In order to investigate the effect of trehalose on the retrogradation properties of rice starch, the molecular structure, water distribution, rheological and thermodynamic properties were studied by means of atomic force microscope, low-field nuclear magnetic resonance, Fourier transform infrared spectroscopy and differential scanning calorimetry analysis. The results showed that the leached straight chain amylose contents of rice starch with the addition of 1.6% trehalose (T-starch), which were stored at 4℃ for 3h and 24h were decreased by 8.6% and 11.5% compared with rice starch, respectively. The arrangement of the starch molecular chains trended to aggregate to an ordered structure with the retrogradation. However, the addition of trehalose could inhibit the aggregation. The order degree of rice starch increased by 19.0%, but the order degree of T-starch only increased by 3.0% after retrogradation for 24h, indicating that the trehalose could significantly decrease the proportion of short-term order structure of rice starch. The binding ability of T-starch to water molecules (strongly and weakly bound water) was significantly higher than that of rice starch (P<0.05). The free water precipitation and recrystallization in T-starch system were blocked, which was one of the reasons for inhibiting the short-term retrogradation of starch. After the addition of trehalose, both regenerative enthalpy and retrogradation degree of the long-term retrogradation of T-starch were significantly reduced. And this advantage brought by trehalose became more and more prominent with the extension of aging time, and the long-term retrogradation enthalpy of T-starch system was significantly reduced. Therefore, trehalose could not only effectively inhibit the short-term retrogradation of starch, but also had a good inhibitory effect on long-term retrogradation.