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Plant and Cell Physiology Advance Access published online on March 25, 2005
Plant and Cell Physiology, doi:10.1093/pcp/pci090
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Plant and Cell Physiology 2005 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received January 30, 2005
Accepted February 13, 2005

Regular Papar

RNAi-Mediated Silencing of OsGEN-L (OsGEN-like), a New Member of the RAD2/XPG Nuclease Family, Causes Male Sterility by Defect of Microspore Development in Rice

Satoru Moritoh 1, Daisuke Miki 1, Masahiro Akiyama 2, Mihoko Kawahara 1, Takeshi Izawa 1, Hisaji Maki 2, and Ko Shimamoto 1*

1 Laboratory of Plant Molecular Genetics, Nara Institute of Science and Technology, Takayama 8916-5, Ikoma, Nara 630-0192, Japan
2 Laboratory of Microbial Molecular Genetics, Nara Institute of Science and Technology, Takayama 8916-5, Ikoma, Nara 630-0192, Japan

* To whom correspondence should be addressed.
Ko Shimamoto, E-mail: simamoto{at}bs.naist.jp


  Abstract

We have cloned a new member of the RAD2/XPG nuclease family, OsGEN-L (OsGEN-like), from rice (Oryza sativa L.). OsGEN-L possesses two domains, the N- and I-regions, that are conserved in the RAD2/XPG nuclease family. Database searches and phylogenetic analyses revealed that OsGEN-L belongs to class 4 of the RAD2/XPG nuclease family, and OsGEN-L homologues were found in animals and higher plants. To elucidate the function of OsGEN-L, we generated rice OsGEN-L-RNAi transgenic plants in which OsGEN-L expression was silenced. Most of the OsGEN-L-RNAi plants displayed low fertility, and some of them were male-sterile. OsGEN-L-RNAi plants lacked mature pollen, resulting from a defect in early microspore development. A OsGEN-L-GFP fusion protein was localized in the nucleus, and the OsGEN-L promoter was specifically active in the anthers. Furthermore, a recombinant OsGEN-L protein possessed flap endonuclease activity and both single-stranded and double-stranded DNA-binding activities. Our results suggest that OsGEN-L plays an essential role in DNA metabolism required for early microspore development in rice.

Keywords: DNA-binding activity; flap endonuclease activity; male sterility; microspore development; RAD2/XPG nuclease family; rice (Oryza sativa L.).
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