卵细胞的分离是进行植物离体受精和研究卵细胞发育的基础。将韭菜的胚珠酶解30MIn后吸打可将其外珠被去掉。将胚珠转移到不含酶的相同溶液中,用解剖针将胚珠从中部切割,然后挤压胚珠的珠孔部位,卵器细胞从胚珠的切口处逸出。用显微操作仪将卵细胞和两个助细胞分开,达到分离韭菜卵细胞的目的。用同样的方法也成功地分离了韭菜的合子和早期原胚。韭菜分离的卵细胞可用于建立韭菜离体受精体系,而分离的合子及早期原胚则可用于探索其受精机制和研究其胚胎发育的过程。Egg cells, zygotes and proembryos of Allium tuberosum Roxb were isolated using an enzymatic digestion.The ovules were incubated in an enzymatic solution for 30 min to remove outer integument.Then they were transferred into an isolated solution without enzymes for mechanical dissection.The ovules were cut from their middle part, and squeezed gently on the micropyle, resulting in the liberation of the egg, zygote and proembryo from the ovules of different stage.About 5-10 egg apparatus could be released from 30 ovules within 1 h.Viable egg cell could be separated with two synergids and collected into a pure population using a micromanipulator.Viable zygotes and proembryos were isolated using the same method.Coupled with our successful isolation of mature sperm cells, the isolation of egg cells of Allium tuberosum Roxb will make in vitro fertilization possible in this plant.The isolation of viable zygotes and proembryos can be used to explore its embryonic development.国家自然科学基金资助项目(No.30670126)---
