Gall-inducing insects produce various types of galls on plants, but little is known about the gall-induction mechanism of these galling insects. The gall wasp Leptocybe invasa Fisher & LaSalle (Hymenoptera: Eulophidae) forms galls of different sizes on several Eucalyptus species. To clarify the physiological responses of Eucalyptus to L. invasa infestation, we measured the dynamics of nitrogen (N), carbon (C), total phenolics, total tannins and four types of phytohormones (zeatin [Z] + zeatin riboside [ZR], gibberellins [GA], indole-3-acetic acid [IAA] and abscisic acid [ABA]) in galled and ungalled leaf tissues of two Eucalyptus horticultural varieties (DH201-2 [Eucalyptus grandis × Eucalyptus camaldulensis] and EA [Eucalyptus exserta]) with different susceptibility to galling throughout the larval developmental stages. Nitrogen, total phenolics, tannins and four kinds of phytohormones strongly accumulated in tissues galled by L. invasa (especially during early larval feeding stages). While N, Z + ZR and GA levels were higher, tannins and ABA levels were lower in the galled tissues on the highly susceptible variety. Nitrogen, total phenolics, GA, Z + ZR and IAA levels in the galled tissues gradually decreased during gall development, but ABA and tannins conversely increased in the galled tissues of the less susceptible variety. Our results suggest that the effects of gall-inducing insects on plants depend not only on the susceptibility of the plant infested but also on the developmental stage of galled tissues. Gall formation process is thus synergistically influenced by both gall-inducing insect and plant genotypes.