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Elicitor-enhanced production of gymnemic acid in cell suspension cultures of Gymnema sylvestre R. Br.

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Abstract

Cell suspension cultures of Gymnema sylvestre treated with four different elicitors, methyl jasmonate (MJ), yeast extract, chitin and pectin were studied for the production of gymnemic acid as gymnemagenin equivalent, that was analyzed by high performance liquid chromatography (HPLC). All the four tested elicitors induced gymnemic acid production in cell suspension cultures. Highest gymnemic acid content was achieved following treatment with yeast extract (100.47 ± 0.28 mg/l), this was followed by MJ (70.43 ± 0.26 mg/l), pectin (64.19 ± 0.23 mg/l) and chitin (62.72 ± 0.13 mg/l). The addition of elicitors has shown a significant influence on cell growth that affected cell growth compared to respective controls. The highest gymnemic acid production was obtained after 20 days of elicitation in cultures treated with 0.5 g l−l yeast extract, it was 5.25-folds greater than in control. These results suggest that the addition of an elicitor to Gymnema sylvestre cell suspension cultures could stimulate and enhance gymnemic acid production. In our present study we could able to overproduce gymnemic acid up to 51.97 ± 0.26 mg l−l (dry weight basis) in yeast extract treated cell suspension cultures.

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References

  • Ahmed ABA, Rao AS, Rao MV (2008) Role of in vivo and in vitro callus of Gymnema sylvestre (Retz) R. Br. Ex. Roemer & Schultes in maintaining the normal levels of blood glucose and lipid profile in diabetic Wistar rats. Biomedicine 28:134–138

    Google Scholar 

  • Ali Ahmed AB, Rao AS, Rao MV (2009) In vitro production of gymnemic acid from gymnema sylvestre (Retz) R. Br. Ex Roemer and Schultes through callus culture under abiotic stress conditions. Methods Mol Biol 547(1):93–105

    Article  PubMed  Google Scholar 

  • Ali MB, Yu KW, Hahn EJ, Paek KY (2006) Methyl jasmonate and salicylic acid elicitation induces ginsenosides accumulation, enzymatic and non-enzymatic antioxidant in suspension culture Panax ginseng roots in bioreactors. Plant Cell Rep 25:613–620

    Article  PubMed  CAS  Google Scholar 

  • Aoyagi H, Kobayashi Y, Yamada K, Yokoyama K, Kusakari K, Tanaka H (2001) Efficient production of saikosaponins in Bupleurum falcatum root fragments combined with signal transducers. Appl Microbiol Biotechnol 57:482–488

    Article  PubMed  CAS  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 

  • Baskaran K, Ahamath B, Shanmugasundaram ER, Shanmugasundaram KR (1990) Antidiabetic effect of a leaf extract from Gymnema sylvestre in non-insulin-dependent diabetes mellitus patients. J Ethnopharmacol 30:295–300

    Article  PubMed  CAS  Google Scholar 

  • Bohlmann J, Gibraltarskaya E, Eilert U (1995) Elicitor induction of furanocoumarin biosynthetic pathway in cell cultures of Ruta graveolens. Plant Cell Tissue Organ Cult 43:155–161

    Article  CAS  Google Scholar 

  • 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

    Article  CAS  Google Scholar 

  • Chakravarti D, Debnath NB (1981) Isolation of gymnemagenin, the sapogenin of Gymnema sylvestre R. Br. (Asclepiadaceae). J Inst Chem (India) 53:155–158

    CAS  Google Scholar 

  • Chattopadhaya RR (1999) A comparative evaluation of some blood sugar lowering agents of plant origin. J Ethanopharmocol 67:367–372

    Article  Google Scholar 

  • Chen H, Chen F (2000) Effect of yeast elicitor on the secondary metabolism of Ti-transformed Salvia miltiorrhiza cell suspension cultures. Plant Cell Rep 19:710–717

    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 

  • Choudhury BP (1988) Assessment and conservation of medicinal plants of Bhubaneswar and its neighbourhood. In: Indigenous medicinal plants, Today and Tomorrow’s Printers and Publishers, New Delhi, p 211

  • Coste A, Viase L, Halmagyi A, Deliu C, Coldea G (2011) Effects of plant growth regulators and elicitors on production of secondary metabolites in shoot cultures of Hypericum hirsutum and Hypericum maculatum. Plant Cell Tissue Organ Cult. doi: 10.1007/s11 240-011-9919-5

  • Dateo GP Jr, Long L Jr (1973) Gymnemic acid the antisaccharine principle of Gymnema sylvestre. Studies on isolation and heterogeneity of gymnemic acid A1. J Agric Food Chem 21:899–903

    Article  PubMed  CAS  Google Scholar 

  • Delauny A, Spasenoski M, Joseph C, Hagege D (2007) Jasmonic acid elicitation of Hypericum perforatum L. cell suspensions and effects on the production of phenylpropanoids and naphtodianthrones. Plant Cell Tiss Org Cult 89:1–13

    Article  Google Scholar 

  • Farmer EE, Ryan CA (1990) Interplant communication-airborne methyl jasmonate induces synthesis of proteinase inhibitors in plant leaves. Proc Natl Acad Sci USA 87:7713–7716

    Article  PubMed  CAS  Google Scholar 

  • Flores-Sánchez IJ, Ortega-López J, del Carmen Montes-Horcasitas M, Ramos-Valdivia AC (2002) Biosynthesis of sterols and triterpenes in cell suspension cultures of Uncaria tomentosa. Plant Cell Physiol 43:1502–1509

