Phomopsis seed decay (PSD) of soybean ( Glycine max L. Merr) is primarily caused by Phomopsis longicolla. Currently, percent seed infection is used as a method for evaluating resistance to PSD. However, researchers need a common system for classifying soybean genotypes for their reaction to P. longicolla that is consistent across environments. The purpose of this research was: (i) to evaluate the reaction of a select set of soybean accessions to P. longicolla, and (ii) determine the effects of irrigation and two harvest regimes (normal and delayed) on seed infection. We propose a classification system for comparing reactions to P. longicolla among accessions based on a Phomopsis seed infection index (PSII), derived by dividing the percent seed infection of each accession by percent seed infection values of susceptible standards selected within early and late maturity groups. Percent seed infection by P. longicolla infection was greater when accessions were irrigated during the growing season and harvested at normal maturity. Some lines such as MO/PSD-0259 were rated as resistant to P. longicolla (7% seed infection) when not irrigated and harvested at maturity, but were rated as susceptible (39% seed infection) with irrigation and delayed harvest. The resistance ratings to P. longicolla using percent seed infection and PSII varied among accessions. SS93-6012, for example, had similar ratings for both (9% seed infection and 16% PSII). PI 416942 exhibited some resistance to P. longicolla based on percent seed infection (8%), but no resistance based on PSII (32%). Accessions PI 594478, Delmar, SS93-6012, SS93-6181, PI 594603A, and PI 594712 were classified by PSII as moderately resistant under irrigation and with delayed harvest. These results indicate that simple comparisons of percent seed infection among accessions without irrigation, without delayed harvest, and without a standard control of a similar maturity group might result in false conclusions about resistance to P. longicolla.