Efficient insertional mutagenesis in rice using the maize En/Spm elements
Efficient insertional mutagenesis in rice using the maize En/Spm elements
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DOI：
10.1111/j.1365-313x.2005.02570.x
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发表时间：
2005-12-01
期刊：
影响因子：
7.2
通讯作者：
Sundaresan, V
中科院分区：
文献类型：
--
作者：
Kumar, CS;Wing, RA;Sundaresan, V
我们已经开发了一个新的系统插入突变水稻（水稻）的基础上，玉米（玉米）增强子/抑制子突变（En/Spm）元件。在该系统中，将具有Spm转座酶和非自主缺陷型抑制突变子（dSpm）元件的单个T-DNA构建体与绿色荧光蛋白（GFP）和Discosoma sp.红色荧光蛋白（DsRed）荧光标记物结合使用，以选择dSpm的非连锁稳定转座。利用这个系统，我们可以证明高频率的非连锁转座水稻dSpm。对来自353个稳定插入系的dSpm侧翼序列的分析表明，dSpm插入似乎广泛分布在水稻染色体上，并偏好基因区域（70%）。的dSpm插入出现不同的激活解离（Ac-Ds）元素的基因组分布和表现出更大的比例相比，Ds元素的非连锁易位。本研究的结果表明，玉米En/Spm元件可以作为水稻功能基因组学的有效工具，并可以补充使用其他插入诱变剂的努力。此外，基于荧光的非侵入性选择系统的功效对于其应用于其他作物是有希望的。
We have developed a novel system for insertional mutagenesis in rice (Oryza sativa) based on the maize (Zea mays) enhancer/suppressor mutator (En/Spm) element. In this system, a single T-DNA construct with Spm-transposase and the non-autonomous defective suppressor mutator (dSpm) element is used in conjunction with green fluorescent protein (GFP) and Discosoma sp. Red Fluorescence Protein (DsRed) fluorescent markers to select unlinked stable transpositions of dSpm. Using this system, we could demonstrate high frequencies of unlinked germinal transposition of dSpm in rice. Analysis of dSpm flanking sequences from 353 stable insertion lines revealed that the dSpm insertions appear to be widely distributed on rice chromosomes with a preference for genic regions (70%). The dSpm insertions appear to differ from Activator-Dissociation (Ac-Ds) elements in genomic distribution and exhibit a greater fraction of unlinked transpositions when compared with Ds elements. The results obtained in this study demonstrate that the maize En/Spm element can be used as an effective tool for functional genomics in rice and can complement efforts using other insertional mutagens. Further, the efficacy of the non-invasive fluorescence-based selection system is promising for its application to other crops.