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

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.

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

2013 ◽  
Vol 170 (6) ◽  
pp. 1253-1262 ◽  
Author(s):  
Pengda Ma ◽  
Jingling Liu ◽  
Chenlu Zhang ◽  
Zongsuo Liang
Keyword(s):  
Phenolic Acid ◽  
Salvia Miltiorrhiza ◽  
Water Soluble ◽  
Salvia Miltiorrhiza Bunge ◽  
Acid Biosynthesis

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

2017 ◽  
Vol 2017 ◽  
pp. 1-10 ◽  
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.

scholarly journals Synergistic Effect of Methyl Jasmonate and Abscisic Acid Co-Treatment on Avenanthramide Production in Germinating Oats

2021 ◽  
Vol 22 (9) ◽  
pp. 4779
Author(s):  
Soyoung Kim ◽  
Tae Hee Kim ◽  
Yu Jeong Jeong ◽  
Su Hyun Park ◽  
Sung Chul Park ◽  
...  
Keyword(s):  
Abscisic Acid ◽  
Synergistic Effect ◽  
Methyl Jasmonate ◽  
Avena Sativa ◽  
Biosynthesis Pathway ◽  
Grass Family ◽  
Expression Of Genes ◽  
Valuable Method ◽  
Genes Encoding ◽  
Anti Oxidant

The oat (Avena sativa L.) is a grain of the Poaceae grass family and contains many powerful anti-oxidants, including avenanthramides as phenolic alkaloids with anti-inflammatory, anti-oxidant, anti-itch, anti-irritant, and anti-atherogenic activities. Here, the treatment of germinating oats with methyl jasmonate (MeJA) or abscisic acid (ABA) resulted in 2.5-fold (582.9 mg/kg FW) and 2.8-fold (642.9 mg/kg FW) increase in avenanthramide content, respectively, relative to untreated controls (232.6 mg/kg FW). Moreover, MeJA and ABA co-treatment synergistically increased avenanthramide production in germinating oats to 1505 mg/kg FW. Individual or combined MeJA and ABA treatment increased the expression of genes encoding key catalytic enzymes in the avenanthramide-biosynthesis pathway, including hydroxycinnamoyl-CoA:hydrocyanthranilate N-hydroxycinnamoyl transferase (HHT). Further analyses showed that six AsHHT genes were effectively upregulated by MeJA or ABA treatment, especially AsHHT4 for MeJA and AsHHT5 for ABA, thereby enhancing the production of all three avenanthramides in germinating oats. Specifically, AsHHT5 exhibited the highest expression following MeJA and ABA co-treatment, indicating that AsHHT5 played a more crucial role in avenanthramide biosynthesis in response to MeJA and ABA co-treatment of germinating oats. These findings suggest that elicitor-mediated metabolite farming using MeJA and ABA could be a valuable method for avenanthramide production in germinating oats.

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

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.

scholarly journals Molecular Characterization and Overexpression of SmJMT Increases the Production of Phenolic Acids in Salvia miltiorrhiza

2018 ◽  
Vol 19 (12) ◽  
pp. 3788 ◽  
Author(s):  
Bin Wang ◽  
Junfeng Niu ◽  
Bin Li ◽  
Yaya Huang ◽  
Limin Han ◽  
...  
Keyword(s):  
Phenolic Acids ◽  
Genetic Manipulation ◽  
Salvia Miltiorrhiza ◽  
Plant Responses ◽  
Biosynthesis Pathway ◽  
Genes Encoding ◽  
Transgenic Lines ◽  
Key Enzyme ◽  
Manipulation Strategy ◽  
Important Medicinal Plant

Jasmonic acid (JA) carboxyl methyltransferase (JMT), a key enzyme in jasmonate-regulated plant responses, may be involved in plant defense and development by methylating JA to MeJA, thus influencing the concentrations of MeJA in plant. In this study, we isolated the JMT gene from Salvia miltiorrhiza, an important medicinal plant widely used to treat cardiovascular disease. We present a genetic manipulation strategy to enhance the production of phenolic acids by overexpresion SmJMT in S. miltiorrhiza. Global transcriptomic analysis using RNA sequencing showed that the expression levels of genes involved in the biosynthesis pathway of phenolic acids and MeJA were upregulated in the overexpression lines. In addition, the levels of endogenous MeJA, and the accumulation of rosmarinic acid (RA) and salvianolic acid (Sal B), as well as the concentrations of total phenolics and total flavonoids in transgenic lines, were significantly elevated compared with the untransformed control. Our results demonstrate that overexpression of SmJMT promotes the production of phenolic acids through simultaneously activating genes encoding key enzymes involved in the biosynthesis pathway of phenolic acids and enhancing the endogenous MeJA levels in S. miltiorrhiza.

scholarly journals ABA-responsive transcription factor bZIP1 is involved in modulating biosynthesis of phenolic acids and tanshinones in Salvia miltiorrhiza

2020 ◽  
Vol 71 (19) ◽  
pp. 5948-5962 ◽  
Author(s):  
Changping Deng ◽  
Min Shi ◽  
Rong Fu ◽  
Yi Zhang ◽  
Qiang Wang ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Phenolic Acids ◽  
Phenolic Acid ◽  
Leucine Zipper ◽  
Salvia Miltiorrhiza ◽  
Market Demand ◽  
Biosynthetic Genes ◽  
Basic Leucine Zipper

