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Simultaneous specific in planta visualization of root-colonizing fungi using fluorescence in situ hybridization (FISH)

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Abstract

In planta detection of mutualistic, endophytic, and pathogenic fungi commonly colonizing roots and other plant organs is not a routine task. We aimed to use fluorescence in situ hybridization (FISH) for simultaneous specific detection of different fungi colonizing the same tissue. We have adapted ribosomal RNA (rRNA) FISH for visualization of common mycorrhizal (arbuscular- and ectomycorrhiza) and endophytic fungi within roots of different plant species. Beside general probes, we designed and used specific ones hybridizing to the large subunit of rRNA with fluorescent dyes chosen to avoid or reduce the interference with the autofluorescence of plant tissues. We report here an optimized efficient protocol of rRNA FISH and the use of both epifluorescence and confocal laser scanning microscopy for simultaneous specific differential detection of those fungi colonizing the same root. The method could be applied for the characterization of other plant–fungal interactions, too. In planta FISH with specific probes labeled with appropriate fluorescent dyes could be used not only in basic research but to detect plant colonizing pathogenic fungi in their latent life-period.

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Acknowledgments

We thank Yoram Kapulnik (ARO Volcani Center, Israel) for kindly providing AMF inoculum. Funding was provided by the Hungarian Scientific Research Fund (OTKA, K72776, and NI72776). GMK is supported by the Bolyai János Research Fellowship (Hungarian Academy of Sciences).

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Authors and Affiliations

  1. Department of Plant Anatomy, Institute of Biology, Eötvös Loránd University, 1117, Budapest, Pázmány Péter sétány 1/c, Hungary

    Pál Vági, Dániel G. Knapp, Annamária Kósa, Diána Seress, Áron N. Horváth & Gábor M. Kovács

  2. Plant Protection Institute, Centre for Agricultural Research, Hungarian Academy of Sciences, Budapest, Hungary

    Pál Vági & Gábor M. Kovács

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  1. Pál Vági
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  2. Dániel G. Knapp
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  6. Gábor M. Kovács
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Correspondence to Gábor M. Kovács.

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Vági, P., Knapp, D.G., Kósa, A. et al. Simultaneous specific in planta visualization of root-colonizing fungi using fluorescence in situ hybridization (FISH). Mycorrhiza 24, 259–266 (2014). https://doi.org/10.1007/s00572-013-0533-8

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