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Detailed observation of the phenotypic changes in rice panicle substantially helps us to understand the yield formation. In recent studies, phenotyping of rice panicles during the heading–flowering stage still lacks comprehensive analysis, especially of panicle development under different nitrogen treatments. In this work, we proposed a pipeline to automatically acquire the detailed panicle traits based on time-series images by using the YOLO v5, ResNet50, and DeepSORT models. Combined with field observation data, the proposed method was used to test whether it has an ability to identify subtle differences in panicle developments under different nitrogen treatments. The result shows that panicle counting throughout the heading–flowering stage achieved high accuracy (R2 = 0.96 and RMSE = 1.73), and heading date was estimated with an absolute error of 0.25 days. In addition, by identical panicle tracking based on the time-series images, we analyzed detailed flowering phenotypic changes of a single panicle, such as flowering duration and individual panicle flowering time. For rice population, with an increase in the nitrogen application: panicle number increased, heading date changed little, but the duration was slightly extended; cumulative flowering panicle number increased, rice flowering initiation date arrived earlier while the ending date was later; thus, the flowering duration became longer. For a single panicle, identical panicle tracking revealed that higher nitrogen application led to earlier flowering initiation date, significantly longer flowering days, and significantly longer total duration from vigorous flowering beginning to the end (total DBE). However, the vigorous flowering beginning time showed no significant differences and there was a slight decrease in daily DBE.
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