Bacterial wilt is a disease caused by Ralstonia solanacearum, which affects over 450 plant species and causes significant reduction in crop yields including of tomato (Solanum lycopersicum) worldwide. Developing and identifying tomato genotypes with Ralstonia solanacearum tolerance and high yield potential, presents an opportunity for improvement of crop productivity. The objective of this study was to explore the heritability of tomato resistance against bacterial wilt and the genetic variation within the population for breeding purposes. A breeding population was created by crossing two bacterial wilt-resistant (MT56 and BL333) and three commercially desirable susceptible (Assila, Rambo and Heinz) tomato varieties, using North Carolina Design II, in a screen house at the Makerere University Agricultural Research Institute, Kabanyolo (MUARIK). Results showed that the Area Under Disease Progress Curve was significant (P<0.001), indicating that cumulative disease progress was less in resistant genotypes of segregating generations. Disease severity increased with days after inoculation (DAI), with Heinz showing the highest level of susceptibility. The General Combining Ability for male parent (GCAm) was significant (P< 0.01), and Specific Combining Ability (SCAf×m) and GCAf were significant (P<0.01) for the F2 generation. Broad-sense heritability was higher than the narrow-sense heritability in both F1 and F2 generations, suggesting that non-additive gene action predominately controlled tomato resistance to bacterial wilt infestation. The genetic diversity ranged from 0.5 to 0.6759, with a mean value of 0.5787. Polymorphism Information Content (PIC) varied from 0.375 to 0.6357, with a mean of 0.4888, indicating a high degree of variation. SLM 12-2 was the most polymorphic marker, with a PIC of 0.6357. The Unweighted Pair-Group Method with Arithmetic Average (UPGMA) classified all tomato genotypes into six clusters, namely Clusters 1 and 2 (Susceptible parents), Cluster 3 (Resistant parents), Clusters 4 and 5 (New source of resistance), and Cluster 6 (F1P1×P5).