The AP2/ERF transcription factor SmERF128 positively regulates diterpenoid biosynthesis in Salvia miltiorrhiza
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The novel AP2/ERF transcription factor SmERF128 positively regulates diterpenoid tanshinone biosynthesis by activating the expression of SmCPS1, SmKSL1, and SmCYP76AH1 in Salvia miltiorrhiza.
Certain members of the APETALA2/ethylene-responsive factor (AP2/ERF) family regulate plant secondary metabolism. Although it is clearly documented that AP2/ERF transcription factors (TFs) are involved in sesquiterpenoid biosynthesis, the regulation of diterpenoid biosynthesis by AP2/ERF TFs remains elusive. Here, we report that the novel AP2/ERF TF SmERF128 positively regulates diterpenoid tanshinone biosynthesis in Salvia miltiorrhiza. Overexpression of SmERF128 increased the expression levels of copalyl diphosphate synthase 1 (SmCPS1), kaurene synthase-like 1 (SmKSL1) and cytochrome P450 monooxygenase 76AH1 (SmCYP76AH1), whereas their expression levels were decreased when SmERF128 was silenced. Accordingly, the content of tanshinone was reduced in SmERF128 RNA interference (RNAi) hairy roots and dramatically increased in SmERF128 overexpression hairy roots, as demonstrated through Ultra Performance Liquid Chromatography (UPLC) and Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) analysis. Furthermore, SmERF128 activated the expression of SmCPS1, SmKSL1, and SmCYP76AH1 by binding to the GCC box, and to the CRTDREHVCBF2 (CBF2) and RAV1AAT (RAA) motifs within their promoters during in vivo and in vitro assays. Our findings not only reveal the molecular basis of how the AP2/ERF transcription factor SmERF128 regulates diterpenoid biosynthesis, but also provide useful information for improving tanshinone production through genetic engineering.
KeywordsSalvia miltiorrhiza Diterpenoid Tanshinones AP2/ERF transcription factor Herbgenomics
4-(Cytidine 5′-diphospho)-2-C-methyl-D-erythritol kinase
Copalyl diphosphate synthase 1
Cytochrome P450 monooxygenase 76AH1
1-Deoxy-D-xylulose-5-phosphate synthase 2
Green fluorescent protein
Geranylgeranyl diphosphate synthase
4-Hydroxy-3-methylbut-2-enyl diphosphate reductase 1
4-Hydroxy-3-methylbut-2-enyl diphosphate synthase
Kaurene synthase-like 1
2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase
2-C-methyl-D-erythritol 2,4-cyclodiphosphate synthase
Nicotinamide adenine dinucleotide phosphate
Terpenoid indole alkaloid
This work was supported by the National Natural Science Foundation of China (Grant No. 81573398), CAMS Innovation Fund for Medical Sciences (CIFMS) (2017-I2M-1-009) and Foundation of Educational Department of Guangdong Province (E1-KFD015181K31).
JS designed the study. YZ and AJ performed experiments. YZ, AJ, ZX and HL analyzed the data. YZ, AJ, JS, and ZX wrote the manuscript. All authors approved the final manuscript.
Compliance with ethical standards
Conflict of interest
All the authors declare that they have no conflict of interests.
- Zhang H, Hedhili S, Montiel G, Zhang Y, Chatel G, Pre M, Gantet P, Memelink J (2011) The basichelix-loop-helix transcription factor CrMYC2 controls the jasmonate-responsive expression of the ORCA genes that regulate alkaloid biosynthesis in Catharanthus roseus. Plant J 67:61–71CrossRefPubMedGoogle Scholar