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Elicitation of diosgenin production in Trigonella foenum-graecum L. seedlings by heavy metals and signaling molecules

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Abstract

Seedlings of Trigonella foenum-graecum were treated with four heavy metal salts (CdCl2, CoCl2, K2Cr2O7 and NiCl2) to study the effect of heavy metals on growth and diosgenin production. It was found that CdCl2 increased diosgenin production up to 40-fold and CoCl2 increased diosgenin production up to 41-fold at concentrations which did not affect growth significantly. But K2Cr2O7 and NiCl2 were toxic to growth and inhibited diosgenin production. Effect of exogenously applied methyl jasmonate (MeJa) and calcium (Ca2+) on diosgenin production in seedlings of T. foenum-graecum was also investigated. MeJa enhanced the production of diosgenin. Maximum increase (10.5-fold) was found at 100 µL L−l concentration of MeJa. To study the role of Ca2+ on diosgenin production, seedlings of T. foenum-graecum were treated with a promoter of Ca2+ influx (calcium ionophore A23187), calcium depleted medium, Ca2+ channel blocker (verapamil) and antagonist (LaCl3), a divalent cation chelator (EGTA) and modulator of calcium release (caffeine). All the treatments were compared with a control containing 220 mg L−l concentration of CaCl2. The results suggest that the increase in cytosolic Ca2+ has an inhibitory role on diosgenin production. However, a calcium chelator or Ca2+ channel inhibitors could be used to elicit diosgenin production in this plant.

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References

  • Aerts RJ, Schafer A, Hesse M, Baumann TW, Slusarenk A (1996) Signalling molecules and the synthesis of alkaloids in Catharanthus roseus seedling. Phytochemistry 42:417–422

    Article  CAS  Google Scholar 

  • Aradhana M, Rao AC, Kale RK (1992) Diosgenin a growth stimulator of mammary gland of ovariectomized mouse. Indian J Exp Biol 30:367–370

    PubMed  CAS  Google Scholar 

  • Asolkar IV, Chadha YR (1979) Diosginin and other steroid drug precursors. Publication & Information Directorate, CSIR, New Delhi

    Google Scholar 

  • Bae KH, Choi YE, Shin CG, Kim YY, Kim YS (2006) Enhanced ginsenoside productivity by combination of ethephon and methyl jasmonate in ginseng (Panax ginseng C.A. Meyer) adventitious root cultures. Biotechnol Lett 28:1163–1166

    Article  PubMed  CAS  Google Scholar 

  • Bhavsar GC, Kapadia NS, Patel NM (1980) Studies of Trigonella foenum-graecum (Linn). Indian J Pharm sci 42:39–40

    CAS  Google Scholar 

  • Bolwell GP, Coulson V, Rodgers MW, Murphy DL, Jones D (1991) Modulation of the elicitation response in cultred fresh bean cells and its implication for the mechanism of signal transduction. Phytochmistry 30:397–405

    Article  CAS  Google Scholar 

  • Cacho M, Moran M, Tarrago JF, Crochete P (1995) Calcium restriction induces cardenolide accumulation in cell suspension cultures of Digitalis thapsi L. Plant Cell Rep 14:786–789

    Article  CAS  Google Scholar 

  • Cheng MS, Wang QL, Tian Q, Song HY, Lin YX, Li Q, Xu X, Miao HD, Yao XS, Yang Z (2003) Total synthesis of methyl protodioscin: a potent agent with antitumour activity. J Org Chem 68:3658–3662

    Article  PubMed  CAS  Google Scholar 

  • Cho HY, Son SY, Rhee HS, Yoon SH, Lee-Parsons CWT, Park JM (2008) Synergistic effects of sequential treatment with methyl jasmonate, salicylic acid and yeast extract on benzophenanthridine alkaloid accumulation and protein expression in Eschscholtzia californica suspension cultures. J Biotechnol 135:117–122

    Article  PubMed  CAS  Google Scholar 

  • 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

    Article  PubMed  CAS  Google Scholar 

  • Crochete MP, Jimenez MA, Moran M, Cacho M, Fernandez-Tarrago J (1991) Effect of calcium, manganese and lithium on growth and cardenolide content in cell suspension cultures of Digitalis thapsi. Plant Cell Rep 10:394–396

    Google Scholar 

  • Curtin C, Zhang W, Franco C (2003) Manipulating anthocyanin composition in Vitis vinifera suspension cultures by elicitation with jasmonic acid and light irradiation. Biotechnol Lett 25:1131–1135

