Botryosphaeria dothidea is one of the most important fungal pathogens associated with trunk diseases of grape that easily infects wounds and usually occurs after pruning wounds. Chitinase III (CHIIII) and β-1,3-glucanase (GLU) are two important defense genes in response to various stresses such as the attack of pathogens. In the present work relative expression of chitinase III (CHIIII) and β-1,3-glucanase (GLU) genes of grape were examined. The experiment was carried out using factorial experiment based on a completely randomized design and LSD test, in three replications and six time periods after inoculation with B. dothidea and three control samples. The obtained results indicates that the defense genes, particularly (CHIIII) and (GLU) were significantly induced at 48, 72 and 48 hours after infection, respectively.