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The Plant Phenome Journal

○ Wiley

Preprints posted in the last 90 days, ranked by how well they match The Plant Phenome Journal's content profile, based on 14 papers previously published here. The average preprint has a 0.01% match score for this journal, so anything above that is already an above-average fit.

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From Field Photosynthesis to Genetic Architecture: Insights from the First Dedicated Photosynthesis Hackathon

Matuszynska, A.; Sansa, O.; Adekoya, F. J.; Akinyemi, O. O.; Anokye, E.; Bashir, O. B.; Boyny, Z. Z. F.; Chukwuka, M. K.; Corvest, E.; Dada, A. O.; DellAcqua, M.; Ehemba, G. L.; Finkbeiner, A. J.; Hamabwe, S.; Hodehou, D. A. T.; Kacheyo, O.; Kamfwa, K.; Mhango, K. J.; Abdullahi, W. M.; Munduwe, G.; Ntukidem, S.; Obisesan, O. K.; Odesina, I. S.; Ogechi, N.-U.; Olaoye, O. D.; Olayinka, M. M.; Osei-Bonsu, I.; Rilwan, K. O.; Stival, L.; Tehar, Z.; Tende, R. M.; To, J.; Ugochukwu, U. K.; Unger, A.; van Aalst, M.; Vrbic, D.; Zhang, C.; Theeuwen, T. P. J. M.; Kramer, D. M.; Kromdijk, J.

2026-08-17 plant biology 10.64898/2026.07.24.740625 medRxiv
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Photosynthesis is among the most consequential yet genetically complex traits in crop plants, and translating its natural variation into actionable genomic targets remains a central challenge for breeding climate-resilient varieties. To start addressing this, researchers are generating increasingly large, multi-environment field photosynthesis datasets. Yet, these data have been structurally under-analysed since their inception. Here we report the outcomes of the first dedicated hackathon focused on computational mining of such field data held in Accra, Ghana, in March 2026. Bringing together data scientists, plant physiologists, geneticists, and breeders from Europe and Africa, these interdisciplinary teams used photosynthetic data collected with hand-held fluorometers to genome-wide marker data across four crop species: cowpea (Vigna unguiculata), barley (Hordeum vulgare), common bean (Phaseolus vulgaris), and potato (Solanum tuberosum). Despite using different species and methods, independent teams identified the same three key findings. First, mechanism-informed feature engineering and dynamic modelling recover genetic signals that are not detected or discarded in standard analysis pipelines, resulting in traits with improved heritability and meaningful associations with yield. Secondly, machine learning methods proved effective at uncovering genetic associations, with temporally resolved features substantially outperforming single time-point measurements. Third, raw chlorophyll fluorescence and absorbance traces consistently contained more information and predictive power than the extracted parameters currently used. A defining feature of this event was having experimentalists and data scientists working together, enabling AI approaches to be grounded in domain knowledge and biological mechanisms rather than relying on data alone.

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User-friendly software for high-throughput processing of chlorophyll fluorescence images to assess plant stress responses: fvfmPy and fvfmR packages

Garen, J. C.; Arnold, P. A.; Crous, K. Y.

2026-10-09 plant biology 10.64898/2026.10.06.757212 medRxiv
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Chlorophyll fluorescence imaging is a powerful tool used to characterize abiotic and biotic stress response of photosynthesis in plants. However, extracting metrics such as Fv/Fm (maximum quantum yield of photosystem II) from chlorophyll fluorescence images remains a substantial bottleneck in large measurement campaigns, often requiring significant manual effort. To resolve this issue, we developed fvfmPy, a user-friendly program which enables rapid analysis of chlorophyll fluorescence images of arrays of leaf discs. fvfmPy uses algorithmic image segmentation to identify regions of interest and extract fluorescence metrics from tens or hundreds of leaf samples per image with little to no manual input. fvfmPy is developed in Python and includes a graphical user interface enabling intuitive parameter adjustment and real-time quality control checking. We additionally developed fvfmR, an R wrapper package for fvfmPy. Fv/Fm values extracted with fvfmPy were indistinguishable from those obtained using manual methods (r2 > 0.99) but required 94% less time to complete (13 s vs. 200 s per 100 measurements). fvfmPy and fvfmR were built to directly integrate with existing fluorescence software packages and can greatly increase throughput of fluorescence measurements to help address key challenges in agriculture and ecology.

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Drought-Spec-Net: Early Tomato Drought Detection and Potential Yield-Impact Assessment Using Vis NIR Data

Hossain, K.; Maiti, A.; Chinnannan, K.; Nimmakayala, P.; Sweeney, E.; Wang, Z.; Chiwele, N.; Khamboo, L.; Nallagari, L. M.; Bucksch, A.

