• Journal of Ginseng Research
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• v.33 no.3
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• pp.206-211
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• 2009

In order to investigate the effects of elicitors on the growth and ginsenoside biosynthesis of ginseng hairy roots, we treated Panax ginseng hairy root with various concentrations of Haliotidis concha according to different time course. Haliotidis concha supplement increased the biomass and ginsenoside accumulation at 10 mg/L concentration. The growth rate of hairy root under a lighter concentration was greater than hairy root treated with a denser concentration. The highest content and productivity of ginsenosides appeared at 2 weeks after the treatment of 10 mg/L Haliotidis concha. The gene expression of squalene synthase, squalene epoxidase, dammarenediol synthase, cycloartenol synthase, $\beta$-amyrin synthase in hairy roots of ginseng were examined by RT-PCR. The Haliotidis concha treatment resulted in the obvious accumulation of the mRNA of triterpene biosynthesis in Panax ginseng hairy root as compared with the control. In this study, Haliotidis concha acts as a kind of elicitor for the production of ginsenosides.

• Journal of Microbiology and Biotechnology
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• v.25 no.5
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• pp.658-661
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• 2015

Genes encoding enzymes with sequence similarity to hopanoids biosynthetic enzymes of other organisms were cloned from the hopanoid (hop) gene cluster of Streptomyces peucetius ATCC 27952 and transformed into Streptomyces venezuelae YJ028. The cloned fragments contained four genes, all transcribed in one direction. These genes encode polypeptides that resemble polyprenyl diphosphate synthase (hopD), squalene-phytoene synthases (hopAB), and squalene-hopene cyclase (hopE). These enzymes are sufficient for the formation of the pentacyclic triterpenoid lipid, hopene. The formation of hopene was verified by gas chromatography/mass spectrometry.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.145.3-146
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• 2003

Transgenic Eleutherococcus senticosus plants were prepared by introducing the genes for squalene synthase (SQS), hygromycin phosphotransferase (HPT) and green fluorescent Protein (GFP) through Agrobacterium-mediated transformation. The enzyme, SQS, represents a putative branch point in the isoprenoid pathway capable of diverting carbon flow specifically to the biosynthesis of phytosterol and oleanolic acid. The full SQS gene was isolated from P. ginseng roots. Early globular embryo clusters developed from embryogenic callus were used as the explant source. (omitted)

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2003.10a
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• pp.90-90
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• 2003

• Mycobiology
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• v.50 no.2
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• pp.121-131
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• 2022

The rare edible and medicinal fungus Antrodia cinnamomea has a substantial potential for development. In this study, Illumina HiSeq 2000 was used to sequence its transcriptome. The results were assembled de novo, and 66,589 unigenes with an N50 of 4413 bp were obtained. Compared with public databases, 6,061, 3,257, and 2,807 unigenes were annotated to the Non-Redundant, Gene Ontology, and Kyoto Encyclopedia of Genes and Genomes databases, respectively. The genes related to terpene biosynthesis in the mycelia of A. cinnamomea were analyzed, and acetyl CoA synthase (ACS2 and ACS4), hydroxymethylglutaryl CoA reductase (HMGR), farnesyl transferase (FTase), and squalene synthase (SQS) were found to be upregulated in XZJ (twig of C. camphora) and NZJ (twig of C. kanehirae). Moreover, ACS5 and 2,3-oxidized squalene cyclase (OCS) were highly expressed in NZJ, while heme IX farnesyl transferase (IX-FIT) and ACS3 were significantly expressed in XZJ. The differential expression of ACS1, ACS2, HMGR, IX-FIT, SQS, and OCS was confirmed by real-time quantitative reverse transcription PCR. This study provides a new concept for the additional exploration of the molecular regulatory mechanism of terpenoid biosynthesis and data for the biotechnology of terpenoid production.

• Journal of Ginseng Research
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• v.46 no.2
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• pp.225-234
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• 2022

Background: Ginseng, officially known as Panax ginseng Meyer, has been traditionally used as a medicinal herb, particularly in Asia. Ginseng is propagated from seeds; however, seed germination is challenging, especially in its natural environment on farms. The seeds typically exhibit morphophysiological dormancy and require release from both morphological and physiological dormancy before germination. Although some studies have proposed methods for increasing seed germination rates, the underlying mechanisms of its dormancy release process remain unclear. Here, we investigated metabolic alterations during dehiscence in P. ginseng to determine their potential roles in dormancy release. Methods: We compared the ginseng seed metabolome before and after dehiscence and the ginsenoside and phytosterol compositions of the seeds in both periods in the presence of related enzymes. Results: After seed dehiscence, the sugar, amino acid, and squalene concentrations were significantly altered, phytosterols associated with the stigmasterol biosynthesis pathway were increased, while ginsenoside and brassinosteroid levels were not significantly altered. In addition, squalene epoxidase, cycloartenol synthase, 24-methylenesterol C-methyltransferase, and the stigmasterol biosynthesis pathway were activated. Conclusion: Overall, our findings suggest that morphological activities that facilitate ginseng seed growth are the primary phenomena occurring during the dehiscence process. This study improves the understanding of P. ginseng germination processes and promotes further research of its germination and cultivation.