    Article  PubMed  CAS  Google Scholar 

  • Dass S, Ramawat KG (2009) Calcium deprivation markedly enhances guggulsterone accumulation in cell cultures of Commiphora wightii. Curr Sci 96:1022–1024

    CAS  Google Scholar 

  • De B (2001) Cell signalling and elicitator induced plant metabolite production. J Med Aromat plants 23:413–428

    CAS  Google Scholar 

  • De Backer-Royer C, Vannereau A, Duret S, Villavigues R, Cosson L (1990) Action de metaux Lourds Cu et cd surdes cellules de Catharanthus Roseus cultivees in vitro adaptation et production alcaloidiqu. Les Colloq de I’INRA 51:247–249

    Google Scholar 

  • De D, De B (2003) Effect of ethephon on antioxidant enzymes and diosgenin production in seedlings of Trigonella foenum-graecum. Food Chem 82:211–216

    Article  CAS  Google Scholar 

  • De D, De B (2005) Elicitation of diosgenin production in Dioscorea floribunda by ethylene generating agent. Fitoterapia 76:153–156

    Article  PubMed  CAS  Google Scholar 

  • Ding J, Shi S, Jiang B, Yang Y, Huang J, Shen S, Xia K, Zhang J, Jiang X (2004) Effects of methyl jasmonate with indole-3-acetic acid and 6-benzylaminopurine on the secondary metabolism of cultured Onosoma paniculatum cells. In Vitro Cell Dev Biol—Plant 40:581–585

    Article  CAS  Google Scholar 

  • Evans WC (1996) Trease and evans pharmacognosy, 14th edn. WB Saunders, Philadalphia

    Google Scholar 

  • Fang Y, Smith MAL, Pépin MF (1999) Effects of exogenous methyl jasmonate in elicited anthocyanin-producing cell cultures of ohelo (Vaccinium phalae). In Vitro Cell Dev Biol—Plant 35:106–113

    Article  CAS  Google Scholar 

  • Franceschi VR, Grimes HD (1991) Induction of soybean vegetative proteins and anthocyanins by low level atmospheric methyl jasmonate. Proc Natl Acad Sci USA 88:6745–6749

    Article  PubMed  CAS  Google Scholar 

  • Gaisser S, Heide L (1996) Inhibition and regulation of shikonin biosynthesis in suspension cultures of Lithospermum. Phytochemistry 41:1065–1072

    Article  CAS  Google Scholar 

  • Guo H, Chang Z, Yang R, Gou D, Zeng J (1998) Anthraquinones from hairy root cultures of Cassia obtusifolia. Phytochemistry 49:1623–1625

    Article  PubMed  CAS  Google Scholar 

  • Hardman R, Fazli FRY (1972) Labelled steroidal sapogenins and hydrocarbons from Trigonella foenum-graecum by acetate, mevalonate and cholesterol feeds to seeds. Planta Med 21:188–195

    Article  PubMed  CAS  Google Scholar 

  • Indrayanta G, Utani M, Santosa MH, Syarani A (1994) In: Florey K (ed) Analytical profiles of drug substances, vol 23. Academic Press, New York

  • Indrayanta G, Utami W, Syahrani A (1996) Agave amaniensis Trel. and Nowell: in vitro culture and the production of phytosteroids. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry, vol 37, Medicinal and aromatic plants IX, Springer, pp 1–15

  • Kim D, Pederson H, Chin C (1991) Stimulation of berberine production of Thalictrum rugosum suspension cultures in response to addition of cupric sulfate. Biotechnol Lett 13:213–216

    Article  CAS  Google Scholar 

  • 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 Tissue Organ Cul 98:25–33

    Article  CAS  Google Scholar 

  • Knight MR, Smith SM, Trewavas AJ (1992) Wind-induced plant motion immediately increases cytosolic calcium. Proc Natl Acad Sci 89:4962–4971

    Article  Google Scholar 

  • Lu MB, Wong HL, Teng WL (2001) Effects of elicitation on the production of saponin in cell culture of Panax ginseng. Plant Cell Rep 20:647–677

    Article  CAS  Google Scholar 

  • Madrid E, Corchete P (2010) Purificación Silymarin secretion and its elicitation by methyl jasmonate in cell cultures of Silybum marianum is mediated by phospholipase D-phosphatidic acid. J Exp Bot 61:747–754

    Article  PubMed  CAS  Google Scholar 

  • Mahady GB, Beecher CWW (1994) Elicitor stimulated benzophenanthridine alkaloid biosynthesis in blood root suspension cultures is mediated by calcium. Phytochemistry 37:415–419

