Efficient Noninvasive FHB Estimation using RGB Images from a Novel Multiyear, Multirater Dataset.

Fusarium head blight (FHB) is one of the most prevalent wheat diseases, causing substantial yield losses and health risks. Efficient phenotyping of FHB is crucial for accelerating resistance breeding, but currently used methods are time-consuming and expensive. The present article suggests a noninvasive classification model for FHB severity estimation using red-green-blue (RGB) images, without requiring extensive preprocessing. The model accepts images taken from consumer-grade, low-cost RGB cameras and classifies the FHB severity into 6 ordinal levels. In addition, we introduce a novel dataset consisting of around 3,000 images from 3 different years (2020, 2021, and 2022) and 2 FHB severity assessments per image from independent raters. We used a pretrained EfficientNet (size b0), redesigned as a regression model. The results demonstrate that the interrater reliability (Cohen's kappa, κ) is substantially lower than the achieved individual network-to-rater results, e.g., 0.68 and 0.76 for the data captured in 2020, respectively. The model shows a generalization effect when trained with data from multiple years and tested on data from an independent year. Thus, using the images from 2020 and 2021 for training and 2022 for testing, we improved the F1w score by 0.14, the accuracy by 0.11, κ by 0.12, and reduced the root mean squared error by 0.5 compared to the best network trained only on a single year's data. The proposed lightweight model and methods could be deployed on mobile devices to automatically and objectively assess FHB severity with images from low-cost RGB cameras. The source code and the dataset are available at https://github.com/cvims/FHB_classification.

The Transferability of Spectral Grain Yield Prediction in Wheat Breeding across Years and Trial Locations.

Grain yield (GY) prediction based on non-destructive UAV-based spectral sensing could make screening of large field trials more efficient and objective. However, the transfer of models remains challenging, and is affected by location, year-dependent weather conditions and measurement dates. Therefore, this study evaluates GY modelling across years and locations, considering the effect of measurement dates within years. Based on a previous study, we used a normalized difference red edge (NDRE1) index with PLS (partial least squares) regression, trained and tested with the data of individual dates and date combinations, respectively. While strong differences in model performance were observed between test datasets, i.e., different trials, as well as between measurement dates, the effect of the train datasets was comparably small. Generally, within-trials models achieved better predictions (max. R2 = 0.27-0.81), but R2-values for the best across-trials models were lower only by 0.03-0.13. Within train and test datasets, measurement dates had a strong influence on model performance. While measurements during flowering and early milk ripeness were confirmed for within- and across-trials models, later dates were less useful for across-trials models. For most test sets, multi-date models revealed to improve predictions compared to individual-date models.

Temporal Dynamics and the Contribution of Plant Organs in a Phenotypically Diverse Population of High-Yielding Winter Wheat: Evaluating Concepts for Disentangling Yield Formation and Nitrogen Use Efficiency.

Enhancing crop nitrogen use efficiency (NUE) is a key requirement for both economic and ecological reasons. Consequently, the genotypic potential for NUE in winter wheat (Triticum aestivum L.) requires further exploitation. Emerging plant phenomic techniques may provide knowledge about traits contributing to grain N uptake (GNup) and grain yield (GY). However, the understanding of beneficial strategies concerning the temporal dynamics of NUE and GY formation and the role of plant organs is still scarce especially under high-yielding European conditions-particularly to discriminate interesting lines in the breeding process. Thus, screening for potentially useful NUE traits in terms of variation, stability, and contribution to target traits will be an essential prerequisite for the development of efficient phenotyping strategies. Therefore, 46 NUE and yield formation traits were assessed in a population of 75 breeding lines over 3 years from 2015 to 2017 in southern Germany, including dry matter (DM), N concentration, and N uptake at anthesis and maturity, both at the aboveground-plant and plant organ levels. Significant genotype and genotypexenvironment effects were observed for all traits. While GY was more related to post-anthesis assimilation, also DM translocation contributed substantially to GY by 31-44%. At maturity, total aboveground DM as opposed to harvest index predominantly determined GY. NUE for GY was better described by N uptake efficiency than by N utilization efficiency. GNup was greatly influenced by variation in GY, but not in grain N concentration, and by total N uptake and not the N harvest index. Post-anthesis N uptake highly depended on the year and was low in comparison to N translocation. However, post-anthesis N uptake was always correlated with GNup, suggesting the need to also consider stay-green strategies under temperate growing conditions. While anthesis traits were only moderately descriptive, GY will be enhanced by increasing total biomass and the N uptake efficiency. Similarly, targeting total N uptake, particularly at post-anthesis, seems to be a rewarding strategy to boost GNup. Thus, high-throughput phenotyping should be targeted rather toward detecting traits related to DM and N acquisition than to the internal allocation and rather to post-anthesis than to anthesis traits.

Temporal and Spectral Optimization of Vegetation Indices for Estimating Grain Nitrogen Uptake and Late-Seasonal Nitrogen Traits in Wheat.

