Journal of Plant Research

, Volume 107, Issue 1, pp 17–21 | Cite as

Isolation and preservation of the living embryo sac ofCrinum asiaticum L. var.japonicum baker

  • Kiyoyuki Ohshika
  • Hideo Ikeda
Original Articles


The enzymatic maceration method was used to isolate an intact embryo sac ofCrinum asiaticum and its component cells. Best results were obtained when using enzyme solutions that contained pectinase hemicellulase, cellulase and pectolyase. Aseptic ovules were incubated in the enzyme solution for 1.5 hr at 25 C. This allowed the isolation of embryo sacs to yield up to 20% of the amount present. An isolated embryo sac usually consists of an egg cell, synergids, antipodals and a central cell. Some embryo sacs can be digested as gametophytic protoplast. The size, shape and position of the isolated embryo sac seemingly possessed similarities with those of the fixed embryo sac in the ovary. An isolated embryo sac can be in a living state when the result of the fluorochromatic reaction (FCR) and protoplasmic streaming is positive. When cultured in proper media, 68% of the isolated gametophytic protoplasts were observed to have sustained their positive FCR for more than 1 month.

Key words

Crinum asiaticum Embryo sac Enzymatic maceration Gametophytic protoplast 


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Copyright information

© The Botanical Society of Japan 1994

Authors and Affiliations

  • Kiyoyuki Ohshika
    • 1
  • Hideo Ikeda
    • 1
  1. 1.Laboratory of Science Education, Faculty of EducationHiroshima UniversityHigashihiroshimaJapan

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