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Abstract：
De novo root regeneration (DNRR) is the process in which adventitious roots are regenerated from damaged plant tissues or organs. We have developed a simple DNRR system in which adventitious roots are formed from detached leaf explants of Arabidopsis (Arabidopsis thaliana) on B5 medium without external hormones. In this chapter, we introduce the methods used to observe gene expression patterns during rooting from leaf explants. Usually, β-glucuronidase (GUS) staining is used to visualize gene expression patterns, since fluorescent proteins are difficult to observe because of the high autofluorescence in leaf explants. Here, we describe the use of the ClearSee technique with Congo red staining for deep imaging to observe fluorescent proteins. This method diminishes autofluorescence in leaf explants and preserves the stability of fluorescent proteins, thus allowing us to investigate the endogenous molecular actions guiding DNRR.
摘要：
从头根再生（DNRR）是从受损的植物组织或器官中再生不定根的过程。我们开发了一种简单的DNRR系统，其中不定根是在没有外部激素的B5培养基上从拟南芥（拟南芥）的离体叶外植体形成的。在这一章中,我们介绍了从叶片外植体生根过程中观察基因表达模式的方法。通常,β-葡糖醛酸酶（GUS）染色用于可视化基因表达模式，因为荧光蛋白是很难观察，因为在叶外植体的高自发荧光。这里,我们描述了使用ClearSee技术与刚果红染色进行深度成像以观察荧光蛋白。这种方法减少了叶片外植体中的自发荧光，并保留了荧光蛋白的稳定性。从而使我们能够研究指导DNRR的内源性分子作用。
