Hundreds of species in one of Australia's dominant plant families, the Myrtaceae, are at risk from the invasive pathogenic fungus Austropuccinia psidii. Since its arrival in Australia in 2010, native plant communities have been severely affected, with highly susceptible species likely to become extinct from recurring infections. While severe impact on Australian native and plantation forestry has been predicted, the lemon myrtle industry is already under threat. Commercial cultivars of lemon myrtle (Backhousia citriodora) are highly susceptible to A. psidii. Detecting and monitoring disease outbreaks is currently only possible by eye, which is costly and subject to human bias. This study aims at developing a proof‐of‐concept for automated, non‐biased classification of healthy (naïve), fungicide‐treated and diseased lemon myrtle trees by means of their spectral reflectance signatures. From a lemon myrtle plantation, spectral signatures of fungicide‐treated and untreated leaves were collected using a portable field spectrometer. A third class of spectra, from naïve lemon myrtle leaves that had not been exposed to A. psidii, was collected from a botanical garden. Reflectance spectra in their primary form and their first‐order derivatives were used to train a random forest classifier resulting in an overall accuracy of 78% (kappa = 0.68) for primary spectra and 95% (kappa = 0.92) for first‐order derivative‐transformed spectra. Thus, an optical sensor‐based discrimination, using spectral reflectance signatures of this as yet uninvestigated pathosystem, seems technically feasible. This study provides a foundation for the development of automated, sensor‐based detection and monitoring systems for myrtle rust.

Disease management in agriculture often assumes that pathogens are spread homogeneously across crops. In practice, pathogens can manifest in patches. Currently, disease detection is predominantly carried out by human assessors, which can be slow and expensive. A remote sensing approach holds promise. Current satellite sensors are not suitable to spatially resolve individual plants or lack temporal resolution to monitor pathogenesis. Here, we used multispectral imaging and unmanned aerial systems (UAS) to explore whether myrtle rust (Austropuccinia psidii) could be detected on a lemon myrtle (Backhousia citriodora) plantation. Multispectral aerial imagery was collected from fungicide treated and untreated tree canopies, the fungicide being used to control myrtle rust. Spectral vegetation indices and single spectral bands were used to train a random forest classifier. Treated and untreated trees could be classified with high accuracy (95%). Important predictors for the classifier were the near-infrared (NIR) and red edge (RE) spectral band. Taking some limitations into account, that are discussedherein, our work suggests potential for mapping myrtle rust-related symptoms from aerial multispectral images. Similar studies could focus on pinpointing disease hotspots to adjust management strategies and to feed epidemiological models.

Epidermal structures (ES) of leaves are known to affect the functional properties and spectral responses. Spectral studies focused mostly on the effect of hairs or wax layers only. We studied a wider range of different ES and their impact on spectral properties. Additionally, we identified spectral regions that allow distinguishing different ES. We used a field spectrometer to measure ex situ leaf spectral responses from 350 nm-2500 nm. A spectral library for 25 species of the succulent family Aizoaceae was assembled. Five functional types were defined based on ES: flat epidermal cell surface, convex to papillary epidermal cell surface, bladder cells, hairs and wax cover. We tested the separability of ES using partial least squares discriminant analysis (PLS-DA) based on the spectral data. Subsequently, variable importance (VIP) was calculated to identify spectral regions relevant for discriminating our functional types (classes). Classification performance was high, with a kappa value of 0.9 indicating well-separable spectral classes. VIP calculations identified six spectral regions of increased importance for the classification. We confirmed and extended previous findings regarding the visible-near-infrared spectral region. Our experiments also confirmed that epidermal leaf traits can be classified due to clearly distinguishable spectral signatures across species and genera within the Aizoaceae.

