Evolutionary diversification of insulin-related peptides (IRPs) in aphids and spatiotemporal distribution in Acyrthosiphon pisum.

Members of the insulin superfamily activate the evolutionarily highly conserved insulin/insulin-like growth factor signaling pathway, involved in regulation of growth, energy homeostasis, and longevity. In the current study we focus on aphids to gain more insight into the evolution of the IRPs and how they may contribute to regulation of the insulin-signaling pathway. Using the latest annotation of the pea aphid (Acyrthosiphon pisum) genome, and combining sequence alignments and phylogenetic analyses, we identified seven putative IRP encoding-genes, with IRP1-IRP4 resembling the classical insulin and insulin-like protein structures, and IRP5 and IRP6 bearing insulin-like growth factor (IGF) features. We also identified IRP11 as a new and structurally divergent IRP present in at least eight aphid genomes. Globally the ten aphid genomes analyzed in this work contain four to 15 IRPs, while only three IRPs were found in the genome of the grape phylloxera, a hemipteran insect representing an earlier evolutionary branch of the aphid group. Expression analyses revealed spatial and temporal variation in the expression patterns of the different A. pisum IRPs. IRP1 and IRP4 are expressed throughout all developmental stages and morphs in neuroendocrine cells of the brain, while IRP5 and IRP6 are expressed in the fat body. IRP2 is expressed in specific cells of the gut in aphids in non-crowded conditions and in the head of aphids under crowded conditions, IRP3 in salivary glands, and both IRP2 and IRP3 in the male morph. IRP11 expression is enriched in the carcass. This complex spatiotemporal expression pattern suggests functional diversification of the IRPs.

Sustainable laser-based technology for insect pest control.

Aphids damage directly or indirectly cultures by feeding and spreading diseases, leading to huge economical losses. So far, only the use of pesticides can mitigate their impact, causing severe health and environmental issues. Hence, innovative eco-friendly and low-cost solutions must be promoted apart from chemical control. Here, we have investigated the use of laser radiation as a reliable solution. We have analyzed the lethal dose required to kill 90% of a population for two major pest aphid species (Acyrthosiphon pisum and Rhopalosiphum padi). We showed that irradiating insects at an early stage (one-day old nymph) is crucial to lower the lethal dose without affecting plant growth and health. The laser is mostly lethal, but it can also cause insect stunting and a reduction of survivors' fecundity. Nevertheless, we did not notice any significant visible effect on the offspring of the surviving irradiated generation. The estimated energy cost and the harmless effect of laser radiation on host plants show that this physics-based strategy can be a promising alternative to chemical pesticides.

Assessment of a 16S rRNA amplicon Illumina sequencing procedure for studying the microbiome of a symbiont-rich aphid genus.

The bacterial communities inhabiting arthropods are generally dominated by a few endosymbionts that play an important role in the ecology of their hosts. Rather than comparing bacterial species richness across samples, ecological studies on arthropod endosymbionts often seek to identify the main bacterial strains associated with each specimen studied. The filtering out of contaminants from the results and the accurate taxonomic assignment of sequences are therefore crucial in arthropod microbiome studies. We aimed here to validate an Illumina 16S rRNA gene sequencing protocol and analytical pipeline for investigating endosymbiotic bacteria associated with aphids. Using replicate DNA samples from 12 species (Aphididae: Lachninae, Cinara) and several controls, we removed individual sequences not meeting a minimum threshold number of reads in each sample and carried out taxonomic assignment for the remaining sequences. With this approach, we show that (i) contaminants accounted for a negligible proportion of the bacteria identified in our samples; (ii) the taxonomic composition of our samples and the relative abundance of reads assigned to a taxon were very similar across PCR and DNA replicates for each aphid sample; in particular, bacterial DNA concentration had no impact on the results. Furthermore, by analysing the distribution of unique sequences across samples rather than aggregating them into operational taxonomic units (OTUs), we gained insight into the specificity of endosymbionts for their hosts. Our results confirm that Serratia symbiotica is often present in Cinara species, in addition to the primary symbiont, Buchnera aphidicola. Furthermore, our findings reveal new symbiotic associations with Erwinia- and Sodalis-related bacteria. We conclude with suggestions for generating and analysing 16S rRNA gene sequences for arthropod-endosymbiont studies.

Genomic insight into the amino acid relations of the pea aphid, Acyrthosiphon pisum, with its symbiotic bacterium Buchnera aphidicola.

The pea aphid genome includes 66 genes contributing to amino acid biosynthesis and 93 genes to amino acid degradation. In several respects, the pea aphid gene inventory complements that of its symbiotic bacterium, Buchnera aphidicola (Buchnera APS). Unlike other insects with completely sequenced genomes, the pea aphid lacks the capacity to synthesize arginine, which is produced by Buchnera APS. However, consistent with other insects, it has genes coding for individual reactions in essential amino acid biosynthesis, including threonine dehydratase and branched-chain amino acid aminotransferase, which are not coded in the Buchnera APS genome. Overall the genome data suggest that the biosynthesis of certain essential amino acids is shared between the pea aphid and Buchnera APS, providing the opportunity for precise aphid control over Buchnera metabolism.

Tests of toxicity and teratogenicity in biphasic vertebrates treated with heavy metals (Cr3+, Al3+, Cd2+).

Developmental toxicity of chromium(III), aluminium(III) and cadmium(II) were evaluated by examining abnormalities and mortality in embryos belonging to different species of amphibians. Cr(III) and Al(III) are lethal at 1.5 mM concentration, and seriously affect the differentiation of central nervous system, skeleton and eye, and cause cephalic and trunk oedemas at lower concentrations, being aluminium significantly more harmful than chromium. Cd(II), tested only in P. waltl, is highly toxic: embryos exposed to concentrations ranging from 0.18 to 50 microM display malformations, delay and arrest of development in a dose dependent manner.

Effect of cadmium(II) on the extent of oxidative DNA damage in primary brain cell cultures from Pleurodeles larvae.

Compounds of cadmium(II) are well-known human and animal carcinogens. Furthermore, they affect development. growth and brain functions at subacute environmental concentrations in experimental animals. We investigated the potential of cadmium(II) to induce oxidative DNA damage in brain cell cultures obtained from larvae of Pleurodeles waltl. As indicators of DNA lesions typical of oxygen free radicals, we determined the frequencies of DNA strand breaks and of DNA base modifications recognized by the bacterial formamidopyrimidine-DNA glycosylase (Fpg protein). DNA strand breaks were generated in a dose-dependent manner at concentrations of 1 microM and greater. In contrast, no significant increase in Fpg-sensitive sites was observed under our experimental conditions. However, the repair of Fpg-sensitive DNA lesions induced by visible light was slightly diminished at 1 microM and inhibited completely at 10 microM of cadmium(II), while the closure of DNA strand breaks was not affected. Our results show that, although cadmium is not able to induce oxidative DNA base modifications in larval brain cells directly, its capability to generate DNA strand breaks and to interfere with the repair of oxidative DNA damage could explain the early life stage neurotoxicity of this metal.