Background Hyperspectral reflectance data in the visible, near infrared and shortwave infrared range (VIS–NIR–SWIR, 400–2500 nm) are commonly used to nondestructively measure plant leaf properties. We investigated the usefulness of VIS–NIR–SWIR as a high-throughput tool to measure six leaf properties of maize plants including chlorophyll content (CHL), leaf water content (LWC), specific leaf area (SLA), nitrogen (N), phosphorus (P), and potassium (K). This assessment was performed using the lines of the maize diversity panel. Data were collected from plants grown in greenhouse condition, as well as in the field under two nitrogen application regimes. Leaf-level hyperspectral data were collected with a VIS–NIR–SWIR spectroradiometer at tasseling. Two multivariate modeling approaches, partial least squares regression (PLSR) and support vector regression (SVR), were employed to estimate the leaf properties from hyperspectral data. Several common vegetation indices (VIs: GNDVI, RENDVI, and NDWI), which were calculated from hyperspectral data, were also assessed to estimate these leaf properties. Results Some VIs were able to estimate CHL and N (R 2 > 0.68), but failed to estimate the other four leaf properties. Models developed with PLSR and SVR exhibited comparable performance to each other, and provided improved accuracy relative to VI models. CHL were estimated most successfully, with R 2 (coefficient of determination) > 0.94 and ratio of performance to deviation (RPD) > 4.0. N was also predicted satisfactorily (R 2 > 0.85 and RPD > 2.6). LWC, SLA and K were predicted moderately well, with R 2 ranging from 0.54 to 0.70 and RPD from 1.5 to 1.8. The lowest prediction accuracy was for P, with R 2 < 0.5 and RPD < 1.4. Conclusion This study showed that VIS–NIR–SWIR reflectance spectroscopy is a promising tool for low-cost, nondestructive, and high-throughput analysis of a number of leaf physiological and biochemical properties. Full-spectrum based modeling approaches (PLSR and SVR) led to more accurate prediction models compared to VI-based methods. We called for the construction of a leaf VIS–NIR–SWIR spectral library that would greatly benefit the plant phenotyping community for the research of plant leaf traits.

Phenotyping plants is an essential component of any effort to develop new crop varieties. As plant breeders seek to increase crop productivity and produce more food for the future, the amount of phenotype information they require will also increase. Traditional plant phenotyping relying on manual measurement is laborious, time-consuming, error-prone, and costly. Plant phenotyping robots have emerged as a high-throughput technology to measure morphological, chemical and physiological properties of large number of plants. Several robotic systems have been developed to fulfill different phenotyping missions. In particular, robotic phenotyping has the potential to enable efficient monitoring of changes in plant traits over time in both controlled environments and in the field. The operation of these robots can be challenging as a result of the dynamic nature of plants and the agricultural environments. Here we discuss developments in phenotyping robots, and the challenges which have been overcome and others which remain outstanding. In addition, some perspective applications of the phenotyping robots are also presented. We optimistically anticipate that autonomous and robotic systems will make great leaps forward in the next 10 years to advance the plant phenotyping research into a new era.

Background Leaf chlorophyll content plays an important role in indicating plant stresses and nutrient status. Traditional approaches for the quantification of chlorophyll content mainly include acetone ethanol extraction, spectrophotometry and high-performance liquid chromatography. Such destructive methods based on laboratory procedures are time consuming, expensive, and not suitable for high-throughput analysis. High throughput imaging techniques are now widely used for non-destructive analysis of plant phenotypic traits. In this study three imaging modules (RGB, hyperspectral, and fluorescence imaging) were, separately and in combination, used to estimate chlorophyll content of sorghum plants in a greenhouse environment. Color features, spectral indices, and chlorophyll fluorescence intensity were extracted from these three types of images, and multiple linear regression models and PLSR (partial least squares regression) models were built to predict leaf chlorophyll content (measured by a handheld leaf chlorophyll meter) from the image features. Results The models with a single color feature from RGB images predicted chlorophyll content with R2 ranging from 0.67 to 0.88. The models using the three spectral indices extracted from hyperspectral images (Ration Vegetation Index, Normalized Difference Vegetation Index, and Modified Chlorophyll Absorption Ratio Index) predicted chlorophyll content with R2 ranging from 0.77 to 0.78. The model using the fluorescence intensity extracted from fluorescence images predicted chlorophyll content with R2 of 0.79. The PLSR model that involved all the image features extracted from the three different imaging modules exhibited the best performance for predicting chlorophyll content, with R2 of 0.90. It was also found that inclusion of SLW (Specific Leaf Weight) into the image-based models further improved the chlorophyll prediction accuracy. Conclusion All three imaging modules (RGB, hyperspectral, and fluorescence) tested in our study alone could estimate chlorophyll content of sorghum plants reasonably well. Fusing image features from different imaging modules with PLSR modeling significantly improved the predictive performance. Image-based phenotyping could provide a rapid and non-destructive approach for estimating chlorophyll content in sorghum.

Many biochemical and physiological properties of plants that are of interest to breeders and geneticists have extremely low throughput and/or can only be measured destructively. This has limited the use of information on natural variation in nutrient and metabolite abundance, as well as photosynthetic capacity in quantitative genetic contexts where it is necessary to collect data from hundreds or thousands of plants. A number of recent studies have demonstrated the potential to estimate many of these traits from hyperspectral reflectance data, primarily in ecophysiological contexts. Here, we summarize recent advances in the use of hyperspectral reflectance data for plant phenotyping, and discuss both the potential benefits and remaining challenges to its application in plant genetics contexts. The performances of previously published models in estimating six traits from hyperspectral reflectance data in maize were evaluated on new sample datasets, and the resulting predicted trait values shown to be heritable (e.g., explained by genetic factors) were estimated. The adoption of hyperspectral reflectance-based phenotyping beyond its current uses may accelerate the study of genes controlling natural variation in biochemical and physiological traits.

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