Applications of KOH-aniline blue fluorescence in the study of plant-fungal interactions
1996
Hood, M.E. | Shew, H.D.
A KOH-aniline blue technique for fluorescent staining of fungi in association with plant tissues was developed. The technique provided rapid, simple, and effective documentation of plant-fungal interactions with specimens representing the Deuteromycotina, Ascomycotina, Basidiomycotina, and Mastigomycotina. Applications of the technique were investigated, including documentation of host-penetration events, characteristics of host colonization, fungal reproduction, and detection of inoculum. In the standard KOH-aniline blue procedure, fresh specimens were autoclaved for 15 min at 121 degrees C in 50 ml of 1 M KOH, rinsed in deionized water, mounted in the stain solution, and examined with ultraviolet fluorescence. The stain solution was prepared as 0.05% aniline blue dye (CI #42755 or CI #42780) in 0.067 M K2HPO4 at pH 9.0. Modifications of the standard procedure also were tested, including use of variously preserved specimens, alteration of the KOH treatment, and moderate variation of the stain solution. The technique produced a high degree of resolution and contrast between hyphae and host-plant tissues. The resulting documentation supported previous observations and, in some cases, provided new information about the nature of specific host-pathogen interactions.
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