Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA, USA.
Department of Agronomy, Iowa State University, Ames, IA, USA.
Plant Biotechnol J. 2015 Sep;13(7):1002-10. doi: 10.1111/pbi.12344. Epub 2015 Feb 3.
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.
转录激活因子样效应物核酸酶（TALEN）技术已广泛应用于靶向基因诱变，特别是在许多生物体中基因失活，包括农业上重要的植物，如水稻、小麦、番茄和大麦。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。TALEN 用于在玉米光泽 2 （gl2）基因座产生稳定的、可遗传的突变。从玉米基因型 Hi-II 中获得含有单等位基因或双等位基因突变的转基因株系的频率约为 10%（91 个转基因事件中有 9 个突变事件）。此外，对三个新等位基因在后代幼苗中的功能进行了测试，它们能够赋予光泽表型。在大多数情况下，整合的 TALEN T-DNA 与新的功能丧失等位基因独立分离，在 T1 代产生突变的纯合子分离后代。我们的结果表明，TALEN 是玉米基因组诱变的有效工具，能够发现基因功能并改善性状。
