Integrating Chlorophyll Fluorescence with Anatomical and Physiological Analyses Reveals Interspecific Variation in...

To investigate interspecific variation in heat tolerance and underlying adaptation mechanisms in Rhododendron, three-year-old potted seedlings of eight taxa, representing four subgenera within the genus Rhododendron, were subjected to 40 °C high-temperature stress. Heat tolerance was comprehensively assessed using phenotypic observation, chlorophyll fluorescence imaging, microscopic examination, and physiological measurements. Results revealed that leaf damage in Rhododendron oldhamii and Rhododendron × pulchrum reached grade III, whereas Rhododendron latoucheae exhibited only grade II injury with rapid recovery. Chlorophyll fluorescence analysis showed a significant decrease in the maximum quantum efficiency of PSII (F_v/F_m) in R. liliiflorum and R. × pulchrum, followed by rapid recovery, while R. latoucheae maintained stable F_v/F_m values. Stomatal closure occurred in all taxa post-stress; stomatal characteristics of R. liliiflorum and R. simiarum remained stable, and leaf tissue structure was least affected in R. kiangsiense. R. × pulchrum demonstrated the most pronounced structural recovery. Physiologically, R. oldhamii exhibited the greatest increases in electrolyte leakage (EL) and malondialdehyde (MDA) content. R. simiarum accumulated the highest proline content under stress, while R. latoucheae showed the most significant proline reduction during recovery. By integrating multiple indicators through principal component analysis (PCA) and a membership function, and assigning weights based on variance contribution, the heat tolerance was comprehensively evaluated and ranked as follows: R. latoucheae > R. simiarum > R. oldhamii > R. ovatum > R. fortunei > R. liliiflorum > R. kiangsiense > R. × pulchrum. These findings demonstrate significant differences in heat tolerance among Rhododendron taxa at the subgenus level, with the subgenus Azaleastrum generally possessing stronger short-term heat tolerance compared to the subgenus Tsutsusi. This study provides a theoretical basis for heat-tolerant cultivar breeding and landscape application of Rhododendron.