Grain nitrogen (N) uptake (GNup) in winter wheat (Triticum aestivum L.) is influenced by multiple components at the plant organ level and by pre- and post-flowering N uptake (Nup). Although spectral proximal high-throughput sensing is promising for field phenotyping, it was rarely evaluated for such N traits. Hence, 48 spectral vegetation indices (SVIs) were evaluated on 10 measurement days for the estimation of 34 N traits in four data subsets, representing the variation generated by six high-yielding cultivars, two N fertilization levels (N), two sowing dates (SD), and two fungicide (F) intensities. Close linear relationships (p < 0.001) were found for GNup both in response to cultivar differences (Cv; R2 = 0.52) and other agronomic treatments (R2 = 0.67 for Cv*F*N, R2 = 0.53 for Cv*SD*N and R2 = 0.57 for the combined treatments), notably during milk ripeness. Especially near-infrared (NIR)/red edge SVIs, such as the NDRE_770_750, outperformed NIR/visible light (VIS) indices. Index rankings and seasonal R2 values were similar for total Nup, while the N harvest index, which expresses the partitioning to the grain, was moderately estimated only during dough ripeness, primarily from indices detecting contrasting senescence between different fungicide intensities. Senescence-sensitive indices, including R787_R765 and TRCARI_OSAVI, performed best for N translocation efficiency and some organ-level N traits at maturity. Even though grain N concentration was best assessed by the red edge inflection point (REIP), the blue/green index (BGI) was more suited for leaf-level N traits at anthesis. When SVIs were quantitatively ranked by data subsets, a better agreement was found for GNup, total Nup, and grain N concentration than for several contributing N traits. The results suggest (i) a good general potential for estimating GNup and total Nup by (ii) red edge indices best used (iii) during milk and early dough ripeness. The estimation of contributing N traits differs according to the agronomic treatment.

Sensitivity of Vegetation Indices for Estimating Vegetative N Status in Winter Wheat.

Precise sensor-based non-destructive estimation of crop nitrogen (N) status is essential for low-cost, objective optimization of N fertilization, as well as for early estimation of yield potential and N use efficiency. Several studies assessed the performance of spectral vegetation indices (SVI) for winter wheat (Triticum aestivum L.), often either for conditions of low N status or across a wide range of the target traits N uptake (Nup), N concentration (NC), dry matter biomass (DM), and N nutrition index (NNI). This study aimed at a critical assessment of the estimation ability depending on the level of the target traits. It included seven years' data with nine measurement dates from early stem elongation until flowering in eight N regimes (0-420 kg N ha-1) for selected SVIs. Tested across years, a pronounced date-specific clustering was found particularly for DM and NC. While for DM, only the R900_970 gave moderate but saturated relationships (R2 = 0.47, p < 0.001) and no index was useful for NC across dates, NNI and Nup could be better estimated (REIP: R2 = 0.59, p < 0.001 for both traits). Tested within growth stages across N levels, the order of the estimation of the traits was mostly Nup ≈ NNI > NC ≈ DM. Depending on the number (n = 1-3) and characteristic of cultivars included, the relationships improved when testing within instead of across cultivars, with the relatively lowest cultivar effect on the estimation of DM and the strongest on NC. For assessing the trait estimation under conditions of high-excessive N fertilization, the range of the target traits was divided into two intervals with NNI values < 0.8 (interval 1: low N status) and with NNI values > 0.8 (interval 2: high N status). Although better estimations were found in interval 1, useful relationships were also obtained in interval 2 from the best indices (DM: R780_740: average R2 = 0.35, RMSE = 567 kg ha-1; NC: REIP: average R2 = 0.40, RMSE = 0.25%; NNI: REIP: average R2 = 0.46, RMSE = 0.10; Nup: REIP: average R2 = 0.48, RMSE = 21 kg N ha-1). While in interval 1, all indices performed rather similarly, the three red edge-based indices were clearly better suited for the three N-related traits. The results are promising for applying SVIs also under conditions of high N status, aiming at detecting and avoiding excessive N use. While in canopies of lower N status, the use of simple NIR/VIS indices may be sufficient without losing much precision, the red edge information appears crucial for conditions of higher N status. These findings can be transferred to the configuration and use of simpler multispectral sensors under conditions of contrasting N status in precision farming.

High-Throughput Field Phenotyping Traits of Grain Yield Formation and Nitrogen Use Efficiency: Optimizing the Selection of Vegetation Indices and Growth Stages.

