Applications of KOH-aniline blue fluorescence in the study of plant-fungal interactions

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 Mastigomy...

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Bibliographic Details
Published in:Phytopathology 1996, Vol.86 (7), p.704-708
Main Authors: Hood, M.E. (North Carolina State University, Raleigh, NC.), Shew, H.D
Format: Article
Language:English
Subjects:
Biological and medical sciences
COLORANT
COLORANTES
DIAGNOSTIC
DIAGNOSTICO
ELYMUS REPENS
ERYSIPHE
ERYSIPHE GRAMINIS
Fundamental and applied biological sciences. Psychology
Fungal plant pathogens
FUSARIUM
Generalities. Techniques
GYMNOSPORANGIUM
HIDROXIDOS
HYDROXYDE
JUNIPERUS VIRGINIANA
LAGERSTROEMIA
MONILINIA
NICOTIANA TABACUM
OIDIUM
OIDIUM (GENRE)
PATOGENICIDAD
PERONOSPORA TABACINA
PHARBITIS
Phytopathology. Animal pests. Plant and forest protection
PIERIS (ERICACEAE)
POTASIO
POTASSIUM
POUVOIR PATHOGENE
PRUNUS
SCLEROTINIA SCLEROTIORUM
SILENE
SOLANUM MELONGENA
TECHNIQUE ANALYTIQUE
TECNICAS ANALITICAS
THIELAVIOPSIS BASICOLA
TRITICUM AESTIVUM
UREDINALES
USTILAGO VIOLACEA
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Summary: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
ISSN:0031-949X
1943-7684
DOI:10.1094/phyto-86-704