Trichomes contribute to plant resistance against herbivory by physical and chemical deterrents. To better understand their role in plant defense, we systemically studied trichome morphology, chemical composition and the response of the insect herbivores Helicoverpa zea and Leptinotarsa decemlineata (Colorado potato beetle = CPB) on the tomato hairless (hl), hairy (woolly) mutants and wild-type Rutgers (RU) and Alisa Craig (AC) plants. Hairless mutants showed reduced number of twisted glandular trichomes (types I, IV, VI and VII) on leaf and stem compared to wild-type Rutgers (RU), while woolly mutants showed high density of non-glandular trichomes (types II, III and V) but only on the leaf. In both mutants, trichome numbers were increased by methyl jasmonate (MeJA), but the types of trichomes present were not affected by MeJA treatment. Glandular trichomes contained high levels of monoterpenes and sesquiterpenes. A similar pattern of transcript accumulation was observed for monoterpene MTS1 (=TPS5) and sesquiterpene synthase SST1 (=TPS9) genes in trichomes. While high density of non-glandular trichome on leaves negatively influenced CPB feeding behavior and growth, it stimulated H. zea growth. High glandular trichome density impaired H. zea growth, but had no effect on CPB. Quantitative real-time polymerase chain reaction (qRT-PCR) showed that glandular trichomes highly express protein inhibitors (PIN2), polyphenol oxidase (PPOF) and hydroperoxide lyase (HPL) when compared to non-glandular trichomes. The SlCycB2 gene, which participates in woolly trichome formation, was highly expressed in the woolly mutant trichomes. PIN2 in trichomes was highly induced by insect feeding in both mutant and wild-type plants. Thus, both the densities of trichomes and the chemical defenses residing in the trichomes are inducible.