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Symbiotic microbes can enable their host to access untapped nutritional resources but may also constrain niche space by promoting specialization. Here, we reconstruct functional changes in the evolutionary history of the symbiosis between a group of (semi-)aquatic herbivorous insects and mutualistic bacteria. Sequencing the symbiont genomes across 26 species of reed beetles (Chrysomelidae, Donaciinae) spanning four genera indicates that the genome-eroded mutualists provide life stage-specific benefits to larvae and adults, respectively. In the plant sap-feeding larvae, the symbionts are inferred to synthesize most of the essential amino acids as well as the B vitamin riboflavin. The adult reed beetles' folivory is likely supported by symbiont-encoded pectinases that complement the host-encoded set of cellulases, as revealed by transcriptome sequencing. However, mapping the occurrence of the symbionts' pectinase genes and the hosts' food plant preferences onto the beetles' phylogeny reveals multiple independent losses of pectinase genes in lineages that switched to feeding on pectin-poor plants, presumably constraining their hosts' subsequent adaptive potential.

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Intracellular bacterial symbionts are known from various insect groups, particularly from those feeding on unbalanced diets, where the bacteria provide essential nutrients to the host. In the case of reed beetles (Coleoptera: Chrysomelidae, Donaciinae), however, the endosymbionts appear to be associated with specialized "glands" that secrete a material used for the beetles' unusual water-tight cocoon. These glands were discovered over a century ago, but the bacteria they contain have yet to be characterized and placed in a phylogenetic context. Here, we describe the ultrastructure of two endosymbiotic species ("Candidatus Macropleicola appendiculatae" and "Candidatus Macropleicola muticae") that reside in cells of the Malpighian tubules of the reed beetle species Macroplea appendiculata and Macroplea mutica, respectively. Fluorescent in situ hybridization using oligonucleotides targeting the 16S rRNA gene specific to Macroplea symbionts verified the localization of the symbionts in these organs. Phylogenetic analysis of 16S rRNA placed "Candidatus Macropleicola" in a clade of typically endosymbiotic Enterobacteriaceae (gamma-proteobacteria). Finally, we discuss the evidence available for the hypothesis that the beetle larvae use a secretion produced by the bacteria for the formation of an underwater cocoon.

The genus Macroplea Samouelle, 1819 is a group of highly specialized aquatic leaf beetles occurring in the Palaearctic. Since the members of this genus are morphologically very similar, we addressed the question of species identification and delimitation by analysing the second half of the mitochondrial gene coding for the cytochrome oxidase I (COI) subunit. Species limits are inferred from the multimodal frequency distribution of genetic distances between specimens: low genetic distances within a species are clearly set apart from distances between species. The species status of the hitherto controversial species M. japana (Jacoby, 1885) is confirmed. The pattern of nucleotide and amino acid substitutions is discussed in the light of functional domains of the COI molecule. Although the data are preliminary, the results provide new data on the distribution of the species. Together with the phylogenetic analysis they allow for a discussion of the phylogeography of the genus.

The present paper reviews the biology of reed beetles (Donaciinae), presents experimental data on the role of specific symbiotic bacteria, and describes a molecular method for the detection of those bacteria. Reed beetles are herbivores living on wetland plants, each species being mono- or oligo-phagous. They lay their eggs on the host plant and the larvae live underwater in the sediment attached to its roots. The larvae pupate there in a water-tight cocoon, which they build using a secretion that is produced by symbiotic bacteria. The bacteria are located in four blind sacs at the foregut of the larvae; in (female) adults they colonize two out of the six Malpighian tubules. Tetracycline treatment of larvae reduced their pupation rate, although the bacteria could not be fully eliminated. When the small amount of bacterial mass attached to eggs was experimentally removed before hatching, symbiont free larvae resulted, showing the external transmission of the bacteria to the offspring. Specific primers were designed to detect the bacteria, and to confirm their absence in manipulated larvae. The pupation underwater enabled the reed beetles to permanently colonize the wetlands and to diversify in this habitat underexploited by herbivorous insects (adaptive radiation).

The elongated labium of rove beetles of the genus Stenus forms an adhesive capture apparatus that enables them to catch fast-fleeing prey such as Collembola. The adhesion is mediated by a secretion produced in glands within the head capsule and secreted onto the paraglossae. Transmission electron microscopy has revealed that these "adhesive glands" are composed of discrete gland units, each consisting of three cells. Two cells are secretorily active, each producing a different secretion, one proteinaceous and the other lipoid. Consequently, a two-phase secretion can be found on the surface of the paraglossae. Adhesive glands and normal epidermal glands share several characteristics and are therefore considered to be homologous. Structural differences can be functionally interpreted. The long glandular ductules themselves serve as a reservoir for the secretion before it is expressed prior to the predatory strike. Van der Waals forces and both the surface tension and the viscosity of the adhesive secretion are discussed as possible mechanisms of adhesion. The adhesion resulting from the viscosity of the fluid is the strongest and exceeds the force theoretically required for catching collemboles.