The AP2/ERF transcription factor SmERF128 positively regulates diterpenoid biosynthesis in Salvia miltiorrhiza
The AP2/ERF transcription factor SmERF128 positively regulates diterpenoid biosynthesis in Salvia miltiorrhiza
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AP2/ERF转录因子SmERF128正向调节丹参二萜生物合成
DOI：
10.1007/s11103-019-00845-7
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发表时间：
2019-05-01
影响因子：
5.1
通讯作者：
Song, Jingyuan
中科院分区：
文献类型：
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作者：
Zhang, Yu;Ji, Aijia;Song, Jingyuan
新型 AP2/ERF 转录因子 SmERF128 通过激活丹参中 SmCPS1、SmKSL1 和 SmCYP76AH1 的表达，正向调节二萜类丹参酮生物合成。 APETALA2/乙烯反应因子 (AP2/ERF) 家族的某些成员调节植物次生代谢。尽管有明确记录表明 AP2/ERF 转录因子 (TF) 参与倍半萜生物合成，但 AP2/ERF 转录因子对二萜生物合成的调节仍然难以捉摸。在这里，我们报道了新型 AP2/ERF TF SmERF128 正向调节丹参中二萜类丹参酮的生物合成。 SmERF128的过表达增加了柯巴基二磷酸合成酶1（SmCPS1）、贝壳杉烯合成酶样1（SmKSL1）和细胞色素P450单加氧酶76AH1（SmCYP76AH1）的表达水平，而当SmERF128被沉默时，它们的表达水平降低。因此，超高效液相色谱 (UPLC) 和液相色谱-串联质谱 (LC-MS/MS) 分析表明，SmERF128 RNA 干扰 (RNAi) 毛状根中丹参酮的含量降低，而 SmERF128 过表达毛状根中丹参酮的含量显着增加。此外，在体内和体外试验中，SmERF128 通过与 GCC 盒以及启动子内的 CRTDREHVCBF2 (CBF2) 和 RAV1AAT (RAA) 基序结合，激活 SmCPS1、SmKSL1 和 SmCYP76AH1 的表达。我们的研究结果不仅揭示了 AP2/ERF 转录因子 SmERF128 如何调节二萜生物合成的分子基础，而且为通过基因工程提高丹参酮生产提供了有用的信息。
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.