Jain, S., Porter, L. D., Kumar, A., Mir, R. R., Eigenbrode, S. D. and McPhee, K. E. 2014. Molecular and phenotypic characterization of variation related to pea enation mosaic virus resistance in lentil (Lens culinaris Medik.). Can. J. Plant Sci. 94: 1333-1344. Identification of genetically diverse lentil germplasm with resistance to pea enation mosaic virus (PEMV) through the combined approach of molecular marker analysis and phenotyping could prove useful in breeding programs. A total of 44 lentil (Lens culinaris Medik.) accessions, were screened for resistance to PEMV. Two accessions (PI 431663 and PI 432028) were identified with resistance to PEMV in field tests while several accessions were found resistant in greenhouse screenings. Thirty-six polymorphic simple sequence repeat (SSR) markers which produced 43 loci with 2 to 12 alleles per locus were used for genetic diversity analysis. The polymorphic information content (PIC) values for these markers ranged from 0.22-0.85 with a mean of 0.55 per marker. Using allelic data of 36 SSR primer pairs, dissimilarity ranging from 0.12 to 0.74 was calculated. Cluster analysis performed using the unweighted pair group method with arithmetic mean (UPGMA) determined that most of PEMV-resistant accessions were grouped in one cluster along with other accessions from Iran, Chile, Ethiopia, India, Pakistan, Turkey, Afghanistan and Lebanon. All the adapted cultivars originating from North and South America were grouped in another cluster along with some European accessions. The 44 accessions were classified into 4 subpopulations using Structure 2.2 software complimenting the results of UPGMA analysis and indicated the effect of geographical origin on the grouping of accessions. The results of this study can be used to select genetically diverse PEMV-resistant accessions for lentil improvement programs.