Isolation of Individual Egg Cells and Zygotes in Alstroemeria Followed by Manual Selection with a Microcapillary-connected Micropump

• HOSHINO, Yoichiro

• MURATA, Naho

• SHINODA, Koichi

Abstract

Aims: To develop a procedure for isolating living egg cells and zygotes from Alstroemeria ovules. Scope: We attempted to isolate egg cells and zygotes from the ovules of Alstroemeria aurea. The ovules were histologically observed using a clearing procedure which revealed the localization and sizes of the embryo sacs and egg apparatus within the ovules. For the isolation of egg cells, ovules were cut into sections with a surgical blade and treated with an enzyme solution. Subsequently, these ovule sections were dissected using a glass needle under an inverted microscope. Egg cells successfully isolated by this procedure were collected using microcapillaries connected to a micropump. For zygote isolation, ovules were excised from ovaries 24 hours after self-pollination. By treating excised ovules with an enzyme solution and subsequently dissecting them using a glass needle, zygotes were successfully isolated from the ovules and collected with a microcapillary. The isolated zygotes were associated with pollen tubes and one of the synergids. Egg cells and zygotes were viable for up to 2 h following isolation, as determined by fluorescein diacetate staining. Conclusions: The procedures for isolating egg cells and zygotes in Alstroemeria were established, and each egg cell and zygote was captured with a microcapillary.

Journal

• Annals of Botany

  Annals of Botany 97 (6), 1139-1144, 2006-06

  Oxford University Press

Keywords

• Alstroemeria aurea
• egg cell
• enzyme treatment
• microdissection
• zygote

Details

• CRID: 1050001338992937600
• NII Article ID: 120000955849
• ISSN: 10958290
• ISSN: 03057364
• HANDLE: 2115/30230
• Text Lang: en
• Article Type: journal article
• Data Source:
  • IRDB
  • CiNii Articles