Abstract: The enzyme shaking technique was used to isolate the viable embryo sac of Paulownia fortunei . Fresh ovules were enzymatically macerated in mixture solution including different content of pictinase(Y 23),cellulase(R 10) and macrozyme(R 10) etc. for 2.5～3?h, at 28～30?℃, under the shaking condition, to isolate the embryo sac. Cytological structure of the whole viable sac was observed under Olympus AHB 2 microscope after aceto carmine staining or DAPI staining.