A plate assay method for the detection of fungal polygalacturonase secretion

Abstract

A method for the detection of polygalacturonase activity has been developed using ruthenium red staining of fungal colonies on polygalacturonate-agarose plates. Ruthenium red was shown to penetrate beneath the surface layers of the gel, in the regions surrounding a fungal colony where degradation of polygalacturonate had occurred. Without degradation of polygalacturonate ruthenium red did not penetrate the medium, was restricted to binding to the surface layers and was easily washed off. The medium containing undegraded polygalacturonate was a colourless clear background and areas of polygalacturonate degradation around the colonies were visualised as an intense purple-red halo. The method has been used to screen yeasts and filamentous fungi for polygalacturonase secretion.