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Metaproteomic analysis of glacial ecosystem

ABSTRACT: Glaciers are populated by a large number of microorganisms including bacteria, archaea and microeukaryotes. From an ecological point of view, three ecosystems can be differentiated in glaciers: the supraglacial ecosystem, the subglacial ecosystem and the englacial ecosystem. Several factors such as solar radiation, nutrient availability and water content greatly determine the diversity and abundance of microbial populations, the type of metabolism and the biogeochemical cycles. Firstly, the supraglacial ecosystem, sunlit and oxygenated, is predominantly populated by autotrophic microorganisms. Secondly, the subglacial ecosystem contains a majority of chemoautrotophs that are fed on the mineral salts of the rocks and basal soil. Lastly, the englacial ecosystem is the less studied and the one that contains the smallest number of microorganisms. However, these unknown englacial microorganisms establish a true trophic chain and appear to have an active metabolism. In order to study their metabolic potentials, samples of englacial ice were taken from an Antarctic glacier. The cells were harvested and their proteins were extracted and analyzed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI/TOF/TOF). Several proteins and enzymes were found that demonstrate the existence of cellular activity at subzero temperatures. In this way it is shown that the englacial microorganisms are not quiescent, but that they maintain an active metabolism and play an important role in the glacial microbial community.

INSTRUMENT(S): autoflex

ORGANISM(S): Environmental Samples

SUBMITTER: Cristina Cid

LAB HEAD: Cristina Cid

PROVIDER: PXD010060 | Pride | 2019-08-29

REPOSITORIES: Pride

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			Action	DRS
	50S_rpL17_Gallibacterium.mzXML	Mzxml
	50S_rpL17_Gallibacterium.xml	Xml
	h_prot_B_aggregatus.mzXML	Mzxml
	h_prot_B_aggregatus.xml	Xml
	kinase_P_fermentans.mzXML	Mzxml

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Project description:Biological nitrogen fixation (BNF) is a primary input of nitrogen to natural and agricultural systems globally. BNF is a temperature-dependant enzymatic process and can be conducted by microbes (including Rhizobia) hosted symbiotically in root nodules of some plants. Heat shock proteins (Hsps) have been implicated in the process of acquiring thermotolerance or acclimating to elevated temperature, as they play a vital role in maintaining cell integrity and homeostasis during heat stress. Although the BNF response to temperature may crucially impact future ecosystem productivity in the face of global climate change, little is known about Hsp expression in nodules of N-fixing non-agricultural species, such as tropical N-fixing trees in the Fabaceae family. This project aimed to characterize small (15-20kDa) Hsp (sHsp) expression in nodule tissue to examine the biochemical mechanisms of heat response in these tissues. To first identify Hsps in nodule tissues, Vachellia farnesiana and Acacia confusa nodules were excised, heat shock was induced, and protein content was isolated via chemical treatment before separation of protein species and analysis with SDS-PAGE. Two polyacrylamide gels yielded bands in the 15-20 kDa region that displayed differential Coomassie staining, which were sent for further characterization by HPLC-MS analysis for protein sequencing. Ten rhizobial sHsps were detected in these samples in addition to seven Acacia sHsps when compared independently to reference rhizobial and plant proteome databases. In an attempt quantify relative expression of Hsps in nodule and root tissue, we performed western blot experiments using Anti-Hsp20 antibodies raised against human and mouse Hsp proteins, with anti-beta actin loading control. While nonuniform beta-actin expression across tissue types (A. confusa nodules versus root control) prevented quantitative analysis, the experiments validated that Hsp20s are expressed in Acacia nodules as well as in root tissue. These experiments could provide a foundation for future studies that aim to determine variation in responses to key stressors predicted to increase with global climate change and help determine the implications of warming across the tropics and beyond.

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Metaproteomic analysis of glacial ecosystem

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