Background In arid environments, plant primary productivity is generally low and highly variable both spatially and temporally. Resources are not evenly distributed in space and time (e.g., soil nutrients, water), and depend on global (El Niño/ Southern Oscillation) and local climate parameters. The launch of the Sentinel2-satellite, part of the European Copernicus program, has led to the provision of freely available data with a high spatial resolution (10 m per pixel). Here, we aimed to test whether Sentinel2-imagery can be used to quantify the spatial variability of a minor tussock grass (Enneapogon spp.) in an Australian arid area and whether we can identify different vegetation cover (e.g., grass from shrubs) along different temporal scenarios. Although short-lasting, the Enneapogon grassland has been identified as a key primary food source to animals in the arid environment. If we are able to identify and monitor the productivity of this species remotely, it will provide an important new tool for examining food resource dynamics and subsequent animal responses to them in arid habitat. Methods We combined field vegetation surveys and Sentinel2-imagery to test if satellite spectral data can predict the spatial variability of Enneapogon over time, through GLMMs. Additionally, a cluster analysis (‘gower’ distance, ‘complete’ method), based on Enneapogon seed-productivity, and total vegetation cover in October 2016, identified three clusters: bare ground, grass dominated and shrub dominated. We compared the vegetation indices between these different clusters from October 2016 to January 2017. Results We found that MSAVI2 and NDVI correlated with the proportion of Enneapogon with seeds across the landscape and this relationship changed over time. Both vegetation indices (MSAVI2 and NDVI) were higher in patches with high seed-productivity of Enneapogon than in bare soil, but only in October, a climatically-favorable period during which this dominant grass reached peak seed-productivity. Discussion MSAVI2 and NDVI provided reliable estimates of the heterogeneity of vegetation type across the landscape only when measured in the Austral spring. This means that grass cover is related to seed-productivity and it is possible to remotely and reliably predict food resource availability in arid habitat, but only in certain conditions. The lack of significant differences between clusters in the summer was likely driven by the short-lasting nature of the vegetation in the study and the sparseness of the grass-dominated vegetation, in contrast to the shrub vegetation cluster that was particularly well measured by the NDVI. Conclusions Overall, our study highlights the potential for Sentinel2-imagery to estimate and monitor the change in grass seed availability remotely in arid environments. However, heterogeneity in grassland cover is not as reliably measured as other types of vegetation and may only be well detected during periods of peak productivity (e.g., October 2016).

Since 2010 Australian ecosystems and managed landscapes have been severely threatened by the invasive fungal pathogen Austropuccinia psidii. Detecting and monitoring disease outbreaks is currently only possible by human assessors, which is slow and labour intensive. Over the last 25 years, spectral vegetation indices (SVIs) have been designed to assess variation in biochemical or biophysical traits of vegetation. However, diagnosis of individual diseases based on classical SVIs is currently not possible because they lack disease specificity. Here, a novel spectral disease index (SDI), the lemon myrtle–myrtle rust index (LMMR), has been developed. The index was designed from hyperspectral leaf‐clip data collected at a lemon myrtle plantation in New South Wales, Australia. A total of 236 fungicide‐treated (disease free) and 228 untreated (diseased) lemon myrtle leaves were sampled and a random forest classifier was used to show that the LMMR discriminates those classes with an overall accuracy of 90%. Compared to three classical SVIs (PRI, MCARI, NBNDVI), commonly applied for stress detection, the LMMR clearly improved classification accuracies (58%, 67%, 60%, respectively). If the LMMR can be validated on independent datasets from similar and different host species, it could enable land managers to reduce disease impact by earlier control. There might also be potential to collect useful data for epidemiology models. Calculating the LMMR based on hyperspectral data collected from aerial platforms (e.g. drones) would allow for rapid and high‐capacity screening for disease outbreaks.

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.

The outwardly directed cell wall and associated plasma membrane of epidermal cells represent the first layers of plant defense against intruding pathogens. Cell wall modifications and the formation of defense structures at sites of attempted pathogen penetration are decisive for plant defense. A precise isolation of these stress-induced structures would allow a specific analysis of regulatory mechanism and cell wall adaption. However, methods for large-scale epidermal tissue preparation from the model plant Arabidopsis thaliana, which would allow proteome and cell wall analysis of complete, laser-microdissected epidermal defense structures, have not been provided. We developed the adhesive tape – liquid cover glass technique (ACT) for simple leaf epidermis preparation from A. thaliana, which is also applicable on grass leaves. This method is compatible with subsequent staining techniques to visualize stress-related cell wall structures, which were precisely isolated from the epidermal tissue layer by laser microdissection (LM) coupled to laser pressure catapulting. We successfully demonstrated that these specific epidermal tissue samples could be used for quantitative downstream proteome and cell wall analysis. The development of the ACT for simple leaf epidermis preparation and the compatibility to LM and downstream quantitative analysis opens new possibilities in the precise examination of stress- and pathogen-related cell wall structures in epidermal cells. Because the developed tissue processing is also applicable on A. thaliana, well-established, model pathosystems that include the interaction with powdery mildews can be studied to determine principal regulatory mechanisms in plant–microbe interaction with their potential outreach into crop breeding.

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