黄花木本曼陀罗卵细胞分离(简报)

ISOLATION OF EGG CELLS FROM Brugmansia aurea Lagerh"Goildens Kornett"

采用分离的精、卵细胞体外融合并诱导人工合子长成植株的离体受精方法可在没有其他组织影响的单细胞水平上探索受精事件的发生过程．为研究高等植物的受精机理、探索配子识别和合子激活等问题提供有效手段。分离的卵细胞不仅可以用来开展离体受精研究．也提供了用分子生物学方法研究被子植物卵细胞发育和合子发育机理的实验基础.

Viable egg cells of Brugmansia aurea Lagerh ＂Goildens Kornett＂ were isolated using mechanical dissection and enzymic digestion. The ovules were cut from middle part and pushed its micropylar position using a dissection needle. Generally the three cells of egg apparatus were released from cut end of ovule. When the ovules put an isolating solution only containing 0.04%CACl2, 1% BSA and 12% mannitol, 7 egg apparatus could be isolated from 40 ovules within 2 h. However, it is some difficult to separate egg cell from two synergids. When ovules were first incubated in an enzymic solution containing 1% Pectinase （Serva）, 1% Cellulase （Onozuka RS） and 12% mannitol for 30 min, and then transferred into the above-mentioned isolating solution to dissect, 8 egg apparatus could be isolated from 40 ovules within 2h and egg cell is easy to separate from two synergids. The isolated egg cells of Brugmansia aurea Lagerh ＂Goildens Kornett＂ could be used in vitro fertilization to explore fertilization mechanism and in egg development using molecular methods.