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AtPAP1 Interacts With and Activates SmbHLH51, a Positive Regulator to Phenolic Acids Biosynthesis in Salvia miltiorrhiza.

ABSTRACT: Phenolic acids from Salvia miltiorrhiza have drawn considerable attention in recent years because of their remarkable pharmacological activities. We previously reported that Arabidopsis thaliana transcription factor production of anthocyanin pigment 1 (AtPAP1) has strong capability to promote the production of phenolic acids in S. miltiorrhiza. However, the responsible molecular mechanism is unclear. Here, we analyzed the transcriptome of transgenic S. miltiorrhiza that over-expressed AtPAP1. Transcriptome analysis revealed 4,152 genes that were differentially expressed due to ectopic AtPAP1 overexpression. SmbHLH51, a novel bHLH gene significantly up-regulated by constitutive expression of AtPAP1, was isolated from S. miltiorrhiza for detailed functional characterization. SmbHLH51 localizes in the nuclei and interacts with AtPAP1, indicating that they probably comprise a regulatory transcription complex. Enhanced or reduced expression of SmbHLH51 was achieved in S. miltiorrhiza by gain- or loss-of-function assays, respectively, revealing that SmbHLH51 is a positive transcriptional regulator of the pathway for phenolic acid biosynthesis. We propose that applying this functional genomics approach through the transcriptomic analyses is an efficient means for identifying novel genes involved in plant secondary metabolism.

SUBMITTER: Wu Y

PROVIDER: S-EPMC6255977 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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AtPAP1 Interacts With and Activates SmbHLH51, a Positive Regulator to Phenolic Acids Biosynthesis in Salvia miltiorrhiza.

Wu Yucui Y Zhang Yuan Y Li Lin L Guo Xiaorong X Wang Bin B Cao Xiaoyan X Wang Zhezhi Z

Frontiers in plant science 20181120

Phenolic acids from Salvia miltiorrhiza have drawn considerable attention in recent years because of their remarkable pharmacological activities. We previously reported that Arabidopsis thaliana transcription factor production of anthocyanin pigment 1 (AtPAP1) has strong capability to promote the production of phenolic acids in S. miltiorrhiza. However, the responsible molecular mechanism is unclear. Here, we analyzed the transcriptome of transgenic S. miltiorrhiza th ...[more]

PMID: 30515184

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Project description:IntroductionSalvia miltiorrhiza is a renowned traditional Chinese medicinal plant with extremely high medicinal value, especially for cardiovascular and cerebrovascular diseases. The jasmonic acid (JA) signaling pathway plays an important role in the improved biosynthesis of secondary metabolites, which is mediated by a major transcriptional regulator, MYC2. However, the JA regulatory mechanism of secondary metabolites biosynthesis in S. miltiorrhiza is still largely unknown.ObjectivesOur work focuses on the dissection of the molecular mechanism of transcriptional regulation in MeJA-mediated biosynthesis of medicinal components of S. miltiorrhiza. We examined the role of MeJA-responsive bHLH transcription factors (TFs) in improving bioactive secondary metabolites accumulation in S. miltiorrhiza.MethodsHairy root transformation based on CRISPR/Cas9 technique was used to decipher gene function(s). Changes in the content of phenolic acids were evaluated by HPLC. Y1H, EMSA and dual-LUC assays were employed to analyze the molecular mechanism of SmbHLH60 in the regulation on the biosynthesis of phenolic acids and anthocyanins. Y2H, BiFC and pull-down affinity assays were used to corroborate the interaction between SmbHLH60 and SmMYC2.ResultsBeing one of the most significantly negatively regulated bHLH genes by MeJA, a new transcription factor SmbHLH60 was discovered and characterized. Over-expression of SmbHLH60 resulted in significant inhibition of phenolic acid and anthocyanin biosynthesis in S. miltiorrhiza by transcriptionally repressing of target genes such as SmTAT1 and SmDFR, whereas CRISPR/Cas9-generated knockout of SmbHLH60 resulted in the opposite effect. In addition, SmbHLH60 and SmMYC2 formed a heterodimer to antagonistically regulate phenolic acid and anthocyanin biosynthesis.ConclusionOur results clarified that SmbHLH60 is a negativeregulator on the biosynthesis of phenolic acids and anthocyanins. SmbHLH60 competed with SmMYC2 in an antagonistic manner, providing new insights for the molecular mechanism of MeJA-mediated regulation on the biosynthesis of secondary metabolites in S. miltiorrhiza.

Dataset Information

AtPAP1 Interacts With and Activates SmbHLH51, a Positive Regulator to Phenolic Acids Biosynthesis in Salvia miltiorrhiza.

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AtPAP1 Interacts With and Activates SmbHLH51, a Positive Regulator to Phenolic Acids Biosynthesis in <i>Salvia miltiorrhiza</i>.

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