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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.

1
A Data-Driven Image Extraction and Analysis Pipeline for Plant Phenotyping in Controlled Environments

Orvati Nia, F.; Peeples, J.; Murray, S. C.; McFarland, A.; Vann, T.; Salehi, S.; Hardin, R.; Baltensperger, D. D.; Ibrahim, A.; Thomasson, J. A.; Fadamiro, H.; Subramanian, N. K.; Oladepo, N.; Vysyaraju, U.

2026-02-27 plant biology 10.64898/2026.02.25.707797 medRxiv
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Advances in automation, imaging, and artificial intelligence have enabled researchers to capture large volumes of high-quality plant data for understanding crop growth, stress, and genotype-by-environment interactions. While genomics has achieved remarkable throughput, phenotypic data acquisition remains a critical bottleneck for accelerating crop improvement and biological discovery. To address this challenge, an integrated multispectral phenotyping framework was developed using imagery from the Texas A&M AgriLife Precision Automated Phenotyping Greenhouse, a fully controlled facility designed for reproducible plant monitoring throughout the entire growth cycle of most crops. The framework expands the Plant Growth and Phenotyping (PGP v2) dataset and establishes a standardized system for continuous image acquisition, segmentation, deep feature extraction, and temporal analysis across multiple crop species. The project was organized around five coordinated areas: Administration and Coordination, Imaging and Sensor Operations, Data Processing and Management, Artificial Intelligence and Analytics, and Plant Science and Discovery. This structure ensured consistent data quality, version-controlled workflows, and communication across disciplines. The analytical pipeline integrates pseudo-RGB generation, deep learning-based detection and segmentation, image stitching, and temporal (longitudinal) tracking to isolate individual plants and analyze changes in morphology, spectral reflectance, and texture over time. Beyond technical innovation, the framework provides a replicable model for interdisciplinary collaboration and administrative integration in plant phenomics. The combined dataset, workflow, and management framework enable scalable, reproducible, and data-driven plant science research that bridges engineering and biological discovery. Plain Language SummaryTemporal imaging of plants in controlled environments helps scientists better understand growth and biological processes. However, analyzing large volumes of images has been limited by a lack of automated tools. Multispectral imagery captures additional information about plant pigments, structure, and stress beyond standard color images. We developed an automated analysis pipeline that identifies individual plants, tracks their growth over time, and measures traits such as height, area, shape, texture, and vegetation indices. Using artificial intelligence, the system efficiently processes thousands of images to provide consistent and repeatable measurements. By integrating engineering and plant biology, this work supports data-driven decisions for crop improvement and agricultural research.

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A Novel Phenotyping Approach for Reconciling Precision and Variance in Disease Severity Estimates from High-resolution Imaging

Zenkl, R.; McDonald, B. A.; Anderegg, J.

2026-02-20 plant biology 10.64898/2026.02.20.707028 medRxiv
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1Accurate quantification of plant disease is essential for resistance breeding, variety testing, and precision agriculture, yet visual ratings are limited by subjectivity, low precision, and restricted throughput. Image-based phenotyping can address these limitations, but field applications face substantial challenges due to spatial heterogeneity, symptom-level diagnostic requirements, and the need for very high-resolution imagery with limited spatial coverage. This introduces a fundamental trade-off: high-resolution images provide precise local measurements of disease, but spot-level estimates can be highly variable within experimental units. We analyzed a large image data set of wheat foliar diseases to characterize the distribution, spatial dependence, and aggregation behavior of spot-level severity estimates in plots. We combined high-resolution macro-scale imaging with focus bracketing to increase the sampled leaf area. Our results highlight focus bracketing as a promising approach for simultaneous diagnosis and quantification of disease in field plots. Autocorrelation in severity estimates both within focal image stacks and across plot positions was comparable, with 10 focal stack images or 10 positions per plot contributing approximately 2.5 independent observations each. Modeling plot-level severity as a latent Beta-distributed variable enabled robust estimation of mean severity and associated uncertainty. This supports both hypothesis testing and efficient sampling across the full range of disease severity associated with genotypic diversity and seasonality of developing epidemics. The proposed imaging approach is non-invasive and, in principle, transferrable to autonomous ground-based phenotyping platforms, offering the potential to shift the dominant source of uncertainty in estimating disease severity from measurement-related limitations toward biologically and environmentally driven variability in disease expression.

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Predicting Lodging Severity in Sorghum Breeding Trials Using UAV-Based Photogrammetrically Derived Height Data

Mothukuri, S. R.; Massey-Reed, S. R.; Potgieter, A.; Laws, K.; Hunt, C.; Amuzu-Aweh, E. N.; Cooper, M.; Mace, E.; Jordan, D.

