The S40 family members delay leaf senescence by promoting cytokinin synthesis.

Leaf senescence is regulated by both endogenous hormones and environmental stimuli in a programmed and concerted way. The members of the S40 family have been reported to play roles in leaf senescence. Here we identified an S40 family member, CiS40-11, from Caragana intermedia. Phylogenetic analysis revealed that the CiS40-11 protein had the highest identity with AtS40-5 (AT1G11700) and AtS40-6 (AT1G61930) of Arabidopsis thaliana. CiS40-11 was highly expressed in leaves and was down-regulated after dark treatment. The subcellular localization analysis showed that CiS40-11 was a cytoplasm-nucleus dual-localized protein. Leaf senescence was delayed in both the CiS40-11 overexpressed A. thaliana and its transiently expressed C. intermedia. Transcriptomic analysis and endogenous hormones assay revealed that CiS40-11 inhibited leaf senescence via promoting the biosynthesis of cytokinins by blocking AtMYB2 expression in the CiS40-11 overexpression lines. Furthermore, overexpression of either AtS40-5 or AtS40-6 showed similar phenotype as the CiS40-11 overexpressing lines, while in the ats40-5a or ats40-6a mutants, the AtMYB2 expression was increased and their leaves exhibited a premature senescence phenotype. These results provide a new molecular mechanism of the S40 family in leaf senescence regulation of plants.