Rhododendron delavayi Franch. is a famous ornamental plant. However, seasonal drought caused by a monsoon climate seriously limits its growth and flowering performance in urban gardens. Our previous study has found that brassinosteroids (BRs) can improve the tolerance of R. delavayi to drought stress. Here, we employed a data-independent acquisition (DIA) approach to compare the protein expression profiles under drought treatment (D) and pre-treatment with BR before drought treatment (BR). With an increase in drought stress, the net photosynthetic rate, stomatal conductance, and transpiration rate in the BR treatment showed more stable changes that were significantly higher than those in the D treatment. However, the contents of malondialdehyde, soluble sugar, soluble protein, and the activity of superoxide dismutase (SOD), peroxidase, and catalase showed opposite trends. The pre-treatment with BR alleviated the negative effect of drought stress on the photosynthetic performance of R. delavayi. A total of 3453 differentially expressed proteins (DEPs) were identified, and 683 DEPs were significantly expressed in the D and BR treatments. The DEPs uniquely expressed in the BR treatment participated in the pathways of “ribosome”, “ether lipid metabolism”, “photosynthesis”, and “oxidative phosphorylation”. The improvement effect of the BR treatment on the drought tolerance of R. delavayi was mainly attributed to improved photosynthesis by alleviating stomatal closure and oxidative stress, maintaining the integrity and stability of the ribosomal complex to mediate protein synthesis and the balance between energy metabolism and carbon metabolism. Our study presents a comprehensive understanding of the key proteins and metabolic pathways related to the response of R. delavayi to drought and will contribute to the breeding of drought-tolerant rhododendrons.

The cytokinin response regulator (RR) gene is essential for cytokinin signal transduction, which plays a crucial role in plant growth and development. Here, we applied bioinformatics to Rhododendron delavayi’s genome to identify its RR gene family and systematically analyzed their gene characteristics, phylogenetic evolution, chromosomal localization, collinearity analysis, promoter cis-elements, and expression patterns. Overall, 33 RdRR genes were distinguished and classified into three types. All these genes harbored motif 5 (YEVTTVNSGLEALELLRENKB), the most conserved one, along with the plant-conserved domain (REC domain), and could be mapped to 10 chromosomes with four gene pairs of segmental replication events but no tandem replication events; 13 RdRR genes showed collinearity with Arabidopsis thaliana genes. Promoter analysis revealed multiple hormone-related cis-elements in the RR genes. After a TDZ (thidiazuron) treatment, 13 genes had higher expression levels than the control, whose magnitude of change depended on the developmental stage of leaves’ adventitious buds. The expression levels of RdRR14, RdRR17, RdRR20, and RdRR24 agreed with the average number of adventitious buds post-TDZ treatment. We speculate that these four genes could figure prominently in bud regeneration from R. delavayi leaves in vitro. This study provides detailed knowledge of RdRRs for research on cytokinin signaling and RdRR functioning in R. delavayi.