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인삼에서 Farnesyl Diphosphate Synthase 과발현이 진세노사이드 생합성에 미치는 영향

Overexpression of Farnesyl Diphosphate Synthase by Introducing CaFPS Gene in Panax ginseng C. A. Mey.

김영창|김옥태|김장욱|방경환|현동윤|차선우|최재을|박홍우
한국약용작물학회|2013.02|한국약용작물학회지 (KOREAN JOURNAL OF MEDICINAL CROP SCIENCE)|제21권 제1호|pp. 32-38|DOI http://bxdoi.org/10.7783/KJMCS.2013.21.1.32

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참고문헌

키워드

Panax ginsengGinsenoside

초록

FPS (farnesyl diphosphate synthase) plays an essential role in organ development in plants. However, FPS has not previously been identified as a key regulatory enzyme in triterpene biosynthesis. In order to investigate the effect of FPS on ginsenosides biosynthesis, we over-expressed FPS of Centella asiatica (CaFPS) in Panax giseng adventitious roots. PCR analysis showed the integrations of the CaFPS and hygromycin phosphotransferase genes and we ultimately selected three lines. The result of Southern blot analysis demonstrated the introduction of the CaFPS gene into genome of ginseng. In addition, the results of RT-PCR analysis revealed that CaFPS gene overexpression induced an accumulation of its transcription in the ginseng adventitious roots. To determine whether or not the overexpression of the CaFPS gene contributes to the downstream gene expression associated with triterpene biosynthesis, the level of mRNAs was analyzed by real-time PCR. The result showed that no differences were detected in any expression of all genes. To determine quantitatively the content of ginsenosides in transgenic ginseng adventitious roots, HPLC analysis was conducted. The content of total 7 ginsenosides was increased to 1.8, 1.4, and 1.7 times than that of the controls, respectively. This indicated that the overexpression of CaFPS in ginseng adventitious roots causes an increase in ginsenoside content, although down stream genes of FPS gene were suppressed by CaFPS overexpression.

Abstract

FPS (farnesyl diphosphate synthase) plays an essential role in organ development in plants. However, FPS has not previously been identified as a key regulatory enzyme in triterpene biosynthesis. In order to investigate the effect of FPS on ginsenosides biosynthesis, we over-expressed FPS of Centella asiatica (CaFPS) in Panax giseng adventitious roots. PCR analysis showed the integrations of the CaFPS and hygromycin phosphotransferase genes and we ultimately selected three lines. The result of Southern blot analysis demonstrated the introduction of the CaFPS gene into genome of ginseng. In addition, the results of RT-PCR analysis revealed that CaFPS gene overexpression induced an accumulation of its transcription in the ginseng adventitious roots. To determine whether or not the overexpression of the CaFPS gene contributes to the downstream gene expression associated with triterpene biosynthesis, the level of mRNAs was analyzed by real-time PCR. The result showed that no differences were detected in any expression of all genes. To determine quantitatively the content of ginsenosides in transgenic ginseng adventitious roots, HPLC analysis was conducted. The content of total 7 ginsenosides was increased to 1.8, 1.4, and 1.7 times than that of the controls, respectively. This indicated that the overexpression of CaFPS in ginseng adventitious roots causes an increase in ginsenoside content, although down stream genes of FPS gene were suppressed by CaFPS overexpression.

참고문헌 (44)

