Synchrotron techniques are powerful tools in material and environmental sciences; however, they are currently underutilized in plant research. The Canadian Light Source synchrotron at the University of Saskatchewan campus is the only such facility in Canada open to academic, government, and industrial clients. This review introduces the potential of synchrotron-based spectroscopic methods and its applications to agriculture and plant sciences. Relative ease of sample preparation, nondestructive analysis, high spatial resolution, and multiple response measurements within a single sample are among its advantages. Synchrotron-based Fourier transform mid-infrared spectromicroscopy, X-ray absorption, and fluorescence spectromicroscopy are included in the several approaches discussed. Examples range from evaluating protein secondary structure and nondestructive compositional analysis of leaf epicuticular wax and pollen surface lipids to cell wall composition and nutrient analyses. Synchrotron technology can help to initially identify key spectra related to plant properties for subsequent higher throughput techniques. One example is the adaptation of synchrotron techniques for lower resolution analysis in the field such as nondestructive elemental analysis for localization of nutrients in fruit crops using handheld high-throughput devices. In addition, interest in creating high-throughput systems based on synchrotron technology itself is driving the development of new hardware to meet these larger challenges.