. euphorbiae (8–70% of sequences depending on the specimen) but were rare in B. crini

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Summary

Introduction

Plants have biochemical and molecular mechanisms to defend themselves from insects attack. Some plant biochemicals are toxic, repellent, or antinutritive for herbivores. Among these compounds, The Generalist Inside the Specialist alkaloids, terpenoids, and complex mixtures of macromolecules such as latex are among the most frequent plant biochemical defense barriers. Terpenes are chemical compounds that are present in large amounts in a large variety of plants: in conifers, for example, they are the main components of resin. Along with alkaloids, natural gum and many other compounds, are present in Euphorbiaceae and other plants exuding latex. The larvae of the sphingid moth Hyles euphorbiae – the “spurge hawk moth” – feed on a broad variety of Euphorbia plants from which they sequester the cytotoxic ingenane diterpene esters (Marsh and Rothschild, 1984)

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