2026-09-25 plant biology 10.64898/2026.09.24.754113 medRxiv
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Drought stress significantly reduces tomato (Solanum lycopersicum L.) productivity, and early detection is critical to minimize yield losses through timely interventions. In this study, we developed Drought-Spec-Net, a hybrid 1D convolutional neural network that integrates local and global spectral feature extraction to detect early drought stress from visible and near infrared (Vis NIR) spectra data of tomato seedlings. The model was trained on 378 samples using an 80:20 train test split, with 20% of the training set reserved for validation. DroughtSpecNet outperformed the evaluated baseline and state of the art models, achieving 97% accuracy, 95% precision, 98% recall, and an F1 score of 97%. To improve the agronomic interpretation of the model outputs, predicted drought probabilities were converted into a literature-informed potential yield impact indicator using a maximum impact level of 60%. On the test set (76 samples), mapped potential yield-impact values ranged from 0% to 60%, with an average reduction of 12.97%. We also conducted an initial experiment using our greenhouse RGB dataset, collected daily from drought treated and well-watered tomato plants at West Virginia State University (WVSU). From this dataset, 44 images were selected for ilastik-based canopy segmentation, producing plant-level drought severity indices (DSI) with a mean of 0.28, median of 0.14, and range of 0.01 to 0.91. Additionally, we trained and fine-tuned a large language model (LLM) based on PLLaMA7BInstruct, called AgriLLaMA, for automated agronomic report generation from Drought-Spec-Net outputs. The generated reports summarize predicted stress levels, mapped potential yield impacts, and preliminary management considerations. This integrated approach not only improves early drought stress detection but also delivers quantitative and interpretable estimates of potential productivity losses, providing a complete framework connecting physiological stress detection to actionable agricultural outcomes.

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Vision Transformers Enable Advanced Plant Phenotyping in Controlled Environments

Milligan, J.; Seethepalli, A.; Tsaris, A.; Wang, X.; York, L.; Lagergren, J. H.

2026-09-10 plant biology 10.64898/2026.09.04.748299 medRxiv
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Reliable plant segmentation in high-throughput phenotyping must transfer across species and imaging conditions without repeated model tuning or extensive reannotation. We compare three segmentation strategies using images from Oak Ridge National Laboratory's Advanced Plant Phenotyping Laboratory: (i) fixed color-based thresholding, (ii) supervised U-Nets trained from scratch, and (iii) pretrained vision transformers fine-tuned for binary segmentation. Models were evaluated on a held-out test set and a generalization set that comprised unseen species. On the held-out test set, thresholding, the best U-Net, and the best vision transformer achieved mean Dice scores of 58.3, 96.6, and 97.3, respectively. On the generalization set, the corresponding Dice scores were 56.5, 86.2, and 95.7. Thresholding remained effective on some datasets but failed when plant appearance changed. Supervised U-Net training resolved within-distribution errors but failed to generalize to novel species and backgrounds. Pretrained vision transformers consistently produced high-accuracy segmentations across the evaluated species, views, soil backgrounds, and tray types. These results benchmark the practical progression from fixed rules to task-specific supervision and pretrained visual representations for controlled-environment plant phenotyping.

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Early-life stage phenomic prediction of field agronomic traits across breeding cycles in intermediate wheatgrass

Harris, Z. N.; Braley, J.; Cassetta, E.; Crain, J.; DeHaan, L.; Van Tassel, D.; Miller, A.; Rubin, M. J.

2026-08-31 plant biology 10.64898/2026.08.28.747871 medRxiv
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Perennial grains represent a promising frontier for sustainable agriculture, but breeding progress is constrained by the accessibility of genotyping and the difficulty of evaluating complex traits expressed for multiple years after establishment across heterogeneous environments. Phenomic selection may help address these challenges by using inexpensive, scalable, high-dimensional phenotypes collected early in development, although the robustness of such predictions across breeding cycles remains uncertain. Here, we compared genomic selection and phenomic selection across two breeding cycles of Thinopyrum intermedium (intermediate wheatgrass; IWG; Kernza(R)), comprising approximately 2,280 individuals from maternal half-sib families evaluated across multiple field sites and years. We constructed relationship matrices from genomic markers and early-life stage phenomic data, including seed and leaf color (HSV), CropReporter multispectral reflectance and indices, and cycle-specific hyperspectral reflectance sensors. Genomic models provided the strongest predictions on average across all field traits in both cycles. Among phenomic predictors, leaf HSV was consistently the most informative, whereas CropReporter and hyperspectral data showed lower and more trait-dependent performance and seed HSV provided little predictive value. Genomic, leaf HSV, and CropReporter models transferred across breeding cycles with little apparent loss of predictive ability relative to within-cycle validation, demonstrating that their predictive signals were not restricted to a single breeding cycle. Early-life stage leaf HSV emerged as a practical, accessible tool for germplasm thinning and early-stage prioritization in perennial breeding programs. Despite limited similarity among relationship matrices, multi-relationship-matrix models rarely improved prediction beyond the stronger constituent single-relationship-matrix model. Together, these results show that early-life stage phenomic data provide reproducible information about agronomic performance expressed years later, but that predictor complexity and data integration do not guarantee improved prediction.

