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

Vági, Pál and Knapp, Dániel G. and Kósa, Annamária and Seress, Diána and Horváth, Áron N. and Kovács, Gábor M. (2014) Simultaneous specific in planta visualization of root-colonizing fungi using fluorescence in situ hybridization (FISH). Mycorrhiza, 24 (4). pp. 259-266. ISSN 0940-6360

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Abstract

ORIGINAL PAPER Simultaneous specific in planta visualization of root-colonizing fungi using fluorescence in situ hybridization (FISH) Pál Vági & Dániel G. Knapp & Annamária Kósa & Diána Seress & Áron N. Horváth & Gábor M. Kovács Received: 29 May 2013 /Accepted: 9 October 2013 /Published online: 13 November 2013 # Springer-Verlag Berlin Heidelberg 2013 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 differen- tial 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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