Performing the genetic mechanism research of internode length in bitter gourd has important guiding significance for making breeding strategy. A cross was made between CN19-1 and Thai4-6, and an F2 segregation population was constructed. The genetic characteristic of the internode length was analyzed through employing the major gene plus polygene mixed genetic model. The results showed that the internode length was continuous distribution in the F2 population. The optimum model for internode length genetic analysis was B-1. The additive effect values of two pairs of major genes controlling the internode length were 3.206 3 and 2.638 3, respectively, which showed a positive effect. The dominant effect values were-3.434 6 and-1.459 6, respectively, which showed a negative effect. The dominant degree of major genes is-1.07 and-0.55, respectively, indicating that the first pair of major genes was over dominant and the second pair of major genes was incompletely dominant. The heritability of the major gene was 82.25%, indicating the genetic stability was high. This study could provide a theoretical reference for the breeding of bitter gourd.