Non-Destructive Monitoring of Foliar Fungicide Efficacy with Hyperspectral Sensing in Grapevine.

Frequent fungicide applications are required to manage grapevine powdery mildew (Erysiphe necator). However, this practice is costly and has led to widespread fungicide resistance. A method of monitoring in-field fungicide efficacy could help growers maximize spray-interval length, thereby reducing costs and the rate of fungicide resistance emergence. The goal of this study was to evaluate if hyperspectral sensing in the visible to shortwave infrared range (400 to 2,400 nm) can quantify foliar fungicide efficacy on grape leaves. Commercial formulations of metrafenone, Bacillus mycoides isolate J (BmJ), and sulfur were applied on Chardonnay grapevines in vineyard or greenhouse settings. Foliar reflectance was measured with handheld hyperspectral spectroradiometers at multiple days post-application. Fungicide efficacy was estimated as a proxy for fungicide residue and disease control measured with the Blackbird microscopy imaging robot. Treatments could be differentiated from the untreated control with an accuracy of 73.06% for metrafenone, 67.76% for BmJ, and 94.10% for sulfur. The change in spectral reflectance was moderately correlated with the cube root of the area under the disease progress curve for metrafenone- and sulfur-treated samples (R2 = 0.38 and 0.36, respectively) and with sulfur residue (R2 = 0.42). BmJ treatment impacted foliar physiology by enhancing the leaf mass/area and reducing the nitrogen and total phenolic content as estimated from spectral reflectance. The results suggest that hyperspectral sensing can be used to monitor in-situ fungicide efficacy, and the prediction accuracy depends on the fungicide and the time point measured. The ability to monitor in-situ fungicide efficacy could facilitate more strategic fungicide applications and promote sustainable grapevine protection. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Scalable Early Detection of Grapevine Viral Infection with Airborne Imaging Spectroscopy.

The U.S. wine and grape industry loses $3B annually due to viral diseases including grapevine leafroll-associated virus complex 3 (GLRaV-3). Current detection methods are labor-intensive and expensive. GLRaV-3 has a latent period in which the vines are infected but do not display visible symptoms, making it an ideal model to evaluate the scalability of imaging spectroscopy-based disease detection. The NASA Airborne Visible and Infrared Imaging Spectrometer Next Generation was deployed to detect GLRaV-3 in Cabernet Sauvignon grapevines in Lodi, CA in September 2020. Foliage was removed from the vines as part of mechanical harvest soon after image acquisition. In September of both 2020 and 2021, industry collaborators scouted 317 hectares on a vine-by-vine basis for visible viral symptoms and collected a subset for molecular confirmation testing. Symptomatic grapevines identified in 2021 were assumed to have been latently infected at the time of image acquisition. Random forest models were trained on a spectroscopic signal of noninfected and GLRaV-3 infected grapevines balanced with synthetic minority oversampling of noninfected and GLRaV-3 infected grapevines. The models were able to differentiate between noninfected and GLRaV-3 infected vines both pre- and postsymptomatically at 1 to 5 m resolution. The best-performing models had 87% accuracy distinguishing between noninfected and asymptomatic vines, and 85% accuracy distinguishing between noninfected and asymptomatic + symptomatic vines. The importance of nonvisible wavelengths suggests that this capacity is driven by disease-induced changes to plant physiology. The results lay a foundation for using the forthcoming hyperspectral satellite Surface Biology and Geology for regional disease monitoring in grapevine and other crop species. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Effects of Nighttime Applications of Germicidal Ultraviolet Light Upon Powdery Mildew (Erysiphe necator), Downy Mildew (Plasmopara viticola), and Sour Rot of Grapevine.

