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Upregulation of phytosterol and triterpene biosynthesis in Centella asiatica hairy roots overexpressed ginseng farnesyl diphosphate synthase

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Abstract

Farnesyl diphosphate synthase (FPS) plays an essential role in organ development in plants. However, FPS has not previously been identified as a key regulatory enzyme in triterpene biosynthesis. To elucidate the functions of FPS in triterpene biosynthesis, C. asiatica was transformed with a construct harboring Panax ginseng FPS (PgFPS)-encoding cDNA coupled to the cauliflower mosaic virus 35S promoter. Higher levels of CaDDS (C. asiatica dammarenediol synthase) and CaCYS (C. asiatica cycloartenol synthase) mRNA were detected in all hairy root lines overexpressing when compared with the controls. However, no differences were detected in any expression of the CaSQS (C. asiatica squalene synthase) gene. In particular, the upregulation of CaDDS transcripts suggests that FPS may result in alterations in triterpene biosynthesis capacity. Squalene contents in the T17, T24, and T27 lines were increased to 1.1-, 1.3- and 1.5-fold those in the controls, respectively. The total sterol contents in the T24 line were approximately three times higher than those of the controls. Therefore, these results indicated that FPS performs a regulatory function in phytosterol biosynthesis. To evaluate the contribution of FPS to triterpene biosynthesis, we applied methyl jasmonate as an elicitor of hairy roots expressing PgFPS. The results of HPLC analysis revealed that the content of madecassoside and asiaticoside in the T24 line was transiently increased by 1.15-fold after 14 days of MJ treatment. This result may indicate that FPS performs a role not only in phytosterol regulation, but also in triterpene biosynthesis.

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References

  • Benveniste P (2004) Biosynthesis and accumulation of sterols. Annu Rev Plant Biol 55:429–457

    Article  CAS  PubMed  Google Scholar 

  • Bonfill M, Mangas S, Cusidó RM, Osuna L, Piñol MT, Palazón J (2006) Identification of triterpenoid compounds of Centella asiatica by thin-layer chromatography and mass spectrometry. Biomed Chromatogr 20:151–153

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Brinkhaus B, Linder M, Schuppan D, Hahn EG (2000) Chemical, pharmacological and clinical profile of the East Asian medical plant Centella asiatica. Phytomedicine 7:427–448

    CAS  PubMed  Google Scholar 

  • Brodelius M, Lundgren A, Mercke P, Brodelius PE (2002) Fusion of farnesyldiphosphate synthase and epi-aristolochene synthase, a sesquiterpene cyclase involved in capsidiol biosynthesis in Nicotiana tabacum. Eur J Biochem 269:3570–3577

    Article  CAS  PubMed  Google Scholar 

  • Chen DH, Ye HC, Li GF (2000) Expression of a chimeric farnesyl diphosphate synthase gene in Artemisia annua L. transgenic plants via Agrobacterium tumefaciens-mediated transformation. Plant Sci 155:179–185

    Article  CAS  PubMed  Google Scholar 

  • Confalonieri M, Cammareri M, Biazzi E, Pecchia P, Fevereiro MPS, Balestrazzi A, Tava A, Conicella C (2009) Enhanced triterpene saponin biosynthesis and root nodulation in transgenic barrel medic (Medicago truncatula Gaertn.) expressing a novel β-amyrin synthase (AsOXA1) gene. Plant Biotechnol J 7:172–182

    Article  CAS  PubMed  Google Scholar 

  • Cunillera N, Arró M, Delourme D, Karst F, Boronat A, Ferrer A (1996) Arabidopsis thaliana contains two differentially expressed farnesyl-diphosphate synthase genes. J Biol Chem 271:7774–7780

    Article  CAS  PubMed  Google Scholar 

  • Cunillera N, Boronat A, Ferrer A (1997) The Arabidopsis thaliana FPS1 gene generates a novel mRNA that encodes a mitochondrial farnesyl-diphosphate synthase isoform. J Biol Chem 272:15381–15388

    Article  CAS  PubMed  Google Scholar 

  • Daudonnet S, Karst F, Tourte Y (1997) Expression of the farnesyl diphosphate synthase gene of Saccharomyces cerevisiae in tobacco. Mol Breed 3:137–145

    Article  CAS  Google Scholar 

  • Francis G, Kerem Z, Makkar HP, Becker K (2002) The biological action of saponins in animal system: a review. Br J Nutr 88:587–605