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Combining 3D-multispectral and hyperspectral imaging to identify environmental stress treatments imposed during plant growth

Stock, F.; Panda, S.; Poire, R.; Brown, T.; Akram, A.; Zheng, L.; Lei, H.; Zha, R.; Zhao, M.; Isabelle, S.; Martel, M.; Comeau, M.-A.; Hamel, L.-P.; Lavoie, P.-O.; D'Aoust, M. A.; Reithinger, H.; Saxena, P.; Stone, E. A.; Li, H.; Way, D. A.; Atkin, O. K.

2026-08-28 plant biology 10.64898/2026.08.28.747774 medRxiv
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Non-invasive, high-throughput phenotyping tools are needed that can identify environmental effects on plant structure and function to diagnose factors responsible for reduced growth in commercial and non-commercial settings. In this study, we explored whether the integration of 3D-multispectral (3D) and 2D-hyperspectral imaging (HSI), aided by machine learning (ML), could be used to identify environmental stress treatments imposed during plant growth. Controlled environment-grown Nicotiana Benthamiana plants were subjected to a range of abiotic treatments - including different growth irradiances, heat treatment and drought stress - with the treatments resulting in differences in shoot height, biomass, leaf area and spectral reflectance. ML models were trained to identify these treatments using morphological and spectral traits measured at 27, 29, 31, and 34 days after sowing (DAS). A 3D-multispectral scanner was used to obtain information on plant height, biomass, and leaf area. A visible and near-infrared (VNIR) HSI camera provided detailed spectral information for deriving spectral indices including the Normalised Difference Vegetation Index (NDVI), Photochemical Reflectance Index (PRI) and Normalized Difference Red Edge (NDRE). Manual measurements provided baseline comparative data. The 3D-multispectral scanner reliably estimated above-ground traits, with high correlations between manual and scanner-derived measurements. The ML models accurately differentiated among environmental stress treatments, with the fused 3D+HSI model achieving the best overall predictive performance across all evaluated metrics compared with models based on either imaging modality alone. Results demonstrated the effectiveness of combining 3D-multispectral and 2D-HSI data with ML analyses for non-destructive, high-throughput phenotyping. The integration of these techniques enabled non-destructive, high-throughput identification of environmental stress treatments imposed during plant growth.

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PhenoStream: A Cyberinfrastructure for Automated and AI-Based Crop Trait Extraction from Aerial Imagery

Varela, S.; Ruhter, J.; Sacks, E.; Zheng, X.; Allen, D.; Hale, A.; Landry, C.; Kuang, X.; Long, B.; Zhu, Y.; Proma, S.; Kaur, S.; Jarquin, D.; Morrison, J.; Leakey, A.

2026-08-30 plant biology 10.64898/2026.08.26.747008 medRxiv
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The integration of digital technologies for high-throughput field phenotyping is critical for accelerating crop improvement in agriculture. However, extracting traits from remote sensing data remains constrained by fragmented workflows, manual intervention, and limited interoperability among existing tools, resulting in delays that hinder timely biological insight and decision-making. To address these challenges, we present PhenoStream (Phenotyping Streaming), a scalable, end-to-end cyberinfrastructure designed to automate the full lifecycle of aerial imagery-based phenotyping, from data acquisition to plot- and genotype-level inference. The framework integrates automated data ingestion from distributed field sites, geospatial processing, and AI-enabled trait extraction within a unified, user-accessible graphical interface. Its modular and extensible architecture supports adaptable trait modeling and seamless integration of new data sources, enabling deployment across diverse crops, environments, and experimental designs. We demonstrate the system across a large multi-location field trial network of bioenergy crops, where it enables high-throughput characterization of spatiotemporal growth dynamics, genotype-by-environment (GxE) interactions, and predictive modeling of key agronomic traits. By significantly reducing processing latency and manual effort, the platform facilitates near-real-time analysis and reproducible workflows. This work establishes a generalizable and scalable pathway for operationalizing very-high-spatial resolution aerial phenotyping in agricultural research. By bridging data acquisition and analytics, the end-to-end cyberinfrastructure provides a foundation for integrating heterogeneous and unstructured data streams--including remote sensing, environmental, and management data--toward data-driven decision making in agriculture.

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Detection of Red Crown Rot of Soybean in Illinois Fields Using High-Resolution Satellite Imagery and Machine Learning

Pugliese, B. D.; Paredes, J. A.; Ruiz, A. F.; Bowman, N. D.; Ersoz, E.; Martin, N. F.; Camiletti, B. X.

