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Plant and Cell Physiology Advance Access first published online on February 2, 2005
This version published online on February 3, 2005
Plant and Cell Physiology, doi:10.1093/pcp/pci032
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Plant and Cell Physiology 2005 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received October 4, 2004
Accepted December 9, 2004

Regular Paper

Identification of Genes That Are Up- or Down-Regulated in the Apical or Basal Cell of Maize Two-Celled Embryos and Monitoring Their Expressions During Zygote Development by a Cell Manipulation- and PCR-Based Approach

Takashi Okamoto 1*, Stefan Scholten 2, Horst Lörz 2, and Erhard Kranz 2

1 Biozentrum Klein Flottbek und Botanischer Garten, Entwicklungsbiologie und Biotechnologie, Universität Hamburg, Ohnhorststr 18 22609 Hamburg, Germany; Department of Biological Sciences, Tokyo Metropolitan University Minami-osawa 1-1, Hachioji, Tokyo, 192-0397 Japan
2 Biozentrum Klein Flottbek und Botanischer Garten, Entwicklungsbiologie und Biotechnologie, Universität Hamburg, Ohnhorststr 18 22609 Hamburg, Germany

* To whom correspondence should be addressed.
Takashi Okamoto, E-mail: okamoto-takashi{at}c.metro-u.ac.jp


  Abstract

In higher plants, a zygote generally divides asymmetrically into a two-celled embryo. Asin planta, maize zygotes produced by in vitro fertilization of an egg cell with a sperm cell also develop into an asymmetrical two-celled embryo that consists of a small plasma-rich apical cell and a large vacuolized basal cell. Subsequently, via zygotic embryogenesis, a proembryo and a transition-phase embryo are formed from the two-celled embryo. In the present study, we focused on identifying genes that were up-or down-regulated only in the apical or basal cell of two-celled maize embryos after fertilization. First, a procedure for isolating the apical and basal cells from two-celled embryos was established, and subsequently cDNAs were synthesized from apical cells, basal cells, egg cells, two-celled embryos and multicellular embryos. These cDNAs were used as templates for randomly amplified polymorphic DNA (RAPD) PCR. Genes with specific expression patterns were identified, and these expression patterns were categorized into six groups:

1. Up-regulated only in the apical cell after gamete fusion

2. Up-regulated only in the basal cell after gamete fusion

3. Up-regulated in both the apical and basal cells after gamete fusion

4. Down-regulated only in the apical cell after gamete fusion

5. Down-regulated only in the basal cell after gamete fusion

6. Constitutively expressed in the egg cell and embryos

In addition, it was revealed that the genes up-regulated in the apical or basal cell (genes in groups 1 and 2) were already expressed in the early zygote, providing the possibility that the transcripts from these genes are localized to the putative apical or basal region of the zygote, or that the transcripts are rapidly degraded in one of the daughter cells after zygotic cell division.

Keywords: apical cell; basal cell; embryogenesis; gene expression; in vitro fertilization; zygote.
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