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The ethylene response factor SmERF6 co-regulates the transcription of SmCPS1 and SmKSL1 and is involved in tanshinone biosynthesis in Salvia miltiorrhiza hairy roots

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Abstract

Main Conclusion

The SmERF6, which recognizes the GCC-box of SmCPS1 and SmKSL1 promoter in nucleus, regulates the tanshinone biosynthesis in Salvia miltiorrhiza hairy roots.

Tanshinone, an important medicinal ingredient in Salvia miltiorrhiza, is best known for its use in medicine. However, the transcription factor regulation of tanshinone biosynthesis is unclear. Here, we isolated and identified a transcription factor in the ERF family of S. miltiorrhiza, SmERF6, which was screened from an S. miltiorrhiza cDNA library by the promoters of two key tanshinone synthesis genes (SmKSL1 and SmCPS1); this factor regulated tanshinone biosynthesis. The gene was highly expressed in the root and responded to ethylene treatment. SmERF6 modulated tanshinone biosynthesis by directly binding to an ethylene-responsive element (GCC-box) of the SmKSL1 and SmCPS1 promoters and activating their transcription. Overexpression of SmERF6 in the hairy roots increased their tanshinone accumulation, and SmERF6 silencing by RNAi led to a lower tanshinone content. Furthermore, tanshinone accumulation maintained homeostasis with the total phenolic acid and flavonoid contents in S. miltiorrhiza. These findings elucidated how SmERF6 directly co-regulates the transcription of SmCPS1 and SmKSL1 and modulates tanshinone synthesis to accelerate the metabolic flux of tanshinone accumulation in S. miltiorrhiza.

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Abbreviations

SmCPS1 :

Salvia miltiorrhiza copalyl diphosphate synthases 1

SmKSL1 :

Salvia miltiorrhiza kaurene synthase like 1

CT:

Cryptotanshinone

DT-I:

Dihydrotanshinone

ERF:

Ethylene response factor

T-I (IIA):

Tanshinone I (IIA)

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Acknowledgements

We are grateful to professors Chao Sun, Shilin Chen and Weibo Jin for kindly providing the genome resources and transcriptome data of S. miltiorrhiza used in this study, respectively. We thanks for the supports from the National Natural Science Foundation of China (81773835, 81373908 and 81373536).

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Authors and Affiliations

  1. College of Life Science, Northwest A&F University, Yangling, 712100, China

    Zhenqing Bai, Yanyan Jia, Jie Jiao, Wenli Huang & Zongsuo Liang

  2. College of Life Science, Yan’an University, Yan’an, China

    Zhenqing Bai

  3. College of Life Science, Zhejiang Sci-Tech University, Hangzhou, China

    Pengguo Xia & Zongsuo Liang

  4. Institute of Soil and Water Conservation, Chinese Academy of Sciences & Ministry of Water Resources, Yangling, China

    Wenrui Li

  5. University of the Chinese Academy of Sciences, Beijing, China

    Wenrui Li

  6. College of Horticulture, Northwest A&F University, Yangling, China

    Zhiyong Yue

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  1. Zhenqing Bai
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Correspondence to Zongsuo Liang.

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Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 Fig. S1 Promoter character analysis of SmCPS1 and SmKSL1 (TIFF 3180 kb)

425_2018_2884_MOESM2_ESM.tif

Supplementary material 2 Fig. S2 The minimal inhibitory concentration of Aureobasidin A (AbA) screens for the bait strain of the SmCPS1 and SmKSL1 promoter (TIFF 3681 kb)

425_2018_2884_MOESM3_ESM.tif

Supplementary material 3 Fig. S3 The Western-blotting analysis of MBP and SmERF6 and the precursor microRNA structure of amiR159b-SmERF6 (TIFF 1532 kb)

Supplementary material 4 Fig. S4 The phylogeny and characteristic analysis of SmERF6 (TIFF 12015 kb)

Supplementary material 5 (DOCX 15 kb)

Supplementary material 6 (DOCX 25 kb)

Supplementary material 7 (DOCX 19 kb)

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Bai, Z., Li, W., Jia, Y. et al. The ethylene response factor SmERF6 co-regulates the transcription of SmCPS1 and SmKSL1 and is involved in tanshinone biosynthesis in Salvia miltiorrhiza hairy roots. Planta 248, 243–255 (2018). https://doi.org/10.1007/s00425-018-2884-z

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