2026-09-23 plant biology 10.64898/2026.09.22.753594 medRxiv
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Red crown rot (RCR), caused by Calonectria ilicicola, is an emerging soybean disease in the U.S. Midwest for which scalable approaches to characterize within-field disease distribution are lacking. This study evaluated high-resolution PlanetScope satellite imagery for mapping RCR-affected soybean canopies across 15 commercial fields in Illinois surveyed during the 2024 and 2025 growing seasons. A total of 2,921 georeferenced canopy plots were classified as asymptomatic or RCR-affected and paired with six multispectral bands and seven vegetation indices. Spectral differences between classes were evaluated using linear mixed-effects models, and seven machine-learning classifiers representing linear, tree-based, neural-network, kernel, and probabilistic approaches were compared using spatially independent leave-one-field-out cross-validation. RCR-affected canopies exhibited increased reflectance in the visible and red-edge regions, reduced near-infrared reflectance, and lower vegetation-index values relative to asymptomatic canopies. All classifiers showed strong discrimination, with ROC-AUC values ranging from 0.963 to 0.982. Regularized logistic regression achieved the highest overall performance, with an accuracy of 0.945, balanced accuracy of 0.945, F1-score of 0.948, and ROC-AUC of 0.982 at the optimized decision threshold. Permutation analysis identified EVI, NDVI, and red reflectance as the most influential predictors across representative model architectures. Satellite-derived probability and classification maps generally corresponded with symptomatic canopy patterns observed in high-resolution UAV imagery, although mixed pixels reduced precision near disease-patch boundaries. These results demonstrate the potential of high-resolution satellite imagery for within-field mapping of RCR-associated canopy symptoms across independent commercial soybean fields.

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A breeder in the sky: scoring flowering with fewer flights

Ji, Z.; DeSalvio, A. J.; Adak, A.; Arik, M. A.; Ertl, D.; Kaeppler, S. M.; Edwards, J.; Flint-Garcia, S.; Hirsch, C. N.; Thompson, A. M.; Washburn, J. D.; de Leon, N.; Maharjan, B.; Santra, D.; Izere, P.; Zhao, B.; Shi, Y.; Murray, S. C.; Ge, Y.; Schnable, J. C.

2026-09-22 plant biology 10.64898/2026.09.19.752900 medRxiv
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Flowering time is a trait of broad interest and importance to both plant breeders and plant biologists. Unlike many other traits, flowering time cannot be scored accurately from a single observation; its measurement requires repeated observations of the same experiments over time. Both classification- and object-detection-based approaches have demonstrated the potential to estimate flowering time from Unmanned aerial vehicle (UAV) imagery rather than by manual observation, but they still require data collection at many time points. Here, we train and deploy a regression-based framework for scoring flowering time. Using a dataset of more than 200,000 UAV images and associated flowering-time records collected across 27 environments, we demonstrate that this approach enables the prediction of flowering time from as few as one observation per field experiment and successfully generalizes to field experiments in environments not represented in the training dataset. These results indicate that regression-based approaches for predicting flowering time from sparse UAV observations have the potential to substantially reduce the data-collection burden in field experiments

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A data-driven approach to automate embolism detection in leaves

Mandelli, L.; Johnson, K. M.; Berretti, S.; Mencuccini, M.

2026-08-03 plant biology 10.64898/2026.07.31.741439 medRxiv
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1O_LIEmbolism, the formation of air bubbles in the plant water transport system, is a mechanistic driver of plant death. The Optical Vulnerability Technique (OVT) is an imaging method for non-invasive quantification of embolism (including P50, a common metric for drought vulnerability), which can also provide detailed spatial and temporal information. Its major cost lies in the post-processing of thousands of images. C_LIO_LIHere we designed, tested, trained, and make publicly available a neural network model to automate post-processing of OVT images. Using a dataset of 65 leaves from Senecio pterophorous, we compared our model predictions to results obtained via traditional post-processing by an expert. C_LIO_LIOur model resolved P50 to within 0.027 MPa of the expert-processed data with training taking 30 minutes to 2.5 hours and model-runtime in the order of seconds to minutes, demonstrating its promise for increasing the efficiency and throughput of P50 calculation. The models performance in replicating the pixels that constitute embolism events was lower (mean event-frame IoU of 0.38). C_LIO_LIWe invite the community to utilise our model but emphasise that it does not replace the expert-processing pipeline and that care must be taken when considering applying this and similar approaches to OVT data. C_LI

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DIRTNet: Enabling Root Phenotyping with Fiber Bragg Grating Sensors

Hossain, K.; Binder, S.; Fok, M.; Bucksch, A.

