The purpose of this study was to investigate and compare root morpho-anatomical traits and physiological responses of susceptible (SP81–3250) and resistant (H. Kawandang) sugarcane genotypes exposed to the attack by nymphs of spittlebug Mahanarva fimbriolata (Stål) (Hemiptera: Cercopidae). Two experiments were conducted to compare the damage caused by spittlebug nymphs on fresh and dry biomass weight; lignin content in stalks; root anatomy; chlorophyll content; photosynthetic rate (A); carboxylation efficiency (A/Ci); stomatal conductance (gS) and transpiration rate (E) of these genotypes. SP81–3250 consistently obtained significantly higher damage scores than H. Kawandang in both experiments, confirming the previously observed level of resistance in each genotype. Attack by spittlebug nymphs had a much higher effect on both fresh and dry biomass weight, chlorophyll content, A, A/Ci, gs and E of SP81–3250, than that on H. Kawandang. Anatomical studies indicated the presence of aerenchyma tissue in the root cortex of SP81–3250, a feature which may facilitate penetration of the nymph's stylet into the vascular cylinder. In contrast, roots of H. Kawandang are characterized by having more dense and compact parenchyma cells. In addition, infested plants of this genotype contained an unidentified mucilaginous compound in the vascular cylinder of the roots. We conclude that resistance of H. Kawandang to spittlebug is related to the ability of this genotype to maintain normal chlorophyll content, as well as stomatal conductance and photosynthesis, thus, allowing for biomass accumulation under spittlebug attack, in contrast to SP81–3250. In addition, the presence of more compact and denser parenchymal cells, as well as that of an induced mucilaginous compound in the root's vascular cylinder, are likely to hinder host-feeding activity in nymphs, causing higher nymph mortality and therefore, reduced damage in plants of this genotype.