2026-03-30 plant biology 10.64898/2026.03.26.713817 medRxiv
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Lodging in sorghum presents a significant challenge for plant breeders due to the trade-off between lodging resistance and grain yield. Manually measuring lodging across thousands of plots is time-consuming, expensive, and error-prone, making selection for lodging resistance challenging in breeding programs. Unmanned Aerial Vehicle (UAV) derived metrics offer a potential high-throughput, cost-effective alternative for lodging phenotyping. This study developed a framework for predicting plot-level lodging from UAV imagery across 2,675 sorghum breeding plots. Multi-temporal canopy height data were collected at two critical time points: maximum crop height and at manual lodging assessment. Height percentiles were extracted from UAV derived point clouds generated using photogrammetric algorithms. These data were used to develop parametric, non-parametric, and ensemble prediction models, which were evaluated using three statistical metrics. The ensemble model, averaging predictions from all models, achieved the highest accuracy with Pearson correlations of r = 0.80-0.84 and lowest residual mean square error (RMSE=16-18), explaining 64-70% of variation in manual lodging counts. Model diagnostics and iterative refinement, including inspection of UAV imagery and dataset curation, had minimal impact on model performance, demonstrating the robustness of the approach. Model performance was consistent across sites, with minimal effects of stratified sampling on accuracy, confirming the ensemble approach as optimal for plot-level lodging assessment. This study demonstrates that integrated multi-temporal UAV imagery offers a practical alternative to labor-intensive manual evaluation methods by enabling high-throughput lodging assessment suitable for implementation in sorghum breeding programs.

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Prediction of late blight severity in a large panel of potato genotypes using low-altitude aerial images and machine learning methods

Loayza, H.; Ninanya, J.; Palacios, S.; Silva, L.; Pujaico Rivera, F.; Rinza, J.; Gastelo, M.; Aponte, M.; Kreuze, J. F.; Lindqvist-Kreuze, H.; Heider, B.; Kante, M.; Ramirez, D. A.

2026-04-09 plant biology 10.64898/2026.04.06.716456 medRxiv
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Potato (Solanum tuberosum L.) is a staple crop crucial to global food security, yet its production is severely threatened by late blight (LB), caused by Phytophthora infestans, one of the most destructive plant diseases worldwide. Breeding programs for LB resistance have traditionally relied on labor-intensive and subjective visual assessments, which limit scalability and consistency, particularly in early-generation trials. Unmanned aerial vehicle (UAV)-based remote sensing combined with machine learning (ML) offers a promising alternative for objective, high-throughput disease phenotyping. This study evaluated the potential of UAV-derived multispectral imagery and ML techniques to estimate LB severity across large and genetically diverse potato breeding populations, comprising 2,745 clones in one trial and 492 accessions in another, conducted in Oxapampa, Pasco, Peru. We compared vegetation index-based approaches with a machine learning framework that integrates K-means clustering and Kernel Ridge Regression (KRR) and assessed their ability to capture genotypic variation and support selection decisions. NDVI consistently showed a strong correlation with visually assessed LB severity, particularly at advanced stages of disease development, enabling objective discrimination between healthy and diseased canopy tissues. However, the KRR-based approach outperformed linear NDVI-based models by capturing nonlinear relationships between spectral responses and disease progression. Estimates of LB severity derived from NDVI and KRR models, expressed as best linear unbiased estimates (BLUEs), showed strong and biologically consistent relationships with the area under the disease progress curve (AUDPC), particularly during later UAV acquisitions. Selection coincidence between UAV-derived estimates and AUDPC-based rankings was substantially higher at intermediate to advanced stages of disease progression, suggesting that UAV assessments at these stages may capture sufficient phenotypic variation to distinguish genotypes. These findings indicate that UAV-based multispectral phenotyping, especially when integrated with ML, provides a practical and scalable approach for assessing LB severity in potato breeding programs while reducing the need for time-consuming field evaluations.

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Spectral Phenotyping Reveals Time-Specific QTLs in Field-Grown Lettuce

Mehrem, S. L.; Zijl, A.; de Haan, M.; Van den Ackerveken, G.; Snoek, B. L.