2026-09-23 plant biology 10.64898/2026.09.22.753535 medRxiv
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Non-destructive methods for real-time phenotyping of root growth and development are essential to accelerate the breeding of climate-resilient crops. We present DIRTNet (Digital Investigations of Root Traits Net), a hybrid deep learning model that interprets in-soil signals collected by Fiber Bragg Grating (FBG) sensors. We first evaluated DIRTNet in a controlled simulation study in which metal rods of varying diameters were inserted into soil to emulate root growth. Using augmented training data with an 80:20 train-test split, DIRTNet surpassed baseline models, achieving 0.97 accuracy for root depth prediction and 0.94 for root diameter prediction. We then applied the best-performing models to maize (Zea mays) roots monitored by six FBG sensors placed at different soil location and depths. A control trial and a drought-stress trial yielded 192 samples, split 80:20 into training and holdout sets; the hold-out portion was divided equally into validation and test sets. Augmentation was applied only to the training data. On real maize root data, DIRTNet achieved 0.92 accuracy for both root depth and root width prediction across variable growth periods, covering root mass lengths up to 20 cm and diameters up to 10 cm. The prediction accuracy is independent of the soil moisture and drought condition. In a fixed-interval analysis, accuracy reached 0.97 for days 25-42 after planting and 0.93 for days 25-48, demonstrating robust performance as root complexity increased.

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One-dimensional CNNs for Near-Infrared Prediction of Protein and Moisture in Cereal Grains: The Effects of Architecture and Input Preparation

Deng, G.; Chi, W.; Yu, P.; WU, F.

2026-09-18 bioinformatics 10.64898/2026.09.12.751160 medRxiv
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Near-infrared (NIR) spectroscopy is widely used for the rapid, non-destructive determination of constituents such as protein and moisture in cereal grains, but model comparisons in this field are often confounded by differences in the inputs received by each model. We benchmark a compact one-dimensional CNN derived by one-factor-at-a-time ablation (CNN-Baseline) and a randomly searched CNN (CNN-RS) against PLSR, SVR, XGBoost on six cereal-grain datasets (n=500-5046) under three input conditions: raw spectra, optimal preprocessing, and preprocessing plus wavelength selection. A single-filter with a kernel size of 11 of convolution and batch normalization proved sufficient. Given a unified raw input, CNN-RS was most accurate on five of the six datasets, while CNN-Baseline came within 0.003-0.017 in test R2 on the four larger datasets. PLSR led only on the smallest protein dataset (XDS, n=500) and was the most stable model. Preprocessing had almost no effect on CNN accuracy on the four larger datasets, but improved accuracy by 0.034-0.047 (CNN-Baseline) and 0.017-0.055 (CNN-RS) on the two smallest, and brought the networks to their optimum in a median of 49% fewer epochs. Therefore, replacing explicit preprocessing with learned filters was only justified when sufficient training data were available. Once each model received its own preprocessing and wavelength subset, SVR became most accurate on four of six datasets and XGBoost rose from the weakest model to within 0.01-0.03 of the best model on four of the six datasets, suggesting that the principal advantage of CNNs lied in robustness to raw input data rather than in attainable accuracy. Overall, model selection for NIR calibration of cereal grains depends more on sample size and the input preparation than on network architecture.

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An open field phenomics resource for multimodal maize yield prediction across divergent environments

DeSalvio, A. J.; Mohseni, P.; Adak, A.; Murray, S. C.; Arik, M. A.; Wong, R. K. W.; Jung, J.; Lima, D. C.; Aviles, A. C.; Buckler, E. S.; Duffield, N.; Edwards, J.; Ertl, D.; Flint-Garcia, S.; Gore, M. A.; Hirsch, C. N.; Holland, J. B.; Kaeppler, S. M.; Miller, J.; Romay, C.; Schnable, J. C.; Singh, M. P.; Sparks, E. E.; Thompson, A.; Washburn, J. D.; Weldekidan, T.; Winans, N. D.; de Leon, N.

2026-09-18 plant biology 10.64898/2026.09.17.752474 medRxiv
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Temporal drone phenotyping captures crop development, but irregular flight schedules complicate comparisons across environments. We release curated imagery from 356 flights across 19 Genomes to Fields environments containing 1,180 maize (Zea mays L.) hybrids. To evaluate its utility, we integrated functional principal components of vegetation index and weather trajectories with genomic information. Combinined genomic and phenomic kernels improved yield prediction, reaching correlations up to r = 0.501 for held-out hybrids in environments represented in training and 0.408 when environments were also withheld. Accumulated growing degree days offered no consistent predictive advantage over days after planting, and weather contributed modest, task-dependent gains. A transformer neural process learned directly from irregular observations, serving as a novel application of neural process models in agriculture. Mapping vegetation index functional principal components identified recurrent quantitative trait loci on chromosomes 3 and 7. This resource and its reproducible analyses guide the use of temporal spectral data for crop prediction and genetic discovery.

