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Canopy temperature (CT) is often interpreted as representing leaf activity traits such as photosynthetic rates, gas exchange rates, or stomatal conductance. This interpretation is based on the observation that leaf activity traits correlate with transpiration which affects leaf temperature. Accordingly, CT measurements may provide a basis for high throughput assessments of the productivity of wheat canopies during early grain filling, which would allow distinguishing functional from dysfunctional stay-green. However, whereas the usefulness of CT as a fast surrogate measure of sustained vigor under soil drying is well established, its potential to quantify leaf activity traits under high-yielding conditions is less clear. To better understand sensitivity limits of CT measurements under high yielding conditions, we generated within-genotype variability in stay-green functionality by means of differential short-term pre-anthesis canopy shading that modified the sink:source balance. We quantified the effects of these modifications on stay-green properties through a combination of gold standard physiological measurements of leaf activity and newly developed methods for organ-level senescence monitoring based on timeseries of high-resolution imagery and deep-learning-based semantic image segmentation. In parallel, we monitored CT by means of a pole-mounted thermal camera that delivered continuous, ultra-high temporal resolution CT data. Our results show that differences in stay-green functionality translate into measurable differences in CT in the absence of major confounding factors. Differences amounted to approximately 0.8°C and 1.5°C for a very high-yielding source-limited genotype, and a medium-yielding sink-limited genotype, respectively. The gradual nature of the effects of shading on CT during the stay-green phase underscore the importance of a high measurement frequency and a time-integrated analysis of CT, whilst modest effect sizes confirm the importance of restricting screenings to a limited range of morphological and phenological diversity., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Anderegg, Kirchgessner, Aasen, Zumsteg, Keller, Zenkl, Walter and Hund.)

Manual assessment of flower abundance of different flowering plant species in grasslands is a time-consuming process. We present an automated approach to determine the flower abundance in grasslands from drone-based aerial images by using deep learning (Faster R-CNN) object detection approach, which was trained and evaluated on data from five flights at two sites. Our deep learning network was able to identify and classify individual flowers. The novel method allowed generating spatially explicit maps of flower abundance that met or exceeded the accuracy of the manual-count-data extrapolation method while being less labor intensive. The results were very good for some types of flowers, with precision and recall being close to or higher than 90%. Other flowers were detected poorly due to reasons such as lack of enough training data, appearance changes due to phenology, or flowers being too small to be reliably distinguishable on the aerial images. The method was able to give precise estimates of the abundance of many flowering plant species. In the future, the collection of more training data will allow better predictions for the flowers that are not well predicted yet. The developed pipeline can be applied to any sort of aerial object detection problem., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Gallmann, Schüpbach, Jacot, Albrecht, Winizki, Kirchgessner and Aasen.)

Robust and automated segmentation of leaves and other backgrounds is a core prerequisite of most approaches in high-throughput field phenotyping. So far, the possibilities of deep learning approaches for this purpose have not been explored adequately, partly due to a lack of publicly available, appropriate datasets. This study presents a workflow based on DeepLab v3+ and on a diverse annotated dataset of 190 RGB (350 x 350 pixels) images. Images of winter wheat plants of 76 different genotypes and developmental stages have been acquired throughout multiple years at high resolution in outdoor conditions using nadir view, encompassing a wide range of imaging conditions. Inconsistencies of human annotators in complex images have been quantified, and metadata information of camera settings has been included. The proposed approach achieves an intersection over union (IoU) of 0.77 and 0.90 for plants and soil, respectively. This outperforms the benchmarked machine learning methods which use Support Vector Classifier and/or Random Forrest. The results show that a small but carefully chosen and annotated set of images can provide a good basis for a powerful segmentation pipeline. Compared to earlier methods based on machine learning, the proposed method achieves better performance on the selected dataset in spite of using a deep learning approach with limited data. Increasing the amount of publicly available data with high human agreement on annotations and further development of deep neural network architectures will provide high potential for robust field-based plant segmentation in the near future. This, in turn, will be a cornerstone of data-driven improvement in crop breeding and agricultural practices of global benefit., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Zenkl, Timofte, Kirchgessner, Roth, Hund, Van Gool, Walter and Aasen.)

Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Grassland-based ruminant livestock production provides a sustainable alternative to intensive production systems relying on concentrated feeds. However, grassland-based roughage often lacks the energy content required to meet the productivity potential of modern livestock breeds. Forage legumes, such as red clover, with increased starch content could partly replace maize and cereal supplements. However, breeding for increased starch content requires efficient phenotyping methods. This study is unique in evaluating a non-destructive hyperspectral imaging approach to estimate leaf starch content in red clover for enabling efficient development of high starch red clover genotypes. We assessed prediction performance of partial least square regression models (PLSR) using cross-validation, and validated model performance with an independent test set under controlled conditions. Starch content of the training set ranged from 0.1 to 120.3 mg g -1 DW. The best cross-validated PLSR model explained 56% of the measured variation and yielded a root mean square error (RMSE) of 17 mg g -1 DW. Model performance decreased when applying the trained model on the independent test set (RMSE = 29 mg g -1 DW, R 2 = 0.36). Different variable selection methods did not increase model performance. Once validated in the field, the non-destructive spectral method presented here has the potential to detect large differences in leaf starch content of red clover genotypes. Breeding material could be sampled and selected according to their starch content without destroying the plant., (Copyright © 2020 Frey, Baumann, Aasen, Studer and Kölliker.)

