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通过RNA干扰对水稻AP3同源基因OsMADS16进行功能分析。

Functional analysis of the rice AP3 homologue OsMADS16 by RNA interference.

作者信息

Xiao Han, Wang Yun, Liu Daofeng, Wang Wemming, Li Xiaobing, Zhao Xianfeng, Xu Jichen, Zhai Wenxue, Zhu Lihuang

机构信息

Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, 917 Bld, Datun Rd, Andingmen-wai, Beijing 100101, China.

出版信息

Plant Mol Biol. 2003 Jul;52(5):957-66. doi: 10.1023/a:1025401611354.

Abstract

The rice OsMADS16 gene is phylogenetically related to the angiosperm B-function MADS-box genes. To investigate if OsMADS16 functions as an AP3/DEF orthologue to regulate the development of lodicules and stamens in rice, we isolated its genomic sequences and characterized its functions in planta by RNA interference. The genomic sequence of the OsMADS16 gene shows that it shares high similarity in genomic structure and the deduced amino acid sequence with the maize B-class gene, Si1. Transgenic lines from the introduced gene expressing double-stranded RNA with the OsMADS16 cDNA fragment were male-sterile and displayed alternations of lodicules and stamens, occasionally depressed palea and overgrown glume. The two lodicules were converted into four palea/lemma-like organs and some stamens into carpels. Further investigations of the transcription of OsMADS16 gene in these transgenic lines by RT-PCR revealed that its transcript was significantly reduced. Transcription of a rice PI homologous gene, OsMADS4, was also reduced remarkably in the transgenic plants. Our results demonstrate that OsMADS16 is an AP3/DEF orthologue to specify the identities of lodicules and stamens in rice flower and also support that OsMADS4 is a PI orthologue. In addition, these results suggest that RNA interference is a useful tool for functional genomics in rice.

摘要

水稻OsMADS16基因在系统发育上与被子植物B功能MADS盒基因相关。为了研究OsMADS16是否作为AP3/DEF同源基因来调控水稻浆片和雄蕊的发育,我们分离了其基因组序列,并通过RNA干扰在植物体内对其功能进行了表征。OsMADS16基因的基因组序列表明,它在基因组结构和推导的氨基酸序列上与玉米B类基因Si1具有高度相似性。导入表达带有OsMADS16 cDNA片段的双链RNA的转基因系是雄性不育的,并且表现出浆片和雄蕊的变化,偶尔内稃凹陷和颖片过度生长。两个浆片转化为四个类似内稃/外稃的器官,一些雄蕊转化为心皮。通过RT-PCR对这些转基因系中OsMADS16基因的转录进行进一步研究发现,其转录本显著减少。水稻PI同源基因OsMADS4在转基因植物中的转录也显著降低。我们的结果表明,OsMADS16是一个AP3/DEF同源基因,用于确定水稻花中浆片和雄蕊的身份,也支持OsMADS4是一个PI同源基因。此外,这些结果表明RNA干扰是水稻功能基因组学的一个有用工具。

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