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Efficient induction of ginsenoside biosynthesis and alteration of ginsenoside heterogeneity in cell cultures of Panax notoginseng by using chemically synthesized 2-hydroxyethyl jasmonate

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Abstract

Chemically synthesized 2-hydroxyethyl jasmonate (HEJA) was for the first time employed to induce the ginsenoside biosynthesis and to manipulate the product heterogeneity in plant cell cultures. The dose response and timing of HEJA elicitation were investigated in cell suspension cultures of Panax notoginseng. The optimal concentration and timing of HEJA addition for both cell growth and ginsenoside accumulation was identified to be 200 μM added on day 4. It was interestingly found that HEJA could stimulate ginsenosides biosynthesis and change their heterogeneity more efficiently than methyl jasmonate (MJA), i.e., the total ginsenoside content and the Rb/Rg ratio increased about 60 and 30% with HEJA elicitation than that by MJA, respectively. The activity of Rb1 biosynthetic enzyme, i.e., UDPG-ginsenoside Rd glucosyltransferase (UGRdGT), was also higher in the former case. A maximal production titer of ginsenoside Rg1, Re, Rb1, and Rd was 47.4±4.8, 52.3±4.4, 190±18, and 12.1±2.5 mg/l with HEJA elicitation, which was about 1.3-, 1.3-, 1.7-, and 2.1-fold than that using MJA, respectively. Early signal events in plant defense response, including oxidative burst and jasmonic acid (JA) biosynthesis, were also examined. Levels of H2O2 and NO in medium and l-phenylalanine ammonia lyase activity in cells were not affected by addition of MJA and HEJA. On the other hand, the JA content in cells was increased with external jasmonates elicitation, and it was inhibited with the addition of JA biosynthesis inhibitors. The results suggest that oxidative burst might not be involved in the jasmonates-elicited signal transduction pathway, and MJA and HEJA may induce the ginsenoside biosynthesis via induction of endogenous JA biosynthesis and key enzymes (such as UGRdGT) in the ginsenoside biosynthetic pathway of P. notoginseng cells. The information is useful for hyperproduction of plant-specific heterogeneous products.

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References

  • Ali MB, Yu KW, Hahn EJ, Paek KY (2005) Differential responses of anti-oxidants enzymes, lipoxygenase activity, ascorbate content and the production of saponins in tissue cultured root of mountain Panax ginseng CA Meyer and Panax quinquefolium L. in bioreactor subjected to methyl jasmonate stress. Plant Sci 169:83–92

    Article  CAS  Google Scholar 

  • Cazalé AC, Rouet-Mayer MA, Barbier-Brygoo H, Mathieu Y, Laurière C (1998) Oxidative burst and hypoosmotic stress in tobacco cell suspensions. Plant Physiol 116:659–669

    Article  Google Scholar 

  • Chen Z, Silva H, Klessig DF (1993) Active oxygen species in the induction of plant systemic acquired resistance by salicylic acid. Science 262:1883–1886

    Article  CAS  Google Scholar 

  • Creelman RA, Mullet JE (1997) Biosynthesis and action of jasmonates in plants. Annu Rev Plant Physiol Plant Mol Biol 48:355–381

    Article  CAS  Google Scholar 

  • Delledonne M, Zeier J, Marocco A, Lamb C (2001) Signal interactions between nitric oxide and reactive oxygen intermediates in the plant hypersensitive disease resistance response. Proc Natl Acad Sci U S A 98:13454–13459

    Article  CAS  Google Scholar 

  • Desikan R, Reynolds A, Hancock JT, Neill SJ (1998) Harpin and hydrogen peroxide both initiate programmed cell death but have differential effects on defence gene expression in Arabidopsis suspension cultures. Biochem J 330:115–120

    Article  CAS  Google Scholar 

  • Dong HD, Zhong JJ (2001) Significant improvement of taxane production in suspension cultures of Taxus chinensis by combining elicitation with sucrose feed. Biochem Eng J 8:145–150

    Article  CAS  Google Scholar 

  • Farmer EE (1994) Fatty acid signaling in plants and their associate microorganisms. Plant Mol Biol 26:1423–1437

    Article  CAS  Google Scholar 

  • Gundlach H, Muller MJ, Kutchan TM, Zenk MH (1992) Jasmonic acid is a signal transducer in elicitor-induced plant cell cultures. Proc Natl Acad Sci U S A 89:2389–2392

