[Isolation of zygotes and apical and basal cells of bicelluare proembryo from Torenia fournieri].

Torenia fournieri is a special plant with embryo sac partly protruding through the micropyle of ovule, and the cells of egg apparatus can be clearly observed using light microscope. Zygotes and the cells of bicellular proembryo could be isolated using enzymatic digestion. In the solution containing 0.05% cellulase and 0.05% pectinase, 14-15 zygotes were isolated from 50 ovules in 1h. In the solution containing 0.2% cellulose, 0.4% hemicellulase and 0.2% pectinase, 19 pairs of apical and basal cells of bicellular proembryo could be isolated from 50 ovules in 1 h. The isolated zygotes and epical and basal cells were collected using a micromanipulator up to a number which will prepare for suing molecular methods to probe the mechanism of early embryogenesis of higher plants.

[Anther development and sperm isolation of Pseudostellaria heterophylla (Miq.)].

Anther wall is general and tapetum is glandular. The process of meiosis of microspore mother cells is simultaneous and the tetrads are tetrahedral. The mature pollen of Pseudostellaria heterophylla (Miq.) is tree-celled. There are 22-30 germ pores on the pollen wall. Many pollen grains could burst in 10% mannitol or 15% sucrose solution and release a pair of sperm cells which could keep alive for 25-50 min by FDA fluorescence. Using micromanipulator the released sperm cells could be collected. When pollen grains were put into a solution containing 0.03% CaCl2, 0.01% H3BO3, 0.01% KH2PO4 and 20% PEG for 2-5 min, they would germinate and the pollen tubes would reach 815 microm at 2h after cultured. A pair of sperms would enter into pollen tube when it grew to 500-600 microm. The fluorescence of both sperms would be observed clearly in pollen tube after DAPI staining. When the pollen tubes were burst in a bursting solution, a pair of sperms would be released from pollen tube.