J Integr Plant Biol. ›› 1998, Vol. 40 ›› Issue (2): -.

• Research Articles •    

Isolation of Egg Cells and Zygotes in Oryza sativa

HAN Hong-Mei,ZHAO Jie, SHI Hua-Zhong, YANG Hong-Yuan and ZHOU Chang   

Abstract: A method of non-enzymatic, manual microdissection was established to isolate egg cells and zygotes in Oryza sativa L. Generally 5 to 8 protoplasts of egg cells or zygotes could be isolated from 20 ovaries in 2 h. Fluorochromatic reaction proved that the isolated cells were viable and could survive in the isolation medium consisting of 0.5 moL/L mannitol and 5 mmol/L CaC12 for at least 2 h. The egg cells and early zygotes usually turned to spherical shape during the course of isolation, whereas the late zygotes often maintained pear-shaped. The rounded zygotes were proved to be true protoplasts by Calcoflour White fluorescent staining. Chlorotetracycine and fluphenanize fluorescence treatment demonstrated that the isolated zygotes were rich in membrane-bound calcium as well as calmodulin.

Key words: Oryza sativa, Egg cell, Zygote, Isolation

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