The role of melanin in the antagonistic interaction between the apple scab pathogen Venturia inaequalis and Microsphaeropsis ochracea.

The objective of this study was to examine the role of melanin in the interaction between the mycoparasite Microsphaeropsis ochracea and the apple scab pathogen Venturia inaequalis. Melanin was extracted from the cell wall of the pathogen and its chemical and physical properties determined on the basis of biochemical tests and visible and infrared spectra. The physical and chemical characteristics of V inaequalis melanin were similar to the those of synthetic dihydroxyphenylalanine (DOPA) melanin. Precursors of the four known melanin biosynthetic pathways were tested for their ability to restore the pigmentation of an albino strain of V inaequalis. Scytalone, an intermediate of the 1,8-dihydroxynaphthalene (DHN) pathway, was the only precursor to restore the dark-brown pigmentation. Tricyclazole and pyroquilon, two antipenetrant fungicides, specific inhibitors of DHN melanin synthesis in Pyricularia oryzae, were used to confirm the melanin pathway in V. inaequalis wild type. A reddish-brown pigment was obtained due to the accumulation of shunt products of the DHN melanin pathway instead of a dark-brown pigment, suggesting that the melanin extracted from V inaequalis was a DHN melanin. Furthermore, growth of an albino mutant of V. inaequalis on scytalone-amended medium resulted in the formation of dark granules similar to those seen in wild-type isolates. Transmission electron microscopic observations of M. ochracea grown in the presence of melanin showed that the granules accumulated gradually along fungal cell walls to form a uniform dark coating.