Around the world, chilli is the most important crop as a vegetable and spice. An essential step in plant breeding procedures to develop and enhance new varieties is the genetic characterization of resources and evaluation of genetic diversity. In the current study, 72 elite chilli genotypes were assessed along with four checks for fruit quality attributes in Augmented Design-II between 2021 and 2022. Ascorbic acid, phenols, oleoresin, dry matter content, peroxidase and capsaicin content were among the fruit quality criteria that were examined. The accessions' average capsaicin content was 0.42 %, with a range of 0.13 to 1.20 %. BCA-26 has maximum amount of capsaicin (1.20 %). For capsaicin concentration, higher estimations of the genotypic and phenotypic coefficients of variation (62.24 % and 67.20 %) were found, along with high heritability (85.79 %) and high GAM (118.93 %). Based on fruit quality and yield attributes, genetic diversity investigation depicted that clusters II and III had the greatest inter-cluster distance, followed by clusters II and V and II and IV, indicating greater genetic divergence. Moreover, 72 genotypes were characterized by molecularly using nine pungency-specific polymorphism SSR markers. The mean allele number was 2.77, with a range of 2 to 4. With an average value of 0.415, the heterozygosity was observed to be lowest for the markers HpmsE005 and HpmsE063 (0) and highest for the primer HpmsE031 (0.444). The PIC value averaged 0.415 and ranged from 0.298 (HpmsE005) to 0.538 (HpmsE022). High and low pungent chilli cultivars might be distinguished with the help of the HpmsE022 marker.