In the present study, molecular (DAMD and ISSR) and chemical (α and β-asarone contents) markers were used to characterize the A. calamus genotypes procured from different parts of India. The cumulative analysis carried out for both DAMD and ISSR markers revealed 24.71 % polymorphism across all genotypes of A. calamus. The clustering patterns of the genotypes in the UPGMA tree showed that the genotypes are diverse, and did not show any specific correlation with their geographical provenances, reflecting the low level of genetic diversity and a high genetic differentiation among the genotypes from the same localities. All the 27 genotypes of A. calamus were also analyzed for α and β-asarone contents, and percentage of essential oil. The genotype (Ac13) from Kullu (Himachal Pradesh) showed maximum (9.5 %) percentage of oil, whereas corresponding minimum (2.8 %) was obtained from the genotypes from Pangthang (Sikkim). Similarly, the highest α and β-asarone contents (16.82 % and 92.12 %) were obtained from genotypes from Renuka (Himachal Pradesh) and Udhampur (Jammu & Kashmir), while lowest α and β-asarone contents (0.83 % and 65.96 %) resulted from Auranwa (Uttar Pradesh) and Pangthang (Sikkim) genotypes, respectively. A. calamus harbours tremendous economic value, and it is therefore, important to identify the genotypes with low α and β-asarone contents for its commercial utilization. Further, this study will help in evaluation and documentation of a large number of diverse genotypes for their value traits.