High-throughput automated image analysis holds great promise for plant breeding by enabling faster, more accurate assessment of traits relevant to crop improvement. Imaging-based systems, such as the CropReporter, allow automated quantification of photosynthetic parameters like PSII efficiency under ambient light from a top-down 2D perspective. However, standard analysis tools average values across the 2D top view, overrepresenting upper leaves and underrepresenting those in the lower canopy. Upper leaves may occlude lower ones, and due to the pinhole projection of the camera, lower leaves of the same size appear smaller in the image. Consequently, vertical heterogeneity in PSII efficiency within the canopy cannot be resolved using a single 2D image. To address these issues, we integrated top-view PSII efficiency data (by CropReporter) with 3D structural data from RGB point clouds (by MaxiMarvin). Alignment accuracy between MaxiMarvin and CropReporter was high, with R² ≥ 0.98 for the x-axis and R² ≥ 0.99 for the y-axis. The method was tested using Chenopodium quinoa, Glycine max, and Solanum tuberosum, exposed to salinity, waterlogging and drought stress respectively. In Chenopodium quinoa, it allowed precise determination of when senescence began in the lower leaves. In Solanum tuberosum, the reduction in PSII efficiency by drought was the same for all leaf layers, while in Glycine max, waterlogging stress most strongly affected the middle layer of the canopy. This framework enables the 3D mapping of PSII efficiency across the vertical plant profile by combining top-view chlorophyll fluorescence imaging (CropReporter) with 3D structural data (MaxiMarvin). It reveals vertical variation in photosynthetic activity across canopy layers. With standard 2D chlorophyll fluorescence imaging it is difficult to distinguish between non-photosynthetic tissues like flower heads and lower layers of leaves, that might have the same PSII values. Using height-based filtering, taking data from the 3D mapping, such distinction can be made with the method presented in this paper. This allows estimating the PSII efficiencies of leaves only. By capturing layer-specific responses to abiotic stress and developmental changes, the method provides physiologically relevant input for crop growth modelling and highlights the importance of accounting for canopy structure in photosynthetic analyses.
Vlaardingen et al. (Wed,) studied this question.