Abstract  [Objectives]  Herbivore induced plant defenses play a key role in protecting plants against herbivore damage. Plants suffer herbivore damage frequently in nature, however, it is unclear if damaged plant have primed defensive mechanisms that allow them to mount a stronger defense against repeated herbivore attacks. [Methods]  Experiments were conducted on tobacco plants Nicotiana tabacum to see if there was a difference in the plant defenses induced by single, or repeated, episodes of attack by Spodoptera litura larvae. Concentrations of jasmonic acid (JA), salicylic acid (SA) and abscisic acid (ABA) were measured to monitor hormone levels. Secondary metabolites were also measured to reveal the key metabolites involved in defense against herbivores. [Results]  Caterpillar damage induced a defensive response from tobacco plants and repeated damage induced a stronger defensive response than a single episode of damage. Compared to healthy plants, plants subject to repeated herbivore damage had significantly higher concentrations of JA (78.9%), but similar levels of SA and ABA. Herbivore damage significantly increased the concentration of nicotine, chlorogenic acid and trypsin inhibitors; plants subject to repeated herbivore attacks had higher concentrations of all three metabolites than those subject to an isolated attack. [Conclusion]  Tobacco plants have evolved primed defense against herbivores which allows them to mount a stronger defensive response to repeated herbivore attacks.