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Model-assisted high-throughput phenotyping of photosynthetic acclimation

Alegria, E. V.; Russell, N. J.; Chen, T.-W. J.

2026-09-22 plant biology 10.64898/2026.09.21.753097 medRxiv
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Photosynthesis is a key determinant of crop productivity, yet conventional gas-exchange measurements are labor-intensive and unsuitable for high-throughput assessment of dynamic photosynthetic acclimation across large genotype panels. Consequently, genetic variation in photosynthetic acclimation remains poorly characterized and difficult to exploit in breeding programs. Here we present a computational pipeline that integrates rapid optical sensing (chlorophyll meters and hyperspectral reflectance) with a mechanistic model of photosynthetic protein-turnover to estimate dynamic nitrogen allocation among photosynthetic components. The pipeline was demonstrated using 60 winter wheat (Triticum aestivum L.) cultivars sampled at 10 time points spanning leaf emergence to senescence. Uncertainties associated with each pipeline component were quantified and benchmarked against gas-exchange ground-truth measurements under three controlled-environment light and temperature regimes. Dynamic nitrogen allocation to light harvesting, electron transport, and carboxylation was characterized using three biologically interpretable parameters: maximum synthesis rate, degradation rate, and age-dependent decline in synthesis. Although hyperspectral models showed moderate predictive accuracy for photosynthetic capacity, prediction errors were predominantly random, allowing robust parameter estimation when observations across the leaf lifespan were integrated. The pipeline successfully resolved genotype-by-environment interactions in photosynthetic acclimation, with environmental and interaction effects contributing more strongly than genotype main effects to nitrogen dynamics. These results demonstrate a scalable, uncertainty-aware sensor-to-trait architecture for dynamic physiological phenotyping that enables high-throughput germplasm screening across diverse environments and provides novel physiological selection targets for improving crop adaptation under variable climates.

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YieldLearn: An interpretable machine-learning pipeline for estimating yield-related traits based on physiological indicators

Zhang, X.; Shaterian, H.; Ashe, P.

2026-09-28 bioinformatics 10.64898/2026.09.22.753232 medRxiv
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Efficient yield prediction under field conditions remains a key challenge for crop breeders and agronomists. YieldLearn is an interpretable machine-learning pipeline developed to estimate yield-related traits in durum wheat (Triticum durum) using physiological indicators measured by the Hansatech Instruments Pocket PEA. The workflow integrates fluorescence parameters (Area, Fo, Fm, Fv/Fm, PI Inst.) with field-measured agronomic traits (Booting, Height, Maturity, Lodging, Yield kg/plot, and Adjusted Yield kg/ha). Five supervised learning algorithms multinomial logistic regression, random forest, support vector machine (radial kernel), k-nearest neighbors, and decision tree were evaluated using five-fold cross-validation. Models achieved average classification accuracy around 0.57 with AUC values exceeding 0.58. SHAP (SHapley Additive exPlanations) visualizations revealed that high Fv/Fm and PI Inst. values were positively associated with yield potential, whereas elevated Fo indicated stress-induced yield reduction. An interactive Shiny App enables real-time prediction and feature-importance exploration. Given the small sample size (165 plots) and absence of environmental covariates such as temperature or soil moisture, YieldLearn should be regarded as a reproducible exploratory framework rather than a production-ready predictor. Nonetheless, it offers a valuable foundation for guiding experimental design, validating physiological indicators, and supporting the next generation of scalable, interpretable crop-yield models.

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Low-cost rhizotron imaging and zero-shot deep-learning resolve temporal, spatial, and genetic variation in grapevine rootstock root systems

Diaz-Garcia, L.; Munoz, J. R.; Torres-Lomas, E.; McElrone, A. J.; Sharma, S.; Lupo, Y.

2026-09-14 plant biology 10.64898/2026.09.10.750709 medRxiv
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Root system architecture shapes how grapevine rootstocks take up water and nutrients, yet roots remain the least phenotyped grapevine organ because they are hidden and hard to image. We present a low-cost phenotyping pipeline that pairs custom acrylic rhizotrons (about US$30 each) with a consumer flatbed scanner and BiRefNet, a general-purpose deep-learning model used without training on root images, followed by automated mask cleaning, skeleton-based trait extraction, and soil moisture mapping. We tested it on nine commercial rootstocks scanned 16 times over 42 days after transplanting (DAT), with half under a ten-day water deficit. From 1,108 images we extracted 21 whole-root, depth-resolved, and topological traits. Genotypes differed in nearly every trait and in how they changed over time. Heritability of size and branching traits peaked at 0.92-0.93 between 21 and 31 DAT and fell for width, depth, and convex hull once roots reached the rhizotron walls, defining the best measurement window. The image-derived soil moisture map accurately tracked the deficit and its recovery. Deficit plants shifted new root growth to deeper soil without growing less overall, and the substrate dried fastest around older and denser roots. Root brightness decreased with root age and local moisture, and transport segments (axes serving several tips) were brighter than terminal laterals in every genotype. Root system size was associated with stomatal conductance in well-watered plants, and stomatal recovery after re-watering correlated with new root growth. The pipeline turns simple hardware into a quantitative, time-resolved root phenotyping platform suitable for breeding.

