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

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Title: Isolation of Individual Egg Cells and Zygotes in Alstroemeria Followed by Manual Selection with a Microcapillary-connected Micropump
Authors: HOSHINO, Yoichiro Browse this author →KAKEN DB
MURATA, Naho Browse this author
SHINODA, Koichi Browse this author
Keywords: Alstroemeria aurea
egg cell
enzyme treatment
microdissection
zygote
Issue Date: Jun-2006
Publisher: Oxford University Press
Journal Title: Annals of Botany
Volume: 97
Issue: 6
Start Page: 1139
End Page: 1144
Publisher DOI: 10.1093/aob/mcl072
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.
Rights: This is a pre-copy-editing, author-produced PDF of an article accepted for publication in Annals of Botany following peer review. The definitive publisher-authenticated version, YOICHIRO HOSHINO, NAHO MURATA and KOICHI SHINODA, Isolation of Individual Egg Cells and Zygotes in Alstroemeria Followed by Manual Selection with a Microcapillary-connected Micropump, Annals of Botany, 97(6), Jun 2006, pp.1139-1144 is available online at: http://aob.oxfordjournals.org/cgi/content/abstract/97/6/1139
Type: article (author version)
URI: http://hdl.handle.net/2115/30230
Appears in Collections:北方生物圏フィールド科学センター (Field Science Center for Northern Biosphere) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 星野 洋一郎

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