Phomopsis seed decay (PSD), caused primarily by the fungal pathogen Phomopsis longicolla, is one of the most important diseases reducing seed quality and yield of soybean. Few cultivars have been identified as resistant. To identify new sources of resistance to PSD, 135 soybean germplasm accessions, originating from 28 countries, were field screened in Arkansas, Mississippi, and Missouri in 2009. Based on seed assays of natural field infection by P. longicolla in 2009, 42 lines, including the most resistant and susceptible lines, were reevaluated in the field in 2010, 2011, and 2012 with P. longicolla-inoculated and noninoculated treatments. Six maturity group (MG) III (PI 189891, PI 398697, PI 417361, PI 504481, PI 504488, and PI 88490), four MG IV (PI 158765, PI 235335, PI 346308, and PI 416779), and five MG V (PI 381659, PI 381668, PI 407749, PI 417567, and PI 476920) lines had significantly lower percent seed infection by P. longicolla than the susceptible checks and other lines in the same test (P ≤ 0.05). They appeared to have some levels of resistance to PSD. These new sources of PSD resistance can be used in developing soybean breeding lines or cultivars with resistance to PSD, and for genetic mapping of PSD resistance genes.