Biological stresses such as soilborne pathogens reduce root growth, health, and thus, the yield potential. We hypothesize that under very low soilborne-pathogen pressure (using soil fumigation), root characteristics will be similar among a wide range of soybean genotypes. The objective is to determine differences in root characteristics of three soybean varieties in Iowa, under a fumigated and a non-fumigated environment. Experiments were conducted in 2004 and 2005 at three locations in Iowa. Field was fumigated with a mixture of 1-3-dichloropropene and chloropicrin (Telone C-35 ™). Experiment was laid out as a randomized complete block design with 4 replications. The varieties used were IA2068, S25J5, and SOI2642. Plants were sampled 21, 42 and 63 days after emergence (DAE) using a golf hole cutter. Roots were extracted from the soil cores with and automatic root washer and roots were characterized using WinRhizo Software and dry weight. A location by variety interaction was observed at 21 DAE. SOI2642 and S25J5 had more root length per volume, and finer roots in DeWitt than in the other two locations. Varieties that responded positively to fumigation were S25J5 in DeWitt and Nevada, and SOI2642 in DeWitt. At 42 DAE, significant main effects of fumigation, variety, and location dominated. Differences between fumigated and non-fumigated sites were more evident at this sampling date, driven by the positive response of some of the varieties at specific locations. No differences were found at 63 DAE. Overall the root growth parameters that were assessed in this study were sensitive enough to reflected differences among varieties and fumigations systems, not obtained with more traditional methods to measure plant growth. Research will continue and help us quantify those differences.