• Mycobiology
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• v.51 no.1
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• pp.49-59
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• 2023

Antrodia cinnamomea, an edible and medicinal fungus with significant economic value and application prospects, is rich in terpenoids, benzenoids, lignans, polysaccharides, and benzoquinone, succinic and maleic derivatives. In this study, the transcriptome of A. cinnamomea cultured on the wood substrates of Cinnamomum glanduliferum (YZM), C. camphora (XZM), and C. kanehirae (NZM) was sequenced using the high-throughput sequencing technology Illumina HiSeq 2000, and the data were assembled by de novo strategy to obtain 78,729 Unigenes with an N50 of 4,463 bp. Compared with public databases, about 11,435, 6,947, and 5,994 Unigenes were annotated to the Non-Redundant (NR), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genome (KEGG), respectively. The comprehensive analysis of the mycelium terpene biosynthesis-related genes in A. cinnamomea revealed that the expression of acetyl-CoA acetyltransferase (AACT), acyl-CoA dehydrogenase (MCAD), 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA), mevalonate pyrophosphate decarboxylase (MVD), and isopentenyl diphosphate isomerase (IDI) was significantly higher on NZM compared to the other two wood substrates. Similarly, the expression of geranylgeranyltransferase (GGT) was significantly higher on YZM compared to NZM and XZM, and the expression of farnesyl transferase (FTase) was significantly higher on XZM. Furthermore, the expressions of 2,3-oxidized squalene cyclase (OCS), squalene synthase (SQS), and squalene epoxidase (SE) were significantly higher on NZM. Overall, this study provides a potential approach to explore the molecular regulation mechanism of terpenoid biosynthesis in A. cinnamomea.

• Korean Journal of Medicinal Crop Science
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• v.13 no.2
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• pp.91-94
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• 2005

Plants have been known to accumulate a very diverse range of triterpene saponins. We have investigated the regulation of saponin biosynthesis in higher plants using Centella asiatica (L.) Urban as a model plant. Effects of a feeding precursor on asiaticoside production from leaves and on the level of two-type OSCs mRNA were investigated. As a feeding precursor, squalene negatively affected the levels of CYS and bAS mRNA, but it also decreased the production of asiaticoside from whole plants. Plant hormones regulate secondary metabolism, and in plant tissue cultures they could affect both culture growth and secondary metabolite production. Although enhancement of asiaticoside production from whole plant cultures by addition of TDZ (thidiazuron) has been reported, the positive effect of TDZ on the levels of OSCs transcripts was not observed.

• Molecules and Cells
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• v.22 no.3
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• pp.269-274
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• 2006

In order to characterize saikosaponin biosynthesis in Bupleurum falcatum, the expression of five isoprenoid pathway genes and their relationship to saikosaponin accumulation in the hairy roots were analyzed. The hairy roots exhibited a rapid accumulation of saikosaponins when incubated in a root culture medium (3XRCM). Homology-based RT-PCR was used to isolate core fragments of five genes, HMGR, IPPI, FPS, SS, and OSC, from the hairy roots. The deduced amino acid sequences exhibited amino acid identities of more than 85% to previously reported genes. Using the fragments as probes, the expression of these five genes in the hairy roots during incubation in 3XRCM medium was examined. Expression of all five genes in the hairy roots increased soon after incubation. In particular, the SS and OSC genes were coordinately induced at 8 days of incubation, and their expression persisted throughout the incubation period. A quantitative HPLC analysis showed that the saikosaponin content of the hairy root culture also began to increase at 8 days of culture. The correlation between SS transcript level and saikosaponin content in the hairy roots suggests that transcriptional regulation plays a regulatory role in saikosaponin biosynthesis.

• Journal of Ginseng Research
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• v.37 no.3
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• pp.332-340
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• 2013

In this study, gamma rays were used to irradiate embryogenic calli induced from cotyledon explants of Panax ginseng Meyer. After the embryogenic calli were irradiated, they were transferred to adventitious roots using an induction medium; next, mutated adventitious root (MAR) lines with a high frequency of adventitious root formations were selected. Two MAR lines (MAR 5-2 and MAR 5-9) from the calli treated with 50 Gy of gamma rays were cultured on an $NH_4NO_3$-free Murashige and Skoog medium with indole-3-butyric acid 3 mg/L. The expression of genes related to ginsenoside biosynthesis was analyzed using reverse transcription polymerase chain reaction with RNA prepared from native ginseng (NG), non-irradiated adventitious root (NAR) and 2 MAR lines. The expression of the squalene epoxidase and dammarenediol synthase genes was increased in the MAR 5-2 line, whereas the phytosterol synthase was increased in the MAR 5-9 line. The content and pattern of major ginsenosides (Rb1, Rb2, Rc, Rd, Re, Rf, and Rg1) were analyzed in the NG, NAR, and 2 MAR lines (MAR 5-2 and MAR 5-9) using TLC and HPLC. In the TLC analysis, the ginsenoside patterns in the NG, NAR, and 2 MAR lines were similar; in contrast, the MAR 5-9 line showed strong bands of primary ginsenosides. In the HPLC analysis, compared with the NG, one new type of ginsenoside was observed in the NAR and 2 MAR lines, and another new type of ginsenoside was observed in the 2 MAR lines irradiated with gamma rays. The ginsenoside content of the MAR 5-9 line was significantly greater in comparison to the NG.