Due to its fungistatic or fungicidal potential, chitosan, a high molecular-weight polymer that is non-toxic and biodegradable, has become an alternative to conventional fungicides. In addition, chitosan is reported to elicit defense mechanisms in plant tissues. In this study, we explored the in vitro fungicidal effect of chitosan on some of the most important grapevine wood fungi, such as Botryosphaeria sp. (dieback and cane blight), Phomopsis sp. (Phomopsis cane and leaf spot), Eutypa lata (eutypa dieback), Neonectria liriodendri (black foot disease), Phaeomoniella chlamydospora (Petri disease and esca) and Fomitiporia sp. (esca). Inhibition of mycelial growth was evaluated at five concentrations 50, 25, 5, 2.5 and 0.5 mg a.i. l-1 of chitosan. Chitosan was effective in reducing mycelial growth of all the fungi. The lowest EC50 values were obtained with Pa. chlamydospora, Fomitiporia sp. and Botryosphaeria sp., and the highest with Neon. liriodendri. All these were inferior to the maximum recommended field rate (8.33 mg a.i. l-1) with exception of the value obtained with Neon. liriodendri. Greenhouse experiments were carried out to evaluate the efficacy of foliar sprays of chitosan on potted grapevine plants (cultivar Castelão) growing in a substrate artificially infested with Pa. chlamydospora or Neon. liriodendri. The effect of chitosan against Neon. liriodendri was similar to that achieved with some selected fungicides (carbendazim+flusilazole, cyprodinil+fludioxonil and tebuconazole). Chitosan significantly improved plant growth (plant height and number of roots) and decreased disease incidence compared with untreated plants. As regards Pa. chlamydospora, chitosan only reduced the disease incidence caused by this fungus.