Downy mildew, caused by Peronosclerospora sorghi is one of the important diseases affecting maize (Zea mays L.) production worldwide. Several downy mildew resistant maize lines have been identified. However, variability in the degree of resistance among maize genotypes to P. sorghi has been reported. In the present study the molecular basis of resistance of maize to P. sorghi was studied by using differential-display reverse transcription PCR (DDRT-PCR) technique. Maize seedlings of downy mildew resistant (MAI 756) and susceptible (CM 500) cultivars at two-leaf stage were inoculated with P. sorghi and leaf samples were collected at 0, 3 and 5 days after inoculation and analyzed for differentially expressed cDNAs using cDNA-RAPD approach. A total of 17 cDNA fragments corresponding to transcripts that showed alterations during the defence response of maize to P. sorghi were identified. Genes involved in signal transduction and several genes with unknown functions were found to be upregulated in maize after infection by P. sorghi. Among 35 random primers tested, OPD-05 has identified a differentially expressed cDNA coding for serine/threonine kinase protein in resistant maize genotype. Constitutive and high level expression of serine/threonine kinase gene was observed in the uninoculated plants of resistant genotype, whereas no expression of this gene was observed in uninoculated plants of susceptible genotype. However, the transcript level was induced 3 days after inoculation in the susceptible genotype and slightly reduced 5 days after inoculation. This study represents the first identification of maize serine/threonine kinase gene that is upregulated following infection by P. sorghi.