A field experiment was conducted in Al-Mu'tasim subdistrict, Samarra district, Salah al-Din Governorate, during the 2024-2025 winter growing season using six barley varieties (Arevat, Normar, Jazira-120, Cleaber, Local Black Barley, and Baghdad-1) and three levels of agricultural sulfur (0, 500, and 1000 kg S ha⁻¹). The experiment was designed according to a randomized complete block design (RCBD) with three replications. The results showed that increasing the sulfur level led to a significant improvement in most growth and yield characteristics. The high application level (1000 kg ha⁻¹) resulted in the highest average plant height (93.2 cm), shoot dry weight (5.12 g plant⁻¹), number of spikes (4.5 spikes plant⁻¹), number of grains spike (46.70 grains), 1000-grain weight (50.32 g), total grain yield (4189 kg ha⁻¹), and biological yield (7642 kg ha⁻¹), in addition to the highest grain protein concentration (12.16%), compared to the no-application treatment, which recorded the lowest values for all studied traits. The varieties also showed significant variation in their response. The Jazira-120 variety had the highest total grain yield (4271 kg ha⁻¹), while the Baghdad-1 variety recorded the highest biological yield (7783 kg ha⁻¹), and the Local Black Barley variety recorded the lowest values for most traits. Interaction analysis confirmed a significant effect between sulfur levels and varieties, with some varieties, particularly Jazira-120 and Baghdad-1, achieving the best performance at the highest application level.