Excess salt induced ionic and osmotic stresses that disturbed metabolism and led to reduction of plant development. Previous studies reported that sugars in stressed plants were involved in stress tolerance. However, the role of sugars in salt-stressed plants against only ionic effects is still unclear. The objective of this research was to investigate accumulation and homeostasis of ions, membrane injury, water content, growth characters and sugar contents in roots, in-response to salt stress under iso-osmotic conditions. Salt-sensitive rice, Pathumthani1 (PT1) was grown on MS culture medium for 7 days and was adjusted to salt stress under iso-osmotic conditions (-1.75 ± 0.20 MPa) by mannitol for 4 days. An increase in NaCl increased Na+ and Na+:K+ in PT1 roots leading to increased membrane injury, while the water content was decreased. Additionally, growth characters, including number, length, fresh weight and dry weight of roots, were inhibited. Sugar accumulations in PT1 roots were enhanced by increases in NaCl. The increase in Na+ was positively related to total soluble sugars, resulting in an osmotic adjustment of the membrane that maintained water availability. The accumulation of sugars in PT1 roots may be a primary salt-defense mechanism and may function as an osmotic control.