Horticultural Sciences Department and Plant Molecular and Cellular Biology Program, University of Florida, Gainesville, FL, USA.
Methods Mol Biol. 2020;2122:25-33. doi: 10.1007/978-1-0716-0342-0_3.
Effective mutagenesis is critical for connecting traits of interest to specific plant genes. The development of site-directed mutagenesis and sequenced-indexed genetics resources in maize allows for targeted analysis of individual genes. These reverse genetics approaches have the potential for confirmation bias by only studying candidate genes for association with traits of interest. Genetic screens of induced, random mutations are important for identifying novel loci as well as interacting factors for known mutant loci. Chemical mutagenesis provides very high mutation rates and can be used for a variety of screen designs. This chapter provides an updated protocol for ethyl methanesulfonate (EMS) mutagenesis of maize pollen using paraffin or mineral oil. Mutagenesis occurs in mature pollen causing nonconcordant endosperm and embryo genotypes as well as sectored M1 plants. Considerations for these factors in genetic screens are discussed.
有效的诱变对于将感兴趣的性状与特定的植物基因联系起来至关重要。玉米中定点诱变和测序索引遗传资源的发展允许对单个基因进行靶向分析。这些反向遗传学方法可能存在确认偏差，因为它们只研究与感兴趣性状相关的候选基因。诱导随机突变的遗传筛选对于鉴定新的基因座以及已知突变基因座的相互作用因子非常重要。化学诱变可提供非常高的突变率，并可用于多种筛选设计。本章提供了使用石蜡或矿物油对玉米花粉进行乙基甲烷磺酸（EMS）诱变的最新方案。诱变发生在成熟花粉中，导致不一致的胚乳和胚胎基因型以及扇形 M1 植物。遗传筛选中对这些因素的考虑进行了讨论。
