Abstract: The ‘Luo Yang Hong’ peony cultured in vitro was studied for effect of basic media, combination of plant growth regulators, incubation condition, and anti-browning material on subculture status, rooting ability, and browning. The results showed that DKW medium was a suitable basic medium for ‘Luo Yang Hong’ Peony’s subculture. Supplying of 2.0～3.0mg/L BA and 0.5mg/L IAA or 0.05mg/L NAA could benefit the subculture, but supplying of ZT did not have positive result. The plantlets incubated at 20~25℃ had no significant difference in terms of the number of propagating buds, but the plantlets were aging and dead faster at 30℃. The 1/2MS basic medium supplied with 2.0mg/L NAA or 0.2mg/L IAA and 2.0mg/L IBA could help the inducing of adventitious roots. The supplying of 0.5g/L PVP in media of subculture and rooting could mitigate browning and promote differentiation of subculture plantlets, but had no significant effect on rooting.