The effect of glyphosate on sudden death syndrome (SDS) of soybean is not clearly understood. In 2011–2013, 14 field experiments were conducted to investigate the impact of glyphosate on SDS, yield, and plant nutrition. Field experiments were conducted in Iowa, Illinois, Indiana, Michigan, Wisconsin, and Ontario, Canada. Six combinations of herbicides with and without glyphosate were compared. Experiments were conducted in fields with histories of SDS, and some locations were also inoculated with local isolates of Fusarium virguliforme and irrigated to create a conducive environment for the disease. Disease, yield, and nutrient content in plant tissue data were collected. In sum, this study demonstrated that glyphosate application did not increase SDS severity or adversely affect soybean yield under field conditions.