Molecular Cloning and Characterization of Genes Involved in Rosmarinic Acid Biosynthesis from &lt;i&gt;Prunella vulgaris&lt;/i&gt;

Kim, Yeon Bok; Shin, YouJin; Tuấn, Phạm Anh; Li, Xiaohua; Park, Yunji; Park, Nam-Il; Park, Sang Un

doi:10.1248/bpb.b14-00139

Cited by 20 publications

(26 citation statements)

References 37 publications

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“…In addition, some literature reported RA contents of the different organs of other Lamiaceae species that were consistent with those in our results; the flowers contained the highest RA levels among all the organs tested from A. rugosa and Prunella vulgaris 13, 20 .…”

Section: Resultssupporting

confidence: 93%

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Transcriptomic analyses reveal biosynthetic genes related to rosmarinic acid in Dracocephalum tanguticum

Sun

et al. 2017

Sci Rep

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Dracocephalum tanguticum Maxim, a Lamiaceae species endemic to the Qinghai-Tibetan Plateau and adjacent regions, is an important ornamental, medicinal and aromatic herb. In this study, a comprehensive transcriptome of 18 libraries from six organs namely, roots, stems, leaves, sepals, flowers and seeds of D. tanguticum were generated. More than 100 Gb of sequence data were obtained and assembled de novo into 187,447 transcripts, including 151,463 unigenes, among which the six organs shared 17.7% (26,841). In addition, all unigenes were assigned to 362 pathways, in which 'biosynthesis of secondary metabolites' is the second enriched pathway. Furthermore, rosmarinic acid (RA) is one of the multifunctional phenolic bioactive compounds produced in some Lamiaceae species. The six organs of D. tanguticum were confirmed to produce RA. A total of 22 predicted biosynthetic genes related to RA from the transcriptome were further isolated. Two of these genes were identified as candidates by evaluating the correlation coefficient between the RA contents and the expression of the predicted biosynthetic genes in the six organs. The new sequence information will improve the knowledge of D. tanguticum, as well as provide a reference tool for future studies of biosynthetic genes related to RA in this species.Dracocephalum tanguticum Maxim, a Lamiaceae species endemic to the Qinghai-Tibetan Plateau (QTP) and adjacent regions, is an important ornamental, medicinal and aromatic herb. This plant is one of the important traditional Tibetan medicines which have been used by locals for a long time. However, no study has been reported on D. tanguticum because of the plant's limited distribution. The high-altitude QTP region has unique climate factors, including low oxygen levels, intense ultraviolet light, and large temperature difference between day and night, as well as prolonged daytime during the short growing seasons, which usually commences on April and ends in October for D. tanguticum. All of these harsh factors may be beneficial for the biosynthesis of protective secondary metabolites in D. tanguticum.Rosmarinic acid (RA) is a multifunctional phenolic bioactive compound that possesses astringent, antimutagen, antibacterial, antiviral, antifungal, anti-inflammatory and anti-oxidative activities [1][2][3][4][5][6] . An increasing number of plant sources have been screened for RA content 7-9 , and biotechnological and metabolic engineering approaches for RA production have been studied in previous reports [10][11][12][13][14][15] . However, plant sources have remained insufficient in meeting the rising demand of RA in recent years.Previous literature reported that the amino acids, L-phenylalanine and L-tyrosine, are the two precursors of the RA biosynthetic pathway in Coleus blumei, a Lamiaceae species 16 . In the phenylalanine-derived pathway of RA biosynthesis, phenylalanine ammonia-lyase (PAL) catalyses the oxidative desamination of phenylalanine, forming t-cinnamic acid, and cinnamic acid; 4-hydroxylase (C4H) introduces the par...

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Section: Resultssupporting

confidence: 93%

“…13 through high-performance liquid chromatography. The determination was conducted with a C18 column (4.6 mm × 150 mm, 5 μm; ZORBAX Eclipse XDB-C18 Analytical, USA) at room temperature.…”

Section: Methodsmentioning

confidence: 99%

Transcriptomic analyses reveal biosynthetic genes related to rosmarinic acid in Dracocephalum tanguticum

Sun

et al. 2017

Sci Rep

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“…Phosphatidylinositol signaling system (20) beta-Alanine metabolism (20) Glycine, serine and threonine metabolism (35) Valine, leucine and isoleucine degradation (22) Glycerophospholipid metabolism (34) Carbon fixation in photosynthetic organisms (23) Glutathione metabolism (20) Oxidative phosphorylation (8) Glycerolipid metabolism (22) Cysteine and methionine metabolism (44) Pyrimidine metabolism (33) Phenylpropanoid biosynthesis (17) Drug metabolism -other enzymes (14) Glycolysis / Gluconeogenesis (28) Purine leaf, stem, and root, respectively (▶ Fig. 5 a).…”

