SmSPL6 Induces Phenolic Acid Biosynthesis and Affects Root Development in Salvia miltiorrhiza

Salvia miltiorrhiza is a renowned model medicinal plant species for which 15 SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) family genes have been identified; however, the specific functions of SmSPLs have not been well characterized as of yet. For this study, the expression patterns of SmSPL6 were determined through its responses to treatments of exogenous hormones, including indole acetic acid (IAA), gibberellic acid (GA3), methyl jasmonic acid (MeJA), and abscisic acid (ABA). To characterize its functionality, we obtained SmSPL6-ovexpressed transgenic S. miltiorrhiza plants and found that overexpressed SmSPL6 promoted the accumulation of phenolic acids and repressed the biosynthesis of anthocyanin. Meanwhile, the root lengths of the SmSPL6-overexpressed lines were significantly longer than the control; however, both the fresh weights and lateral root numbers decreased. Further investigations indicated that SmSPL6 regulated the biosynthesis of phenolic acid by directly binding to the promoter regions of the enzyme genes Sm4CL9 and SmCYP98A14 and activated their expression. We concluded that SmSPL6 regulates not only the biosynthesis of phenolic acids, but also the development of roots in S. miltiorrhiza.

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The methyl jasmonate-responsive transcription factor SmMYB1 promotes phenolic acid biosynthesis in Salvia miltiorrhiza

Horticulture Research ◽

10.1038/s41438-020-00443-5 ◽

2021 ◽

Vol 8 (1) ◽

Author(s):

Wei Zhou ◽

Min Shi ◽

Changping Deng ◽

Sunjie Lu ◽

Fenfen Huang ◽

...

Keyword(s):

Transcription Factor ◽

Methyl Jasmonate ◽

Phenolic Acids ◽

Phenolic Acid ◽

Salvia Miltiorrhiza ◽

Water Soluble ◽

Biosynthesis Pathway ◽

Acid Biosynthesis ◽

Expression Of Genes ◽

Genes Encoding

AbstractWater-soluble phenolic acids are major bioactive compounds in the medicinal plant species Salvia miltiorrhiza. Phenolic acid biosynthesis is induced by methyl jasmonate (MeJA) in this important Chinese herb. Here, we investigated the mechanism underlying this induction by analyzing a transcriptome library of S. miltiorrhiza in response to MeJA. Global transcriptome analysis identified the MeJA-responsive R2R3-MYB transcription factor-encoding gene SmMYB1. Overexpressing SmMYB1 significantly promoted phenolic acid accumulation and upregulated the expression of genes encoding key enzymes in the phenolic acid biosynthesis pathway, including cytochrome P450-dependent monooxygenase (CYP98A14). Dual-luciferase (dual-LUC) assays and/or an electrophoretic mobility shift assays (EMSAs) indicated that SmMYB1 activated the expression of CYP98A14, as well as the expression of genes encoding anthocyanin biosynthesis pathway enzymes, including chalcone isomerase (CHI) and anthocyanidin synthase (ANS). In addition, SmMYB1 was shown to interact with SmMYC2 to additively promote CYP98A14 expression compared to the action of SmMYB1 alone. Taken together, these results demonstrate that SmMYB1 is an activator that improves the accumulation of phenolic acids and anthocyanins in S. miltiorrhiza. These findings lay the foundation for in-depth studies of the molecular mechanism underlying MeJA-mediated phenolic acid biosynthesis and for the metabolic engineering of bioactive ingredients in S. miltiorrhiza.

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Multiplexed CRISPR/Cas9-Mediated Knockout of Laccase Genes in Salvia miltiorrhiza Revealed Their Roles in Growth, Development, and Metabolism

Frontiers in Plant Science ◽

10.3389/fpls.2021.647768 ◽

2021 ◽

Vol 12 ◽

Author(s):

Zheng Zhou ◽

Qing Li ◽

Liang Xiao ◽

Yun Wang ◽

Jingxian Feng ◽

...

Keyword(s):

Phenolic Acids ◽

Hairy Roots ◽

Cross Sections ◽

Phenolic Acid ◽

Lignin Content ◽

Salvia Miltiorrhiza ◽

Acid Biosynthesis ◽

Disease Treatment ◽

In The Wild ◽

Laccase Genes

Laccases are multicopper-containing glycoproteins related to monolignol oxidation and polymerization. These properties indicate that laccases may be involved in the formation of important medicinal phenolic acid compounds in Salvia miltiorrhiza such as salvianolic acid B (SAB), which is used for cardiovascular disease treatment. To date, 29 laccases have been found in S. miltiorrhiza (SmLACs), and some of which (SmLAC7 and SmLAC20) have been reported to influence the synthesis of phenolic acids. Because of the functional redundancy of laccase genes, their roles in S. miltiorrhiza are poorly understood. In this study, the CRISPR/Cas9 system was used for targeting conserved domains to knockout multiple genes of laccase family in S. miltiorrhiza. The expressions of target laccase genes as well as the phenolic acid biosynthesis key genes decrease dramatically in editing lines. Additionally, the growth and development of hairy roots was significantly retarded in the gene-edited lines. The cross-sections examination of laccase mutant hairy roots showed that the root development was abnormal and the xylem cells in the edited lines became larger and looser than those in the wild type. Additionally, the accumulation of RA as well as SAB was decreased, and the lignin content was nearly undetectable. It suggested that SmLACs play key roles in development and lignin formation in the root of S. miltiorrhiza and they are necessary for phenolic acids biosynthesis.

