AbstractDisassortative mating is a rare form of mate preference that promotes the persistence of polymorphism. While the evolution of assortative mating and its consequences for trait variation and speciation have been extensively studied, the conditions enabling the evolution of disassortative mating are still poorly understood. Mate preferences increase the risk of missing mating opportunities, a cost that can be compensated by a greater fitness of offspring. Heterozygote advantage should therefore promote the evolution of disassortative mating, which maximizes the number of heterozygous offspring. From the analysis of a two-locus diploid model with one locus controlling the mating cue under viability selection and the other locus coding for the level of disassortative preference, we show that heterozygote advantage and negative frequency-dependent viability selection acting at the cue locus promote the evolution of disassortative preferences. We predict conditions of evolution of disassortative mating coherent with selection regimes acting on traits observed in the wild. We also show that disassortative mating generates sexual selection, which disadvantages heterozygotes at the cue locus, limiting the evolution of disassortative preferences. Altogether, our results partially explain why this behavior is rare in natural populations.