Abstract Currently, the main control strategy for arthropod-pests in tomato cropping is the use of synthetic chemical insecticides and miticides. The study aimed to select tomato genotypes of Santa Cruz type, with determinate growth habit, which were rich in acylsugar and presented resistance to spider mite. The evaluated segregating populations were derived from interspecific crossing and were in the advanced generation F2BC2. Genotypes were assessed for acylsugars content, present in leaflets, trichomes at abaxial and adaxial parts and a repellence bioassay to the mite was performed. It was found that the genotype UFU-102-F2BC2#13 stood out in comparison to the others, showing determinate growth habit, high acylsugar content and resistance to spider mite. It was also possible to obtain genotypes that were similar to the wild specie S. pennellii, regarding the acylsugar content.