    Article  CAS  Google Scholar 

  • Heide L, Nishioka N, Fukuki H, Tabata M (1989) Enzymatic regulation of shikonin biosynthesis in Lithospermum erythrorhizon cell cultures. Phytochemistry 28:1873–1877

    Article  CAS  Google Scholar 

  • Hu XY, Neill S, Cai WM, Tang Z (2003a) Hydrogen peroxide and jasmonic acid mediate oligogalacturonic acid-induced saponin accumulation in suspension-cultured cells of Panax ginseng. Physiol Plant 118:414–421

    Article  CAS  Google Scholar 

  • Hu XY, Neill S, Cai WM, Tang Z (2003b) Nitric oxide mediates elicitor-induced saponin synthesis in cell cultures of Panax ginseng. Funct Plant Biol 30:901–907

    Article  CAS  Google Scholar 

  • Ketchum REB, Gibson DM, Croteau RB, Shuler ML (1999) The kinetics of taxoid accumulation in cell suspension cultures of Taxus following elicitation with methyl jasmonate. Biotechnol Bioeng 62:97–105

    Article  CAS  Google Scholar 

  • Koda Y, Kikuta Y, Tazaki H, Tsujino Y, Sakamura S, Yoshihara T (1991) Potato tuber-inducing activities of jasmonic acid and related compounds. Phytochemistry 30:1435–1438

    Article  CAS  Google Scholar 

  • Lobler M, Lee J (1998) Jasmonate signaling in barley. Trends Plant Sci 3:8–9

    Article  Google Scholar 

  • Melan MA, Dong X, Endara ME, Davis KR, Ausubel FM, Peterman TK (1993) An Arabidopsis thaliana lipoxygenase gene can be induced by pathogens, abscisic acid, and methyl jasmonate. Plant Physiol 101:441–450

    Article  CAS  Google Scholar 

  • Miersch O, Ksamell R, Parthier B, Wasternack C (1999) Structure activity relations of substituted, deleted or stereospecifically altered jasmonic acid in gene expression of barley leaves. Phytochemistry 50:353–361

    Article  CAS  Google Scholar 

  • Muller MJ, Brodschelm W, Spannagl E, Zenk MH (1993) Signaling in the elicitation process in mediated through the octadecanoid pathway leading to jasmonic acid. Proc Natl Acad Sci U S A 90:7490–7494

    Article  Google Scholar 

  • Murphy ME, Noack E (1994) Nitric oxide assay using hemoglobin method. Methods Enzymol 233:240–250

    Article  CAS  Google Scholar 

  • Orozco-Cárdenas ML, Ryan CA (2002) Nitric oxide negatively modulates wound signaling in tomato plants. Plant Physiol 130:487–493

    Article  Google Scholar 

  • Qian ZG, Zhao ZJ, Xu Y, Qian X, Zhong JJ (2004a) Novel chemically synthesized hydroxyl-containing jasmonates as powerful inducing signals for plant secondary metabolism. Biotechnol Bioeng 86:809–816

    Article  CAS  Google Scholar 

  • Qian ZG, Zhao ZJ, Tian WH, Xu Y, Zhong JJ, Qian X (2004b) Novel synthetic jasmonates as highly efficient elicitors for taxoid production by suspension cultures of Taxus chinensis. Biotechnol Bioeng 86:595–599

    Article  CAS  Google Scholar 

  • Rijhwani S, Shanks JV (1998) Effect of elicitor dosage and exposure time on biosynthesis of indole alkaloids by Catharanthus roseus hairy root cultures. Biotechnol Prog 14:442–449

    Article  CAS  Google Scholar 

  • Staniszewska I, Królicka A, Maliñski E, Łojkowska E, Szafranek J (2003) Elicitation of secondary metabolites in in vitro cultures of Ammi majus L. Enzyme Microb Technol 33:565–568

    Article  CAS  Google Scholar 

  • Staswick PE, Huang JF, Rhee Y (1991) Nitrogen and methyl jasmonate induction of soybean vegetative storage protein genes. Plant Physiol 96:130–136

    Article  CAS  Google Scholar 

  • Sticher O (1998) Getting to the root of ginseng. Chemtech 28:26–32

    CAS  Google Scholar 

  • Swiatek A, van Dongen W, Esmans EL, van Onckelen H (2004) Metabolic fate of jasmonates in tobacco Bright Yellow-2 cells. Plant Physiol 135:161–172

