Molecular Mechanisms of Tungsten Toxicity Differ for Glycine max Depending on Nitrogen Regime.

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Since W shares certain chemical properties with the essential plant micronutrient molybdenum (Mo), it is proposed to inhibit enzymatic activity of molybdoenzymes [e.g., nitrate reductase (NR)] by replacing the Mo-ion bound to the co-factor. Recent studies suggest that W, much like other heavy metals, also exerts toxicity on its own. To create a comprehensive picture of tungsten stress, this study investigated the effects of W on growth and metabolism of soybean (Glycine max), depending on plant nitrogen regime [nitrate fed (N fed) vs. symbiotic N2 fixation (N fix)] by combining plant physiological data (biomass production, starch and nutrient content, N2 fixation, nitrate reductase activity) with root and nodule proteome data. Irrespective of N regime, NR activity and total N decreased with increasing W concentrations. Nodulation and therefore also N2 fixation strongly declined at high W concentrations, particularly in N fix plants. However, N2 fixation rate (g N fixed g-1 nodule dwt) remained unaffected by increasing W concentrations. Proteomic analysis revealed a strong decline in leghemoglobin and nitrogenase precursor levels (NifD), as well as an increase in abundance of proteins involved in secondary metabolism in N fix nodules. Taken together this indicates that, in contrast to the reported direct inhibition of NR, N2 fixation appears to be indirectly inhibited by a decrease in nitrogenase synthesis due to W induced changes in nodule oxygen levels of N fix plants. Besides N metabolism, plants exhibited a strong reduction of shoot (both N regimes) and root (N fed only) biomass, an imbalance in nutrient levels and a failure of carbon metabolic pathways accompanied by an accumulation of starch at high tungsten concentrations, independent of N-regime. Proteomic data (available via ProteomeXchange with identifier PXD010877) demonstrated that the response to high W concentrations was independent of nodule functionality and dominated by several peroxidases and other general stress related proteins. Based on an evaluation of several W responsive proteotypic peptides, we identified a set of protein markers of W stress and possible targets for improved stress tolerance.", "claims": [{"id": "claim-method", "statement": "An analysis of variance (ANOVA) followed by a Tukey HSD post hoc test, indicating a statistical significant difference at p < 0.05, was performed using R-studio.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:31001297", "locator": "sections[14] \"Statistical Analysis and Data Mining\" paragraph 3", "quote": "An analysis of variance (ANOVA) followed by a Tukey HSD post hoc test, indicating a statistical significant difference at p < 0.05, was performed using R-studio."}], "state": "complete"}, {"id": "claim-data-access", "statement": "The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE (Vizcaíno et al., 2016) partner repository (https://www.ebi.ac.uk/pride/archive/) with the dataset identifier PXD010877.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:31001297", "locator": "sections[38] \"Data Availability\" paragraph 2", "quote": "The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE (Vizcaíno et al., 2016) partner repository (https://www.ebi.ac.uk/pride/archive/) with the dataset identifier PXD010877."