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Spatial distribution and catalytic mechanisms of β-glucosidase activity at the root-soil interface

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Abstract

We compared modifications of soil zymography, a new in situ technique to visualize enzyme activities, based on contact of fluorgenic substrate-saturated membranes with soil either through the gel layer (gel zymography) or without gel application (direct zymography). We coupled zymography with quantitative measurements of enzyme kinetics to characterize catalytic mechanisms of β-glucosidase activity at the plant-soil interface including root surface (rhizoplane), rhizosphere, and bulk soil. Direct zymography refined and focused image resolution. The area of hotspots (i.e., spots with most intensive enzyme activity) as well as color intensity ratios estimated using direct zymography exceeded by a factor of 2 the corresponding values obtained with gel zymography. As determined by direct zymography, the percentage of hotspots associated to root surfaces was 58–68 % of total hotspot area. Hotspot area comprised only 6.8 ± 0.1 % of the total area of an image and 9.0 ± 3 % of the root surface area. The intensity of β-glucosidase activity, however, was up to 20 times higher in the hotspots versus bulk soil. The contribution of rhizosphere to β-glucosidase activity of the whole image (77–82 %) was four times higher than the contribution of the root surface. Enzyme kinetic parameters indicated different enzyme systems in bulk and rhizosphere soil. Higher substrate affinity and catalytic efficiency in bulk than in rhizosphere soil suggested relative domination of microorganisms with more efficient enzyme systems in the former. Coupling direct zymography and kinetic assays enabled mapping the two-dimensional (2D) distribution of enzyme activity at the root-soil interface and estimating the catalytic properties of root-associated and soil-associated enzymes.

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Acknowledgments

We acknowledge the Alexander von Humboldt Foundation for financial support of MS and the DAAD for BSR. The contribution of EB was supported by the Russian Scientific Foundation (project No. 14-14-00625). This study was supported by the German Research Foundation (DFG) within the Research Unit “Carbon flux through the soil animal food web” (FOR 918; KU1184/3-2). We highly acknowledge the guidance and input of the editor and the reviewers for the improvement of this manuscript.

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Authors and Affiliations

  1. Department of Agricultural Soil Science, Georg August University of Göttingen, Büsgenweg 2, 37077, Göttingen, Germany

    Muhammad Sanaullah, Bahar S. Razavi, Evgenia Blagodatskaya & Yakov Kuzyakov

  2. Institute of Soil and Environmental Sciences, University of Agriculture, Faisalabad, Pakistan

    Muhammad Sanaullah

  3. Institute of Physicochemical and Biological Problems in Soil Science, 142290, Pushchino, Russia

    Evgenia Blagodatskaya

  4. Institute of Environmental Sciences, Kazan Federal University, Kazan, Russia

    Yakov Kuzyakov

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  1. Muhammad Sanaullah
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  2. Bahar S. Razavi
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  3. Evgenia Blagodatskaya
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  4. Yakov Kuzyakov
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Correspondence to Muhammad Sanaullah or Evgenia Blagodatskaya.

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Sanaullah, M., Razavi, B.S., Blagodatskaya, E. et al. Spatial distribution and catalytic mechanisms of β-glucosidase activity at the root-soil interface. Biol Fertil Soils 52, 505–514 (2016). https://doi.org/10.1007/s00374-016-1094-8

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