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Trehalose is an α-α linked glucose disaccharide. Trehalose supplied to the growth medium of Arabidopsis seedlings inhibits root growth, primary leaf development and carbon allocation from shoot to root. In the current study, interaction between trehalose and ascorbic acid on some growth parameters and photosynthetic gene expression was investigated in Arabidopsis seedlings. For this purpose, Arabidopsis seeds were grown on MS medium, MS medium supplemented with 0.1 mM ascorbic acid, MS medium supplemented with 100 mM trehalose and MS medium supplemented with 0.1 mM ascorbic acid combined with 100 mM trehalose for 15 days. The results showed that trehalose decreased root length, and dry and fresh weight as well. Meanwhile, the amount of soluble sugar, starch, ascorbic acid, dehydroascorbate, total ascorbic acid and trehalase activity were increased by trehalose treatment. On the other hand, the combination of trehalose and ascorbic acid treatment increased root length, dry and fresh weight and decreased soluble sugar, starch, ascorbic acid, dehydroascorbate, total ascorbic acid and trehalase activity. Gene expression profiling showed that trehalose suppressed SUS, INV and SUC expression. But ascorbic acid partially lowered gene expression suppression mediated by trehalose. In summary, the current results showed that the inhibitory effect of 100 mM trehalose on Arabidopsis seedlings growth can be because of its inhibitory effect on sucrose transport from source tissue (leaf) to sink tissue (root).