The aim of this study was to investigate the biochemical, physiological and morphological responses of eucalypts to iso-osmotic salt and water deficit stress. Chlorophyll a, chlorophyll b, total chlorophyll (TC), total carotenoids, maximum quantum yield of PSII (F v /F m ), photon yield of PSII and non-photochemical quenching, in severely osmotic-stressed plantlets (−1.20 MPa) decreased significantly when compared to control plantlets, leading to low net photosynthetic rate (P n ) and growth reduction. In addition, the photosynthesis and growth parameters of salt stressed plantlets declined to a greater degree than those cultivated in water-deficit conditions. On the other hand, the proline content of the osmotic-stressed leaves increased significantly, especially under iso-osmotic salt stress conditions. The TC content of iso-osmotic stressed leaves was significantly degraded, with low F v /F m , leading to P n reduction and growth inhibition. Those parameters in salt stressed plantlets were significantly lower and to a greater degree than in plantlets subjected to water-deficit. The basic knowledge of biochemical, physiological and morphological changes in responses to iso-osmotic salt and water-deficit stresses will be further applied as effective indices for salt and water-deficit tolerant screening in large population of eucalyptus breeding program.