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Mucuna pruriens, a widely available but underutilized legume in India, stands out for its nutritional and medicinal potential. Understanding the genetic diversity of M. pruriensis vital for its conservation and utilization in breeding programs aimed at enhancing its nutritional and agronomic traits. In this regard, molecular markers like ISSRs offer a reliable and informative tool for assessing genetic diversity. In the present study, 23 M. pruriensaccessions from northeastern India were evaluated using ISSR markers, revealing high levels of polymorphism and significant genetic diversity. A total of 512 amplicons were generated across 33 ISSR primers, with an average polymorphic percentage of 98.05 %. PIC values ranged from 0.22 to 0.50, indicating substantial genetic variation within the accessions. Supplementary indicators such as EMR and MI were employed to evaluate the marker system's efficacy. EMR ranged from 6 to 22, averaging 14.95, while MI ranged from 2.64 to 9.02, averaging 6.04, with a positive correlation observed between EMR and MI. Cluster analysis based on Jaccard's similarity coefficient classified the accessions into two major groups, with the northeastern Indian accessions displaying higher diversity compared to those from other regions of India. Interestingly, geographical proximity did not consistently correlate with genetic similarity, suggesting complex patterns of gene flow and historical germplasm exchange. Overall, this study provides valuable insights into the genetic diversity of M. pruriensand highlights the importance of conserving and utilizing this underutilized legume for addressing global food security challenges.