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Accurate acquisition of plant phenotypic information has raised long-standing concerns in support of crop breeding programs. Different methods have been developed for high throughput plant phenotyping, while they mainly focused on the canopy level without considering the spatiotemporal heterogeneity at different canopy layers and growth stages. This study aims to phenotype spatiotemporal heterogeneity of chlorophyll (Chl) content and fluorescence response within rice leaves and canopies. Multipoint Chl content and high time-resolved Chl  a  fluorescence (ChlF) transient (OJIP transient) of rice plants were measured at different nitrogen levels and growth stages. Results showed that the Chl content within the upper leaves exhibited an increasing trend from the basal to the top portions but a decreasing pattern within the lower leaves at the most growth stages. Leaf Chl content within the rice canopy was higher in the lower leaves in the vegetative phase, while from the initial heading stage the pattern gradually reversed with the highest Chl content appearing in the upper leaves. Nitrogen supply mainly affects the occurrence time of the reverse vertical pattern. This could be the result of different nutritional demands of leaves transforming from sinks to sources, and it was further confirmed by the fall of the JI phase of OJIP transient in the vegetative phase and the rise in the reproductive phase. We further deduced that the vertical distribution of Chl content could have a defined pattern at a specific growth stage. Furthermore, the reduction of end acceptors at photosystem I (PSI) electron acceptor side per cross section (RE 0 /CS) was found to be a potential sensitive predictor for identifying the vertical heterogeneity of leaf Chl content. These findings provide prior knowledge on the vertical profiles of crop physiological traits, which explore the opportunity to develop more efficient plant phenotyping tools for crop breeding.

frontiers in plant science

Spatiotemporal Heterogeneity of Chlorophyll Content and Fluorescence Response Within Rice (Oryza sativa L.) Canopies Under Different Nitrogen Treatments