Mitogen-activated protein kinase (MAPK) cascade-mediated signalling is essential in the establishment of resistance towards pathogens. The present study compared MAPK activities in a compatible and incompatible interaction between pearl millet [Pennisetum glaucum (L.) R. Br.] and downy mildew pathogen Sclerospora graminicola. Differential expression was observed with rapid and increased activation of MAPKs, PgMPK1 (48kDa) and PgMPK2 (44kDa), in the incompatible interaction; with a weak activity of only PgMPK1 in the compatible interaction. Immunoblot analysis showed PgMPK1 and PgMPK2 to be orthologs of salicylic acid-induced protein kinase and wound-induced protein kinase, respectively. Immunocytochemical analysis revealed pathogen-induced accumulation and nuclear localisation of PgMPKs only in the incompatible interaction with highest signals in the vascular tissues. Maximum PgMPKs activation correlated with the activation of several defence-related enzymes. In addition, inhibition of MAPK-activation by kinase cascade inhibitors correlated with the suppression of defence-related enzyme activities and pathogen-induced H 2 O 2 accumulation. Treatment of pearl millet seedlings with abiotic and biotic elicitors led to a strong early induction of only PgMPK1. β-Amino butyric acid and H 2 O 2 were found to be best activators of PgMPK1. These results suggest that in pearl millet MAPK signalling is involved in mediating several defence mechanisms in response to pathogen infection.