  1. 안인옥, "인삼 신품종의 뿌리부위별 진세노사이드 함량 및 패턴비교", Journal of Ginseng Research, 32(1), pp.15~18 (2008), 고려인삼학회
  2. Chen D, "Expression of a chimeric farnesyl diphosphate synthase gene in Artemisia annua L. transgenic plants via Agrobacterium tumefaciens-mediated transformation", Plant Science, 155, pp.179~185 (2000)
  3. Choi KT., "Botanical characteristics, pharmacological effects and medicinal components of Korean Panax ginseng C. A. Meyer", Acta Pharmacologica Sinica, 29, pp.1109~1118 (2008)
  4. Chen DT, "Expression of a chimeric farnesyl diphosphate synthase gene in Artemisia annua L. transgenic plants via Agrobacterium tumefaciens-mediated transformation", Plant Science, 155, pp.179~185 (2000)
  5. Chung YY, "Comparison of plant growth and morphological characteristics among the Korean ginseng, the American ginseng and the bamboo ginseng", Journal of Ginseng Research, 22, pp.147~153 (1998)
  6. Cunillera N, "The Arabidopsis thaliana FPS1 gene generates a novel mRNA that encodes a mitocondrial farnesyl-diphosphate synthase isoform", Journal of Biological Chemistry, 272, pp.15381~15388 (1997)
  7. Cunillera N, "Spatial and temporal patterns of GUS expression directed by 5’regions of the Arabidopsis thaliana farnesyl diphosphate synthase genes FPS1 and FPS2", Plant Molecular Biology, 44, pp.747~758 (2000)
  8. 도은수, "발효인삼의 Ginsenoside 변화와 항산화 활성", 한국약용작물학회지, 18(4), pp.255~265 (2010), 한국약용작물학회
  9. Ehlting J, "An extensive (co-)expression analysis tool for the cytochrome P450 superfamily in Arabidopsis thaliana", BMC Plant Biology, 8, pp.1~1994 (2008)
  10. Gaffe J, "LEFPS1, a thomato farnesyl pyrophosphate gene highly expressed during early fruit development", Plant Physiology, 123, pp.137~1362 (2000)
  11. Gillis CN., "Panax gineng pharmacology: Anitricoxidelink?", Biochemical Pharmacology, 54, pp.1~8 (1997)
  12. Han JY, "Regulation of ginsenoside and phytosterol biosynthesis by RNA interferences of squalene epoxidase gene in Panax ginseng", PHYTOCHEMISTRY, 71, pp.36~46 (2010), PERGAMON-ELSEVIER SCIENCE LTD
  13. Hayashi H, "Up-regulation of soyasaponin biosynthesis by methyl jasmonate in cultured cells of Glycyrrhiza glabra", Plant and Cell Physiology, 44, pp.404~411 (2003)
  14. Hu SY., "The genus Panax(Ginseng) in Chinese medicine", Economic Botany, 30, pp.11~28 (1976)
  15. Kim MW, "Distribution of saponin various sections of Panax ginseng root and change so fits contents according to root age", Journal of Ginseng Research, 11, pp.10~16 (1987)
  16. Kim OT, Kim MY, Huh SM, Bai DG, Ahn JC and Hwang B., "Cloning of a cDNA probably encoding oxidosqualene cyclase associated with asiaticoside biosynthesis from Centella asiatica (L.) Urban", PLANT CELL REPORTS, 24(5), pp.304~311 (2005), SPRINGER
  17. Kim OT, "Upregulation of phytosterol and triterpene biosynthesis in Centella asiatica hairy roots overexpressed ginseng farnesyl diphosphate synthase", PLANT CELL REPORTS, 29(4), pp.403~411 (2010), SPRINGER
  18. Kim YS, "Ginseng metabolic engineering: Regulation of genes related to ginsenoside biosynthesis", JOURNAL OF MEDICINAL PLANTS RESEARCH, 3, pp.1270~1276 (2009), Academic Journal
  19. Kwon YD, "Analysis of international competitiveness of Chinese ginseng in comparison with Korean ginseng", Research Institute for Agricultural Sciences and Technology, 33, pp.43~56 (2006), Chungnam National University
  20. Lairson LL, "Glycosyl-transferases: Structures, functions, and mechanisms", Annual Review Biochemistry, 77, pp.521~555 (2008)
  21. Lee MH, "Enhanced triterpene and phytosterol biosynthesis in Panax ginseng overexpressing squalene synthase gene", PLANT AND CELL PHYSIOLOGY, 45, pp.976~984 (2004), OXFORD UNIV PRESS
  22. Livak KJ, "Analysis of relative gene expression data using real-time quantitative PCR and the 2CT method", Methods, 25, pp.402~408 (2001)
