Abstract: The enzymatic isolation of megagametophyte from fixed and fresh of Pinus yunnanensis Fr. were macerated in a mixture solution of 3% pectinase and 3% cellulase at the 30℃ for 10–13 hours, and the archegonium has been isolated. Materials were stained with A-S, Feulgen and PAS, and following procedure was cleared in the lactophenol. The ventral canal and egg cell which from division of central cell, the functional sperm nucleus becomes pressed against the egg nucleus, and the new cytoplasm round the zygote nucleus were observated under the microscope of usual light. The four proembryonal nuclei divide simultaneously, resulting in eight nuclei arranged in two tiers of four cells each, and following development were formed wall. In the end, the proembryo of six- teen cells were formed. Two methods on enzymatic and dimethyl sulfoxide with potassium hydroxide to isolate the female sexual cells of gymnosperms have been compared, and several problems are discussed.

Key words: Enzymatic isolation, Pinus yunnanensis, Archegonium, Proembryo Development