Dataset Information

Drosophila hemolymph infected with Spiroplasma poulsonii

ABSTRACT: We investigated the effect of Spiroplasma infection on Drosophila hemolymph protein content using Liquid Chromatography-tandem Mass Spectrometry (LC-MS/MS). To this end, we extracted total hemolymph from uninfected and infected 10 days old females. At this age, Spiroplasma is already present at high titers in the hemolymph but does not cause major deleterious phenotypes to the fly. Extraction was achieved by puncturing the thorax and drawing out with a microinjector. Four replicates were made

INSTRUMENT(S): Q Exactive

ORGANISM(S): Drosophila Melanogaster (fruit Fly)

TISSUE(S): Hemolymph

SUBMITTER: Samuel Rommelaere

LAB HEAD: Bruno Lemaitre EPFL Switzerland

PROVIDER: PXD024340 | Pride | 2021-05-04

REPOSITORIES: Pride

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Publications

Dual proteomics of Drosophila melanogaster hemolymph infected with the heritable endosymbiont Spiroplasma poulsonii.

Masson Florent F Rommelaere Samuel S Marra Alice A Schüpfer Fanny F Lemaitre Bruno B

PloS one 20210429 4

Insects are frequently infected with heritable bacterial endosymbionts. Endosymbionts have a dramatic impact on their host physiology and evolution. Their tissue distribution is variable with some species being housed intracellularly, some extracellularly and some having a mixed lifestyle. The impact of extracellular endosymbionts on the biofluids they colonize (e.g. insect hemolymph) is however difficult to appreciate because biofluid composition can depend on the contribution of numerous tissu ...[more]

PMID: 33914801

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Project description:Pollinators are of crucial importance for maintaining biodiversity in ecosystems and for agriculture, including the Buff-tailed bumblebee Bombus terrestris. The health decline in these populations is thought to be linked to various abiotic and biotic stressors. In the hope of protecting these populations, deciphering their immune response in stress conditions represent a critical issue. To assess this metric, we analysed the bumblebee hemolymph, as a readout of the immune status. Here, we show in laboratory conditions that bacterial infections with two entomopathogenic strains and a well-recognised inducer of defence reactions in insects, impact the systemic immune response. Analysis of the hemolymph was carried out using a two-stage approach of mass spectrometry that combines MALDI molecular mass fingerprinting (MALDI MFP or MALDI-BeeTyping®) for its effectiveness in assessing the immune status of the bumblebee through a basic “blood” test and LC-ESI-MS/MS to measure the impact of our infectious models on the “haemoproteome”. By carrying out three different types of bacterial infections, we found that the bumblebee reacts in a specific way to bacterial attacks. Indeed, bacteria impact mortality and stimulate an immune response in infected individuals that is visible through changes in the molecular composition of their hemolymph. The characterisation and label-free quantification of proteins involved in specific signalling pathways in bumblebees by LC-ESI-MS/MS revealed differences in the protein expression between the non-experimentally infected and the infected bumblebees. Our results highlights differences in the molecular composition of their hemolymph to distinguish these different infections from non-experimentally infected bumblebees, and an alteration of pathways involved in (i) immunity and defences, (ii) the metabolism of carbohydrates (glycolysis), (iii) oxidative stress and (iv) amino acid biosynthesis. To conclude, we established bioinformatics models based on molecular markers reflecting the health status of these pollinators to enable diagnosis/prognosis at the population level in response to environmental stress.

Project description:Citrus greening disease, also known as Huanglongbing (HLB), is the most serious disease of citrus plants. It is associated with the Gram-negative bacterium ‘Candidatus Liberibacter asiaticus’ (CLas), which is transmitted between host plants by the hemipteran insect vector Diaphorina citri in a circulative, propagative manner involving specific interactions with various insect tissues, including the hemolymph, fluid that occupies the body cavity akin to insect blood. High resolution quantitative mass spectrometry was performed to investigate the effect of CLas exposure on D. citri hemolymph at the proteome level. In contrast to the broad proteome effects on hundreds of proteins and a diverse array of metabolic pathways previously reported in gut and whole insect proteome analyses, the effect of CLas on the hemolymph was observed to be highly specific, restricted to key immunity and metabolism pathways, and lower in magnitude than that previously observed in the whole insect body and gut. Vitellogenins were abundantly expressed and CLas-responsive. Gene-specific RNA expression analysis suggests that these proteins are expressed in both male and female insects, and may have roles outside of reproductive vitellogenesis. Proteins for fatty acid synthesis were found to be up-regulated, along with metabolic proteins associated with energy production, supported at the organismal level by the previously published observation that D. citri individuals experience a higher level of hunger when reared on CLas-infected plants. Prediction of post-translational modifications identified hemolymph proteins with phosphorylation and acetylation upon CLas exposure. Proteins derived from the three most prominent bacterial endosymbionts of the psyllid were also detected in the hemolymph, and several of these have predicted secretion signals. A DNAK protein, the bacterial HSP70, detected in the hemolymph expressed from Wolbachia pipientis was predicted to encode a eukaryotic nuclear localization signal. Taken together, these data show specific changes to immunity and metabolism in D. citri hemolymph involving host and endosymbiont proteins. These data provide a novel context for proteomic changes seen in other D. citri tissues in response to CLas and align with organismal data on the effects of CLas on D. citri metabolism and reproduction.

Dataset Information

Drosophila hemolymph infected with Spiroplasma poulsonii

Publications

Dual proteomics of Drosophila melanogaster hemolymph infected with the heritable endosymbiont Spiroplasma poulsonii.

Similar Datasets

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