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Background Meloidae (blister beetles) are known to synthetize cantharidin (CA), a toxic and defensive terpene mainly stored in male accessory glands (MAG) and emitted outward through reflex-bleeding. Recent progresses in understanding CA biosynthesis and production organ(s) in Meloidae have been made, but the way in which self-protection is achieved from the hazardous accumulation and release of CA in blister beetles has been experimentally neglected. To provide hints on this pending question, a comparative de novo assembly transcriptomic approach was performed by targeting two tissues where CA is largely accumulated and regularly circulates in Meloidae: the male reproductive tract (MRT) and the haemolymph. Differential gene expression profiles in these tissues were examined in two blister beetle species, Lydus trimaculatus (Fabricius, 1775) (tribe Lyttini) and Mylabris variabilis (Pallas, 1781) (tribe Mylabrini). Upregulated transcripts were compared between the two species to identify conserved genes possibly involved in CA detoxification and transport. Results Based on our results, we hypothesize that, to avoid auto-intoxication, ABC, MFS or other solute transporters might sequester purported glycosylated CA precursors into MAG, and lipocalins could bind CA and mitigate its reactivity when released into the haemolymph during the autohaemorrhaging response. We also found an over-representation in haemolymph of protein-domains related to coagulation and integument repairing mechanisms that likely reflects the need to limit fluid loss during reflex-bleeding. Conclusions The de novo assembled transcriptomes of L. trimaculatus and M. variabilis here provided represent valuable genetic resources to further explore the mechanisms employed to cope with toxicity of CA in blister beetle tissues. These, if revealed, might help conceiving safe and effective drug-delivery approaches to enhance the use of CA in medicine.

Larvae of the two European species of the genus Pyrochroa (P. coccinea and P. serraticornis) are redescribed, for the first time based on Italian samples, and new relevant and distinctive characters have emerged from P. serraticornis. Particularly, the presence of a single urogomphal pit between urogomphi (representing a new condition for the entire family), and new characters of mandibles, open new questions about the current taxonomic status of the European species.

The Rosalia longicorn (Rosalia alpina) is a strictly protected saproxylic beetle, widely distributed in Central and Southern Europe and mainly associated with ancient beech forests. To improve knowledge about the conservation status of R. alpina in Italy, available molecular markers (microsatellites and mitochondrial cytochrome c oxidase I(COI)) were tested for the first time on Italian populations. The study was performed in four sampling sites distributed in two areas placed in Northern ("Foreste Casentinesi" National Park) and Central Apennines ("Abruzzo, Lazio and Molise" National Park) where populational data about Rosalia longicorn were collected in the framework of the European LIFE MIPP Project. The genetic relationship among Apennine and Central/South-eastern European populations was explored by a comparison with mitochondrial DNA (mtDNA) data from literature. Microsatellite markers were only partially informative when applied to R. alpina Italian individuals, although providing some preliminary indication on an extensive gene flow among populations from the Apennines and local ongoing processes of genetic erosion. Genetic data are consistent with previous ecological data suggesting that the maintenance of variability in this species could be related to both habitat continuity and preservation of large senescent or standing dead trees in forests. Finally, a peculiar origin of the Apennine populations of R. alpina from a putative "Glacial Refugium" in Italy was inferred through COI data. The high genetic distance scored among the analysed populations and those from Central and Southeastern Europe indicates that the R. alpina deme from Apennine Mountains might represent a relevant conservation unit in Europe. Further genetic analyses will allow assessing other possible conservation units of R. alpina and, thus, defining large-scale conservation strategies to protect this endangered longhorn beetle in Europe.

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