Coating of Leaf Surfaces with Agarose to Retain Fungal Inoculum in Situ for Staining

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Abstract
To avoid removing spores of Rhynchosporium secalis from barley leaves during decolorizing with aceticethanol (1:l) the inoculated leaves were placed on a horizontal sheet of chromatography paper through which the solvent drained and removed pigments from the leaves. Decolorized leaves were placed on water-soaked filter paper for 30 min to remove excess acetic-ethanol and then placed on microscope slides, chilled with ice or a thermoelectric microtome stage and sprayed with molten 0.5% agamse. Agamsemated leaves were stained with periodic add-Schiff reagent. Spores were not removed dnring clearing, coating and staining. Staining and visibility of spores, germ-tubes and subcuticular hyphae were not affected by the agarose film.

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