Predator-prey interactions are crucial components of populations and communities. Their dynamics depend on the covariation of traits of the interacting organisms, and there is increasing evidence that intraspecific trade-off relationships between defense and competitive traits are important drivers of trophic interactions. However, quantifying the relevant traits forming defense-competitiveness trade-offs and how these traits determine prey and predator fitness remains a major challenge. Here, we conducted feeding and growth experiments to assess multiple traits related to defense and competitiveness in six different strains of the green alga Chlamydomonas reinhardtii exposed to predation by the rotifer Brachionus calyciflorus. We found large differences in defense and competitive traits among prey strains and negative relationships between these traits for multiple trait combinations. Because we compared trait differences among strains whose ancestors evolved previously in controlled environments where selection favored either defense or competitiveness, these negative correlations suggest the presence of a trade-off between defense and competitiveness. These differences in traits and trade-offs translated into differences in prey and predator fitness, demonstrating the importance of intraspecific trade-offs in predicting the outcome of predator-prey interactions.