Soybean rust (SBR) caused byPhakopsora pachyrhiziSyd. and P. Syd. is one of the most important foliar diseases of soybean. SBR has the potential to cause major economic damage to global and U.S. soybean production. Analysis of reactions of soybean genotypes toP. pachyrhiziis an important step towards breeding for resistance to SBR. Fifty-four diverse soybean genotypes with both known and unknownRppresistance genes were tested for their reactions to a MississippiP. pachyrhiziisolate. PI 567102B (Rpp6) had a near-immune reaction with the lowest disease severity score and no sporulation. Among seventeen genotypes with resistant or incomplete resistant reddish-brown (RB) reactions, eight are improved breeding lines that are available to researchers through material transfer agreements (MTAs). Thirty-six genotypes had the susceptible TAN reaction. Four soybean lines (RN06-32-1(7-b, GC 00138-29, G01-PR16, and GC 84051-9-1) had RB reactions and significantly lower SBR severity and sporulation than three of the six resistant checks, PI 230970 (Rpp2), PI 462312 (Rpp3), and PI 459025B (Rpp4). G01-PR16 is a publicly released germplasm. This research provides new information about reactions of different soybean genotypes to a midsouthern USA isolate ofP. pachyrhiziand thereby aids in breeding for resistance to SBR.