Heritable site-specific mutagenesis using TALENs in maize
Heritable site-specific mutagenesis using TALENs in maize
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DOI：
10.1111/pbi.12344
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发表时间：
2015-09-01
影响因子：
13.8
通讯作者：
Yang, Bing
中科院分区：
文献类型：
--
作者：
Char, Si Nian;Unger-Wallace, Erica;Yang, Bing
转录激活因子样效应核酸酶（Transcription activator-like effector nuclease， TALEN）技术已广泛应用于许多生物体的靶向基因诱变，特别是基因失活，包括水稻、小麦、番茄和大麦等重要农业植物。本报告描述了该技术在玉米中产生可遗传基因组修饰的应用。利用TALENs在玉米gl2位点产生稳定的、可遗传的突变。从玉米基因型Hi-II中获得含有单或双等位基因突变的转基因品系，频率约为10%（91个转基因事件中有9个突变事件）。此外，三个新的等位基因在后代幼苗中进行了功能测试，在那里它们能够赋予光滑的表型。在大多数事件中，整合的TALEN T-DNA独立于新的功能等位基因的缺失，在T1代产生突变的零分离后代。研究结果表明，TALENs是玉米基因组诱变的有效工具，有助于发现基因功能和发展性状改良。
Transcription activator-like effector nuclease (TALEN) technology has been utilized widely for targeted gene mutagenesis, especially for gene inactivation, in many organisms, including agriculturally important plants such as rice, wheat, tomato and barley. This report describes application of this technology to generate heritable genome modifications in maize. TALENs were employed to generate stable, heritable mutations at the maize glossy2 (gl2) locus. Transgenic lines containing mono- or di-allelic mutations were obtained from the maize genotype Hi-II at a frequency of about 10% (nine mutated events in 91 transgenic events). In addition, three of the novel alleles were tested for function in progeny seedlings, where they were able to confer the glossy phenotype. In a majority of the events, the integrated TALEN T-DNA segregated independently from the new loss of function alleles, producing mutated null-segregant progeny in T1 generation. Our results demonstrate that TALENs are an effective tool for genome mutagenesis in maize, empowering the discovery of gene function and the development of trait improvement.