Drought stress is one of the most important environmental stresses that has the greatest impact on wheat fields. Therefore, in order to investigate the drought stress on the defense system of several wheat genotypes, firstly, the top varieties and lines of the Dryland Agricultural Research Institute (DARI) were investigated during three crop years (2017-2020), in the form of a simple lattice design. Finally, genotypes No. 4 and 40 were selected as superior cultivars under stress conditions and genotypes No. 30 and 33 under supplementary irrigation conditions. According to the results, the highest activity of guaiacol peroxidase and proline was recorded in genotype number 4 under rain conditions and the lowest activity of these enzymes was recorded in genotype number 40 under supplementary irrigation conditions. The highest and lowest chlorophyll content was obtained from line number 4 under supplementary irrigation conditions and genotype number 40 under stress conditions, respectively.