Abstract The North East Hill Region (NEHR) of India is home to diverse maize landraces including pigmented accessions rich in antioxidants and nutritional properties. The present study attempted to characterize a representative collection of this mostly unexplored diversity. Altogether eighty-three local maize landraces from the seven hill states of the NEHR were studied with special emphasis on pigmentation diversity. For the morphological traits, a significant ANOVA indicated the presence of substantial genetic variability for which selection would be fruitful. A number of these accessions were found to have traits that help cope with moisture stress, improve stalk strength and optimize photosynthesis. Principal component analysis studies for the yield attributing traits indicated that ear weight was most variable. The bleaching/histological studies confirmed that anthocyanin pigment when present was always restricted to the aleurone layer of the kernels, typical of blue maize. Quantitative analysis for kernel anthocyanin/phlobaphene content also revealed genetic differences among the accessions. Genetic analysis using the model-based STRUCTURE indicated significant population structuring among the accessions. Specifically, for the pigmentation diversity studies both principal coordinate analysis and neighbour joining methods revealed near concurrent population structuring due in part to the high differentiation of seven of the twenty-one pigmentation specific loci studied. The results obtained provide comprehensive evidence of a significant amount of genetic differentiation among the landraces under study. Landraces are valuable reservoirs of favourable alleles for which selection can be made and, as in this study, identify accessions for breeding maize with enhanced levels of beneficial secondary metabolites.