Journal Article

Identification of Genes that are Up- or Down-regulated in the Apical or Basal Cell of Maize Two-celled Embryos and Monitoring their Expression During Zygote Development by a Cell Manipulation- and PCR-based Approach

Takashi Okamoto, Stefan Scholten, Horst Lörz and Erhard Kranz

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 46, issue 2, pages 332-338
Published in print February 2005 | ISSN: 0032-0781
Published online February 2005 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pci032
Identification of Genes that are Up- or Down-regulated in the Apical or Basal Cell of Maize Two-celled Embryos and Monitoring their Expression During Zygote Development by a Cell Manipulation- and PCR-based Approach

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In higher plants, a zygote generally divides asymmetrically into a two-celled embryo. As in 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 polymerase chain reaction (PCR) with randomly amplified polymorphic DNA (RAPD) primers. 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; and (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; EST, expressed sequence tag; FTase, farnesyltransferase; LCM, laser capture microdissection; RAPD, randomly amplified polymorphic DNA

Journal Article.  5328 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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