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
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
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DOI：
10.1093/pcp/pci090
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发表时间：
2005-05-01
影响因子：
4.9
通讯作者：
Shimamoto, K
中科院分区：
文献类型：
--
作者：
Moritoh, S;Miki, D;Shimamoto, K
我们从水稻（Oryza sativa L.）中克隆了RAD 2/XPG核酸酶家族的一个新成员OsGEN-L（OsGEN-like）。OsGEN-L具有在RAD 2/XPG核酸酶家族中保守的两个结构域，N-区和I-区。数据库检索和系统发育分析表明，OsGEN-L属于RAD 2/XPG核酸酶家族的第4类，在动物和高等植物中发现了OsGEN-L的同源物。为了阐明OsGEN-L的功能，我们获得了OsGEN-L表达沉默的水稻OsGEN-L-RNAi转基因植株。大部分OsGEN-L-RNAi植株表现出低育性，其中一些植株为雄性不育。OsGEN-L-RNAi植物缺乏成熟花粉，这是由于早期小孢子发育缺陷所致。OsGEN-L-绿色荧光蛋白（GFP）融合蛋白定位于细胞核中，OsGEN-L启动子在花药中特异表达。此外，重组OsGEN-L蛋白具有瓣状内切酶活性和单链和双链DNA结合活性。这些结果表明OsGEN-L在水稻小孢子发育过程中的DNA代谢中起着重要作用。
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.