Hunan Provincial Key Laboratory of Phytohormones and Growth Development, Hunan Agricultural University, Changsha, China.
Department of Botany and Plant Sciences, University of California, Riverside, CA, USA.
Methods Mol Biol. 2023;2686:131-162. doi: 10.1007/978-1-0716-3299-4_6.
The flower is a hallmark feature that has contributed to the evolutionary success of land plants. Diverse mutagenic agents have been employed as a tool to genetically perturb flower development and identify genes involved in floral patterning and morphogenesis. Since the initial studies to identify genes governing processes such as floral organ specification, mutagenesis in sensitized backgrounds has been used to isolate enhancers and suppressors to further probe the molecular basis of floral development. Here, we first describe two commonly employed methods for mutagenesis (using ethyl methanesulfonate (EMS) or T-DNAs as mutagens), and then describe three methods for identifying a mutation that leads to phenotypic alterations: traditional map-based cloning, modified high-efficiency thermal asymmetric interlaced PCR (mhiTAIL-PCR), and deep sequencing in the plant model Arabidopsis thaliana.
花是一个标志性特征，它促进了陆地植物的进化成功。多种诱变剂已被用作工具，用于遗传干扰花发育并鉴定参与花模式形成和形态发生的基因。自最初鉴定控制花器官特化等过程的基因的研究以来，在敏感背景下进行诱变已被用于分离增强子和抑制剂，以进一步探究花发育的分子基础。在这里，我们首先描述了两种常用的诱变方法（使用乙基甲烷磺酸酯（EMS）或 T-DNAs 作为诱变剂），然后描述了三种用于鉴定导致表型改变的突变的方法：传统的基于图谱的克隆、改良的高效热不对称交错 PCR（mhiTAIL-PCR）和植物模式拟南芥中的深度测序。
