Isolation of egg cells and zygotes in Oryza sativa

Hong Mei Han, Jie Zhao, Hua Zhong Shi, Hong Yuan Yang, Chang Zhou

Research output: Contribution to journalArticlepeer-review

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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 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.

Original languageEnglish
Pages (from-to)186-188
Number of pages3
JournalActa Botanica Sinica
Volume40
Issue number2
StatePublished - 1998

Keywords

  • Egg cell
  • Isolation
  • Oryza sativa
  • Zygote

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