2026-03-18 plant biology 10.64898/2026.03.16.711173 medRxiv
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Lettuce (Lactuca sativa) is an important field crop, but our understanding of its phenotypic variation and underlying genetics under natural field conditions remains limited, posing challenges for identifying effective crop breeding targets. Longitudinal hyperspectral phenotyping allows for non-invasive monitoring of crop performance under diverse agricultural conditions. In this study, we used hyperspectral imaging to assess the phenotypic variation of almost 200 different field-grown lettuce varieties, following the same plants from just after seedling- to flowering-stage. With automated image processing, we extracted a wide range of spectral phenotypes related to metabolite content, growth efficiency, and environmental stress responses, creating a multi-dimensional time-resolved data set. Principal component analysis (PCA) revealed the major axes of spectral variation over time, and highlighted differences in spectral patterns among lettuce genotypes. Integrating on-site weather data, we modelled GxE interactions of reflectance, revealing regions of the lettuce vegetation spectrum that are primarily shaped by genotype and/or environment. We estimated phenotypic plasticity in response to time, temperature and rainfall using best linear unbiased predictions (BLUPs), capturing genotype-specific developmental trajectories and responses to the environment. We used genome-wide association studies (GWAS) to identify quantitative trait loci (QTLs) of PC-based, single and BLUP-based phenotypes, disentangling the genetic architecture of spectral lettuce phenotypes from major axes of variation down to single wavelength spectral plasticity. These findings provide new insights into the genome-wide genetic regulation and dynamics of spectral phenotypes in field grown lettuce.

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Leaf and cluster spectral signatures reveal trait-dependent prediction performance for grapevine cluster architecture and juice quality

Robles-Zazueta, C. A.; Strack, T.; Schmidt, M.; Callipo, P.; Robinson, H.; Vasudevan, A.; Voss-Fels, K.

2026-03-31 plant biology 10.64898/2026.03.27.714894 medRxiv
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Grapevine cluster architecture is a key selection target in breeding programs because it influences disease susceptibility, yield stability and juice quality. High-throughput phenotyping offers a rapid and non-destructive approach to capture biochemical and structural variation in these traits, yet the influence of plant organ reflectance and data partitioning strategies on trait prediction remains poorly understood. In this study, we evaluated how hyperspectral reflectance from different grapevine organs contributes to the prediction of cluster architecture and juice quality traits in two clonal populations of Riesling and Pinot. Using partial least squares regression (PLSR), we assessed the prediction accuracy of eight cluster architecture and six juice quality traits under two data partitioning strategies. Models based on cluster reflectance outperformed those using dry leaf reflectance for most traits, except for pH. Partitioning the dataset by cluster type increased trait variance and improved predictions for number of berries (R{superscript 2} = 0.53), berry diameter (R{superscript 2} = 0.79), and total acidity (R{superscript 2} = 0.48). Visible, red-edge and NIR spectra were most informative regions to predict the traits studied. Together, our results highlight the importance of organ-specific data and appropriate calibration strategies to improve phenomic models for the development of scalable proxies for grapevine improvement. HighlightSpectral phenomics reveals that prediction accuracy in grapevine depends on organ spectral signatures and traits, with cluster reflectance outperforming leaves, informing new phenotyping strategies for breeding improvement.

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Presymptomatic plant disease detection with PSNet: A low-cost hyperspectral imaging and RGB fusion framework.

Crabb, G. U.; Cevik, V.; Chen, X.; Priest, N. K.; Zhao, Y.

2026-03-04 plant biology 10.64898/2026.03.02.709086 medRxiv
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Plant pathogens cause major yield losses worldwide, threatening food security and livelihoods. Because early infection is difficult to diagnose, management often relies on prophylactic pesticide use, increasing costs and environmental impact. Here we present PSNet, a multimodal framework that fuses hyperspectral imaging with RGB information for presymptomatic plant disease detection, together with a low-cost, portable hyperspectral camera incorporating a 3D-printed housing and optical mounts, costing under {pound}500. We validate the approach using Arabidopsis thaliana infected with the oomycete Albugo candida. Imaging at 2 and 4 days post inoculation, prior to visible symptoms, revealed consistent spectral signatures that distinguished infected from healthy plants, while imaging at 6 days post inoculation captured the transition toward early symptom emergence. The most discriminative spectral regions overlapped wavelengths previously associated with plant responses to biotic stress, supporting the biological plausibility of these signatures. On a four-class task (healthy, 2 dpi, 4 dpi, 6 dpi), PSNet achieved 92.7% overall accuracy and 97.1% accuracy for binary healthy versus infected classification. Together, these results demonstrate that presymptomatic detection is feasible under controlled conditions using low-cost hardware and multimodal learning, underscoring the potential of scalable, multimodal systems for early disease monitoring.

8
Reconstructing coniferous tree crown shape from incomplete point clouds using deep learning

Bornand, A.; Abegg, M.; Morsdorf, F.; Puliti, S.; Astrup, R.; Rehush, N.

