Sorghum genes responsive to purple witchweed parasitism were isolated, and their expression was analyzed. Using the suppression-subtractive hybridization strategy, 30 genes that were up-regulated in response to purple witchweed parasitism were isolated from the roots of a susceptible sorghum cultivar ‘Abu 70’. The changes in the expression of each gene were investigated in the roots and leaves of the sorghum cultivars ‘Wad Ahmed’ and ‘Tabat’ were parasitized by purple witchweed. Tabat is more susceptible and Wad Ahmed is less susceptible to purple witchweed than Abu 70. Further, the changes in the gene expression and host susceptibility to purple witchweed were studied in the roots of the three sorghum cultivars after treatment with salicylic acid (SA) or methyl jasmonate (MeJA). Purple witchweed parasitism induced jasmonic acid (JA)-responsive genes and suppressed SA-responsive genes in the roots of the cultivars Abu 70 and Tabat. In contrast, purple witchweed parasitism in the less-susceptible cultivar Wad Ahmed induced SA-responsive genes and induced JA-responsive genes to a small extent. SA, but not MeJA, decreased the susceptibility of all the sorghum cultivars to purple witchweed. Systemic expression of a few genes was observed in the leaves of the purple witchweed–parasitized sorghum cultivars.