SmCSN5 is a synergist in the transcription factor SmMYB36-mediated biosynthesis of tanshinones and phenolic acids in Salvia miltiorrhiza

Abstract

The ubiquitin-26S proteasome system (UPS) is associated with protein stability and activity, regulation of hormone signaling, and the production of secondary metabolites in plants. Though the mechanism of action of SmMYB36 on the tanshinone and phenolic acid biosynthesis is well understood, its regulation through post-translational modifications is unclear. A constitutive photomorphogenesis 9 (COP9) signalosome subunit 5 (SmCSN5), which interacted with SmMYB36 and inhibited its ubiquitination-based degradation, was identified in Salvia miltiorrhiza. SmCSN5 promoted tanshinone biosynthesis but inhibited phenolic acid biosynthesis in the hairy roots of S. miltiorrhiza. SmMYB36 also activated the transcription of the target genes SmDXS2 and SmCPS1 but repressed that of SmRAS in a SmCSN5-dependent manner. SmCSN5 acts as a positive regulator in MeJA-induced biosynthesis of tanshinones and phenolic acids. Specifically, SmCSN5 alone, when expressed transiently in tobacco and rice protoplasts, was localized to the cytoplasm, cell membrane, and nucleus, whereas when coexpressed with SmMYB36, it was detected only in the nucleus. Additionally, the degradation of SmMYB36^1-153 by ubiquitination was lowered after truncation of the self-activating structural domain of SmMYB36^154-160. Collectively, these results suggest that SmCSN5 affected the transcriptional activation of SmMYB36 and stabilized SmMYB36, providing insights into the SmMYB36-based regulation of the accumulation of tanshinone and phenolic acid at the transcriptional and post-translational levels.

Acknowledgements

This work was supported by the National Natural Science Foundation of China (Project No. 32270278) and Key project at central government level: the ability establishment of sustainable use for valuable Chinese medicine resources (2060302). We thank Yu Tang and Shuang Wang from the College of Life Sciences, Sichuan Agricultural University, for their help in the tissue-specific expression experiments.

Author contributions

P.D.M. and J.E.D. designed the research project. Q.L. conducted majority of the experiments, data processing, and manuscript drafting. X.J.W., Y.S., Y.Y.G., and J.Y. performed a few of the experiments. Z.Y.X. and J.Y.L. participated in the editing of the manuscript. P.D.M. provided constructive comments for the modification of the manuscript.

Data availability

Detailed data supporting the findings of this study can be found in the supplementary material.

Conflict of interest statement

The authors have no conflicts of interest to declare.

Supplementary data

Supplementary data is available at Horticulture Research online.

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