An efficient field screening procedure for identifying transposants for constructing an Ac/Ds-based insertional-mutant library of rice

文献类型： 外文期刊

第一作者： He, Cheng-Kun

作者： He, Cheng-Kun;Dey, Moul;Duan, Faping;Garg, Ajay;Wu, Ray;Luan, Wei-Jiang;Hu, Guo-Cheng;Fu, Ya-Ping;Si, Hua-Min;Zhu, L.;Liu, Wen-Zhen;Zhang, Hong;Liu, Wen-Ying;Zhuo, Ren-Ying;Sun, Zong-Xiu

作者机构：

关键词： rice;Ac/Ds transposon;large-scale screening;mutant library;gene trap

期刊名称：GENOME （ 影响因子：2.166； 五年影响因子：2.474 ）

ISSN： 0831-2796

年卷期： 2008 年 51 卷 1 期

页码：

收录情况： SCI

摘要： An efficient system was developed, and several variables tested, for generating a large-scale insertional-mutagenesis population of rice. The most important feature in this improved Ac/Ds tagging system is that one can conveniently carry out large-scale screening in the field and select transposants at the seedling stage. Rice was transformed with a plasmid that includes a B asta-re si stance gene (bar). After the Ds element is excised during transposition, bar becomes adjacent to the ubiquitin promoter, and the rice plant becomes resistant to the herbicide Basta. In principle, one can plant up to one million plants in the field and select those plants that survive after spraying with Basta. To test the utility of this system, 4 Ds starter lines were crossed with 14 different Ac plants, and many transposants were successfully identified after planting 134 285 F-2 plants in the field. Over 2 800 of these transposants were randomly chosen for PCR analysis, and the results fully confirmed the reliability of the field screening procedure.

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