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.

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.

Non-destructive in-season grain yield (GY) prediction would strongly facilitate the selection process in plant breeding but remains challenging for phenologically and morphologically diverse germplasm, notably under high-yielding conditions. In recent years, the application of drones (UAV) for spectral sensing has been established, but data acquisition and data processing have to be further improved with respect to efficiency and reliability. Therefore, this study evaluates the selection of measurement dates, sensors, and spectral parameters, as well as machine learning algorithms. Multispectral and RGB data were collected during all major growth stages in winter wheat trials and tested for GY prediction using six machine-learning algorithms. Trials were conducted in 2020 and 2021 in two locations in the southeast and eastern areas of Germany. In most cases, the milk ripeness stage was the most reliable growth stage for GY prediction from individual measurement dates, but the maximum prediction accuracies differed substantially between drought-affected trials in 2020 (R² = 0.81 and R² = 0.68 in both locations, respectively), and the wetter, pathogen-affected conditions in 2021 (R² = 0.30 and R² = 0.29). The combination of data from multiple dates improved the prediction (maximum R² = 0.85, 0.81, 0.61, and 0.44 in the four-year*location combinations, respectively). Among the spectral parameters under investigation, the best RGB-based indices achieved similar predictions as the best multispectral indices, while the differences between algorithms were comparably small. However, support vector machine, together with random forest and gradient boosting machine, performed better than partial least squares, ridge, and multiple linear regression. The results indicate useful GY predictions in sparser canopies, whereas further improvements are required in dense canopies with counteracting effects of pathogens. Efforts for multiple measurements were more rewarding than enhanced spectral information (multispectral versus RGB).

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.

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.