Using map-based cloning, we delimited the dialytic gene to an approximately 109-kb fragment, which controls multicellular trichome formation and stamen development in tomato. Trichomes exist in the epidermis of nearly all terrestrial plants, including unicellular and multicellular types. The molecular mechanism of unicellular trichomes in Arabidopsis is well characterized. However, knowledge about the regulatory pathway of multicellular trichomes in tomato (Solanum lycopersicum) is limited. Phenotypic analysis of the dialytic (dl) mutant LA3724 demonstrated that the trichomes are forked and the stamens are unclosed. To clone and characterize dl, we mapped this gene to an approximately 109-kb fragment using two F2 populations derived from the two crosses of dl mutant: LA3724 × IL8-1 and LA3724 × LA1589 (Solanum pimpinellifolium). Two types of molecular markers were utilized in this study, including cleaved amplified polymorphic sequences and insertion-deletion events. Sequence analysis predicted the presence of seven putative open reading frames, including two unknown proteins, two phospholipase Ds, glycosyl hydrolase family 5 protein/cellulose, choline/ethanolamine kinase, and aquaporin-like protein. The aquaporin-like protein gene was evidently upregulated in dl mutant. Thus, we inferred that this gene is a potential candidate for the phenotypes. The results provide a basis to elucidate the regulatory pathway responsible for trichome formation and stamen development in tomato.