This study was conducted during the summer seasons of 2019 and 2020 at the greenhouse of National Research Centre; Giza, Egypt, to study the effect of foliar spraying with zinc in improvement negative effects of irrigation water regime in Sorghum. Three levels of water regime (1, 1.5 and 2 Liter) were used interacting with three concentrations of zinc as foliar spraying (00, 50 and 100 ppm). Generally, obtained results showed significant increment in all vegetative growth criteria, yield and chemical constituents by increasing both quantity of irrigation and zinc spraying. Moreover, using high concentrations of zinc leaded to reduce and alleviate the harmful effect of irrigation water deficiency. Inter simple sequence repeats (ISSR) molecular marker was applied using three different ISSR primers, and results recorded 61 amplified bands with different molecular weights ranged from 183 – 1278 bp detected by these primers. Moreover, these amplified fragments were distributed between polymorphic bands (37 bands) with an average 12.33, unique bands (9 bands) with an average 3 and monomorphic bands (15 bands) with an average 5, so, polymorphism percentage was 60.82%.