Analyzing Subcellular Reorganization During Early Arabidopsis Embryogenesis Using Fluorescent Markers.

Abstract

Virtually all growth, developmental, physiological, and defense responses in plants are accompanied by reorganization of subcellular structures to enable altered cellular growth, differentiation or function. Visualizing cellular reorganization is therefore critical to understand plant biology at the cellular scale. Fluorescently labeled markers for organelles, or for cellular components are widely used in combination with confocal microscopy to visualize cellular reorganization. Early during plant embryogenesis, the precursors for all major tissues of the seedling are established, and in Arabidopsis, this entails a set of nearly invariant switches in cell division orientation and directional cell expansion. Given that these cellular reorganization events are genetically regulated and coupled to formative events in plant development, they offer a good model to understand the genetic control of cellular reorganization in plant development. Until recently, it has been challenging to visualize subcellular structures in the early Arabidopsis embryo for two reasons: embryos are deeply embedded in seed coat and fruit, and in addition, no dedicated fluorescent markers, expressed in the embryo, were available. We recently established both an imaging approach and a set of markers for the early Arabidopsis embryo. Here, we describe a detailed protocol to use these new tools in imaging cellular reorganization.

Plant phenotyping relevance

初期シロイヌナズナ胚の細胞内構造を可視化する専用イメージング手法と蛍光マーカーを開発し、その使用プロトコルを提示しているため、植物表現型取得法が中心である。

Code and data availability

This is a methods/protocol chapter describing embryo extraction and confocal imaging procedures. No public phenotype/trait datasets, plant images, analysis code, trained models, or supplements with such assets are mentioned; no availability or deposit statements for paper-specific data or code appear in the supplied.

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