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 CaCl2 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.
|Number of pages||3|
|Journal||Acta Botanica Sinica|
|State||Published - 1998|
- Egg cell
- Oryza sativa