Phomopsis longicolla is a major seed pathogen of soybean (Glycine max) in hot, humid environments. The objective of this study was to monitor the infection and development of P. longicolla on vegetative and reproductive tissues of six cultivars and to determine the relationship between this infection and subsequent seed infection and seed germination. Cultivars were grown for 3 years (2002 to 2004) without irrigation or with irrigation applied at pre- plus postflowering or at postflowering growth stages. P. longicolla was isolated most frequently from leaf, stem, pod, root, and seed. Diaporthe phaseolorum and three unidentified Phomopsis sp. were also isolated. Diaporthe aspalathi, which previously has not been reported on soybean, was also recovered from leaf samples. These isolates, however, were recovered very infrequently. Recovery of P. longicolla from roots was much lower than from leaves, stems, and pods in all years and irrigation environments. The recovery of P. longicolla from seed was affected by irrigation environments. Seed from irrigated plots had more P. longicolla than that from nonirrigated plots. Isolation of P. longicolla from seed was negatively correlated with percentage of seed germination in irrigated environments but not in the nonirrigated environment. Pod infection was correlated with seed infection in all three irrigation environments. Even though infection of leaves and stems increased with increasing moisture availability, such infection did not consistently correlate with seed infection. Seed germination and seed infection were negatively correlated with percent hard seed. This study provided the first demonstration of the seasonal progression of P. longicolla on soybean cultivars grown under three irrigation environments.