    Article  CAS  Google Scholar 

  • Mangas S, Bonfill M, Osuna L, Moyano E, Tortoriello J, Cusido RM M, Piñol T, Palazón J (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

    Article  PubMed  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Ning W, Wang J-Xi, Liu Y-M, Li N, Cao R-Q (1998) The effects of CaSUP2+/SUP during the elicitation of shikonin derivatives in Onosma Paniculatum cells. In Vitro Cell Dev Biol–Plant 34:261–265

    Article  CAS  Google Scholar 

  • Ohlsson AB, Berglund T (1989) Effects of high MnSO4 levels on cardenolide accumulation by Digitalis lanata tissue cultures in light and darkness. J Plant Physiol 135:505–507

    CAS  Google Scholar 

  • Ortuno A, Oncina R, Botia JM, Del Rio JA (1999) Regulation of the diosgenin expression in Trigonella foenum-graceum plants by different plant growth regulators. Food Chem 65:227–232

    Article  Google Scholar 

  • Palazo′n J, Cusido′ RM, Bonfill M, Mallol A, Moyano E, Morales C, Pin˜ol MT (2003) Elicitation of different Panax ginseng transformed root phenotypes for an improved ginsenoside production. Plant Physiol Biochem 41:1019–1025

    Article  Google Scholar 

  • Reed PW, Lardy HA (1972) A23187: a divalent cation ionophore. J Biol Chem 247:6970–6977

    PubMed  CAS  Google Scholar 

  • Roman ID, Thewles A, Coleman R (1995) Fractionation of livers following diosgenin treatment to elevate biliary cholesterol. Biochem Biophys Acta 1255:77–81

    PubMed  Google Scholar 

  • Sauvaire Y, Ribes G, Bacccou JC (1991) Implication of steroid saponins and sapogenins in the hypocholesterolemic effect of fenugreek. Lipids 26:191–197

    Article  PubMed  CAS  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 Russian. J Plant Physiol 56:621–626

    CAS  Google Scholar 

  • Sierra MI, Dagnino D, Van der Heijden R, Verpoorte R (1991) Influence of Ca on peroxidase activity and alkaloid formation in Tabernaemontana divaricata cell suspension cultures. In: Lobarzewski J, Greppin H, Panel C, Th Gaspar (eds) Biochemical molecular and physiological aspects of plant peroxidases. University of Geneva, Geneva, pp 295–304

    Google Scholar 

  • Solichatun NN, Anggarwulan E (2008) The reserpine production and callus growth of indian snake root (Rauvolfia serpentina (L.) Benth. Ex Kurz) culture by addition of Cu2+. Biodiversitas 9:177–179

    Google Scholar 

  • Sudha G, Ravishankar GA (2002) Involvement and interaction of various signaling compounds on the plant metabolic events during defense response, resistance to stress factors, formation of secondary metabolites and their molecular aspects. Plant Cell Tissue Organ Cult 71:181–212

    Article  CAS  Google Scholar 

  • Sudha G, Ravishankar GA (2003a) The role of calcium channels in anthocyanin production in callus cultures of Daucus carota. Plant growth Regul 40:163–169

    Article  CAS  Google Scholar 

  • Sudha G, Ravishankar GA (2003b) Elicitation of anthocyanin production in callus cultures of Daucus carota and the involvement of methyl jasmonate and salicylic acid. Acta Physiol Plant 25:249–256

    Article  CAS  Google Scholar 

  • Sung SL, Huang YS (2000) Headspace ethylene accumulation on Stizolobium hassjoo hairy root culture producing 1–3, 4 dihydroxyphenylalanine. Biotechnol Lett 22:875–878

    Article  CAS  Google Scholar 

  • Vitrac X, Larronde F, Krisa S, Decendit A, Deffieux G, Merillon JM (2000) Sugar sensing and Ca2+-calmodulin requirement in Vitis vinifera cells producing anthocyanin. Phytochemistry 53:659–665

    Article  PubMed  CAS  Google Scholar 

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The authors gratefully acknowledge financial assistance by Department of Science and Technology, Government of India.

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Correspondence to Bratati De.

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Communicated by M. Hajduch.

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De, D., De, B. Elicitation of diosgenin production in Trigonella foenum-graecum L. seedlings by heavy metals and signaling molecules. Acta Physiol Plant 33, 1585–1590 (2011). https://doi.org/10.1007/s11738-010-0691-7

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  • DOI: https://doi.org/10.1007/s11738-010-0691-7

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