We examined the effect of conservation conditions on restricting the conservation of Acer rubrum L. cv. ‘Somerset’, including photoperiod, light intensity, temperature and diurnal temperature variation. In the 182-day conservation period, the in vitro materials were mainly for differentiation in the early period and vegetative growth in the later period, also gradually adapted to the low-temperature environment. The effect of temperature on the growth of in vitro materials reached extremely significant level (P<0.01); the effect of photoperiod and light intensity lasted for a long time, and their interaction reached significant level (P<0.05). Diurnal temperature variation had a significant effect on average number of shoots and rate of rooting. In all of the tests, 25°C, illumination 12 h, 62.50 μmol·m ^-2·s ^-1 and 25°C, illumination 12 h, 31.25 μmol·m ^-2·s ^-1 conditions were both able to restrict conservation. The selection criteria for the best conservation conditions was not unique, and the core was keeping the differentiation ability of the in vitro materials.