Department of Plant Sciences, University of Saskatchewan, Saskatoon, SK, Canada.
Canadian Light Source, Saskatoon, SK, Canada.
Methods Mol Biol. 2020;2156:141-159. doi: 10.1007/978-1-0716-0660-5_11.
Despite the extensive use of synchrotron radiation in material and biomedical sciences, it has only recently been utilized to expand our understanding of plant responses to environmental stress. Recent advances have led to the development of phenotyping platforms to identify chemical and morphological differences in breeding plant material. While these methodologies are applicable for and tested with a variety of abiotic and biotic stresses, they are particularly useful as a first step to identify cold-induced chemical and morphological changes in plants. Here, we describe two methods to determine cold acclimation-induced changes at the cellular and tissue levels. First, we illustrate how to quantify and visualize changes in tissue chemistry using Fourier-transform infrared spectroscopy. Second, we describe how to nondestructively prepare, analyze, and interpret X-ray phase contrast images and render this data as two- or three-dimensional models. While these techniques utilize synchrotron radiation, the methodology and standard practices are applicable for handheld and laboratory bench-top equipment operating with conventional light sources.
尽管同步辐射在材料和生物医学科学中得到了广泛应用，但它最近才被用于扩展我们对植物环境胁迫响应的理解。最近的进展导致了表型平台的发展，以识别在植物繁殖材料中的化学和形态差异。虽然这些方法适用于各种非生物和生物胁迫，并经过了测试，但它们特别有用，可作为识别植物冷诱导化学和形态变化的第一步。在这里，我们描述了两种确定细胞和组织水平冷驯化诱导变化的方法。首先，我们说明了如何使用傅里叶变换红外光谱定量和可视化组织化学变化。其次，我们描述了如何非破坏性地准备、分析和解释 X 射线相衬图像，并将这些数据渲染为二维或三维模型。虽然这些技术利用了同步辐射，但该方法和标准实践适用于使用传统光源的手持式和实验室台式设备。
