花粉生殖细胞的大量分离与纯化

MASS ISOLATION AND PURIFICATION OF GENERATIVE CELLS FROM POLLEN GRAINS

被子植物的花粉生殖细胞是精子的前体,在生殖过程中占有重要地位。由于它被营养细胞与花粉壁包围,一般只能以花粉粒为单位进行研究。1986年以来,我们用压片法分离出花粉生殖细胞,作了有关细胞生物学研究,同时指出分离的生殖细胞用于离体培养与遗传操作的意义。作者参考了 Russell 分离精子的一步“渗透压冲击法”,建立了适于大量分离与纯化生殖细胞的“二步渗透压冲击法”(以下简称“二步法”)。此外。

A technique named“two-step osmotic shock”was developed for the isolation of large number of generative cells from pollen grains in Vicia faba and Zephyranthes candida.The procedure included:1.Pollen hydration:mature pollen grains were incubated in adequate sucrose solution for 30—50 min.2.Osmotic shock:when the pollen grains swelled and show- ed the tendency of germination,a sudden osmotic shock by adding equal volume of water made them bursting.The contents carrying generative cells spurted from the germ pores.By su- bsequent filtration and centrifugation,purified generative cell population was obtained.FDA test proved that the isolated and purified generative cells were viable.