    Article  CAS  PubMed  Google Scholar 

  • Gaffe J, Bru JP, Causse M, Vidal A, Stamitti-Bert L, Carde JP, Gallusci P (2000) LEFPS1, a tomato farnesyl pyrophosphate gene highly expressed during early fruit development. Plant Phsiol 123:1351–1362

    Article  CAS  Google Scholar 

  • Han JY, In JG, Kwon YS, Choi YE (2010) Regulation of ginsenoside and phytosterol biosynthesis by RNA interferences of squalene epoxidase gene in Panax ginseng. Phytochemistry 71:36–46

    Article  CAS  PubMed  Google Scholar 

  • Harker M, Holmberg N, Clayton JC, Gibbard CL, Wallace AD, Rawlins S, Hellyer SA, Lanot A, Safford R (2003) Enhancement of seed phytosterol levels by expression of an N-terminal truncated Hevea brasiliensis (rubber tree) 3-hydroxy-3-methylglutaryl-CoA reductase. Plant Biotechnol J 1:113–121

    Article  CAS  PubMed  Google Scholar 

  • Hartmann MA, Benveniste P (1987) Plant membrane sterols: isolation, identification and biosynthesis. Methods Enzymol 148:632–650

    Article  CAS  Google Scholar 

  • Hayashi H, Hirota A, Hiraoka N, Ikeshiro Y (1999) Molecular cloning and characterization of two cDNAs for Glycyrrhiza glabra squalene synthase. Biol Pharm Bull 22:947–950

    CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Hey SJ, Powers SJ, Beale MH, Hawkins ND, Ward JL, Halford NG (2006) Enhanced seed phytosterol accumulation through expression of a modified HMG-CoA reductase. Plant Biotechnol J 4:219–229

    Article  CAS  PubMed  Google Scholar 

  • Kim OT, Bang KH, Shin YS, Lee MJ, Jung SJ, Hyun DY, Kim YC, Seong NS, Cha SW, Hwang B (2007) Enhanced production of asiaticoside from hairy root cultures of Centella asiatica (L.) Urban elicited by methyl jasmonate. Plant Cell Rep 26:1941–1949

    Article  CAS  PubMed  Google Scholar 

  • Kim OT, Bang KH, Kim YC, Hyun DY, Kim MY, Cha SW (2009a) 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

    Article  CAS  Google Scholar 

  • Kim OT, Lee JW, Bang KH, Kim YC, Hyun DY, Cha SW, Choi YE, Jin ML, Hwang B (2009b) Characterization of a dammarenediol synthase in Centella asiatica (L.) Urban. Plant Physiol Biochem 47:998–1002

    Article  CAS  PubMed  Google Scholar 

  • Kim OT, Bang KH, Jung SJ, Kim YC, Hyun DY, Kim SH, Cha SW (2010) Molecular characterization of ginseng farnesyl diphosphate synthase gene and its up-regulation by methyl jasmonate. Biol Plant 54:47–53

    Google Scholar 

  • Lee MH, Jeong JH, Seo JW, Shin CG, Kim YS, In JG, Yang DC, Yi JS, Choi YE (2004) Enhanced triterpene and phytosterol biosynthesis in Panax ginseng overexpressing squalene synthase gene. Plant Cell Physiol 45:976–984

    Article  CAS  PubMed  Google Scholar 

  • Mangas S, Bonfill M, Osuna L, Moyano E, Tortoriello J, Cusido RM, Piñol MT, Palazón (2006) The effect of methyl jasmonate on triterpene and sterol metabolisms of Centella asiatica, Ruscus aculeatus and Galphimia glauca cultured plants. Phytochemistry 67:2041–2049

  • Manzano D, Fernández-Busquets X, Schaller H, González V, Boronat A, Arrό M, Ferrer A (2004) The metabolic imbalance underlying lesion formation in Arabidopsis thaliana overexpressing farnesyl diphosphate synthase (isoform 1S) leads to oxidative stress and is triggered by the developmental decline of endogenous HMGR activity. Planta 219:982–992

    Article  CAS  PubMed  Google Scholar 

  • Masferrer A, Arrό M, Manzano D, Schaller H, Fernández-Busquets X, Moncaleán P, Fernández B, Cunillera N, Boronat A, Ferrer A (2002) Overexpression of Arabidopsis thaliana farnesyl diphosphate synthase (FPS1S) in transgenic Arabidopsis induces a cell death/senescence-like response and reduced cytokinin levels. Plant J 30:123–132

