Proteomic analysis of the soil filamentous fungus Aspergillus nidulans exposed to a Roundup formulation at a dose causing no macroscopic effect: a functional study
- 285 Downloads
Roundup® is a glyphosate-based herbicide (GBH) used worldwide both in agriculture and private gardens. Thus, it constitutes a substantial source of environmental contaminations, especially for water and soil, and may impact a number of non-target organisms essential for ecosystem balance. The soil filamentous fungus Aspergillus nidulans has been shown to be highly affected by a commercial formulation of Roundup® (R450), containing 450 g/L of glyphosate (GLY), at doses far below recommended agricultural application rate. In the present study, we used two-dimensional gel electrophoresis combined to mass spectrometry to analyze proteomic pattern changes in A. nidulans exposed to R450 at a dose corresponding to the no-observed-adverse-effect level (NOAEL) for macroscopic parameters (31.5 mg/L GLY among adjuvants). Comparative analysis revealed a total of 82 differentially expressed proteins between control and R450-treated samples, and 85% of them (70) were unambiguously identified. Their molecular functions were mainly assigned to cell detoxification and stress response (16%), protein synthesis (14%), amino acid metabolism (13%), glycolysis/gluconeogenesis/glycerol metabolism/pentose phosphate pathway (13%) and Krebs TCA cycle/acetyl-CoA synthesis/ATP metabolism (10%). These results bring new insights into the understanding of the toxicity induced by higher doses of this herbicide in the soil model organism A. nidulans. To our knowledge, this study represents the first evidence of protein expression modulation and, thus, possible metabolic disturbance, in response to an herbicide treatment at a dose that does not cause any visible effect. These data are likely to challenge the concept of “substantial equivalence” when applied to herbicide-tolerant plants.
KeywordsRoundup® NOAEL Aspergillus nidulans Proteomics Metabolism Herbicide tolerance Substantial equivalence
This work was supported by the non-governmental organization “Générations Futures” and the Committee for Independent Research and Information on Genetic Engineering (CRIIGEN), in the framework of a participatory research project.
It received funding from the Regional Council Ile-de-France and the University Paris-Sud.
- Binder S (2010) Branched-chain amino acid metabolism in Arabidopsis thaliana. Arabidopsis Book. https://doi.org/10.1199/tab.0137
- Braconi D, Sotgiu M, Millucci L, Paffetti A, Tasso F, Alisi C, Martini S, Rappuoli R, Lusini P, Sprocati AR, Rossi C, Santucci A (2006) Comparative analysis of the effects of locally used herbicides and their active ingredients on a wild-type wine Saccharomyces cerevisiae strain. J Agric Food Chem 54:3163–3172CrossRefGoogle Scholar
- Cerqueira GC, Arnaud MB, Inglis DO, Skrzypek MS, Binkley G, Simison M, Miyasato SR, Binkley J, Orvis J, Shah P, Wymore F, Sherlock G, Wortman JR (2014) The Aspergillus genome database: multispecies curation and incorporation of RNA-Seq data to improve structural gene annotations. Nucleic Acids Res 42:705–710CrossRefGoogle Scholar
- Duke SO, Baerson SR, Rimando AM (2003) Herbicides: glyphosate. In: Plimmer JR, Gammon DW, Ragsdale NN (eds) Encyclopedia of agrochemicals. John Wiley & Sons, New York, pp 708–869Google Scholar
- Hickey PC, Swift SR, Roca MG, Read ND (2005) Live-cell imaging of filamentous fungi using vital fluorescent dyes and confocal microscopy. In: Savidge T, Pothoulakis C (eds) Methods in microbiology, Microbial Imaging, vol 35. Elsevier, London, pp 63–87Google Scholar
- James C (2011) Global status of commercialized biotech/GM crops. ISAAA brief no 43. ISAAA, Ithaca, NYGoogle Scholar
- Kelly JM, Hynes MJ (1981) The regulation of phosphoenolpyruvate carboxykinase and the NADP-linked malic enzyme in Aspergillus nidulans. J Gen Microbiol 123:371–375Google Scholar
- Martinelli SD, Kinghorn JR (1994) Aspergillus: 50 years on—progress in industrial microbiology. Volume 29. Elsevier, Amsterdam—London—New York—TokyoGoogle Scholar
- Mesnage R, Defarge N, Spiroux de Vendômois J, Séralini GE (2014) Major pesticides are more toxic to human cells than their declared active principles. Biomed Res Int. https://doi.org/10.1155/2014/179691
- Navarro RE, Aguirre J (1998) Posttranscriptional control mediates cell type-specific localization of catalase A during Aspergillus nidulans development. J Bacteriol 180:5733–5738Google Scholar
- Sabatier P, Poulenard J, Fanget B, Reyss JL, Develle AL, Wilhelm B, Ployon E, Pignol C, Naffrechoux E, Dorioz JM, Montuelle B, Arnaud F (2014) Long-term relationships among pesticide applications, mobility, and soil erosion in a vineyard watershed. Proc Natl Acad Sci U S A 111:15647–15652CrossRefGoogle Scholar
- Steffen W, Richardson K, Rockström J, Cornell SE, Fetzer I, Bennett EM, Biggs R, Carpenter SR, de Vries W, de Wit CA, Folke C, Gerten D, Heinke J, Mace GM, Persson LM, Ramanathan V, Reyers B, Sörlin S (2015) Sustainability. Planetary boundaries: guiding human development on a changing planet. Science 347:1259855CrossRefGoogle Scholar