Abstract Phenolic acids and tanshinones are major bioactive ingredients in Salvia miltiorrhiza, which possess pharmacological activities with great market demand. However, transcriptional regulation of phenolic acid and tanshinone biosynthesis remains poorly understood. Here, a basic leucine zipper transcription factor (TF) named SmbZIP1 was screened from the abscisic acid (ABA)-induced transcriptome library. Overexpression of SmbZIP1 positively promoted phenolic acid biosynthesis by enhancing expression of biosynthetic genes such as cinnamate-4-hydroxylase (C4H1). Furthermore, biochemical experiments revealed that SmbZIP1 bound the G-Box-like1 element in the promoter of the C4H1 gene. Meanwhile, SmbZIP1 inhibited accumulation of tanshinones mainly by suppressing the expression of biosynthetic genes including geranylgeranyl diphosphate synthase (GGPPS) which was confirmed as a target gene by in vitro and in vivo experiments. In contrast, the phenolic acid content was reduced and tanshinone was enhanced in CRISPR/Cas9 [clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein 9]-mediated knockout lines. In addition, the previously reported positive regulator of tanshinone biosynthesis, SmERF1L1, was found to be inhibited in SmbZIP1 overexpression lines indicated by RNA sequencing, and was proven to be the target of SmbZIP1. In summary, this work uncovers a novel regulator and deepens our understanding of the transcriptional and regulatory mechanisms of phenolic acid and tanshinone biosynthesis, and also sheds new light on metabolic engineering in S. miltiorrhiza.

scholarly journals SmMYC2a and SmMYC2b played similar but irreplaceable roles in regulating the biosynthesis of tanshinones and phenolic acids in Salvia miltiorrhiza

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Yangyun Zhou ◽  
Wei Sun ◽  
Junfeng Chen ◽  
Hexin Tan ◽  
Ying Xiao ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Phenolic Acids ◽  
Phenolic Acid ◽  
Biosynthetic Pathway ◽  
Salvia Miltiorrhiza ◽  
Loss Of Function ◽  
Bioactive Components ◽  
Jaz Proteins ◽  
E Box ◽  
Inducible Genes

Abstract Salvia miltiorrhiza Bunge, which contains tanshinones and phenolic acids as major classes of bioactive components, is one of the most widely used herbs in traditional Chinese medicine. Production of tanshinones and phenolic acids is enhanced by methyl jasmonate (MeJA). Transcription factor MYC2 is the switch of jasmontes signaling in plants. Here, we focused on two novel JA-inducible genes in S. miltiorrhiza, designated as SmMYC2a and SmMYC2b, which were localized in the nucleus. SmMYC2a and SmMYC2b were also discovered to interact with SmJAZ1 and SmJAZ2, implying that the two MYC2s might function as direct targets of JAZ proteins. Ectopic RNA interference (RNAi)-mediated knockdown experiments suggested that SmMYC2a/b affected multiple genes in tanshinone and phenolic acid biosynthetic pathway. Besides, the accumulation of tanshinones and phenolic acids was impaired by the loss of function in SmMYC2a/b. Meanwhile, SmMYC2a could bind with an E-box motif within SmHCT6 and SmCYP98A14 promoters, while SmMYC2b bound with an E-box motif within SmCYP98A14 promoter, through which the regulation of phenolic acid biosynthetic pathway might achieve. Together, these results suggest that SmMYC2a and SmMYC2b are JAZ-interacting transcription factors that positively regulate the biosynthesis of tanshinones and Sal B with similar but irreplaceable effects.

scholarly journals Application of 1H-NMR combined with qRT-PCR technology in the exploration of rosmarinic acid biosynthesis in hair roots of Salvia miltiorrhiza Bunge and Salvia castanea f. tomentosa Stib

Planta ◽  
2020 ◽  
Vol 253 (1) ◽  
Author(s):  
Zhuoni Hou ◽  
Yuanyuan Li ◽  
Feng Su ◽  
Jipeng Chen ◽  
Xiaodan Zhang ◽  
...  
Keyword(s):  
Methyl Jasmonate ◽  
Phenolic Acids ◽  
Rosmarinic Acid ◽  
Salvia Miltiorrhiza ◽  
Water Soluble ◽  
H Nmr ◽  
Qrt Pcr ◽  
Medicinal Value ◽  
Hair Roots ◽  
Pcr Technology

Abstract Main conclusion Methyl jasmonate promotes the synthesis of rosmarinic acid in Salvia miltiorrhiza Bunge and Salvia castanea f. tomentosa Stib, and it promotes the latter more strongly. Abstract Salvia miltiorrhiza Bunge (SMB) is a traditional Chinese medicinal material, its water-soluble phenolic acid component rosmarinic acid has very important medicinal value. Salvia castanea f. tomentosa Stib (SCT) mainly distributed in Nyingchi, Tibet. Its pharmacological effects are similar to SMB, but its rosmarinic acid is significantly higher than the former. Methyl jasmonate (MJ) as an inducer can induce the synthesis of phenolic acids in SMB and SCT. However, the role of MJ on rosmarinic acid in SMB is controversial. Therefore, this study used SMB and SCT hair root as an experimental material and MJ as a variable. On one hand, exploring the controversial reports in SMB; on the other hand, comparing the differences in the mechanism of action of MJ on the phenolic acids in SMB and SCT. The content of related metabolites and the expression of key genes in the synthesis pathway of rosmarinic acid was analyzed by 1H-NMR combined with qRT-PCR technology. Our research has reached the following conclusions: first of all, MJ promotes the accumulation of rosmarinic acid and related phenolic acids in the metabolic pathways of SMB and SCT. After MJ treatment, the content of related components and gene expression are increased. Second, compared to SMB, SCT has a stronger response to MJ. It is speculated that the different responses of secondary metabolism-related genes to MJ may lead to different metabolic responses of salvianolic acid between the two.

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

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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