Many scientific disciplines are currently experiencing a 'reproducibility crisis' because numerous scientific findings cannot be repeated consistently. A novel but controversial hypothesis postulates that stringent levels of environmental and biotic standardization in experimental studies reduce reproducibility by amplifying the impacts of laboratory-specific environmental factors not accounted for in study designs. A corollary to this hypothesis is that a deliberate introduction of controlled systematic variability (CSV) in experimental designs may lead to increased reproducibility. To test this hypothesis, we had 14 European laboratories run a simple microcosm experiment using grass (Brachypodium distachyon L.) monocultures and grass and legume (Medicago truncatula Gaertn.) mixtures. Each laboratory introduced environmental and genotypic CSV within and among replicated microcosms established in either growth chambers (with stringent control of environmental conditions) or glasshouses (with more variable environmental conditions). The introduction of genotypic CSV led to 18% lower among-laboratory variability in growth chambers, indicating increased reproducibility, but had no significant effect in glasshouses where reproducibility was generally lower. Environmental CSV had little effect on reproducibility. Although there are multiple causes for the 'reproducibility crisis', deliberately including genetic variability may be a simple solution for increasing the reproducibility of ecological studies performed under stringently controlled environmental conditions.

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This study benefited from the Centre Nationnal de la Recherche Scientifique human and technical resources allocated to the Ecotrons research infrastructures, the state allocation ‘Investissement d’Avenir’ ANR-11-INBS-0001 and financial support from the ExpeER (grant 262060) consortium funded under the EU-FP7 research programme (FP2007–2013). Brachypodium seeds were provided by R. Sibout (Observatoire du Végétal, Institut Jean-Pierre Bourgin) and Medicago seeds were supplied by J.-M. Prosperi (Institut National de la Recherche Agronomique Biological Resource Centre). We further thank J. Varale, G. Hoffmann, P. Werthenbach, O. Ravel, C. Piel, D. Landais, D. Degueldre, T. Mathieu, P. Aury, N. Barthès, B. Buatois and R. Leclerc for assistance during the study. For additional acknowledgements, see the Supplementary Information.

Author information


  1. Ecotron (Unité Propre de Service 3248), Centre National de la Recherche Scientifique, Campus Baillarguet, Montferrier-sur-Lez, France

    • Alexandru Milcu
    • , Sebastien Devidal
    •  & Jacques Roy
  2. Centre d’Ecologie Fonctionnelle et Evolutive, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5175, Université de Montpellier/Université Paul Valéry – École Pratique des Hautes Études, Montpellier, France

    • Alexandru Milcu
    • , Grégoire T. Freschet
    •  & Katherine Urban-Mead
  3. Institut de l’Ecologie et des Sciences de l’Environnement de Paris, Université Paris-Est Créteil, Créteil, France

    • Ruben Puga-Freitas
    • , Manuel Blouin
    •  & Agnès Gigon
  4. Harvard Forest, Harvard University, Petersham, MA, USA

    • Aaron M. Ellison
  5. Tropical Forests and People Research Centre, University of the Sunshine Coast, Maroochydore DC, Queensland, Australia

    • Aaron M. Ellison
  6. Agroécologie, AgroSup Dijon, Institut National de la Recherche Agronomique, Université Bourgogne Franche-Comté, Dijon, France

    • Manuel Blouin
    •  & Anne Pando
  7. Johann-Friedrich-Blumenbach Institute for Zoology and Anthropology, Georg August University Göttingen, Göttingen, Germany

    • Stefan Scheu
    •  & Olaf Butenschoen
  8. Department of Geobotany, Faculty of Biology, University of Freiburg, Freiburg, Germany

    • Laura Rose
    • , Anna Greiner
    • , Paula Rotter
    •  & Michael Scherer-Lorenzen
  9. Institut de Recherche pour le Développement, Institut de l’Ecologie et des Sciences de l’Environnement de Paris, Université Pierre et Marie Curie, Paris, France

    • Sebastien Barot
  10. German Centre for Integrative Biodiversity Research, Halle-Jena-Leipzig, Leipzig, Germany

    • Simone Cesarz
    • , Nico Eisenhauer
    •  & Alexandra Weigelt
  11. Institute of Biology, Leipzig University, Leipzig, Germany

    • Simone Cesarz
    • , Nico Eisenhauer
    •  & Alexandra Weigelt
  12. Institut Jean-Pierre Bourgin, INRA, AgroParisTech, Centre National de la Recherche Scientifique, Université Paris-Saclay, Versailles, France

    • Thomas Girin
  13. Department of Agricultural, Forest and Food Sciences, University of Turin, Grugliasco, Italy

    • Davide Assandri
    • , Carlo Grignani
    •  & Laura Zavattaro
  14. Cluster of Excellence on Plant Sciences, Terrestrial Ecology Group, Institute for Zoology, University of Cologne, Cologne, Germany

    • Michael Bonkowski
  15. Institute of Agricultural Sciences, ETH Zurich, Zurich, Switzerland

    • Nina Buchmann
    •  & Neringa Mannerheim
  16. Senckenberg Biodiversität und Klima Forschungszentrum, Frankfurt, Germany

    • Olaf Butenschoen
  17. Max Planck Institute for Biogeochemistry, Postfach 100164, Jena, Germany

    • Gerd Gleixner
    •  & Markus Lange
  18. Leibniz Centre for Agricultural Landscape Research, Institute of Landscape Biogeochemistry, Müncheberg, Germany

    • Arthur Gessler
    •  & Marina E. H. Müller
  19. Swiss Federal Research Institute, Zürcherstrasse 111, Birmensdorf, Switzerland

    • Arthur Gessler
    •  & Zachary Kayler
  20. Département de Biologie, Ecole Normale Supérieure, Université de recherche Paris Sciences & Lettres Research University, Centre National de la Recherche Scientifique, Unité Mixte de Service 3194 (Centre de Recherche en Écologie Expérimentale et Prédictive-Ecotron IleDeFrance), Saint-Pierre-lès-Nemours, France

    • Amandine Hansart
    •  & Jean-François Le Galliard
  21. Department of Soil and Water Systems, University of Idaho, Moscow, ID, USA

    • Zachary Kayler
  22. Institut de l’Ecologie et des Sciences de l’Environnement de Paris, Sorbonne Universités, Paris, France

    • Jean-Christophe Lata
    • , Jean-François Le Galliard
    •  & Rahme Seyhun
  23. School of Agriculture, Policy and Development, University of Reading, Reading, UK

    • Martin Lukac
  24. Faculty of Forestry and Wood Sciences, Czech University of Life Sciences, Prague, Czech Republic

    • Martin Lukac


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A.M. and J.R. designed the study with input from M. Blouin, S.B., M. Bonkowski and J.-C.L. Substantial methodological contributions were provided by S.S., T.G., L.R. and M.S.-L. Conceptual feedback on an early version was provided by G.T.F., N.E., J.R. and A.M.E. Data were analysed by A.M. with input from A.M.E. A.M. wrote the manuscript with input from all authors. All authors were involved in carrying out the experiments and/or analyses.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Alexandru Milcu.

Supplementary information

  1. Supplementary information

    Supplementary Tables 1–3; Supplementary Figures 1–5; Model outputs; Supplementary Acknowledgements.

  2. Life Sciences Reporting Summary