This experiment was conducted to investigate the growth inhibition effects caused by continuous cropping soil in peony(Paeonia lactiflora Pallas). The effect of extracts from continuous cropping soil of peony was tested with bio-assay method using callus cells induced from peony filament tissues and seedlings derived from peony zygotic embryos. The cell viability and seedling growth were significantly inhibited by methanol extract in continuous cropping soil. Methanol extract from continuous cropping soil was successively fractionated with solvents such as n-hexane, ethyl acetate, n-butanol and water. The seedling growth was inhibited by ethyl acetate fraction obtained in methanol extract.