Author/Authors :
NOWICKI، Marcin نويسنده Research Institute of Horticulture Konstytucji 3 Maja 1/3, 96-100 Skierniewice, Poland , , LICHOCKA، Ma?gorzata نويسنده Institute of Biochemistry and Biophysics, Polish Academy of Sciences Pawi?skiego 5a, 02-106 Warszawa, Poland , , NOWAKOWSKA، Marzena نويسنده Research Institute of Horticulture Konstytucji 3 Maja 1/3, 96-100 Skierniewice, Poland , , K?OSI?SKA، Urszula نويسنده Research Institute of Horticulture Konstytucji 3 Maja 1/3, 96-100 Skierniewice, Poland , , GOLIK، Paulina نويسنده Research Institute of Horticulture Konstytucji 3 Maja 1/3, 96-100 Skierniewice, Poland , , KOZIK، El?bieta U. نويسنده Institute of Biochemistry and Biophysics, Polish Academy of Sciences Pawi?skiego 5a, 02-106 Warszawa, Poland ,
Abstract :
Dramatic increase in confocal microscopy observation output has been gained by optimization of a simple trypan blue and aniline blue dual-stain and its application to two model pathosystems: Pseudoperonospora cubensis-cucumber and Phytophthora infestans-tomato. Comparison of two dual-stain methods for confocal microscopy studies of P. cubensis-challenged cucumber leaves indicated the ʹmildʹ approach most successful. This methodology provides simultaneous detection of different pathogen structures layered with the plant defense reactions. Moreover, ImageJ-assisted quantification of plant defense responses renders this method useful for addressing the host plant resistance reactions, as well as investigating the given isolateʹs pathogenicity. Application of this method for the P. infestans-challenged tomato leaf samples resulted in detection of several fungal infection structures, along with plant defense responses. The dual-stain also enabled detection of a peculiar aniline blue-sensitive material in the pathogen cell walls at the area of its hyphae emerging through the leaf stomata. Results presented herein indicate this method is applicable for detailed (possibly quantitative) investigations of multiple plant-fungal pathosystems.
