[Isolation of egg cells from Brugmansia aurea Lagerh "Goildens Kornett"].
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.
Liu
Huan Huan
HH
School of Life Sciences, Xiamen University, Xiamen, China.
Jian
Ming Xia
MX
Wang
Ya Ying
YY
Tian
Hui Qiao
HQ
D035264:Flowers / Q000166:cytology*; D019657:Solanaceae / Q000166:cytology*