2026-01-21 plant biology 10.64898/2026.01.18.700158 medRxiv
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Individual tree structure plays a key role in forest monitoring, biomass estimation, and ecological assessment. However, ground-based remote sensing methods such as terrestrial and mobile laser scanning frequently produce incomplete point clouds due to occlusion, particularly in the upper canopy. This limits the accuracy of derived structural metrics such as tree height or crown volume. In this study, we present a novel deep learning-based method to reconstruct the outer crown shape of coniferous trees from incomplete point clouds. Instead of completing the full tree structure, we focus on predicting the alpha-shape of the crown, enabling a more efficient and generalizable approach for structural reconstruction. We train a geometry-aware transformer model (AdaPoinTr) on synthetically generated partial tree crowns and evaluate its performance across three independent datasets encompassing different forest types and acquisition conditions. The model consistently improved crown shape similarity metrics and reduced height estimation errors compared to using partial data alone (reduced bias from -11% to -3.5%). Our results demonstrate that this shape-based strategy enables the extraction of key tree-level parameters from incomplete data, offering a practical solution for gaining improved 3D forest structural information from cost-sensitive or logistically constrained forest monitoring acquisitions.

9
Field and lab phenomics facilitate detection of genetic variation for iron deficiency chlorosis tolerance in sorghum

Cerimele, G.; Kent, M.; Miller, M.; Best, R.; Franks, C.; Kakar, N.; Felderhoff, T.; Sexton-Bowser, S.; Morris, G. P.

2026-04-05 genetics 10.64898/2026.04.01.715717 medRxiv
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Bioavailability of iron, an essential micronutrient to plants, is low in alkaline or calcareous soils, which are prevalent across semi-arid production regions. Breeding efforts to increase tolerance to iron deficiency chlorosis (IDC) in sorghum, a major crop of semi-arid regions, are confounded by spatial variation of stress severity in field trials. Here we developed and validated two high-throughput phenotyping approaches to address this challenge, with multi-spectral aerial imaging in the field and a controlled-environment assay to isolate the effects of iron bioavailability. In the field, severity and uniformity of stress are highly predictive of genetic signals for IDC tolerance (R2 > 0.6 for soil pH metrics and H2). Plot-level data filtering for stress conditions based on control genotypes successfully addresses field spatial variation (unfiltered H2 = 0.18 vs. filtered H2 = 0.4). The controlled-environment assay proxies field stress using iron sources with differential bioavailability, evidenced by high heritability ( H2 = 0.98) and phenotypic differential for hybrid control genotypes that matches field performance. Finally, we show that assay phenotypes are suitable for genome-wide association studies in global germplasm. Together, these field and lab phenomic approaches can be deployed to understand genetics of IDC tolerance and develop crops resilient to alkaline soils. HIGHLIGHTStress severity and uniformity greatly impact detection of genetic signals underlying iron deficiency chlorosis tolerance in sorghum. A controlled-environment assay reduces spatial heterogeneity and improves assessment of tolerance genetics.

10
Inter-variety competition dynamics in US inbred and hybrid maize

Schulz, A. J.; Bohn, M. O.; Bradbury, P.; Lima, D. C.; De Leon, N.; Flint-Garcia, S.; Holland, J. B.; Lepak, N.; Lorenz, A. J.; Romay, M. C.; Hirsch, C. N.; Buckler, E. S.; Robbins, K. R.

2026-02-28 plant biology 10.64898/2026.02.26.708322 medRxiv
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Variety mixtures provide a potential avenue in US cropping systems to improve yield stability and disease resistance. However, implementation of variety mixtures requires an understanding of the competitive dynamics of the crop. In this study, we examine the effects of plant competition both between and within plots through five unique experiments: 1) 5,000 diverse inbred lines in single-row plots, 2) hybrids in two-row plots developed from the above inbred lines, 3) over 4,000 hybrids measured in 141 locations in two-row plots as part of Genomes to Fields, 4) mixtures of two hybrids within a two-row plot planted across two years and five locations, and 5) mixtures of up to twenty hybrids in four-row plots in three locations. Across all experiments, we find that competitive interactions are extremely limited. Within inbred lines, height of the neighboring plot accounts for 1.2% of the variance in focal plot height. Similarly, neighbor height explains 1.7% of the variance in focal plot yield in hybrids developed from the inbred lines. The genetics of neighboring plots explains 1.55% of the variation in yield across 141 location-year environments, reinforcing the generally modest impacts of neighbor competition. In evaluating mixtures of hybrids in both two and four-row plots, we observe no yield penalty compared to conventional single hybrid plots, even with large height differentials of the hybrids included in the mixture or in mixtures of up to 20 hybrids within a plot. Finally, we observe that mixtures have more yield stability compared to conventional plots, highlighting a new avenue for increased stability in higher risk environments. The lack of yield penalty and stability benefits are promising for future investigations of mixtures that may complement each other in disease resistance or abiotic stress tolerance and increase overall yield stability in the field.

