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Plant and Cell Physiology Advance Access originally published online on November 28, 2007
Plant and Cell Physiology 2008 49(1):58-67; doi:10.1093/pcp/pcm167
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© The Author 2007. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Ultrastructural Characterization of Exine Development of the transient defective exine 1 Mutant Suggests the Existence of a Factor Involved in Constructing Reticulate Exine Architecture from Sporopollenin Aggregates

Tohru Ariizumi1,4,5, Takahiro Kawanabe1,5, Katsunori Hatakeyama2, Shusei Sato3, Tomohiko Kato3,6, Satoshi Tabata3 and Kinya Toriyama1,*

1Laboratory of Environmental Biotechnology, Graduate School of Agricultural Science, Tohoku University, Sendai, 981-8555 Japan
2Vegetable Breeding Research Team, National Institute of Vegetable and Tea Science, Ano, 514-2392 Japan
3Department of Plant Genome Research, Kazusa DNA Research Institute, Kisarazu, 292-0818 Japan

*Corresponding author: E-mail, torikin{at}bios.tohoku.ac.jp; Fax, +81-22-717-8834.


  Abstract

A male-sterile mutant of Arabidopsis thaliana, in which filament elongation was defective although pollen fertility was normal, was isolated by means of T-DNA tagging. Transmission electron microscopy (TEM) analysis revealed that primexine synthesis and probacula formation, which are thought to be the initial steps of exine formation, were defective, and that globular sporopollenin aggregation was randomly deposited onto the microspore at the early uninucleate microspore stage. Sporopollenin aggregation, which failed to anchor to the microspore plasma membrane, was deposited on the locule wall and in the locule at the uninucleate microspore stage. However, visually normal exine with a basic reticulate structure was observed at the middle uninucleate microspore stage, indicating that the exine formation was restored in the mutant. Thus, the mutant was designated transient defective exine 1 (tde1). These results indicated that tde1 mutation affects the initial process of the exine formation, but does not impair any critical processes. Our results also suggest the existence of a certain factor responsible for exine patterning in A. thaliana. The TDE1 gene was found to be identical to the DE-ETIOLATED 2 gene known to be involved in brassinosteroid (BR) biosynthesis, and the tde1 probacula-defective phenotypes were recovered in the presence of BR application. These results suggest that BRs control the rate or efficiency of initial process of exine pattern formation.

Keywords: Arabidopsis thaliana - Exine formation - Male sterility - Probacula

Abbreviations: BAC, bacterial artificial chromosome; BR, brassinosteroid; CV, coated vesicle; ER, endoplasmic reticulum; FAA, formalin/alcohol/acetic acid; qRT–PCR, quantitative RT–PCR; SEM, scanning electron microscopy; TEM, transmission electron microscopy.

4Present address: Department of Crop and Soil Science, Washington State University, Pullman, WA 99164-6420, USA.

5These authors contributed equally to this work.

6Present address: Forestry Research Institute, Oji Paper Company Co. Ltd, Kameyama, 519-0212 Japan.

(Received September 23, 2007; Accepted November 25, 2007)
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