cDNAs encoding prothoracicotropic hormone (PTTH) and diapause hormone (DH) were isolated from the tobacco budworm, Heliothis virescens (Hvi). Hvi-PTTH cDNA reveals key structural features known from other PTTHs, yet there is a > 30% amino acid difference between the PTTH sequences found in this noctuid when compared with the PTTHs known from Bombycoidea. Hvi-DH-PBAN cDNA encodes a precursor protein including a DH-like peptide, pheromone biosynthesis-activating neuropeptide (PBAN) and three additional neuropeptides with an FXPRL sequence at the C terminus. PTTH and DH-PBAN transcripts are most highly expressed in the brain and subesophageal ganglion (SG), respectively. Expression of both the PTTH and the DH-PBAN transcripts is high in larvae and remains high in nondiapausing pupae, but in larvae programmed for pupal diapause, expression declines sharply at the onset of larval wandering behaviour and remains low during pupal diapause. This pattern implies that both of these genes are shut down during diapause. These results are not consistent with a role for DH in promoting the entry into diapause as noted in Bombyx mori. Instead, the higher expression of DH in nondiapausing pupae suggests a possible role for DH in promoting continuous development. The injection of DH into diapausing pupae did indeed successfully terminate diapause, thus suggesting a possible new role for this neuropeptide.