Chilli (Capsicum annuum L.) is an economically important spice and vegetable crop grown widely across tropical and subtropical regions. It is valued not only for its pungency and flavour but also for its nutritional and therapeutic properties. For enhancing its productivity and quality it requires systematic breeding strategies, however, breeding efforts are often challenged by the complex inheritance of yield and quality traits, genotype x environment interactions, and the limited availability of elite parental lines. The present investigation entitled “Combining Ability Analysis for Morphological, Biochemical and Yield Traits in Chilli (Capsicum annuum L.) Using Line × Tester Design” was conducted at the Vegetable Research Farm, SKUAST-Jammu using 8 lines and 3 testers in a Line × Tester mating design, generating 24 hybrids. The experiment was laid out in a Randomized Complete Block Design with three replications during Kharif 2023–24. Observations were recorded for 15 morphological, yield and quality traits to assess the general combining ability (gca) of 8 lines and 3 testers in chilli (Capsicum annuum L.). Combining ability plays a crucial role in identifying promising parents for hybrid breeding. Results revealed significant gca effects, indicating the importance of additive gene action for several traits. Parents L4(DKC-8), L3(Pusa Sadabahar), L5(SJC-1) and T2(Surai mukhi) emerged as superior general combiners for yield and yield attributing traits, while L2(TNAU-9), L4(DKC-8), L6(Hot Queen) and T3(TNAU-6) were identified as promising general combiners for the majority of quality and growth-related traits. These findings offer valuable insights for selecting parents in future chilli improvement programs.