Reef coral fluorescent proteins for visualizing fungal pathogens

The fluorescent proteins AmCyan, ZsGreen, ZsYellow, and AsRed, modified versions of proteins identified recently from several Anthozoa species of reef corals, were expressed for the first time in a heterologous system and used for imaging two different fungal plant pathogens. When driven by strong c...

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Bibliographic Details
Published in:Fungal genetics and biology 2002-12, Vol.37 (3), p.211-220
Main Authors: Bourett, Timothy M, Sweigard, James A, Czymmek, Kirk J, Carroll, Anne, Howard, Richard J
Format: Article
Language:English
Subjects:
Animals
Anthozoa - metabolism
Apical growth
Filamentous fungi
Fusarium - genetics
Fusarium - metabolism
Fusarium - pathogenicity
Fusarium - ultrastructure
Fusarium verticillioides
GFP
Hordeum - microbiology
Hyphal tip growth
Live cell imaging
Luminescent Proteins - genetics
Luminescent Proteins - metabolism
Magnaporthe - genetics
Magnaporthe - metabolism
Magnaporthe - pathogenicity
Magnaporthe - ultrastructure
Magnaporthe grisea
Microscopy - methods
Microscopy, Confocal
Mitosis
Multi-photon microscopy
Photons
Plant Diseases - microbiology
Transformation, Genetic
Two-photon microscopy
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Summary:The fluorescent proteins AmCyan, ZsGreen, ZsYellow, and AsRed, modified versions of proteins identified recently from several Anthozoa species of reef corals, were expressed for the first time in a heterologous system and used for imaging two different fungal plant pathogens. When driven by strong constitutive fungal promotors, expression of these reef coral fluorescent proteins yielded bright cytoplasmic fluorescence in Fusarium verticillioides and Magnaporthe grisea, and had no detectable effect on either growth rate or the ability to cause disease. Differential intracellular localization of the fluorescent proteins resulted in the discrimination of many subcellular organelles by confocal and multi-photon microscopy, and facilitated monitoring of such details as organelle dynamics and changes in the permeability of the nuclear envelope. AmCyan and ZsGreen were sufficiently excited at 855 and 880 nm, respectively, to allow for time-resolved in planta imaging by two-photon microscopy.
ISSN:1087-1845
1096-0937
DOI:10.1016/S1087-1845(02)00524-8