Phoma sclerotioides, causal agent of brown root rot of alfalfa, causes severe root and crown lesions on alfalfa and other perennial forage legumes in regions with harsh winters. Isolates of P. sclerotioides exhibit diverse cultural morphologies on potato dextrose agar (PDA), suggesting that they may exhibit a high degree of genetic diversity. To investigate the genetic relatedness of P. sclerotioides isolates, 154 isolates from North America were sequenced at 10 loci. Maximum parsimony and maximum likelihood analyses of the complete 10-locus data set placed isolates into multiple strongly supported clades, and analyses of gene-jackknife and single-gene partitions of the data set indicated robust support for six major clades and three subclades. Genetic differences corresponded closely to differences in conidial size and septation, pycnidial neck length, mycelial pigmentation, and growth rate in axenic culture at 18 and 25°C. Isolates exhibited morphologies broadly consistent with the species description of P. sclerotioides, and new species were not designated. On the basis of genetic and morphological differences, we propose establishing seven infraspecific varieties within P. sclerotioides: P. sclerotioides var. sclerotioides, champlainii, viridis, obscurus, steubenii, macrospora, and saskatchewanii. All varieties of P. sclerotioides caused brown root rot of alfalfa and grew well at low temperatures.