High-throughput, non-invasive phenotyping is promising for evaluating crop nitrogen (N) use efficiency (NUE) and grain yield (GY) formation under field conditions, but its application for genotypes differing in morphology and phenology is still rarely addressed. This study therefore evaluates the spectral estimation of various dry matter (DM) and N traits, related to GY and grain N uptake (Nup) in high-yielding winter wheat breeding lines. From 2015 to 2017, hyperspectral canopy measurements were acquired on 26 measurement dates during vegetative and reproductive growth, and 48 vegetation indices from the visible (VIS), red edge (RE) and near-infrared (NIR) spectrum were tested in linear regression for assessing the influence of measurement stage and index selection. For most traits including GY and grain Nup, measurements at milk ripeness were the most reliable. Coefficients of determination (R²) were generally higher for traits related to maturity than for those related to anthesis canopy status. For GY (R² = 0.26-0.51 in the three years, p < 0.001), and most DM traits, indices related to the water absorption band at 970 nm provided better relationships than the NIR/VIS indices, including the normalized difference vegetation index (NDVI), and the VIS indices. In addition, most indices including RE bands, notably NIR/RE combinations, ranked above the NIR/VIS group. Due to index saturation, the index differentiation was most apparent in the highest-yielding year. For grain Nup and total Nup, the RE/VIS index MSR_705_445 and the simple ratio R780_R740 ranked highest, followed by other RE indices. Among the vegetative organs, R² values were mostly highest and lowest for leaf and spike traits, respectively. For each trait, index and partial least squares regression (PLSR) models were validated across years at milk ripeness, confirming the suitability of optimized index selection. PLSR improved the prediction errors of some traits but not consistently the R² values. The results suggest the use of sensor-based phenotyping as a useful support tool for screening of yield potential and NUE and for identifying contributing plant traits-which, due to their expensive and cumbersome destructive determination are otherwise not readily available. Water band and RE indices should be preferred over NIR/VIS indices for DM traits and N-related traits, respectively, and milk ripeness is suggested as the most reliable stage.

Evaluating RGB Imaging and Multispectral Active and Hyperspectral Passive Sensing for Assessing Early Plant Vigor in Winter Wheat.

Plant vigor is an important trait of field crops at early growth stages, influencing weed suppression, nutrient and water use efficiency and plant growth. High-throughput techniques for its evaluation are required and are promising for nutrient management in early growth stages and for detecting promising breeding material in plant phenotyping. However, spectral sensing for assessing early plant vigor in crops is limited by the strong soil background reflection. Digital imaging may provide a low-cost, easy-to-use alternative. Therefore, image segmentation for retrieving canopy cover was applied in a trial with three cultivars of winter wheat (Triticum aestivum L.) grown under two nitrogen regimes and in three sowing densities during four early plant growth stages (Zadok's stages 14⁻32) in 2017. Imaging-based canopy cover was tested in correlation analysis for estimating dry weight, nitrogen uptake and nitrogen content. An active Greenseeker sensor and various established and newly developed vegetation indices and spectral unmixing from a passive hyperspectral spectrometer were used as alternative approaches and additionally tested for retrieving canopy cover. Before tillering (until Zadok's stage 20), correlation coefficients for dry weight and nitrogen uptake with canopy cover strongly exceeded all other methods and remained on higher levels (R² > 0.60***) than from the Greenseeker measurements until tillering. From early tillering on, red edge based indices such as the NDRE and a newly extracted normalized difference index (736 nm; ~794 nm) were identified as best spectral methods for both traits whereas the Greenseeker and spectral unmixing correlated best with canopy cover. RGB-segmentation could be used as simple low-cost approach for very early growth stages until early tillering whereas the application of multispectral sensors should consider red edge bands for subsequent stages.

Nitrogen Use Efficiency and Carbon Traits of High-Yielding European Hybrid vs. Line Winter Wheat Cultivars: Potentials and Limitations.

In contrast to allogamous crops, hybrid wheat has only recently been fostered by breeding companies in Europe. Hybrid cultivars are often associated with higher stress resistance, e.g. under drought conditions, but little is known about the nitrogen (N) use efficiency of modern hybrid wheat cultivars. Therefore, four high-yielding European hybrid and nine line winter wheat (Triticum aestivum L.) cultivars were grown under three N regimes in a high-yielding German environment and compared over 3 years at anthesis and maturity for 53 direct and indirect traits of yield formation and N allocation. Dry matter and N uptake were determined on the plant and plant organ levels. Commercial heterosis, expressing the performance of hybrid in comparison to line cultivars, was positive for about one-third of the 53 direct and indirect N and carbon traits. On average, hybrid cultivars yielded more grain (+5.5%), mainly due to a higher harvest index (+3.5%) together with higher post-anthesis assimilation and more grains per spike. However, grain N content was lower for hybrids (-8.5%), so their grain N uptake was not higher. This went along with comparable trait values for N translocation and the temporal N uptake of the different plant organs. Current wheat hybrids seem to be more efficient in overall N use because they are better at converting (higher N utilization efficiency) comparable amounts of N uptake (N uptake efficiency) into grain biomass. The results suggest that given increased seed costs for hybrids, the yield advantage of hybrid cultivars over locally adapted line cultivars will have to be further increased for establishing hybrids in low-stress, high-yielding environments.