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Unlocking the potential of Capsicum Germplasm Collections for Climate Resilience and Fruit Quality

Halpin-McCormick, A.; Nalla, M. K.; Radlicz, Z.; Zhang, A.; Fumia, N.; Lin, T.-h.; Lin, S.-w.; Wang, Y.-w.; Zohoungbogbo, H. P. F.; Wang, D. R.; Runck, B.; Gore, M. A.; Kantar, M. B.; Barchenger, D. W.

2026-03-28 plant biology 10.64898/2026.03.25.714358 medRxiv
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Climate change increasingly threatens global Capsicum (pepper) production. Accelerating the deployment of climate-resilient cultivars requires effective use of genetic diversity conserved in genebanks. We implement a "turbocharging" strategy in Capsicum by integrating genome-wide association studies and genomic prediction in a core collection (n = 423), followed by genomic prediction across the global collection (n = 10,250) using the core as a training population. We generated genomic estimated breeding values (GEBVs) for 31 high-accuracy traits (r > 0.5) encompassing hyperspectral phenotypes (heat/control), agronomic performance (heat/control) and fruit quality. To enhance accessibility and decision-making, we developed a large language model (LLM) integrated application that enables flexible, preference-based selection of candidates. By narrowing the parental decision space, this framework streamlines screening of large germplasm collections while balancing climate resilience, quality attributes and market demands. Our approach provides a scalable decision-support system to accelerate climate-resilient Capsicum breeding and maximize global genetic resources.

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Radiation-Driven Prediction of Daily Irrigation Demand under Different Electrical Conductivity Scenarios in Greenhouse Tomato

Xiao, L.

2026-01-24 plant biology 10.64898/2026.01.23.701235 medRxiv
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In soilless greenhouse tomato cultivation, daily transpiration and irrigation demand are largely governed by solar radiation, while irrigation-solution electrical conductivity (EC) used for salinity management may further modulate plant water use. This study developed a low-input, radiation-driven modeling approach to predict daily irrigation demand under contrasting water-salt management scenarios. Two tomato cultivars were grown under four treatments: conventional baselines (CK1, CK2) and regulated scenarios combining irrigation volume with solution EC (low-water high-EC, TK; high-water moderate-EC, TC). Daily irrigation volume (I) and drainage were recorded, and daily cumulative radiation (G) was derived from photosynthetically active radiation (PAR). Within each treatment, we compared a radiation-only baseline model with an EC-adjusted model and evaluated predictive performance using 5-fold blocked time-series cross-validation. Results showed strong positive correlations between G and I across all treatments (p < 0.001). The EC-adjusted models achieved cross-validated root-mean-square errors (RMSE) of 0.815-1.393 L d-1 per trough and Nash-Sutcliffe efficiencies (NSE) of 0.407-0.730. Incorporating EC yielded a small but consistent improvement under the TK scenario ({Delta}RMSE = -0.014 L d-1; {Delta}NSE = +0.019), whereas its effect was negligible or slightly negative under CK1, CK2, and TC, highlighting scenario dependence. Our radiation-driven framework, with an optional EC correction, offers a practical and scalable tool for daily irrigation forecasting and supports integrated water-salt management in soilless greenhouse tomato production.

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Potato yield can be predicted by using drone-captured and environmental measurements early in the growing season

Vizintin, A.; Zagorscak, M.; Turk, E.; Kriznik, M.; Petek, M.; Stare, K.; Wurzinger, B.; Shaikh, M. A.; Heselmans, G.; Sollinger, J.; Lindenbergh, P.-J.; Graveland, R.; Oome, S.; Prat, S.; Bachem, C.; Teige, M.; Doevendans, B.; Ribarits, A.; Zrimec, J.; Gruden, K.

