利用基因组编辑技术定点突变IPA1基因创制水稻新株型材料  

Morphological Alternation of Rice Plant by Site-Directed Mutagenesis on IPA1 Gene

作　　者：刘华清[1] 孙庆山[1] 杨绍华[1] 周淑芬[1] 王锋[1] LIU Hua-qing;SUN Qing-shan;YANG Shao-hua;ZHOU Shu-fen;WANG Feng(Institute of Biotechnology,Fujian Academy of Agricultural Sciences/Fujian Key Laboratory of Agricultural Genetic Engineering,Fuzhou,Fujian 350003,China)

机构地区：[1]福建省农业科学院生物技术研究所/福建省农业遗传工程重点实验室

出　　处：《福建农业学报》2019年第8期867-872,共6页Fujian Journal of Agricultural Sciences

基　　金：福建省科技计划项目——省属公益类科研院所基本科研专项(2015R1019-1);福建省科技重大专项(2015NZ0002-3)

摘　　要：【目的】水稻株型改良是提高水稻产量的一个有效途径。水稻理想株型基因IPA1是一个调控水稻株型的关键基因,利用基因组编辑技术(TALENs技术)定点突变水稻IPA1基因,了解IPA1基因不同序列变异的株型效应,为进一步利用IPA1基因创制实用型水稻新株型材料奠定基础。【方法】利用TALENs技术定点突变优良恢复系明恢86的IPA1基因,通过测序鉴定突变体,种植于标准小区,调查分析其株型相关性状。【结果】利用TALENs技术获得了8种不同序列突变的水稻ipa1突变体,并通过转基因植株自交结合PCR分析筛选到去除了TALENs表达框,获得4种不含外源转基因成分的纯合突变体材料(IPA1基因表达区分别缺失2、4、16、23 bp)。表型分析发现,IPA1基因突变能够显著改变水稻的株高、有效穗数、穗长及穗粒数等性状。与野生型比较,缺失移码突变体株高降低7.9%~11.4%,有效穗数增加46.9%~68.4%,穗长短24.2%~29.3%,穗粒数减少31%~34%,结实率和千粒重差异不明显。【结论】利用TALENs技术定点突变水稻IPA1基因能够明显改变水稻株高、有效穗数、穗长及穗粒数等主要性状,产生水稻新株型。【Objective】The site-directed mutagenesis on grain-yield-related IPA1 to modifying the morphological characteristics of a rice plant for breeding purpose was studied.【Method】Minghui 86,a rice elite restorer,was used in the study.The primary gene that governs the gain yield of the rice plant,IPA1,was edited using the transcription activator-like effector nucleases(TALENs)to generate mutated plants for subsequent identification by gene sequencing and cultivation on standard plots to verify results of the mutagenesis.Eight transgenic rice plants that differed in ipa1 sequences had the TALENs boxes cleaved for self-crossing followed by screening with PCR analysis to arrive at 4 mutants with 2bp,4bp,16bp or 23bp segment deleted in their IPA1 genes.【Result】A phenotypic analysis on the 4 mutants free of exogenous transgenic elements confirmed that the site-directed mutagenesis on IPA1 had significantly altered the plant height,effective panicle number,panicle length,and grain number per panicle of the mutated rice plants.Compared with the wild-type Minghui 86,the plant height of the mutants was decreased by 7.9% to 11.4%,the effective panicle number increased by 46.9% to 68.4%,the panicle length shortened by 24.2% to 29.3%,the number of grains per panicle reduced by 31% to 34%,but the seed setting rate and 1000-grain weight were not changed significantly.【Conclusion】The site-directed mutagenesis applying TALENs on IPA1 could significantly modify the plant height,effective panicle number,panicle length,and grain number per panicle on the new rice plant.The technique could be applied for the breeding program on rice to increase grain yield.

关 键 词：水稻 IPA1基因 基因组编辑 突变体 新株型 

分 类 号：S511[农业科学—作物学] S330