太子参花药发育及精细胞分离

太子参花药壁发育为基本型，腺质绒毡层。小孢子母细胞减数分裂为同时型，小孢子四分体为四面体型，成熟花粉具两个精细胞，为3胞花粉。在花粉表面具散孔，孔数22—30个，均匀分布于花粉粒表面上。花粉在10％甘露醇或15％蔗糖溶液中可直接爆破，精细胞易被释放并散开，通过显微操作仪可收集到一定数目的精细胞。FDA染色荧光显示释放出来的精细胞活力可维持25—50min。花粉在舍O．03％CaCl2、0．01％H3803、0．01％KH2P04和20％PEG、pH5．8的培养液中2—5min即萌发花粉管．花粉管生长2h可达815μm。一般花粉管伸长500—600μm时，一对精细胞才进入花粉管。DAPI染色后荧光观察．可观察到精细胞和营养细胞核在花粉管中的移动状况。爆破花粉管后可释放出一对精细胞。

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.