Fungal infections trigger defense or signaling responses in plants, leading to various changes in plant metabolites. The changes in metabolites, for example chlorophyll or flavonoids, have long been detectable using time-consuming destructive analytical methods including high-performance liquid chromatography or photometric determination. Recent plant phenotyping studies have revealed that hyperspectral imaging (HSI) in the UV range can be used to link spectral changes with changes in plant metabolites. To compare established destructive analytical methods with new nondestructive hyperspectral measurements, the interaction between sugar beet leaves and the pathogens Cercospora beticola, which causes Cercospora leaf spot disease (CLS), and Uromyces betae, which causes sugar beet rust (BR), was investigated. With the help of destructive analyses, we showed that both diseases have different effects on chlorophylls, carotenoids, flavonoids, and several phenols. Nondestructive hyperspectral measurements in the UV range revealed different effects of CLS and BR on plant metabolites resulting in distinct reflectance patterns. Both diseases resulted in specific spectral changes that allowed differentiation between the two diseases. Machine learning algorithms enabled the differentiation between the symptom classes and recognition of the two sugar beet diseases. Feature importance analysis identified specific wavelengths important to the classification, highlighting the utility of the UV range. The study demonstrates that HSI in the UV range is a promising, nondestructive tool to investigate the influence of plant diseases on plant physiology and biochemistry.
Rhizoctonia crown and root rot (RCRR), caused by Rhizoctonia solani, can cause severe yield and quality losses in sugar beet. The most common strategy to control the disease is the development of resistant varieties. In the breeding process, field experiments with artificial inoculation are carried out to evaluate the performance of genotypes and varieties. The phenotyping process in breeding trials requires constant monitoring and scoring by skilled experts. This work is time demanding and shows bias and heterogeneity according to the experience and capacity of each individual person. Optical sensors and artificial intelligence have demonstrated great potential to achieve higher accuracy than human raters and the possibility to standardize phenotyping applications. A workflow combining red–green–blue and multispectral imagery coupled to an unmanned aerial vehicle (UAV), as well as machine learning techniques, was applied to score diseased plants and plots affected by RCRR. Georeferenced annotation of UAV-orthorectified images was carried out. With the annotated images, five convolutional neural networks were trained to score individual plants. The training was carried out with different image analysis strategies and data augmentation. The custom convolutional neural network trained from scratch together with pretrained MobileNet showed the best precision in scoring RCRR (0.73 to 0.85). The average per plot of spectral information was used to score the plots, and the benefit of adding the information obtained from the score of individual plants was compared. For this purpose, machine learning models were trained together with data management strategies, and the best-performing model was chosen. A combined pipeline of random forest and k-nearest neighbors has shown the best weighted precision (0.67). This research provides a reliable workflow for detecting and scoring RCRR based on aerial imagery. RCRR is often distributed heterogeneously in trial plots; therefore, considering the information from individual plants of the plots showed a significant improvement in UAV-based automated monitoring routines.
In crop protection, disease quantification parameters such as disease incidence (DI) and disease severity (DS) are the principal indicators for decision making, aimed at ensuring the safety and productivity of crop yield. The quantification is standardized with leaf organs, defined as individual scoring units. This study focuses on identifying and segmenting individual leaves in agricultural fields using unmanned aerial vehicle (UAV), multispectral imagery of sugar beet fields, and deep instance segmentation networks (Mask R-CNN). Five strategies for achieving network robustness with limited labeled images are tested and compared, employing simple and copy-paste image augmentation techniques. The study also evaluates the impact of environmental conditions on network performance. Metrics of performance show that multispectral UAV images recorded under sunny conditions lead to a performance drop. Focusing on the practical application, we employ Mask R-CNN models in an image-processing pipeline to calculate leaf-based parameters including DS and DI. The pipeline was applied in time-series in an experimental trial with five varieties and two fungicide strategies to illustrate epidemiological development. Disease severity calculated with the model with highest Average Precision (AP) shows the strongest correlation with the same parameter assessed by experts. The time-series development of disease severity and disease incidence demonstrates the advantages of multispectral UAV-imagery in contrasting varieties for resistance, as well as the limits for disease control measurements. This study identifies key components for automatic leaf segmentation of diseased plants using UAV imagery, such as illumination and disease condition. It also provides a tool for delivering leaf-based parameters relevant to optimize crop production through automated disease quantification by imaging tools.