2026-03-11 plant biology 10.64898/2026.03.09.709817 medRxiv
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Accurate pre-harvest prediction of crop yield informs variety selection, optimizes management, and accelerates breeding. As potato is the worlds leading non-grain staple, here we evaluate a diverse panel of varieties in a three-year field trial across five European locations. Canopy development and environmental parameters are monitored throughout the growing season using drone-based imaging, in-field sensors and gene expression measurements, while tuber yield and quality traits are quantified at harvest. We show that these data enable the identification of climate-resilient, high-yielding genotypes and support the development of machine learning models that explain over 80% of yield variance in independent test sets. Strikingly, measurements collected within the first two months after planting achieve predictive performance comparable to models trained on full-season data. Model interrogation further shows that simplified five-parameter linear equations capture over 70% of yield variability. Our framework thus demonstrates the potential of integrative field phenotyping and data-driven modeling to improve variety selection across heterogeneous environments. Significance statementThe ability to predict harvest crop yields from pre-harvest measurements can enable farmers and growers to make informed decisions on variety selection and management practices, while breeders can benefit from accelerated breeding cycles. We perform a panel of field trials with potato, the no. 1 global non-grain staple, across varying conditions and locations, recording various growth- and climate-related data, including gene expression, and post-harvest yield and quality of tubers. We demonstrate the potential of the field trial data to facilitate the analysis and selection of best-performing varieties across diverse conditions and locations, and to revolutionize farming by enabling early (already within 2 months) and straightforward (only a couple of key measured variables) yield predictions with high accuracy.

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A standard area diagram for potato common scab: comparable performance of image- and object-based validation

Cazon, L. I.; Paredes, J. A.; Quiroga, M.; Guzman, F.

2026-03-20 plant biology 10.64898/2026.03.18.712681 medRxiv
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Potato common scab (Streptomyces sp.) is an economically important disease that reduces the quality and market value of tubers. A key aspect in developing management strategies involves accurately quantifying the disease. Due to the three-dimensional nature of the tuber and the heterogeneous distribution of lesions across its surface, visual estimates of severity can be challenging. Therefore, the objectives of this study were to develop and validate a standard area diagram (SAD) for estimating common scab severity on potato tubers and to compare validation outcomes obtained using real tubers and digital images. A SAD comprising six severity levels (from 1.3 to 66.8%) was developed based on image analysis of naturally infected tubers. Validation was conducted using two complementary approaches in which inexperienced raters evaluated either real potato tubers or digital images of the same tubers under unaided and aided conditions. Accuracy, bias components, and inter-rater reliability were quantified using absolute error metrics, Lins concordance correlation coefficient, intraclass correlation coefficients, and overall concordance correlation coefficients. Use of the SAD significantly improved accuracy, reduced systematic bias, and increased inter-rater reliability across both validation approaches. No significant differences were detected between assessments conducted on real tubers and images, although image-based evaluations showed a slight, non-significant tendency toward reduced scale and location bias under aided conditions. These results demonstrate that a dimension-aware SAD integrating information across the full tuber surface enhances the reliability and reproducibility of visual severity assessments and supports the use of image-based evaluations for training, large-scale surveys, and remote or collaborative applications involving three-dimensional plant organs.

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Transformers Outperform ConvNets for Root Segmentation: A Systematic Comparison Across Nine Datasets

Smith, A. G.; Lamprinidis, S.; Seethepalli, A.; York, L. M.; Han, E.; Mohl, P.; Boulata, K.; Thorup-Kristensen, K.; Petersen, J.

2026-02-19 plant biology 10.64898/2026.02.18.706562 medRxiv
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Root segmentation is a fundamental yet challenging task in image-based plant phenotyping. We present the first systematic comparison of Transformer and Convolutional Neural Network (ConvNet) architectures for root segmentation, evaluating 21 architectures across nine diverse datasets and comparing pre-trained models to training from scratch. Transformer-based models significantly outperform ConvNets for segmentation accuracy and root-diameter agreement. Pre-training significantly improves mean Dice from 0.623 to 0.666 (p = 3.3 x 10-10). We also find that Transformers benefit more from pre-training than ConvNets, with Dice improvements of +0.072 versus +0.022 (p = 3.7 x 10-4), supporting the hypothesis that fine-tuned Transformers transfer more effectively across large domain gaps. Among evaluated models, MobileSAM achieved the highest Dice score while maintaining computational efficiency. Dataset choice explained far more performance variance (70.9%) than model architecture (6.7%), suggesting that data curation matters more than model selection.

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Spectral network analysis illuminates coordinated planttraits across a climate gradient

Ray, R.; Quarles-Chidyagwai, B.; Ashlock, S.; Lyons, J.; Gremer, J. R.; Maloof, J.; Magney, T.

