Abstract
It has been recognized that ginsenoside Rg3 is not naturally produced in ginseng although this ginsenoside can accumulate in red ginseng as the result of a thermal process. In order to determine whether or not Rg3 is synthesized in ginseng, hairy roots were treated with methyl jasmonate (MJ). From HPLC analysis, no peak for Rg3 was observed in the controls. However, Rg3 did accumulate in hairy roots that were MJ-treated for 7 days. Rg3 content was 0.42 mg/g (dry weight). To gain more insight into the effects of MJ on UDP-glucosyltransferase (UGT) activity, we attempted to evaluate ginsenoside Rg3 biosynthesis by UGT. A new peak for putative Rg3 was observed, which was confirmed by LC-MS/MS analysis. Our findings indicate that the proteins extracted from our hairy root lines can catalyze Rg3 from Rh2. This suggests that our ginseng hairy root lines possess Rg3 biosynthesis capacity.
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Bonfill M, Mangas S, Moyano E, Cusido RM, Palazon J (2011) Production of centellosides and phytosterols in cell suspension cultures of Centella asiatica. Plant Cell Tiss Organ Cult 104:61–67
Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Briskin DP (2000) Medicinal plants and phytomedicines. Linking plant biochemistry and physiology to human health. Plant Physiol 124:507–514
Choi DW, Jung JD, Ha YI, Park HW, In DS, Chung HJ, Liu JR (2005) Analysis of transcripts in methyl jasmonate-treated ginseng hairy roots to identify genes involved in the biosynthesis of ginsenosides and other secondary metabolites. Plant Cell Rep 23:557–566
Han JY, Kim HJ, Kwon YS, Choi YE (2011) The Cyt P450 enzyme CYP716A47 catalyzes the formation of protopanaxadiol from dammarenediol-II during ginsenoside biosynthesis in Panax ginseng. Plant Cell Physiol 52:2062–2073
Hayashi H, Huang PY, Inoue K (2003) Up-regulation of soyasaponin biosynthesis by methyl jasmonate in cultured cells of Glycyrrhiza glabra. Plant Cell Physiol 44:404–411
Hu FX, Zhong JJ (2007) Role of jasmonic acid in alteration of ginsenoside heterogeneity in elicited cell cultures of Panax notoginseng. J Biosci Bioeng 104:513–516
Hu YH, Yu YT, Piao CH, Liu JM, Yu HS (2011) Methyl jasmonate- and salicylic acid-induced d-chiro-inositol production in suspension cultures of buckwheat (Fagopyrum esculentum). Plant Cell Tiss Organ Cult 106:419–424
Hwang B, Ko KM (1989) Induction and culture of hairy roots from ginseng (Panax ginseng C.A. Meyer) roots discs by Agrobacterium rhizogenes. Korean J Biotechnol Bioeng 4:288–292
Jung JD, Park HW, Hahn Y, Hur CG, In DS, Chung HJ, Liu JR, Choi DW (2003) Discovery of genes for ginsenoside biosynthesis by analysis of ginseng expressed sequence tags. Plant Cell Rep 22:224–230
Kim YS, Hahn EJ, Yeung EC, Paek KY (2004) Adventitious root growth and ginsenoside accumulation in Panax ginseng cultures as affected by methyl jasmonate. Biotechnol Lett 26:1619–1622
Kim KM, Lee BS, In JG, Sun H, Yoon JH, Yang DC (2006) Comparative analysis of expressed sequence tags (ESTs) of ginseng leaf. Plant Cell Rep 25:599–606
Kim OT, Bang KH, Kim YC, Hyun DY, Kim MY, Cha SW (2009) Upregulation of ginsenoside and gene expression related to triterpene biosynthesis in ginseng hairy root cultures elicited by methyl jasmonate. Plant Cell Tiss Organ Cult 98:25–33
Ko SR, Suzuki Y, Suzuki K, Choi KJ, Cho BG (2007) Marked production of ginsenoside Rd, F2, Rg3, and compound K by enzymatic method. Chem Pharm Bull 55:1522–1527
Liu WK, Xu SX, Che CT (2000) Anti-proliferative effect of ginseng saponins on human prostate cancer cell line. Life Sci 67:1297–1306
Lu MB, Wong HL, Teng WL (2001) Effects of elicitation on the production of saponin in cell culture of Panax ginseng. Plant Cell Rep 20:674–677
Ma XQ, Liang XM, Xu Q, Zhang XZ, Xiao HB (2005) Identification of ginsenosides in roots of Panax ginseng by HPLC-APCI/MS. Phytochem Anal 16:181–187
Min JK, Kim JH, Cho YL, Maeng YS, Lee SJ, Pyun BJ, Kim YM, Park JH, Kwon YG (2006) 20(S)-Ginsenoside Rg3 prevents endothelial cell apoptosis via inhibition of a mitochondrial caspase pathway. Biochem Biophys Res Commun 349:987–994
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue. Physiol Plant 15:473–497
Popovich DG, Kitts DD (2004) Generation of ginsenosides Rg3 and Rh2 from North American ginseng. Phytochemistry 65:337–344
Qu JG, Zhang W, Yu XJ (2011) A combination of elicitation and precursor feeding leads to increased anthocyanin synthesis in cell suspension cultures of Vitis vinifera. Plant Cell Tiss Organ Cult 107:261–269
Sun CS, Li Y, Wu Q, Luo H, Sun Y, Song J, Lui E, Chen S (2010a) De novo sequencing and analysis of the American ginseng root transcriptome using a GS FLX Titanium platform to discover putative genes involved in ginsenoside biosynthesis. BMC Genomics 11:262
Sun S, Wang CZ, Tong R, Li XL, Fishbein A, Wang Q, He TC, Du W, Yuan CS (2010b) Effects of steaming the root of Panax notoginseng on chemical composition and anticancer activities. Food Chem 118:307–314
Wang W, Zhong JJ (2002) Manipulation of ginsenoside heterogeneity in cell cultures of Panax notoginseng by addition of jasmonate. J Biosci Bioeng 93:48–53
Wasternack C (2007) Jasmonates: an update on biosynthesis, signal transduction and action in plant stress response, growth and development. Ann Bot 100:681–697
Yue CJ, Zhong JJ (2005) Purification and characterization of UDPG: ginsenoside Rd glucosyltransferase from suspended cells of Panax notoginseng. Process Biochem 40:3742–3748
Zhang QH, Wu CF, Duan L, Yang JY (2008) Protective effects of ginsenoside Rg(3) against cyclophosphamide-induced DNA damage and cell apoptosis in mice. Arch Toxicol 82:117–123
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This work was supported by the Cooperative Research Program for Agricultural Science & Technology Development (Project No. 200901AFT143782304), RDA, Republic of Korea.
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Ok Tae Kim and Nam Hee Yoo equally contributed to this paper.
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Kim, O.T., Yoo, N.H., Kim, G.S. et al. Stimulation of Rg3 ginsenoside biosynthesis in ginseng hairy roots elicited by methyl jasmonate. Plant Cell Tiss Organ Cult 112, 87–93 (2013). https://doi.org/10.1007/s11240-012-0218-6
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DOI: https://doi.org/10.1007/s11240-012-0218-6