Abstract
Panax notoginseng is a traditional Chinese medicine for the treatment of blood diseases, in which saponins were the main active components. Dammarenediol synthase (DS) is a key enzyme in the saponin synthesis pathway of P. notoginseng. The promoter is an important region to regulate gene expression, and the study of the promoter sequence provides important evidence for revealing the mechanism of gene expression regulation. However, there was still little research on the promoter function of P. notoginseng. In this study, the 1382 bp promoter upstream of DS from P. notoginseng was cloned and sequenced. The promoter sequence was analyzed by online databases. The plant expression vector fused with the β-glucuronidase gene was constructed and transferred into Agrobacterium tumefaciens. Then tobacco was injected, and its response to exogenous hormones (gibberellin and abscisic acid) was studied by transient expression to verify its unique action elements. The results showed that the tobacco leaves transferred with DS promoter had significantly increased GUS protease activity after spraying GA and ABA, indicating that both DS promoter can specifically and significantly respond to exogenous GA and ABA signal. These findings will help us to better understand the regulatory mechanisms of the upstream region of the DS gene and provide a basis for future research on the interaction of cis-acting elements of promoters with related transcription factors.
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The datasets used in this study are available from the corresponding author upon reasonable request.
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Funding
This research was funded by National Natural Science Foundation of China (81703641), key project at central government level: the ability establishment of sustainable use for valuable Chinese medicine resources (2060302), China Postdoctoral Science Foundation (2020T130601).
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Yujie Zheng and Rongyu Shao performed the experiments and data analysis, wrote the manuscript. Pengguo Xia conceived the study and contributed to the study design, discussion, and revision of the manuscript. Zongsuo Liang contributed to study design and revision of the manuscript. Kaijing Yan contributed to data verification, revision, and proofreading of the manuscript. All authors have read and approved the final manuscript and, therefore, have full access to all the data in the study and take responsibility for the integrity and security of the data.
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Zheng, Y., Shao, R., Xia, P. et al. Activity and function studies of the promoter cis-acting elements of the key enzymes in saponins biosynthesis of DS from Panax notoginseng. Protoplasma 259, 163–171 (2022). https://doi.org/10.1007/s00709-021-01653-x
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DOI: https://doi.org/10.1007/s00709-021-01653-x