A field experiment was conducted in an experimental field affiliated with the College of Agricultural Engineering Sciences, University of Baghdad, Al-Jadriya, during the fall season of 2022 to investigate the effect of bio-fertilizer and agricultural sulfur levels on the availability of nutrients and broccoli’s productivity. Split plot order in the randomized complete block design (RCBD) of three replicates was used comprising three levels of sulfur (0, 500, and 1000kg.ha-1) symbolized by S0, S1, and S2 forming the main plots, added to the soil before planting by two months, while the subplots were the bio-fertilizers represented by B0 and B1. Results showed the superiority of the third level of agricultural sulfur, giving the highest traits involving the sulfur concentration in the soil after 30, 60, and 90 days from planting, reaching 45.4, 50.9, and 58.0 mg S.kg soil-1, nitrogen concentration after 45 and 90 day from planting that was 41.50 and 26.52 mg N. kg soil-1, phosphorous concentration after 45 days from planting that was 29.21 mg P. kg soil-1, and potassium concentration 45 and 90 days after planting that was 255.2 and 235.3 mg K. kg soil-1. The bio-fertilization was also superior in increasing elements in the soil, including sulfur concentration after 30, 60, and 90 days after planting was 44.7, 44.9, and 51.2 mg S. kg soil-1, nitrogen concentration 45 and 90 days after planting was 35.3 and 22.53 mg N. kg soil-1, phosphorous concentration 45 and 90 days after planting was 24.15 and 15.55 mg P. Kg soil-1, and potassium concentration 45 and 90 days after planting was 227.6 and 207.3 mg K. kg soil-1 as well as in the total yield in a value of 43.6 t.ha-1. For the binary interaction, the treatment of bio-fertilization with the third level of agriculture sulfur was superior, giving the highest values of element concentrations in the soil, including sulfur 30, 60, and 90 days after planting was 47.4, 55.5, and 66.1 mg S. kg soil-1, nitrogen concentration 45 and 90 days after planting was 43.6 and 27.9mg N.kg soil-1, phosphorous concentration after 45 days from planting that was 30.12 mg P.kg soil-1, and potassium concentration 45 and 90 days after planting was 271.9 and 246.4 mg K. kg soil-1 in addition to the total yield that was 45.7t.ha-1.