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MIRA: an open source and user-friendly software to automate counting and sizing of fungal spores

Mejias, J.; Adreit, H.; Blanc, A.; Lubin, N.; Jolivet, C.; Guyot, V.; Brayle, O.; Poncelet, N.; Fournier, E.; Wicker, E. P.; Carlier, J.; Tharreau, D.; Ravel, S.

2026-08-07 plant biology 10.64898/2026.08.06.743221 medRxiv
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BackgroundThe quantification of fungal spores constitutes a fundamental metric in phytopathology, serving as the primary variable for inoculum standardization and being used as a proxy for disease severity. Historically, spore quantification has relied on manual hemocytometry, which remains the most precise counting process to date, where chambers such as the Malassez slide are used to count a subsample of the inoculum. However, this method applied manually is highly labor-intensive, time-consuming, and can be prone to operator-dependent variability. To overcome these limitations, we introduce MIRA (Microscopy Image Recognition & Analysis), a novel open-source software integrating You Only Look Once (YOLO) deep learning algorithms. Featuring a user-friendly graphical interface, MIRA is adaptable to multiple camera systems and supports advanced object detection models, including YOLOv11 and YOLOv26. ResultsWe demonstrate that MIRA can be used to accurately detect and count spores from several phytopathogenic fungi, automatically measure spore surface area, and to differentiate spores across different genera. In an exhaustive comparative analysis using Pyricularia oryzae spores as an example, MIRA was benchmarked against manual gold-standard counting slides (Malassez and Kova) and indirect spectrophotometric methods (SPARK). The P. oryzae model loaded via MIRA achieved a strong correlation (R = 0.96) with manual gold standards while reducing processing time by over 90% for high-concentration samples (10 spores/mL). Beyond this benchmark, we also successfully tested specific YOLO models designed to recognize macro- and microconidia of Fusarium oxysporum f. sp. cubense, a model for Pseudocercospora fijiensis, and a single multiclass model capable of identifying six different rice pathogenic fungi. We provide comprehensive tutorials for operating the software and training custom detection models for free using Roboflow and Google Colab. MIRA is available both as open-source Python code and as standalone executables for Windows and Linux. ConclusionsMIRA provides a rapid, accurate, and highly reproducible alternative to manual spore counting, effectively removing a major bottleneck in phytopathology workflows. By combining advanced YOLO-based deep learning with an accessible interface and comprehensive training resources, MIRA makes accessible automated image analysis for researchers without programming expertise. Moreover, MIRA drastically improves the efficiency of high-throughput disease phenotyping and can be adapted for a wide range of microscopic quantification tasks across various biological disciplines.

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Time-resolved volatile organic compound profiling enables non-invasive detection of phenological progression in soybean

Nakata, R.; Hiraga, S.; Ishimoto, M.

2026-08-28 plant biology 10.64898/2026.08.28.747781 medRxiv
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Background and aims Plant volatile organic compounds (VOCs) change dynamically with plant development and in response to environmental conditions. However, their potential as non-invasive indicators of phenological progression remains poorly explored. In this study, we developed a framework integrating automated VOC sampling, time-resolved VOC profiling, and machine-learning analysis for the non-invasive assessment of plant phenology. Using soybean (Glycine max (L.) Merr.), we investigated whether development-associated temporal variation in VOC emissions could delineate and predict developmental phases. Methods We collected VOCs daily under controlled environmental conditions from 16 to 43 days after sowing, spanning the transition from vegetative to reproductive stages, using an automated sampling system coupled with thermal desorption-gas chromatograph-mass spectrometer (TD-GC-MS). To characterise temporal changes in VOC profiles associated with phenological progression, we analysed the daily VOC data using a multi-step pipeline combining statistical filtering and similarity-based network analysis. We defined VOC-derived developmental phases from similarity patterns in the VOC profiles, then developed and evaluated machine-learning models to predict these phases. Key results Seven VOCs exhibited distinct phase-dependent dynamics, including green leaf volatiles and monoterpenes showing characteristic temporal changes during phenological progression. Network-based clustering of VOC profiles resolved five developmental phases closely aligned with conventional developmental stages. A machine-learning model predicted these phases from the VOC profiles with high predictive accuracy on independent test data, demonstrating that phenological progression could be quantitatively inferred from VOC emission patterns. Conclusions Our findings support VOC profiling as a reliable and non-invasive approach for assessing phenological progression in soybean. By extracting temporally structured VOC signals, this framework captures developmental information that may be difficult to obtain through visual observation alone, particularly after canopy closure. VOC profiling offers a practical tool for monitoring crop developmental dynamics and has broader potential for plant phenotyping and precision crop management.

