Ahmad, S., Singh, M., Lamb-Palmer, N. D., Lefsrud, M. and Singh, J. 2012. Assessment of genetic diversity in 35Pisum sativumaccessions using microsatellite markers. Can. J. Plant Sci. 92: 1075-1081. Field pea is an important Canadian pulse crop and therefore developing high-performing cultivars is critical for Canadian pea growers. Information about genetic diversity is a key component for the creation of novel and desirable germplasm to develop elite pea breeding lines. The objective of the present study is to assess genetic diversity in 35 diverse Pisum accessions using 15 polymorphic microsatellites located on different pea chromosomes. Microsatellites were found to be polymorphic, amplifying a total of 41 alleles and were able to differentiate all 35 Pisum genotypes. These markers were scored by their polymorphic information content (PIC), ranging from 0.055 (AA206) to 0.660 (AB72) with an average of 0.460, and by their discriminating power (D), which varied from 0.057 (AA206) to 0.679 (AB 72) with an average of 0.475. Genetic similarity values ranged from 0.074 (between Maple pea NZ and Line 45760) to 0.875 (between Galena and Dakota) with an average of 0.336. Unweighted pair group method with arithmetic averages (UPGMA) cluster analysis grouped the 35 pea accessions into two major clusters and eight sub-clusters. The majority of Canadian and European genotypes were grouped separately, suggesting both these groups are from genetically distinct gene pools. The genetically diverse groups identified in this study can be used to derive parental lines for pea breeding.