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": null, "steps": [{"id": "read_published_estimate", "order": 1, "name": "Read the published estimate from the source", "kind": "other", "state": "complete", "blocked_on": null, "description": "The claim, method and evidence span published with the source article.", "duration_hours": null, "parameters": {}, "inputs": ["source:pmid:31001297"], "outputs": ["the published estimate and its evidence"], "instruments": [], "reagents": []}, {"id": "obtain_source_data", "order": 2, "name": "Obtain the source data from the named repository", "kind": "data-preparation", "state": "complete", "blocked_on": null, "description": "The GEO series accession resolves and its sample accessions are listed. 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partner repository (https://www.ebi.ac.uk/pride/archive/) with the dataset identifier PXD010877.", "structure": {"repository": "GEO", "access_route": "GEO accession URL", "access_statement_verbatim": "The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE (Vizcaíno et al., 2016) partner repository (https://www.ebi.ac.uk/pride/archive/) with the dataset identifier PXD010877.", "access_statement_kind": "data_availability_statement", "sample_accessions": [], "sample_count": null, "group_sizes": null, "group_sizes_basis": null, "platform": null, "taxon": null, "title": "Tungsten stress response of soy bean (glycine max), LC-MSMS", "resolution_checked_at": "2026-10-31T18:35:55Z", "resolution_check": "identifier_resolution_only_no_download_no_licence_read", "downloaded": false, "licence_read": false}}, {"id": "asset:table:pmid:31001297:1", "role": "table", "format": "jats-table", "media_type": "application/vnd.jats.table+json", "uri": null, 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(control, 0.1, 0.5 mM W supplied as sodium tungstate) and under differing nitrogen supply regimes (N fix: week 2&3 0. 25 mM KNO3 & week 4-7 zero N; N fed: week 2–7 10 mM KNO3).", "rows": [["", "unit", "N fix", "N fix", "N fix", "N fed", "N fed", "N fed"], ["", "", "Control", "0.1 mM W", "0.5 mM W", "Control", "0.1 mM W", "0.5 mM W"], ["(A) SHOOT"], ["W", "mg kg−1", "< LOQ", "B", "379 ± 80.1", "A", "535 ± 108", "A", "< LOQ", "B", "446 ± 92.2", "A", "704 ± 137", "A"], ["Mo", "mg kg−1", "2.12 ± 0.82", "B", "5.14 ± 2.11", "A", "18.60 ± 8.28", "A", "1.32 ± 0.50", "B", "1.95 ± 0.75", "B", "10.28 ± 4.41", "A"], ["Cu", "mg kg−1", "9.5 ± 1.75", "A", "2.6 ± 2.47", "B", "11.5 ± 1.7", "A", "6.4 ± 1.95", "B", "4.8 ± 2.10", "B", "6.5 ± 1.94", "B"], ["Cr", "mg kg−1", "3.4 ± 1.06", "B", "13.2 ± 4.86", "A", "40.8 ± 17.0", "A", "2.2 ± 0.66", "B", "2.7 ± 0.82", "B", "22.0 ± 8.6", "A"], ["Mn", "mg kg−1", "66.5 ± 9.94", "A", "59.1 ± 9.27", "A", "27.8 ± 5.92", "B", "42.5 ± 7.62", "B", "31.66 ± 6.4", "B", 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1.27", "A", "2.47 ± 1.69", "A", "7.19 ± 2.31", "A", "2.21 ± 1.07", "A", "3.48 ± 1.05", "A", "4.88 ± 2.23", "A"], ["Cu", "mg kg−1", "14.2 ± 4.49", "B", "11.9 ± 4.48", "B", "72.1 ± 14.3", "A", "20.7 ± 5.71", "B", "32.7 ± 7.67", "B", "36.4 ± 8.22", "B"], ["Cr", "mg kg−1", "33.4 ± 7.21", "A", "25.1 ± 4.85", "A", "56.9 ± 20.7", "A", "14.6 ± 1.66", "B", "24.9 ± 4.28", "A", "50.9 ± 15.2", "A"], ["Mn", "mg kg−1", "48.2 ± 5.98", "B", "56.9 ± 7.91", "B", "60.2 ± 8.39", "B", "42.1 ± 5.20", "B", "101 ± 12.8", "A", "52.9 ± 6.57", "B"], ["Fe", "g kg−1", "2.20 ± 0.34", "A", "1.27 ± 0.16", "B", "2.12 ± 0.36", "A", "0.95 ± 0.09", "B", "1.02 ± 0.10", "B", "1.43 ± 0.17", "B"], ["P", "g kg−1", "2.56 ± 0.17", "C", "4.64 ± 0.44", "A", "3.71 ± 0.31", "B", "1.58 ± 0.08", "D", "3.16 ± 0.24", "B", "3.50 ± 0.28", "B"], ["Mg", "g kg−1", "5.54 ± 0.56", "A", "2.95 ± 0.39", "B", "2.31 ± 0.35", "B", "5.65 ± 0.57", "A", "4.06 ± 0.45", "A", "2.93 ± 0.38", "B"], ["S", "g kg−1", "6.03 ± 0.44", "A", "7.14 ± 0.55", "A", 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