Díez Alba Rodríguez, Duque Paula, Henriques Rossana
Instituto Gulbenkian de Ciência, Oeiras, Portugal.
School of Biological, Earth and Environmental Sciences, University College Cork, Cork, Ireland.
Methods Mol Biol. 2022;2494:217-227. doi: 10.1007/978-1-0716-2297-1_15.
Plants live in highly dynamic surroundings and need to cope with constant environmental challenges. In order to do so, they developed quick reactions to stress that allow them to gain time while mounting a major response. This first line of defense includes the stomata, leaf epidermal pores in charge of regulating water loss and photosynthesis. Stomatal movements are controlled by the stress phytohormone abscisic acid (ABA), which induces fast closure of the stomata upon perception of stress conditions. By modulating plasma membrane ion channels, ABA leads to loss of water from the guard cells surrounding the stomatal pore and a consequent reduction of its aperture. Here, we provide a microscopy-based method to assess the plant's response to ABA through measurements of the stomatal aperture. This protocol describes a simple, quick, and unexpensive method to prepare high-quality impressions of leaves from Arabidopsis thaliana seedlings from long-lasting silicone-based casts, allowing detailed imaging and accurate determination of the aperture of stomatal pores.
植物生长在高度动态的环境中，需要应对不断的环境挑战。为此，它们对压力产生快速反应，以便在做出主要反应时赢得时间。这第一道防线包括气孔，即负责调节水分流失和光合作用的叶片表皮孔。气孔运动受应激植物激素脱落酸（ABA）控制，ABA在感知应激条件时会诱导气孔快速关闭。通过调节质膜离子通道，ABA导致围绕气孔孔的保卫细胞失水，从而使其孔径减小。在这里，我们提供一种基于显微镜的方法，通过测量气孔孔径来评估植物对ABA的反应。本方案描述了一种简单、快速且经济的方法，用于从持久的硅基铸型中制备拟南芥幼苗叶片的高质量印记，从而实现对气孔孔孔径的详细成像和准确测定。