Nighttime applications of germicidal ultraviolet were evaluated as a means to suppress three diseases of grapevine. In laboratory studies, UV-C light (peak 254 nm, FWHM 5 nm) applied during darkness strongly inhibited the germination of conidia of Erysiphe necator, and at a dose of 200 J/m2, germination was zero. Reciprocity of irradiance and duration of exposure with respect to conidial germination was confirmed for UV-C doses between 0 and 200 J/m2 applied at 4 or 400 s. When detached grapevine leaves were exposed during darkness to UV-C at 100 J/m2 up to 7 days before they were inoculated with zoospores of Plasmopara viticola, infection and subsequent sporulation was reduced by over 70% compared to untreated control leaves, indicating an indirect suppression of the pathogen exerted through the host. A hemicylindrical array of low-pressure discharge UV-C lamps configured for trellised grapevines was designed and fitted to both a tractor-drawn carriage and a fully autonomous robotic carriage for vineyard applications. In 2019, in a Chardonnay research vineyard with a history of high inoculum and severe disease, weekly nighttime applications of UV-C suppressed E. necator on leaves and fruit at doses of 100 and 200 J/m2. In the same vineyard in 2020, UV-C was applied once or twice weekly at doses of 70, 100, or 200 J/m2, and severity of E. necator on both leaves and fruit was significantly reduced compared to untreated controls; twice-weekly applications at 200 J/m2 provided suppression equivalent to a standard fungicide program. None of the foregoing UV-C treatments significantly reduced the severity of P. viticola on Chardonnay vines compared to the untreated control in 2020. However, twice-weekly applications of UV-C at 200 J/m2 to the more downy mildew-resistant Vitis interspecific hybrid cultivar Vignoles in 2021 significantly suppressed foliar disease severity. In commercial Chardonnay vineyards with histories of excellent disease control in Dresden, NY, E. necator remained at trace levels on foliage and was zero on fruit following weekly nighttime applications of UV-C at 200 J/m2 in 2020 and after weekly or twice-weekly application of UV-C at 100 or 200 J/m2 in 2021. In 2019, weekly nighttime applications of UV-C at 200 J/m2 also significantly reduced the severity of sour rot, a decay syndrome of complex etiology, on fruit of 'Vignoles' but not the severity of bunch rot caused by Botrytis cinerea. A similar level of suppression of sour rot was observed on 'Vignoles' vines treated twice-weekly with UV-C at 200 J/m2 in 2021. Nighttime UV-C applications did not produce detectable indications of metabolic abnormalities, phytotoxicity, growth reduction, or reductions of fruit yield or quality parameters, even at the highest doses and most frequent intervals employed.

Contact Reflectance Spectroscopy for Rapid, Accurate, and Nondestructive Phytophthora infestans Clonal Lineage Discrimination.

Populations of Phytophthora infestans, the oomycete causal agent of potato late blight in the United States, are predominantly asexual, and isolates are characterized by clonal lineage or asexual descendants of a single genotype. Current tools for clonal lineage identification are time consuming and require laboratory equipment. We previously found that foliar spectroscopy can be used for high-accuracy pre- and postsymptomatic detection of P. infestans infections caused by clonal lineages US-08 and US-23. In this work, we found subtle but distinct differences in spectral responses of potato foliage infected by these clonal lineages in both growth-chamber time-course experiments (12- to 24-h intervals over 5 days) and naturally infected samples from commercial production fields. In both settings, we measured continuous visible to shortwave infrared reflectance (400 to 2,500 nm) on leaves using a portable spectrometer with contact probe. We consistently discriminated between infections caused by the two clonal lineages across all stages of disease progression using partial least squares (PLS) discriminant analysis, with total accuracies ranging from 88 to 98%. Three-class random forest differentiation between control, US-08, and US-23 yielded total discrimination accuracy ranging from 68 to 76%. Differences were greatest during presymptomatic infection stages and progressed toward uniformity as symptoms advanced. Using PLS-regression trait models, we found that total phenolics, sugar, and leaf mass per area were different between lineages. Shortwave infrared wavelengths (>1,100 nm) were important for clonal lineage differentiation. This work provides a foundation for future use of hyperspectral sensing as a nondestructive tool for pathovar differentiation.

