Integrated Metabonomic−Proteomic Analysis of an Insect−Bacterial Symbiotic System

Abstract

The health of animals, including humans, is dependent on their resident microbiota, but the complexity of the microbial communities makes these associations difficult to study in most animals. Exceptionally, the microbiology of the pea aphid Acyrthosiphon pisum is dominated by a single bacterium Buchnera aphidicola (B. aphidicola). A 1 H NMR-based metabonomic strategy was applied to investigate metabolic profiles of aphids fed on a low essential amino acid diet and treated by antibiotic to eliminate B. aphidicola. In addition, differential gel electrophoresis (DIGE) with mass spectrometry was utilized to determine the alterations of proteins induced by these treatments. We found that these perturbations resulted in significant changes to the abundance of 15 metabolites and 238 proteins. Ten (67%) of the metabolites with altered abundance were amino acids, with nonessential amino acids increased and essential amino acids decreased by both perturbations. Over-represented proteins in the perturbed treatments included catabolic enzymes with roles in amino acid degradation and glycolysis, various cuticular proteins, and a C-type lectin and regucalcin with candidate defensive roles. This analysis demonstrates the central role of essential amino acid production in the relationship and identifies candidate proteins and processes underpinning the function and persistence of the association.

Metaorganisms as the new frontier

“…Consistent with the holobiont theory of evolution (Zilber-Rosenberg et al, 2008) this implies that aphids acquire resistance to natural enemies by picking up bacterial symbionts, rather than having changes in the aphids' genes. A truly transdisciplinary analysis performed by experts from a Department of Entomology, a Department of Biology, two laboratories of Magnetic Resonance and Atomic and Molecular Physics, an Institute of Physics and Mathematics, a School of Biology and Environmental Science, a Department of Biomolecular Medicine, and a Department of Surgery and Cancer, deepened our understanding of the interaction between aphids and B. aphidicola by uncovering the central role of amino acid metabolism in the aphid – Buchnera symbiosis (Wang et al, 2010). …”

Entometabolomics: applications of modern analytical techniques to insect studies

“…Although many applications are currently only represented by relatively few studies, entometabolomics has contributed to the understanding of such topics as hypoxia (Coquin et al., ; Feala et al., ), insect–bacterial symbiosis (Wang et al., ), behavioural ecology (Lenz et al., ), parasitism (Thompson et al., ), development (Phalaraksh et al., ; Wu et al., ), infectious diseases (Aliferis et al., ), the effects of commercial pesticides (Derecka et al., ), and temperature‐dependent stresses (Michaud & Denlinger, ; Michaud et al., ; Koštál et al., ,b) (Table ). Simultaneously, metabolomic investigations have indirectly generated information about insect life histories; particularly work focusing on plant‐insect interactions (Hunt et al., , ; Faria et al., ; Gattolin et al., ; Jansen et al., ; Leiss et al., ).…”

“…Metabolomics has been applied to a number of insect study systems in an effective manner, generating new insights into the mechanisms underlying aspects of biology including behaviour (Lenz et al., ), infection (Chambers et al., ), temperature stress responses (Malmendal et al., ; Li et al., ; Colinet et al., ), CO 2 sedation (Colinet & Renault, ), and bacteria–insect symbiosis (Wang et al., ). We are sure that this list of topics will expand in the near future.…”

Genetics Coupled to Quantitative Intact Proteomics Links Heritable Aphid and Endosymbiont Protein Expression to Circulative Polerovirus Transmission

“…In the present study, we coupled two-dimensional (2-D) fluorescence difference gel electrophoresis (DIGE), one of the most widely used platforms for quantitative proteomics (68), and tandem mass spectrometry with genetic analyses to quantitatively compare the proteomes of S. graminum genotypes that differ in the ability to transmit CYDV-RPV. The recently published genome sequence of the aphid species A. pisum has provided additional resources for protein discovery in our S. graminum genotypes and has propelled proteomics to the forefront of biological discovery technologies for S. graminum and other aphid species (10,14,25,26,50,51,74).…”