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SmMYB111 Is a Key Factor to Phenolic Acid Biosynthesis and Interacts with Both SmTTG1 and SmbHLH51 in Salvia miltiorrhiza

Journal of Agricultural and Food Chemistry ◽

10.1021/acs.jafc.8b02548 ◽

2018 ◽

Vol 66 (30) ◽

pp. 8069-8078 ◽

Cited By ~ 16

Author(s):

Shasha Li ◽

Yucui Wu ◽

Jing Kuang ◽

Huaiqin Wang ◽

Tangzhi Du ◽

...

Keyword(s):

Phenolic Acid ◽

Salvia Miltiorrhiza ◽

Acid Biosynthesis ◽

Key Factor

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Regulation of Water-Soluble Phenolic Acid Biosynthesis in Salvia miltiorrhiza Bunge

Applied Biochemistry and Biotechnology ◽

10.1007/s12010-013-0265-4 ◽

2013 ◽

Vol 170 (6) ◽

pp. 1253-1262 ◽

Cited By ~ 34

Author(s):

Pengda Ma ◽

Jingling Liu ◽

Chenlu Zhang ◽

Zongsuo Liang

Keyword(s):

Phenolic Acid ◽

Salvia Miltiorrhiza ◽

Water Soluble ◽

Salvia Miltiorrhiza Bunge ◽

Acid Biosynthesis

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Comparative RNA-Sequence Transcriptome Analysis of Phenolic Acid Metabolism in Salvia miltiorrhiza, a Traditional Chinese Medicine Model Plant

International Journal of Genomics ◽

10.1155/2017/9364594 ◽

2017 ◽

Vol 2017 ◽

pp. 1-10 ◽

Cited By ~ 11

Author(s):

Zhenqiao Song ◽

Linlin Guo ◽

Tian Liu ◽

Caicai Lin ◽

Jianhua Wang ◽

...

Keyword(s):

Chinese Medicine ◽

Traditional Chinese Medicine ◽

Phenolic Acid ◽

Expression Profiles ◽

Salvia Miltiorrhiza ◽

Lignin Biosynthesis ◽

Biosynthesis Pathway ◽

Acid Biosynthesis ◽

The Core ◽

Phenylpropanoid Biosynthesis

Salvia miltiorrhiza Bunge is an important traditional Chinese medicine (TCM). In this study, two S. miltiorrhiza genotypes (BH18 and ZH23) with different phenolic acid concentrations were used for de novo RNA sequencing (RNA-seq). A total of 170,787 transcripts and 56,216 unigenes were obtained. There were 670 differentially expressed genes (DEGs) identified between BH18 and ZH23, 250 of which were upregulated in ZH23, with genes involved in the phenylpropanoid biosynthesis pathway being the most upregulated genes. Nine genes involved in the lignin biosynthesis pathway were upregulated in BH18 and thus result in higher lignin content in BH18. However, expression profiles of most genes involved in the core common upstream phenylpropanoid biosynthesis pathway were higher in ZH23 than that in BH18. These results indicated that genes involved in the core common upstream phenylpropanoid biosynthesis pathway might play an important role in downstream secondary metabolism and demonstrated that lignin biosynthesis was a putative partially competing pathway with phenolic acid biosynthesis. The results of this study expanded our understanding of the regulation of phenolic acid biosynthesis in S. miltiorrhiza.

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R2R3-MYB Transcription Factor SmMYB52 Positively Regulates Biosynthesis of Salvianolic Acid B and Inhibits Root Growth in Salvia miltiorrhiza

International Journal of Molecular Sciences ◽

10.3390/ijms22179538 ◽

2021 ◽

Vol 22 (17) ◽

pp. 9538

Author(s):

Rao Yang ◽

Shengsong Wang ◽

Haolan Zou ◽

Lin Li ◽

Yonghui Li ◽

...

Keyword(s):

Transcription Factor ◽

Root Growth ◽

Phenolic Acid ◽

Salvia Miltiorrhiza ◽

Salvianolic Acid B ◽

Myb Transcription Factor ◽

Acid Biosynthesis ◽

Key Enzyme ◽

R2r3 Myb ◽

Encoding Genes

The dried root of Salvia miltiorrhiza is a renowned traditional Chinese medicine that was used for over 1000 years in China. Salvianolic acid B (SalB) is the main natural bioactive product of S. miltiorrhiza. Although many publications described the regulation mechanism of SalB biosynthesis, few reports simultaneously focused on S. miltiorrhiza root development. For this study, an R2R3-MYB transcription factor gene (SmMYB52) was overexpressed and silenced, respectively, in S. miltiorrhiza sterile seedlings. We found that SmMYB52 significantly inhibited root growth and indole-3-acetic acid (IAA) accumulation, whereas it activated phenolic acid biosynthesis and the jasmonate acid (JA) signaling pathway. Quantitative real-time polymerase chain reaction (qRT-PCR) analyses revealed that SmMYB52 suppressed the transcription levels of key enzyme-encoding genes involved in the IAA biosynthetic pathway and activated key enzyme-encoding genes involved in the JA and phenolic acid biosynthesis pathways. In addition, yeast one-hybrid (Y1H) and dual-luciferase assay showed that SmMYB52 directly binds to and activates the promoters of several key enzyme genes for SalB biosynthesis, including SmTAT1, Sm4CL9, SmC4H1, and SmHPPR1, to promote the accumulation of SalB. This is the first report of a regulator that simultaneously affects root growth and the production of phenolic acids in S. miltiorrhiza.

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