2026-02-21 plant biology 10.1101/2025.09.18.676927 medRxiv
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O_LIUnderstanding how plant populations respond to environmental variation through functional leaf traits remains challenging due to limitations of traditional phenotyping approaches. Hyperspectral reflectance offers a rapid, non-destructive and high-throughput method to capture functional trait variation and detect signatures of local adaptation across populations. C_LIO_LIWe combined hyperspectral data, inverse modeling, and network analysis to investigate population-level variation in Streptanthus tortuosus. Using a common garden experiment with four geographically distinct populations, we applied partial least square discriminant analysis (PLS-DA) and ridge regression for population discrimination, inverse PROSPECT modeling to estimate leaf biochemical traits, and canonical correlation analysis to examine trait-climate relationships across historical (1900-1994) and recent (1995-2024) periods. We developed a spectral network approach treating wavelength correlations as biologically meaningful trait networks. C_LIO_LIPopulations showed distinct, heritable spectral signatures with high classification accuracy. Significant population differences emerged in anthocyanins, carotenoids, chlorophyll, and water content. Trait-climate correlations shifted between time periods, consistent with historical climate adaptation. Network analysis revealed population-specific integration patterns, with more variable environments displaying greater spectral modularity. C_LIO_LIHyperspectral signatures provide a high-throughput tool for detecting population-level adaptation and trait coordination. Our findings provide a framework to investigate how plant populations respond to climate change through evolved shifts in trait networks rather than isolated traits alone. C_LI

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Reliable quantification of multiplexed genetically encoded biosensors responsiveness in plant tissues

Levak, V.; Zupanic, A.; Pogacar, K.; Marondini, N.; Stare, K.; Arnsek, T.; Fink, K.; Gruden, K.; Lukan, T.

2026-03-16 plant biology 10.64898/2026.03.13.711581 medRxiv
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Genetically encoded biosensors are one of essential tools in biological research. They enable visualization of molecules of interest from the subcellular level to entire organism level in vivo and can be used to monitor presence of small molecules, gene expression, protein activity, and protein degradation. However, multiplexing fluorescent biosensors in plants is notoriously difficult due to signal bleed-through and strong autofluorescence from chlorophyll. In this study, we investigated the potential of multiplexing biosensors based on the selection of reporter fluorescent proteins. We characterized the emission spectra, fluorescence lifetimes, and relative brightness of diverse fluorescent proteins in plant leaves. We show that selected proteins exhibit comparable brightness, supporting their use in co-expression experiments and reliable quantification of individual signals. To separate overlapping signals, we applied two different linear unmixing approaches and compared them to results obtained without unmixing. We identified channel separation unmixing approach as the most suitable for biosensors. Additionally, we show how unmixing with the selected approach can be applied to separate autofluorescence and we validated this approach in virus-infected cells by following organelle dynamics in vivo. Overall, our work demonstrates that biosensors can be multiplexed, even when their emission spectra overlap. Significance statementMultiplexing genetically encoded biosensors in plants has been limited by overlapping fluorescent signals and strong autofluorescence. This study presents an optimized framework for linear unmixing and provides a MATLAB-based organelle segmentation tool, allowing precise quantification of multiple fluorescent reporters in vivo and advancing real-time visualization of complex cellular processes in plants.

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Dissecting genetic variance structure and evaluating genomic prediction models for single-cross hybrids derived from Stiff Stalk and Non-Stiff Stalk maize heterotic groups

Godoy, J. C.; Edwards, J.; Lee, E. C.; Mikel, M. A.; Fernandes, S. B.; Hirsch, C. N.; Berry, S. P.; Lipka, A. E.; Bohn, M. O.

2026-03-13 genetics 10.64898/2026.03.11.710575 medRxiv
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The early 20th-century discovery of heterosis and the establishment of heterotic groups transformed maize (Zea mays L.) into a keystone of global agriculture. However, maize breeding faces two significant challenges: the gradual decline of general combining ability (GCA) variance within heterotic groups and the impracticality of testing all possible single crosses in the early stages of a breeding program. Here, we developed genomic best linear unbiased prediction (GBLUP)-based multi-kernel models, using additive and two alternative non-additive genomic relationship matrices, to estimate the variance components associated with the GCA of Stiff Stalk (SS) and Non-Stiff Stalk (NSS) heterotic groups and the specific combining ability (SCA) arising from their crosses. We further applied these models to predict the performance of untested single-cross combinations under varying levels of parental information. We showed that the SS and NSS groups retained significant GCA variance across traits in both early- and late-maturity groups. The SS group, in contrast, exhibited no detectable GCA variance in grain yield for the intermediate-flowering subset of hybrids, highlighting a limitation for future genetic improvement. Furthermore, our results showed that GBLUP-based multi-kernel models effectively identified superior hybrids when parental information was available. In the absence of this information, however, these models underperformed compared to covariance-based approaches. Both types of non-additive matrices produced similar results, affirming the robustness of the inferred genetic architecture. Overall, this study sheds light on the future use of US maize commercial germplasm and demonstrates how GBLUP-based multi-kernel models can improve the efficiency of hybrid breeding programs.

