Skip Navigation



AOBPreview published online on April 18, 2006
Annals of Botany, doi:10.1093/aob/mcl072
This Article
Right arrow FREE Full Text (PDF) Freely available
Right arrow Content Select
Right arrow All Versions of this Article:
97/6/1139    most recent
mcl072v1
Right arrow E-letters: Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when E-letters are posted
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Google Scholar
Right arrow Articles by HOSHINO, Y.
Right arrow Articles by SHINODA, K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by HOSHINO, Y.
Right arrow Articles by SHINODA, K.
Agricola
Right arrow Articles by HOSHINO, Y.
Right arrow Articles by SHINODA, K.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2006. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received November 4, 2005
Revised December 8, 2005
Accepted February 11, 2006

Technical Article

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

YOICHIRO HOSHINO 1 *, NAHO MURATA 2, and KOICHI SHINODA 2

1 Field Science Center for Northern Biosphere, Hokkaido University, Kita 11, Nishi 10, Kita-Ku, Sapporo 060-0811, Japan; Division of Innovative Research, Creative Research Initiative ‘Sousei’ (CRIS), Hokkaido University, Kita 21, Nishi 10, Kita-Ku, Sapporo 001-0021, Japan
2 National Agricultural Research Center for Hokkaido Region, Hitsujigaoka 1, Toyohira-Ku, Sapporo 062-8555, Japan

* To whom correspondence should be addressed.
YOICHIRO HOSHINO, E-mail: hoshino{at}exfarm.agr.hokudai.ac.jp


  Abstract

• Aims To develop a procedure for isolating living egg cells and zygotes from Alstroemeria ovules.

• Scope An attempt was made 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 h 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.

Keywords: Alstroemeria aurea, egg cell, enzyme treatment, microdissection, zygote.
Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.