Understanding the interaction of plant growth with environmental conditions is crucial to increase the resilience of current cropping systems to a changing climate. Here, we investigate PhenoCams as a high-throughput approach for field phenotyping experiments to assess growth dynamics of many different genotypes simultaneously in high temporal (daily) resolution. First, we develop a method that extracts a daily phenological signal that is normalized for the different viewing geometries of the pixels within the images. Second, we investigate the extraction of the in season traits of early vigor, leaf area index (LAI), and senescence dynamic from images of a soybean (Glycine max) field phenotyping experiment and show that it is possible to rate early vigor, senescence dynamics, and track the LAI development between LAI 1 and 4.5. Third, we identify the start of green up, green peak, senescence peak, and end of senescence in the phenological signal. Fourth, we extract the timing of these points and show how this information can be used to assess the impact of phenology on harvest traits (yield, thousand kernel weight, and oil content). The results demonstrate that PhenoCams can track growth dynamics and fill the gap of high temporal monitoring in field phenotyping experiments., (Copyright © 2020 Aasen, Kirchgessner, Walter and Liebisch.)

Canopy temperature (CT) has been related to water-use and yield formation in crops. However, constantly (e.g., sun illumination angle, ambient temperature) as well as rapidly (e.g., clouds) changing environmental conditions make it difficult to compare measurements taken even at short time intervals. This poses a great challenge for high-throughput field phenotyping (HTFP). The aim of this study was to i) set up a workflow for unmanned aerial vehicles (UAV) based HTFP of CT, ii) investigate different data processing procedures to combine information from multiple images into orthomosaics, iii) investigate the repeatability of the resulting CT by means of heritability, and iv) investigate the optimal timing for thermography measurements. Additionally, the approach was v) compared with other methods for HTFP of CT. The study was carried out in a winter wheat field trial with 354 genotypes planted in two replications in a temperate climate, where a UAV captured CT in a time series of 24 flights during 6 weeks of the grain-filling phase. Custom-made thermal ground control points enabled accurate georeferencing of the data. The generated thermal orthomosaics had a high spatial accuracy (mean ground sampling distance of 5.03 cm/pixel) and position accuracy [mean root-mean-square deviation (RMSE) = 4.79 cm] over all time points. An analysis on the impact of the measurement geometry revealed a gradient of apparent CT in parallel to the principle plane of the sun and a hotspot around nadir. Averaging information from all available images (and all measurement geometries) for an area of interest provided the best results by means of heritability. Correcting for spatial in-field heterogeneity as well as slight environmental changes during the measurements were performed with the R package SpATS. CT heritability ranged from 0.36 to 0.74. Highest heritability values were found in the early afternoon. Since senescence was found to influence the results, it is recommended to measure CT in wheat after flowering and before the onset of senescence. Overall, low-altitude and high-resolution remote sensing proved suitable to assess the CT of crop genotypes in a large number of small field plots as is required in crop breeding and variety testing experiments., (Copyright © 2020 Perich, Hund, Anderegg, Roth, Boer, Walter, Liebisch and Aasen.)

The ability of a genotype to stay green affects the primary target traits grain yield (GY) and grain protein concentration (GPC) in wheat. High throughput methods to assess senescence dynamics in large field trials will allow for (i) indirect selection in early breeding generations, when yield cannot yet be accurately determined and (ii) mapping of the genomic regions controlling the trait. The aim of this study was to develop a robust method to assess senescence based on hyperspectral canopy reflectance. Measurements were taken in three years throughout the grain filling phase on >300 winter wheat varieties in the spectral range from 350 to 2500 nm using a spectroradiometer. We compared the potential of spectral indices (SI) and full-spectrum models to infer visually observed senescence dynamics from repeated reflectance measurements. Parameters describing the dynamics of senescence were used to predict GY and GPC and a feature selection algorithm was used to identify the most predictive features. The three-band plant senescence reflectance index (PSRI) approximated the visually observed senescence dynamics best, whereas full-spectrum models suffered from a strong year-specificity. Feature selection identified visual scorings as most predictive for GY, but also PSRI ranked among the most predictive features while adding additional spectral features had little effect. Visually scored delayed senescence was positively correlated with GY ranging from r = 0.173 in 2018 to r = 0.365 in 2016. It appears that visual scoring remains the gold standard to quantify leaf senescence in moderately large trials. However, using appropriate phenotyping platforms, the proposed index-based parameterization of the canopy reflectance dynamics offers the critical advantage of upscaling to very large breeding trials., (Copyright © 2020 Anderegg, Yu, Aasen, Walter, Liebisch and Hund.)