Digital plant pathology: a foundation and guide to modern agriculture.

Over the last 20 years, researchers in the field of digital plant pathology have chased the goal to implement sensors, machine learning and new technologies into knowledge-based methods for plant phenotyping and plant protection. However, the application of swiftly developing technologies has posed many challenges. Greenhouse and field applications are complex and differ in their study design requirements. Selecting a sensor type (e.g., thermography or hyperspectral imaging), sensor platform (e.g., rovers, unmanned aerial vehicles, or satellites), and the problem-specific spatial and temporal scale adds to the challenge as all pathosystems are unique and differ in their interactions and symptoms, or lack thereof. Adding host-pathogen-environment interactions across time and space increases the complexity even further. Large data sets are necessary to enable a deeper understanding of these interactions. Therefore, modern machine learning methods are developed to realize the fast data analysis of such complex data sets. This reduces not only human effort but also enables an objective data perusal. Especially deep learning approaches show a high potential to identify probable cohesive parameters during plant-pathogen-environment interactions. Unfortunately, the performance and reliability of developed methods are often doubted by the potential user. Gaining their trust is thus needed for real field applications. Linking biological causes to machine learning features and a clear communication, even for non-experts of such results, is a crucial task that will bridge the gap between theory and praxis of a newly developed application. Therefore, we suggest a global connection of experts and data as the basis for defining a common and goal-oriented research roadmap. Such high interconnectivity will likely increase the chances of swift, successful progress in research and practice. A coordination within international excellence clusters will be useful to reduce redundancy of research while supporting the creation and progress of complementary research. With this review, we would like to discuss past research, achievements, as well as recurring and new challenges. Having such a retrospect available, we will attempt to reveal future challenges and provide a possible direction elevating the next decade of research in digital plant pathology.

Deep semantic segmentation for the quantification of grape foliar diseases in the vineyard.

Plant disease evaluation is crucial to pathogen management and plant breeding. Human field scouting has been widely used to monitor disease progress and provide qualitative and quantitative evaluation, which is costly, laborious, subjective, and often imprecise. To improve disease evaluation accuracy, throughput, and objectiveness, an image-based approach with a deep learning-based analysis pipeline was developed to calculate infection severity of grape foliar diseases. The image-based approach used a ground imaging system for field data acquisition, consisting of a custom stereo camera with strobe light for consistent illumination and real time kinematic (RTK) GPS for accurate localization. The deep learning-based pipeline used the hierarchical multiscale attention semantic segmentation (HMASS) model for disease infection segmentation, color filtering for grapevine canopy segmentation, and depth and location information for effective region masking. The resultant infection, canopy, and effective region masks were used to calculate the severity rate of disease infections in an image sequence collected in a given unit (e.g., grapevine panel). Fungicide trials for grape downy mildew (DM) and powdery mildew (PM) were used as case studies to evaluate the developed approach and pipeline. Experimental results showed that the HMASS model achieved acceptable to good segmentation accuracy of DM (mIoU > 0.84) and PM (mIoU > 0.74) infections in testing images, demonstrating the model capability for symptomatic disease segmentation. With the consistent image quality and multimodal metadata provided by the imaging system, the color filter and overlapping region removal could accurately and reliably segment grapevine canopies and identify repeatedly imaged regions between consecutive image frames, leading to critical information for infection severity calculation. Image-derived severity rates were highly correlated (r > 0.95) with human-assessed values, and had comparable statistical power in differentiating fungicide treatment efficacy in both case studies. Therefore, the developed approach and pipeline can be used as an effective and efficient tool to quantify the severity of foliar disease infections, enabling objective, high-throughput disease evaluation for fungicide trial evaluation, genetic mapping, and breeding programs.

Plant Disease Sensing: Studying Plant-Pathogen Interactions at Scale.

