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Plant and Cell Physiology Advance Access originally published online on March 25, 2005
Plant and Cell Physiology 2005 46(5):699-715; doi:10.1093/pcp/pci090
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JSPP © 2005

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 Moritoh1,3, Daisuke Miki1, Masahiro Akiyama2, Mihoko Kawahara1, Takeshi Izawa1,4, Hisaji Maki2 and Ko Shimamoto1,5

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

5 Corresponding author: E-mail: simamoto{at}bs.naist.jp; Fax: +81-743-72-5509.

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 homologs 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–green fluorescent protein (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.

3 Present address: Division of Molecular Genetics, National Institute for Basic Biology, National Institute of Natural Sciences, 38 Nishigonaka, Myodaiji, Okazaki, Aichi, 444-8585 Japan.

4 Present address: Laboratory of Applied Plant Genomics, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, 305-8602 Japan.

The nucleotide sequences reported in this paper have been submitted to EMBL/GenBank/DDBJ under accession numbers AB158320 and AB194139 for OsGEN-L cDNA and OsGEN-L genomic DNA, respectively.

(Received November 5, 2004; Accepted February 13, 2005 )
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