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An industry perspective on whole genome-informed hybrid maize disease resistance characterization to improve breeding decisions

Check, J. C.; Technow, F.; Totir, L. R.

2026-08-05 pathology 10.64898/2026.07.30.741863 medRxiv
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Characterizing hybrid maize disease resistance is a costly and labor-intensive effort in commercial breeding programs. Field trials are carefully inoculated and managed but remain error-prone due to spatial variability in disease pressure, microclimatic conditions and inter-rater variability. Quantitative ordinal disease rating scales are used to increase scoring speed at the expense of resolution, accuracy, and the ability to use conventional statistical methods. To improve traditional methods of disease resistance characterization, we propose to leverage readily available low-density SNP marker profiles to create genome-informed disease scores. Specifically, a whole genome ordered probit regression (WGOPR) model is used to deconstruct field-observed disease phenotypes into marker effects and reconstruct genome-informed disease scores. This approach is demonstrated in hybrid maize using data from Exserohilum turcicum-inoculated field trials across the central and northern U.S. and Canadian Corn Belt in 2024. Resulting Genomic Estimated Categorical Probabilities (GECPs) are compared to observed frequencies of disease scores to validate the methodology and evaluate the accuracy of regional hybrid maize disease resistance characterization. The benefit of a probabilistic output is demonstrated through two use cases: a comparison of hybrids with highly variable observed disease resistance scores at a single location, and a comparison of breeding selection schemes from a regional analysis. Because GECPs are the product of estimated marker effects, they better represent the expected behavior of a genotype independent of location-, rater- and plot-specific noise, and will therefore offer a step towards improving hybrid maize characterization and better informing breeding decisions.

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Near-infrared phenomic and genomic prediction for seed protein in winter legume white lupin (Lupinus albus L.): A utility comparison

Castillo, M. P.; Oyebode, O. G.; Lenahan, A.; Orloski, A.; Wolfe, M.

2026-08-11 genomics 10.64898/2026.08.05.743001 medRxiv
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White lupin (Lupinus albus L.) is a cool-season grain legume with seed crude protein of 33-47%, competitive with soybean (Glycine max L.) meal. It also fixes nitrogen and mobilizes soil phosphorus. Because soybean is a summer crop, white lupin can occupy Southeastern winter fields as a complementary protein source. Breeding for seed protein is limited by the cost and throughput of reference phenotyping. To determine how each is best deployed, we compared the utility of near-infrared spectroscopy (NIRS)-based phenomic selection with genomic selection based on 246,847 SNPs from low-pass, whole genome sequencing in a panel of Auburn University breeding lines and USDA National Plant Germplasm System germplasm. A handheld NIR calibration against Dumas reference protein reached screening-grade accuracy (R2 = 0.81). Under common cross-validation, phenomic predictive ability was 0.93 and genomic was 0.12. The low genomic value was consistent with moderate heritability (H2 = 0.33) and strong genotype-by-year interaction. Beyond predictive ability, NIRS recovered superior accessions the strictest selection intensity, and 40 to 60 reference assays sufficed to calibrate the model. Handheld NIRS is a low-cost tool for protein calibration and early-generation screening, while genomic prediction remains suited to parental selection, together supporting a complementary strategy for legume breeding Plain Language SummarySoybean meal is the main protein source for livestock and fish farms in the United States. Because soybean is a summer crop, many Southeastern fields sit idle or grow low-value cover crops in winter. White lupin, a cool-season legume whose seeds are as protein-rich as soybean meal, makes a good complementary winter crop: it yields high-protein grain while serving as a cover crop that fixes nitrogen and frees up soil phosphorus for later crops. In our early-stage lupin breeding program, measuring seed protein by standard lab methods is slow and costly. We built a calibration that lets a handheld scanner estimate protein from light, and compared it with predicting protein from the plants DNA. The scanner gave accurate, low-cost protein screening from only about 40-60 lab tests, while DNA-based prediction remains suited to guiding parent selection. Used together, these tools offer breeders a practical path to develop high-protein white lupin. Core ideasO_LIHandheld NIRS provides screening-grade prediction of white lupin seed crude protein. C_LIO_LISpectra carried more usable protein signal than markers by measuring seed chemistry directly. C_LIO_LINIRS and genomic prediction serve different stages of a white lupin breeding program. C_LIO_LIAbout 40 to 60 reference assays sufficed to calibrate NIRS to near-full accuracy. C_LI