    Article  PubMed  Google Scholar 

  • Gandzovska S, Maury S, Delaunay A, Spasenoski M, Joseph C, Hagège D (2007) Jasmonic acid elicitation of Hypericum perforatum L. cell suspensions and effects on the production of phenylpropanoids and napthodianthrones. Plant Cell Tiss Organ Cult 89:1–13

    Article  Google Scholar 

  • Gholap S, Kar A (2005) Gymnemic acids from Gymnema sylvestre potentially regulates dexamethasone induced hyperglycemia in mice. Pharm Biol 43:192–195

    Article  CAS  Google Scholar 

  • Gopi C, Vatsala TM (2006) In vitro studies in effects of plant growth regulators on callus and suspension culture biomass yield from Gymnema sylvestre R. Br. Afr J Biotechnol 5(12):1215–1219

    CAS  Google Scholar 

  • Hayashi H, Huang P, Inoue K (2003) Up-regulation of soyasaponin biosynthesis by methyl jasmonate in cultured cell of Glycyrrhiza glabra. Plant Cell Physiol 44:404–411

    Article  PubMed  CAS  Google Scholar 

  • Hirata S, Abe T, Imoto T (1992) Effect of crude gymnemic acid on oral glucose tolerance test in human being. J Yonago Med Assoc 43:392–396

    CAS  Google Scholar 

  • Holden MA, Holden PR, Yeoman MM (1988) Elicitation of cell cultures. In: Robins RJ, Rhodes MJC (eds) Manipulating secondary metabolism in culture. Cambridge University Press, Cambridge, p 57

    Google Scholar 

  • Hooper D (1889) Gymnemic acid. Chemical News 59:158–160

    Google Scholar 

  • Hu Y, Yu Y, Piao C, Liu J, Yu H (2011) Methyl jasmonate and salicylic acid induced D-chiro-inositol production in suspension cultures of buckwheat (Fagopyrum esculentum). Plant Cell Tiss Organ Cult doi: 10.1007/s11 2340-011-9938-2

  • Junge Qu, Wei Z, Xingju Yu (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. doi: 10.1007/s11240-011-9977.8

  • Kar A, Choudhary BK, Bandyopadhyay NG (1999) Preliminary studies of the inorganic constituents of some indigenous hypoglycaemic herbs on oral glucose tolerance test. J Ethnopharmacol 64:179–184

    Article  PubMed  CAS  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  PubMed  CAS  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, Bang KH, Kim YC, Hyun DY, Kim MY, Cha SW (2009b) 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, Kim SH, Ohyama K, Muranaka T, Choi YE, Lee HY, Kim MY, Hwang B (2010) Upregulation of phytosterol and triterpene biosynthesis in Centella asiatica hairy roots overexpressed ginseng farnesyl diphosphate synthase. Plant Cell Rep 29:403–411

    Article  PubMed  CAS  Google Scholar 

  • Kim YS, Cho JH, Park S, Han JY, Back K, Choi YE (2011) Gene regulation patterns in triterpene biosynthetic pathway driven by overexpression of squalene synthase and methyl jasmonate elicitation in Bupleurum falcatum. Planta 233:343–355

    Article  PubMed  CAS  Google Scholar 

  • Kittipongpatana N, Davis DL, Porter JR (2002) Methyl jasmonate increases the production of valepotriates by transformed root cultures of Valerianella locusta. Plant Cell Tiss Organ Cult 71:65–75

    Article  CAS  Google Scholar 

  • Kiuchi F, Liu HM, Tsuda Y (1992) Two new gymnemic acid congeners containing a hexulopyoanoside moiety. Chem Pharm Bull 38:2326–2328

    Google Scholar 

  • Korsangruang S, Soonthornchareonnon N, Chintapakorn Y, Saralamp P, Prathanturarug S (2010) Effects of abiotic and biotic elicitors on growth and isoflavonoid accumulation in Pueraria candollei var. candollei and P. candollei var. mirifica cell suspension cultures. Plant Cell Tiss Organ Cult 103:333–342

    Article  CAS  Google Scholar 

  • Kumar U, Singh I, Priyanka VY (2010) In vitro salt stress induced production of gymnemic acid in callus cultures of Gymnema sylvestre R. Br. Afr J Biotechnol 9(31):4904–4909

    CAS  Google Scholar 

  • Kurihara K (1992) Characteristics of antisweet substances, sweet proteins, and sweetness-inducing proteins. Crit Rev Food Sci Nutr 32:231–252

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Liu HM, Kiuchi F, Tsuda Y (1992) Isolation and structure elucidation of gymnemic acids, anti-sweet principles of Gymnema sylvestre. Chem Pharm Bull 40:1366–1375

    PubMed  CAS  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 20:674–677

    CAS  Google Scholar 

  • Lystvan K, Belokurova V, Sheludko Y, Ingham JL, Prykhodko V, Kishchenko O, Paton E, Kuchuk M (2009) Production of bakuchiol by in vitro systems of Psoralea drupacea Bge. Plant Cell Tiss Organ Cult 101:99–103

    Article  Google Scholar 

  • Maeda M, Iwashita T, Kurihara Y (1989) Studies on taste modifiers II. Purification and structure determination of Gymnemic acids, anti-sweet active principle from Gymnema sylvestre leaves. Tetrahedron Lett 30:1547–1550

    Article  CAS  Google Scholar 

  • Mangas S, Bonfill M, Osuna L, Moyano E, Tortoriello J, Cusido RM, Pinol MT, Palaźon 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 

  • Masayuki Y, Toshiyuki M, Hisashi M (1997) