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, G. Kovács

Research output: Contribution to journalArticle

12 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)259-266
Number of pages8
JournalMycorrhiza
Volume24
Issue number4
DOIs
Publication statusPublished - 2014

Fingerprint

fluorescence in situ hybridization
Plantae
Fluorescence In Situ Hybridization
visualization
Fungi
fluorescence
fungus
fungi
Plant Roots
Ribosomal RNA
fluorescent dyes
Fluorescent Dyes
RNA
dye
probe
ribosomal RNA
Mycorrhizae
ectomycorrhiza
ectomycorrhizae
confocal laser scanning microscopy

Keywords

  • Cadophora
  • CLSM
  • Dark septate
  • Endophyte
  • FISH
  • Glomus
  • Microscopy
  • Rhizophagus
  • Root
  • rRNA

ASJC Scopus subject areas

  • Plant Science
  • Ecology, Evolution, Behavior and Systematics
  • Genetics
  • Molecular Biology
  • Medicine(all)

Cite this

Simultaneous specific in planta visualization of root-colonizing fungi using fluorescence in situ hybridization (FISH). / Vági, Pál; Knapp, Dániel G.; Kósa, Annamária; Seress, Diána; Horváth, Áron N.; Kovács, G.

In: Mycorrhiza, Vol. 24, No. 4, 2014, p. 259-266.

Research output: Contribution to journalArticle

Vági, Pál ; Knapp, Dániel G. ; Kósa, Annamária ; Seress, Diána ; Horváth, Áron N. ; Kovács, G. / Simultaneous specific in planta visualization of root-colonizing fungi using fluorescence in situ hybridization (FISH). In: Mycorrhiza. 2014 ; Vol. 24, No. 4. pp. 259-266.
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