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Plant and Cell Physiology Advance Access originally published online on October 6, 2009
Plant and Cell Physiology 2009 50(11):1911-1922; doi:10.1093/pcp/pcp135
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© The Author 2009. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

High Temperatures Cause Male Sterility in Rice Plants with Transcriptional Alterations During Pollen Development

Makoto Endo1,6, Tohru Tsuchiya2, Kazuki Hamada3, Shingo Kawamura3, Kentaro Yano3, Masahiro Ohshima1, Atsushi Higashitani4,5, Masao Watanabe4,5 and Makiko Kawagishi-Kobayashi1,*

1Rice Biotechnology Research Team, National Institute of Crop Science, NARO, Tsukuba, Ibaraki, 305-8518 Japan
2Life Science Research Center, Mie University, Tsu, 514-8507 Japan
3Faculty of Agriculture, Meiji University, Kawasaki, 214-8571 Japan
4Graduate School of Life Sciences, Tohoku University, Katahira, Sendai, 980-8577 Japan
5Faculty of Science, Tohoku University, Aramaki-Aoba, Sendai, 980-8578 Japan

*Corresponding author: E-mail, makikokk{at}affrc.go.jp; Fax, +81-29-838-8484.


  Abstract

Plant male reproductive development is highly organized and sensitive to various environmental stressors, including high temperature. We have established an experimental procedure to evaluate high temperature injury in japonica rice plants. High temperature treatment (39°C/30°C) starting at the microspore stage repeatedly reduced spikelet fertility in our system. Morphological observations revealed that pollen viability in plants exposed to high temperatures was lower than that in control plants. Most pollen grains in high temperature-treated plants displayed a normal round shape and stained reddish purple with Alexander’s reagent; however, the pollen grains were very poorly attached and displayed limited germination on the stigma. To investigate gene regulatory mechanisms in the anther in high temperature environments, DNA microarray analysis was performed by comparing non-treated samples with samples treated with 2–4 d of high heat. Genes responsive to high temperatures were identified from clustering of microarray data. Among these, at least 13 were designated as high temperature-repressed genes in the anther. Expression analyses revealed that these genes were expressed specifically in the immature anther mainly in the tapetum at the microspore stage and down-regulated after 1 d of high temperature. The expression levels of Osc6, OsRAFTIN and TDR, which are tapetum-specific genes, were unaffected by high temperatures. These results suggest that not all tapetal genes are inhibited by increased temperatures and the tapetum itself is not degraded in such an environment. However, high temperatures may disrupt some of the tapetum functions required for pollen adhesion and germination on the stigma.

Keywords: High temperature stress - Male sterility - Microarray - Rice - Tapetum

Abbreviations: AD, auricle distance; GO, gene ontology; RT–PCR, reverse transcription–PCR.

6Present address: Takii Plant Breeding Experiment Station, Konan, Shiga, 520-3231 Japan.

(Received July 15, 2009; Accepted September 29, 2009)
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