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Plant and Cell Physiology Advance Access originally published online on May 17, 2008
Plant and Cell Physiology 2008 49(6):981-993; doi:10.1093/pcp/pcn076
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© The Author 2008. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Characterization of novel genes induced by sexual adhesion and gamete fusion and of their transcriptional regulation in Chlamydomonas reinhardtii

Takeaki Kubo1,5, Jun Abe2,6, Takeshi Oyamada1, Makoto Ohnishi1, Hideya Fukuzawa3, Yoshihiro Matsuda4 and Tatsuaki Saito1,*

1Department of Applied Science, Faculty of Science, Okayama University of Science, Okayama, 700-0005 Japan
2Department of Molecular Science, Graduate School of Science and Technology, Kobe University, Nada-ku, Kobe, 657-8501 Japan
3Division of Integral Life Science, Graduate School of Biostudies, Kyoto University, Kyoto, 606-8502 Japan
4Department of Biology, Faculty of Science, Kobe University, Kobe, 657-8501 Japan

*Corresponding author: E-mail, saito{at}das.ous.ac.jp; Fax, +81-86-256-9408.


  Abstract

When mating type plus and minus gametes of Chlamydomonas are mixed, they agglutinate with each other via their flagella, fuse, then initiate the zygote formation program which includes synthesis of the zygote cell wall, fusion of nuclei and chloroplasts, and the digestion of chloroplast DNA from the minus parent. The mRNAs from gamete and zygote cells was isolated and hybridized to cDNA-macroarray filters both to identify new genes expressed during the mating reaction and the early zygote formation process and to analyze the gene expression programs that underlie these sexual processes. Twenty-one novel genes were identified in this screen, designated as EZY (early zygote expressed) genes. The EZY genes included genes encoding proteins whose function is unknown, and genes encoding proteins that appear to be involved in processes such as cell wall synthesis, gene expression, intracellular trafficking or secretion, and vesicular transport in zygotic cells. All of the EZY genes were strongly induced within 1 h during the mating process, including early zygote formation. The transcriptional characteristics of EZY genes were analyzed by using the fusion-defective mutant fus mt+. Among the EZY genes, 12 genes were not activated in fusion-defective conditions, suggesting that cell fusion is required for their expression. The remaining nine that were transcribed in fusion-defective fus matings were also inducible by cell wall removal in either vegetative or gametic cells, indicating that these genes were induced only indirectly by the cAMP signaling pathway initiated by flagellar agglutination as a result of mating-induced cell wall loss.

Keywords: Cell–cell recognition - Chlamydomonas reinhardtii - Transcriptional regulation - Zygote formation

Abbreviations: CHX, cycloheximide; cpDNA, chloroplast DNA; DRP, dynamin-related protein; ECM, extracellular matrix; ER, endoplasmic reticulum; EST, expressed sequence tag; EZY, early zygotic expressed; miRNA, microRNA; ORF, open reading frame; RACE, rapid amplification of cDNA ends; UTR, untranslated region.

5Present address: Division of Integral Life Science, Graduate School of Biostudies, Kyoto University, Kyoto, 606-8502 Japan.

6Present address: Department of Chemical and Biological Sciences, Faculty of Science, Japan Women's University, Tokyo, 112-8681 Japan.

(Received March 14, 2008; Accepted May 12, 2008)
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