This chapter provides a methodology for evaluating plant health and leaf characteristics using spectral reflectance. It provides a step-by-step guide to using spectrometers for high-resolution point measurements of leaf spectral reflectance and multispectral imaging for capturing spatial data, emphasizing the importance of consistent measurement conditions. The chapter further explores the intricacies of multispectral imaging, including calibration, data collection, and image processing. Finally, this chapter delves into the application of various spectral indices for the quantification of key traits such as pigment content, the status of the xanthophyll cycle, water content, and how to identify spectral regions of interest for further research and development. Serving as a guide for researchers and practitioners in plant science, this chapter provides a straightforward framework for plant health assessment using spectral reflectance.
Plant Leaves , Spectrum Analysis , Plant Leaves/chemistry , Spectrum Analysis/methods , Image Processing, Computer-Assisted/methods , Water/chemistry , Calibration , Plants , Xanthophylls
Infrared thermography offers a rapid, noninvasive method for measuring plant temperature, which provides a proxy for stomatal conductance and plant water status and can therefore be used as an index for plant stress. Thermal imaging can provide an efficient method for high-throughput screening of large numbers of plants. This chapter provides guidelines for using thermal imaging equipment and illustrative methodologies, coupled with essential considerations, to access plant physiological processes.
Infrared Rays , Phenotype , Thermography , Thermography/methods , Plants , High-Throughput Screening Assays/methods , Plant Physiological Phenomena , Temperature , Plant Stomata/physiology
Hyperspectral imaging is a remote sensing technique that enables remote, noninvasive measurement of plant traits. Here, we outline the procedures for camera setup, scanning, and calibration, along with the acquisition of black and white reference materials, which are the key steps in collecting hyperspectral imagery. We also discuss the development of predictive models such as partial least-squares regression, using both large and small datasets, which are used to predict plant traits from hyperspectral data. To ensure practical applicability, we provide code examples that allow readers to immediately implement these techniques in real-world scenarios. We introduce these topics to beginners in an accessible and understandable manner.
Data Analysis , Hyperspectral Imaging , Remote Sensing Technology , Remote Sensing Technology/methods , Hyperspectral Imaging/methods , Least-Squares Analysis , Plants , Calibration , Image Processing, Computer-Assisted/methods
BACKGROUND: Spectral imaging is a key method for high throughput phenotyping that can be related to a large variety of biological parameters. The Normalised Difference Vegetation Index (NDVI), uses specific wavelengths to compare crop health and performance. Increasing the accessibility of spectral imaging systems through the development of small, low cost, and easy to use platforms will generalise its use for precision agriculture. We describe a method for using a dual camera system connected to a Raspberry Pi to produce NDVI imagery, referred to as NDVIpi. Spectral reference targets were used to calibrate images into values of reflectance, that are then used to calculated NDVI with improved accuracy compared with systems that use single references/standards. RESULTS: NDVIpi imagery showed strong performance against standard spectrometry, as an accurate measurement of leaf NDVI. The NDVIpi was also compared to a relatively more expensive commercial camera (Micasense RedEdge), with both cameras having a comparable performance in measuring NDVI. There were differences between the NDVI values of the NDVIpi and the RedEdge, which could be attributed to the measurement of different wavelengths for use in the NDVI calculation by each camera. Subsequently, the wavelengths used by the NDVIpi show greater sensitivity to changes in chlorophyll content than the RedEdge. CONCLUSION: We present a methodology for a Raspberry Pi based NDVI imaging system that utilizes low cost, off-the-shelf components, and a robust multi-reference calibration protocols that provides accurate NDVI measurements. When compared with a commercial system, comparable NDVI values were obtained, despite the fact that our system was a fraction of the cost. Our results also highlight the importance of the choice of red wavelengths in the calculation of NDVI, which resulted in differences in sensitivity between camera systems.