    Article  CAS  Google Scholar 

  • Tabata H (2004) Paclitaxel production by plant-cell-culture technology. Adv Biochem Eng Biotechnol 87:1–23

    CAS  PubMed  Google Scholar 

  • Wang JW, Wu JY (2005) Nitric oxide is involved in methyl jasmonate-induced defense responses and secondary metabolism activities of Taxus cells. Plant Cell Physiol (adv access)

  • Wang W, Zhong JJ (2002) Manipulation of ginsenoside heterogeneity in cell cultures of Panax notoginseng by addition of jasmonates. J Biosci Bioeng 93:48–53

    Article  CAS  Google Scholar 

  • Wang W, Zhang ZY, Zhong JJ (2005) Enhancement of ginsenoside biosynthesis in high density cultivation of Panax notoginseng cells by various strategies of methyl jasmonate elicitation. Appl Microbiol Biotechnol 67:752–758

    Article  CAS  Google Scholar 

  • Woragidbumrung K, Sae-Tang P, Yao H, Han J, Chauvatcharin S, Zhong JJ (2001) Impact of conditioned medium on cell cultures of Panax notoginseng in an air-lift bioreactor. Process Biochem 37:209–213

    Article  CAS  Google Scholar 

  • Yu KW, Gao WY, Hahn EJ, Paek KY (2002) Jasmonic acid improving ginsenoside accumulation in adventitious root culture of Panax ginseng CA Meyer. Biochem Eng J 11:211–215

    Article  CAS  Google Scholar 

  • Yu KW, Murthy HN, Hahn EJ, Paek KY (2005) Ginsenoside production by hairy root cultures of Panax ginseng: influence of temperature and light quality. Biochem Eng J 23:53–56

    Article  CAS  Google Scholar 

  • Yue CJ, Zhong JJ (2005a) Impact of external calcium and calcium sensors on ginsenoside Rb1 biosynthesis by Panax notoginseng cells. Biotechnol Bioeng 89:444–452

    Article  CAS  Google Scholar 

  • Yue CJ, Zhong JJ (2005b) Purification and characterization of UDPG: ginsenoside Rd glucosyltransferase from suspended cells of Panax notoginseng. Process Biochem (in press)

  • Zhang ZY, Zhong JJ (2004) Scale-up of centrifugal impeller bioreactor for hyperproduction of ginseng saponin and polysaccharide by high-density cultivation of Panax notoginseng cells. Biotechnol Prog 20:1076–1081

    Article  CAS  Google Scholar 

  • Zhao J, Sakai K (2003) Multiple signaling pathways mediate fungal elicitor induced β-thujaplicin biosynthesis in Cupressus lusitanica cell cultures. J Exp Bot 54:647–656

    Article  CAS  Google Scholar 

  • Zhao J, Davis L, Verpoorte R (2005) Elicitor signal transduction leading to production of plant secondary metabolites. Biotechnol Adv 23:283–333

    Article  CAS  Google Scholar 

Download references

Acknowledgements

Financial support from the National Natural Science Foundation of China (NSFC project nos. 20225619, 20236040, and 20376023), the National Key Project for Basic Research, the Ministry of Science and Technology of China (2003CB114400), and Shanghai Science and Technology Commission (Project no. 04QMH1410) is gratefully acknowledged.

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Authors and Affiliations

  1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China

    Wei Wang & Jian-Jiang Zhong

  2. School of Pharmacy, Shanghai Jiao-Tong University, 1954 Huashan Road, Shanghai, 200030, China

    Wei Wang

  3. Shanghai Key Laboratory of Chemical Biology and Institute of Pesticides and Pharmarceuticals, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China

    Zhen-Jiang Zhao, Yufang Xu & Xuhong Qian

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  1. Wei Wang
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  2. Zhen-Jiang Zhao
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  3. Yufang Xu
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  4. Xuhong Qian
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  5. Jian-Jiang Zhong
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Correspondence to Wei Wang or Jian-Jiang Zhong.

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Wang, W., Zhao, ZJ., Xu, Y. et al. Efficient induction of ginsenoside biosynthesis and alteration of ginsenoside heterogeneity in cell cultures of Panax notoginseng by using chemically synthesized 2-hydroxyethyl jasmonate. Appl Microbiol Biotechnol 70, 298–307 (2006). https://doi.org/10.1007/s00253-005-0089-4

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