Abstract Increasing global warming is severely affecting tree growth and development. However, research on the sex-specific responses of dioecious trees to warming is scarce. Here, male and female Salix paraplesia were selected for artificial warming (an increase of 4 °C relative to ambient temperature) to investigate the effects on morphological, physiological, biochemical and molecular responses. The results showed that warming significantly promoted the growth of female and male S. paraplesia, but females grew faster than males. Warming affected photosynthesis, chloroplast structures, peroxidase activity, proline, flavonoids, nonstructural carbohydrates (NSCs) and phenolic contents in both sexes. Interestingly, warming increased flavonoid accumulation in female roots and male leaves but inhibited it in female leaves and male roots. The transcriptome and proteome results indicated that differentially expressed genes and proteins were significantly enriched in sucrose and starch metabolism and flavonoid biosynthesis pathways. The integrative analysis of transcriptomic, proteomic, biochemical and physiological data revealed that warming changed the expression of SpAMY, SpBGL, SpEGLC and SpAGPase genes, resulting in the reduction of NSCs and starch and the activation of sugar signaling, particularly SpSnRK1s, in female roots and male leaves. These sugar signals subsequently altered the expression of SpHCTs, SpLAR and SpDFR in the flavonoid biosynthetic pathway, ultimately leading to the differential accumulation of flavonoids in female and male S. paraplesia. Therefore, warming causes sexually differential responses of S. paraplesia, with females performing better than males.

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