    Article  CAS  PubMed  Google Scholar 

  • Matsuda H, Morikawa T, Ueda H, Yoshikawa M (2001) Medicinal Foodstuffs XXVII: saponin constituents Gotu Kola (2)Structures of new ursane- and oleanane-type triterpene oligoglycosides, centellasaponin B, C, and D, from Centella asiatica cultivated in Sri Lanka. Chem Pharm Bull 49:1368–1371

    Article  CAS  PubMed  Google Scholar 

  • Mook-Jung IH, Shin JE, Yun SH, Huh K, Koh JY, Park HK, Jew SS, Jung MW (1999) Protective effects of asiaticoside derivatives against beta-amyloid neurotoxicity. J Neurosci Res 58:417–425

    Article  CAS  PubMed  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Osbourn AE (1996) Performed antimicrobial compounds and plant defense against fungal attack. Plant Cell 8:1821–1831

    Article  CAS  PubMed  Google Scholar 

  • Randriamampionona D, Diallo B, Rakotoniriana F, Rabemanantsoa C, Cheuk K, Corbisier AM, Mahillon J, Ratsimamanga S, Jaziri ME (2007) Comparative analysis of active constituents in Centella asiatica samples from Madegascar: application for ex situ conservation and clonal propagation. Fitoterapia 78:482–489

    Article  CAS  PubMed  Google Scholar 

  • Schaller H, Grausem B, Benveniste P, Chye ML, Tan CT, Song YH, Chua NH (1995) Expression of the Hevea barsiliensis 3-hydroxy-3-methylglutaryl-coenzyme A reductase in tobacco results in sterol overproduction. Plant Physiol 109:761–770

    CAS  PubMed  Google Scholar 

  • Seo JW, Jeong JH, Shin CG, Lo SC, Han SS, Yu KW, Harada E, Han JY, Choi YE (2005) Overexpression of squalene synthase in Eleutherococcus senticosus increases phytosterol and triterpene accumulation. Phytochemistry 66:869–877

    Article  CAS  PubMed  Google Scholar 

  • Shabani L, Ehsanpour AA, Asghari G, Emami J (2009) Glycyrrhizin production by in vitro cultured Glycyrrhiza glabra elicited by methyl jasmonate and salicylic acid. Russ J Plant Physiol 56:621–626

    Article  CAS  Google Scholar 

  • Suzuki M, Muranaka T (2007) Molecular genetics of plant sterol backbone synthesis. Lipids 42:47–54

    Article  CAS  PubMed  Google Scholar 

  • Suzuki H, Srinivasa Reddy MS, Naoumkina M, Aziz N, May GD, Huhman DV, Sumner LW, Blount JW, Mendes P, Dixon RA (2005) Methyl jasmonate and yeast elicitor induce differential transcriptional and metabolic re-programming in cell suspension cultures of the model legume Medicago truncatula. Planta 220:696–707

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by a Korea Research Foundation Grant funded by the Korean Goverment (MOEHRD) (KRF-2007-C00271).

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Authors and Affiliations

  1. Department of Herbal Crop Research, National Institute of Horticultural and Herbal Science, RDA, Eumseong, 369-873, South Korea

    Ok Tae Kim

  2. Department of Biology, Chonnam National University, Gwangju, 500-757, South Korea

    Sun Hee Kim & Baik Hwang

  3. Department of Chemistry and Materials Science, Tokyo Institute of Technology, Meguro-ku, Tokyo, 152-8551, Japan

    Kiyoshi Ohyama

  4. Kihara Institute for Biological Research, Yokohama City University, Yokohama, Kanagawa, 244-0813, Japan

    Toshiya Muranaka

  5. Division of Forest Resources, College of Forest Sciences, Kangwon National University, Chunchon, 200-701, South Korea

    Yong Eui Choi

  6. College of Bioscience and Biotechnology, Kangwon National University, Chunchon, 200-701, South Korea

    Hyeon Yong Lee

  7. Jeonnam Agricultural Research and Extension Service, Naju, 520-715, South Korea

    Min Young Kim

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  1. Ok Tae Kim
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Correspondence to Baik Hwang.

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Communicated by J. R. Liu.

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Kim, O.T., Kim, S.H., Ohyama, K. et al. Upregulation of phytosterol and triterpene biosynthesis in Centella asiatica hairy roots overexpressed ginseng farnesyl diphosphate synthase. Plant Cell Rep 29, 403–411 (2010). https://doi.org/10.1007/s00299-010-0831-y

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  • DOI: https://doi.org/10.1007/s00299-010-0831-y

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