Organosulfur compounds in onion extracts inhibit the aggregation of human blood platelets. Antiplatelet activity is important to human cardiovascular health. We hypothesized that modification of sulfur fertility may increase organosulfur compound concentration and thereby affect platelet inhibitory activity in onion. Four contrasting onion genotypes were grown at four sulfur levels in a hydroponic system in the greenhouse and in contrasting sulfur environments in seven field locations in Wisconsin, Oregon, and New York. The contrasting field sites were comprised of sandy soils with a mean sulfate level of 5.4 ppm and muck soils with a mean sulfate level of 20.3 ppm. Onions grown in field environments with increased soil sulfur concentrations had significantly higher antiplatelet activity (33% higher than sand-grown onions; P < 0.001). The greenhouse experiment was conducted in hydroponics with nutrient solutions containing four sulfur levels ranging from 0.8 mM to 15 mM sulfate. The 10-mM sulfur treatment resulted in onion bulbs with 10% higher antiplatelet activity over those grown in the 0.8-mM sulfur treatment (P < 0.06). These data suggest that sulfur concentration in nutrient solution and in soil may be directly responsible for the increased antiplatelet activity in onion extracts observed in this study.