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Development and evaluation of a cost-effective, mid-density SNP array as a sorghum community genotyping resource

Kumar, V.; Klein, R. R.; Kaufman, B.; Winans, N. D.; Crozier, D.; Rooney, W. L.; Harrison, M.; Hayes, C.; Tello-Ruiz, M. K.; Gladman, N. P.; Olson, C.; Burow, G.; Sexton-Bowser, S.; Punnuri, S.; Knoll, J.; Dahlberg, J.; Ware, D.

2026-02-23 plant biology 10.64898/2026.02.20.706663 medRxiv
Top 0.1%
2.9%
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The development of accessible and cost-effective genotyping platforms is essential to accelerate genetic gain in crop improvement. To address the U.S. sorghum communitys need for a standardized, mid-density genotyping resource, we developed and validated a targeted single-nucleotide polymorphism (SNP) array using the PlexSeq next-generation sequencing (NGS) platform. The resulting genotyping array includes 2,421 SNPs spanning all ten Sorghum bicolor chromosomes and integrates trait-linked and quality control markers selected by public and private stakeholders. Genotyping 2,726 diverse accessions, including the Sorghum Association Panel (SAP), demonstrated high call rates (>90% for most samples and markers), low missing data, and accurate resolution of population structure consistent with prior whole-genome studies. In comparative genomic prediction analyses, the mid-density array performed equivalently to high-density genotype-by-sequencing (GBS) platforms for key traits such as grain yield and plant height across multi-environment trials. Designed for broad utility in breeding pipelines, the array enables marker-assisted selection, genomic prediction, identity verification, and germplasm quality control. Moreover, its adoption by the USDA National Plant Germplasm System facilitates the curation of genebanks and the management of core collections. This community-driven genotyping platform offers a scalable, reproducible, and customizable tool to support molecular breeding in sorghum and underscores the value of targeted marker systems in resource-optimized crop improvement programs.

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How important is the intra-regional soil heterogeneity for the design of future stress-avoidant wheat ideotypes? A modeling study in central France

Blanchet, G.; Semenov, M. A.; Allard, V.

2026-02-12 plant biology 10.64898/2026.02.11.705307 medRxiv
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2.8%
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Accurate projections of crop adaptation to climate change require accounting for the spatial heterogeneity of soils, which modulates both water availability and the effectiveness of genetic adaptation. Using the process-based crop model Sirius, we investigated how intra-regional variability in soil available water capacity (AWC) influences wheat yields and the adaptive value of stress-avoidant ideotypes under future climates in central France (Limagne plain). Detailed soil databases were aggregated across five representative sites and combined with multiple climate projections (CMIP6), two emission pathways (SSP2-4.5 and SSP5-8.5), and three time horizons (2031-2050, 2051-2070 and 2071-2090). Variance decomposition revealed that soil AWC accounted for 23% of the simulated yield variability, significantly exceeding the contribution of local climate contrasts (10%), a pattern consistent across current and future periods. Deep soils (>80 mm AWC) buffered drought effects whereas yields stagnated in shallow soils (<80 mm AWC) where water deficits persisted despite phenology hastening. On average, the reference cultivar showed earlier anthesis by 8-21 days under future climates, leading to higher yields mainly in deep soils. Optimization of flowering timing through stress-avoidant ideotypes provided mean yield gains of +6.33 dt{middle dot}ha-1 in deep soils, but limited benefits (+1.71 dt.ha-1) in shallow ones, highlighting pedological dependence of breeding efficiency. Advancing anthesis also increased exposure to early-spring frost: frost probability rose from <0.1 to >0.4 when flowering occurred more than 250 {degrees}C.days earlier, particularly in the frost-prone part of the study area. Hence, frost risk remains a critical constraint for early ideotypes, even under strong warming. Overall, our results demonstrate that intra-regional soil heterogeneity remains a dominant driver of wheat yield variability and adaptation potential under climate change. Designing stress-avoidant ideotypes without explicit consideration of local soil AWC could lead to maladaptation, especially in regions with shallow soils represent a significant portion of cropped areas. In such situation, breeding for terminal stress avoidance may offer only limited benefit. We advocate that breeding and modeling frameworks integrate high-resolution soil data to refine regional ideotype design, reconcile terminal-stress avoidance with frost tolerance, and better capture the spatial realism required for sustainable crop adaptation strategies. Highlights- Local soil water capacity limits wheat adaptation to climate change. - Deep soils favor earlier, stress-avoidant ideotypes. - Shallow soils restrict the benefits of phenological adjustment for stress avoidance. - Frost exposure remains a key risk when shifting phenology toward earliness.