Plant disease threatens the environmental and financial sustainability of crop production, causing $220 billion in annual losses. The dire threat disease poses to modern agriculture demands tools for better detection and monitoring to prevent crop loss and input waste. The nascent discipline of plant disease sensing, or the science of using proximal and/or remote sensing to detect and diagnose disease, offers great promise to extend monitoring to previously unachievable resolutions, a basis to construct multiscale surveillance networks for early warning, alert, and response at low latency, an opportunity to mitigate loss while optimizing protection, and a dynamic new dimension to agricultural systems biology. Despite its revolutionary potential, plant disease sensing remains an underdeveloped discipline, with challenges facing both fundamental study and field application. This article offers a perspective on the current state and future of plant disease sensing, highlights remaining gaps to be filled, and presents a bold vision for the future of global agriculture.

Plant pest surveillance: from satellites to molecules.

Plant pests and diseases impact both food security and natural ecosystems, and the impact has been accelerated in recent years due to several confounding factors. The globalisation of trade has moved pests out of natural ranges, creating damaging epidemics in new regions. Climate change has extended the range of pests and the pathogens they vector. Resistance to agrochemicals has made pathogens, pests, and weeds more difficult to control. Early detection is critical to achieve effective control, both from a biosecurity as well as an endemic pest perspective. Molecular diagnostics has revolutionised our ability to identify pests and diseases over the past two decades, but more recent technological innovations are enabling us to achieve better pest surveillance. In this review, we will explore the different technologies that are enabling this advancing capability and discuss the drivers that will shape its future deployment.

Investigating potato late blight physiological differences across potato cultivars with spectroscopy and machine learning.

Understanding plant disease resistance is important in the integrated management of Phytophthora infestans, causal agent of potato late blight. Advanced field-based methods of disease detection that can identify infection before the onset of visual symptoms would improve management by greatly reducing disease potential and spread as well as improve both the financial and environmental sustainability of potato farms. In-vivo foliar spectroscopy offers the capacity to rapidly and non-destructively characterize plant physiological status, which can be used to detect the effects of necrotizing pathogens on plant condition prior to the appearance of visual symptoms. Here, we tested differences in spectral response of four potato cultivars, including two cultivars with a shared genotypic background except for a single copy of a resistance gene, to inoculation with Phytophthora infestans clonal lineage US-23 using three statistical approaches: random forest discrimination (RF), partial least squares discrimination analysis (PLS-DA), and normalized difference spectral index (NDSI). We find that cultivar, or plant genotype, has a significant impact on spectral reflectance of plants undergoing P. infestans infection. The spectral response of four potato cultivars to infection by Phytophthora infestans clonal lineage US-23 was highly variable, yet with important shared characteristics that facilitated discrimination. Early disease physiology was found to be variable across cultivars as well using non-destructively derived PLS-regression trait models. This work lays the foundation to better understand host-pathogen interactions across a variety of genotypic backgrounds, and establishes that host genotype has a significant impact on spectral reflectance, and hence on biochemical and physiological traits, of plants undergoing pathogen infection.

A multi-omics approach to solving problems in plant disease ecology.

The swift rise of omics-approaches allows for investigating microbial diversity and plant-microbe interactions across diverse ecological communities and spatio-temporal scales. The environment, however, is rapidly changing. The introduction of invasive species and the effects of climate change have particular impact on emerging plant diseases and managing current epidemics. It is critical, therefore, to take a holistic approach to understand how and why pathogenesis occurs in order to effectively manage for diseases given the synergies of changing environmental conditions. A multi-omics approach allows for a detailed picture of plant-microbial interactions and can ultimately allow us to build predictive models for how microbes and plants will respond to stress under environmental change. This article is designed as a primer for those interested in integrating -omic approaches into their plant disease research. We review -omics technologies salient to pathology including metabolomics, genomics, metagenomics, volatilomics, and spectranomics, and present cases where multi-omics have been successfully used for plant disease ecology. We then discuss additional limitations and pitfalls to be wary of prior to conducting an integrated research project as well as provide information about promising future directions.