Abstract：
Plants must adapt to environmental constraints. For this, they are able to perceive several types of stress in isolation or in combination manner. At the cellular level, after the perception of stress, cell signaling is set up to allow the establishment of the specific response. The calcium ion is known to be one of the ubiquitous second messengers which is involved in most of the stresses perceived by the plant. Changes of free cytosolic calcium but also in other cellular compartments are able to activate or inactivate several mechanisms involved in the cell to cope with the changes of environmental conditions. Several calcium reporters have been intensively used to visualize calcium signals in different conditions. In this chapter, we will present only genetically encoded fluorescent reporters for calcium imaging in living plant tissues to measure variations in calcium at several scales. The FRET (fluorescence resonance energy transfer) YC3.60 and the intensiometric GCamP3 sensors will be used in this method chapter. The image analyses will be also detailed for fluorescence quantification of calcium variation.
摘要：
植物必须适应环境的限制。为此，他们能够单独或组合地感知几种类型的压力。在细胞层面,在感知到压力之后，建立小区信令以允许建立特定响应。已知钙离子是普遍存在的第二信使之一，它与植物感知的大多数胁迫有关。游离胞质钙的变化以及其他细胞区室中的变化能够激活或失活细胞中涉及的几种机制，以应对环境条件的变化。几种钙报告基因已被广泛用于在不同条件下可视化钙信号。在这一章中,我们将只提供基因编码的荧光报告，用于活体植物组织中的钙成像，以测量不同尺度的钙变化。FRET（荧光共振能量转移）YC3.60和增强式GCamP3传感器将用于本方法章节。图像分析也将详细用于钙变化的荧光定量。
