The present investigation was undertaken at the Hybrid Rice plot of TCA, Dholi, and Muzaffarpur, and molecular analysis was conducted in the Molecular Laboratory of Postgraduate Dept. of Genetics & Plant Breeding, RPCAU, and Bihar to generate heterosis studies for 18 traits and molecular characterization using SSR markers. The experimental material comprised 31 three-line rice hybrids and 3 commercial checks evaluated in the RBD design. Among the tested varieties, Rajendra Sweta performed best in terms of grain yield per plant. Two rice hybrid genotypes, namely, IR68897A × KMR-3R and Rajendra-3A × RRR–4, exhibited superior standard heterosis over all three tests for trait grain yield per plant. By utilizing 12 primer pairs, a total of 33 shared alleles and 13 unique alleles were produced as amplified products. Among the 12 primers, seven primers were found to be comparatively informative for all nineteen hybrids and eleven parents. Only five primers, namely, MRG2894, RM515, RM520, RM538, and RM555, were able to confirm the hybridity (F1) with the respective parental lines.