Analysis of intercellular washing fluids of potato tubers and detection of increased proteolytic activity upon fungal infection

Abstract

We present the first procedure for extracting intercellular fluids of potato (Solanum tuberosum L. cv. Spunta) tubers. Intercellular washing fluids were isolated from healthy and Fusarium‐infected potato tissue. The electrophoretic pattern using SDS‐PAGE indicated differences between the fluids from the two tissues. A significant extracellular proteolytic activity was accumulated during the infection with Fusarium solani f. sp. eumartii. A major proteolytic band with an apparent molecular mass of 70 kDa and another of approximately 30 kDa were detected after separation of intercellular fluids by casein gel electrophoresis. Proteolytic activity was principally inhibited by diisopropylfluorophosphate, which is indicative of the involvement of serine protease(s). In vitro degradation assay indicated that specific potato proteins from healthy tubers were hydrolyzed by fluid proteases from infected tubers. The biological role of such activity in potato‐Fusarium interaction is still unknown. Our results suggest that the intercellular serine protease has a fungal origin.