López-Juez Enrique, Hills Alison
School of Biological Sciences, Royal Holloway, University of London, Egham, Surrey, UK.
Methods Mol Biol. 2011;774:3-18. doi: 10.1007/978-1-61779-234-2_1.
The power of Arabidopsis as a model organism lies in the depth and breadth of genetic tools available for its study. This also applies to the study of chloroplast biology. Although vast numbers of mutants have been identified in Arabidopsis, the continued use of forward-genetic screening approaches remains valuable for the isolation and study of previously overlooked mutants and novel mutations in sensitised backgrounds (i.e., suppressors or enhancers of previously known mutants). In addition, reverse-genetic collections of insertional mutants are now extensive and provide unique opportunities for gene function discovery. Here, we describe methods for the chemical mutagenesis of Arabidopsis, the screening of mutants visually, on the basis of gene-expression phenotypes (scored as reduced or enhanced activity of reporter genes), and the use of databases to select for existing mutations from historic collections or insertional mutagenesis programmes.
拟南芥作为模式生物的优势在于可用于其研究的遗传工具的深度和广度。这同样适用于叶绿体生物学的研究。尽管在拟南芥中已鉴定出大量突变体,但继续使用正向遗传筛选方法对于在敏感背景(即先前已知突变体的抑制子或增强子)中分离和研究先前被忽视的突变体及新突变仍然很有价值。此外,插入突变体的反向遗传文库现在已经很丰富,为基因功能发现提供了独特机会。在这里,我们描述了拟南芥化学诱变、基于基因表达表型(以报告基因活性降低或增强来评分)进行突变体视觉筛选以及利用数据库从历史文库或插入诱变计划中选择现有突变的方法。