  23. Lichtenthaler HK, "The1-deoxy-D-xylulose-5-phosphate pathway of isoprenoid biosynthesis in plants", Plant Molecular Biology, 50, pp.47~65 (1999)
  24. Agrobacterium tumefaciens-mediated transformation of Narcissus tazzeta var. chinensis, "Lu G", Plant Cell Reports, 1585~1593 (2007)
  25. Manzano D, "The metabolic imbalance under lying lesion for mationin Arabidopsis thaliana farnesyl diphosphate synthase(isoform1s) leads to oxidative stress and is triggered by the developmental decline of endogenous HMGR activity", Planta, 219, pp.982~992 (2004)
  26. Masferrer A Arró M, "Overexpression of Arabidosis thaliana farnesyl diphosphate synthase(FPS1S) in transgenic Arabidopsis induces a cell death", senescence-like response and reduced cytokinin levels, 2002, pp.30, 123~132
  27. Meesapyodsuk D, "Saponin biosynthesis in Saponaria vaccaria: cDNAs encoding ß-amyrin synthase and a triterpene carboxylic acid glucosyltransferase", Plant Physiology, 143, pp.959~969 (2007)
  28. Murashige T, "A revised medium for rapid growth and bioassays with tobacco tissues", Physiologia Plantarum, 15, pp.473~497 (1962)
  29. 남기열, "고려인삼의 방사선 방어효과에 대한 연구현황과 전망", 한국약용작물학회지, 19(4), pp.287~299 (2011), 한국약용작물학회
  30. Proctor JTA, "Ginseng: Industry, botany and culture", Horticultural Reviews, 9, pp.187~236 (1987)
  31. Rohmer M., "Mevalonate-independent methylerythritol phosphate pathway for isoprenoid biosynthesis. Elucidation and distribution", Pure and Applied Chemistry, 75, pp.375~387 (2003)
  32. Sambrook J, "Molecular cloning. A Laboratory Manual", pp.931~958 (1989), Cold Spring Harbor Laboratory Press
  33. Scaglione F, "Efficacy and safety of the standardized ginseng extract G 115 for potentiating vaccination against common cold and", or influenza syndrome, 1996, pp.22, 65~72
  34. See DM, "In vitro effects of Echinacea and ginseng on natural killer and antibodydependent cell cytotoxicity in healthy subjects and chronicfatique syndrome or acquired immunodeficiency efficiency syndrome patients", Immunopharmacology, 35, pp.229~235 (1997)
  35. Schuler MA, "Functional genomics of P450s", Annual Review of Plant Biology, 54, pp.629~637 (2003)
  36. Suzuki H, "A genomic approach to the early stages of triterpene saponin biosynthesis in Medicago truncatula", The Plant Journal, 32, pp.1033~1048 (2002)
  37. Takahashi M., "Anti-stress effect of ginseng on the inhibition of the development of morphine tolerance in stressed mice", Japanese Journal of Pharmacology, 59, pp.399~404 (1992)
  38. Yu KW, "Improvement of ginsenoside production bt jasmonic acid and some other elicitors in hairy root culture of ginseng(Panax ginseng C. A. Meyer)", In Vitro Cellular and Developmental Biology, 36, pp.424~428 (2000)
  39. Verpoorte R, "Metabolic engineering for the improvement of plant secondary metabolite production", Plant Tissue Culture and Biotechnology, 46, pp.3~20 (1998)
  40. Vogler BK, "The efficacy of ginseng. A systematic review of randomized clinical trials", European Journal of Clinical Pharmacology, 55, pp.567~575 (1999)
  41. Wen J, "Phylogeny and biogeography of Panax L. (the ginseng genus, Araliaceae): Inferences from ITS sequences of nuclear ribosomal DNA", Molecular Phylogenetics and Evolution, 6, pp.167~177 (1996)
  42. Wu Y, "An experimental assessment of the factors influencing Agrobacterium-mediated transformation in tomato", Russian Journal of Plant Physiology, 53, pp.252~284 (2006)
  43. Yamunadevi M, "Phytochemical studies on the terpenoids of medicinally important plant Aerva lanata L. using HPTLC", Asian Pacific Journal of Tropical Biomedicine, 1, pp.220~225 (2011)
  44. Zwenger S, "Plant terpenoids: Applications and future potentials", Biotechnology and Molecular Biology Reviews, 37, pp.1~79 (2008)

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