Section: Assemblermentioning

confidence: 99%

De Novo Transcriptome Assembly and Characterization of Lithospermum officinale to Discover Putative Genes Involved in Specialized Metabolites Biosynthesis

et al. 2018

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is a valuable source of bioactive metabolites with medicinal and industrial values. However, little is known about genes involved in the biosynthesis of these metabolites, primarily due to the lack of genome or transcriptome resources. This study presents the first effort to establish and characterize transcriptome assembly resource for and expression analysis for three of its tissues, namely leaf, stem, and root. Using over 4Gbps of RNA-sequencing datasets, we obtained transcriptome assembly of, consisting of 77,047 unigenes with assembly N50 value as 1524 bps. Based on transcriptome annotation and functional classification, 52,766 unigenes were assigned with putative genes functions, gene ontology terms, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. KEGG pathway and gene ontology enrichment analysis using highly expressed unigenes across three tissues and targeted metabolome analysis showed active secondary metabolic processes enriched specifically in the root of . Using co-expression analysis, we also identified 20 and 48 unigenes representing different enzymes of lithospermic/chlorogenic acid and shikonin biosynthesis pathways, respectively. We further identified 15 candidate unigenes annotated as cytochrome P450 with the highest expression in the root of as novel genes with a role in key biochemical reactions toward shikonin biosynthesis. Thus, through this study, we not only generated a high-quality genomic resource for but also propose candidate genes to be involved in shikonin biosynthesis pathways for further functional characterization.

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“…In addition, Kim et al. cloned the genes involved in the phenylpropanoid branched‐chain of the RA biosynthetic pathway in P. vulgaris , so we also investigated the relationship of genes expressed in both branches of the pathway and RA biosynthesis.…”

Section: Discussionmentioning

confidence: 99%

“…Given this, we decided to use these two elicitors to induce the hairy root cultures of P. vulgaris, and further characterized the relationship between the expression of these genes (PvTAT, PvHPPR, PvRAS, and PvCYP) and RA biosynthesis. In addition, Kim et al [37] cloned the genes involved in the phenylpropanoid branched-chain of the RA biosynthetic pathway in P. vulgaris, so we also investigated the relationship of genes expressed in both branches of the pathway and RA biosynthesis.…”

Section: Discussionmentioning

confidence: 99%

Prunella vulgaris L. hairy roots: Culture, growth, and elicitation by ethephon and salicylic acid

Wang

et al. 2016

Engineering in Life Sciences

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Prunella vulgaris L. hairy roots: Culture, growth, and elicitation by ethephon and salicylic acidHairy roots have gained popularity as an alternative source for secondary metabolites not only because of their biosynthetic and bioactive potentials, but also because of studies of experimental systems aiming to understand secondary metabolic pathways. While many secondary metabolites have been identified in Prunella vulgaris, a comprehensive understanding of the metabolic pathways has remained elusive. Here, Agrobacterium rhizogenes (ATCC15834)-mediated hairy root induction of P. vulgaris resulted in 15-30 times more rosmarinic acid (RA) than in the intact plant. After the growth and RA biosynthesis of P. vulgaris, hairy roots were investigated over a 1-month period, we studied elicitation by ethephon (Eth) and salicylic acid (SA) and found 200 μg L −1 Eth and 6.9 mg L −1 SA exhibited the optimum induction efficiency. These concentrations were further selected for a time-course assay of hairy root cultures to assess exposure to these compounds over 8 days. During that period, RA reached maximum accumulations of 1.66-fold 8 days after Eth elicitation and 1.48-fold 2 days post-SA addition. We also examined the transcripts of genes involved in the RA biosynthesis pathway, and found the expression of tyrosine aminotransferase (TAT) and phenylalanine ammonia-lyase (PAL) were increased by Eth, and SA heightened the expression of TAT, 4-hydroxyphenylpyruvate reductase (HPPR), PAL, 4-coumaric acid CoA-ligase 1 (4CL1), and cytochrome P450-dependent monooxygenase (CYP). Altogether, our results indicate that the convenient hairy root culture system of P. vulgaris can provide an alternate source for RA production and would be conducive for the engineering of secondary metabolic pathways in P. vulgaris.

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Molecular Cloning and Characterization of Genes Involved in Rosmarinic Acid Biosynthesis from <i>Prunella vulgaris</i>

Cited by 20 publications

References 37 publications

Transcriptomic analyses reveal biosynthetic genes related to rosmarinic acid in Dracocephalum tanguticum

Transcriptomic analyses reveal biosynthetic genes related to rosmarinic acid in Dracocephalum tanguticum

De Novo Transcriptome Assembly and Characterization of Lithospermum officinale to Discover Putative Genes Involved in Specialized Metabolites Biosynthesis

Prunella vulgaris L. hairy roots: Culture, growth, and elicitation by ethephon and salicylic acid

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