Nucleic acids were extracted from in vitro shoot tip cultures of 31 grapevine cultivars affected with rupestris stem pitting (RSP) disease and from cultures of 11 RSPfree cultivars, 4 of which were disease-free, 2 of which were fleck-affected and 5 of which were grapevine leafroll disease-affected. Analysis of the extracts by polyacrylamide slab gel electrophoresis showed that 21 of the 31 RSP-affected cultivars contained a previously unreported nucleic acid which was absent from the RSP-free controls. Nuclease digestions showed that the nucleic acid was double-stranded RNA (dsRNA). The apparent size of the dsRNA was inconsistent with that expected for either a viroid or a closterovirus. The observation that 10 of the 31 RSP-affected cultivars lacked this dsRNA is consistent with the view that there may be more than one 'RSP-like' disease. The dsRNA detected in this investigation may be associated with one of these diseases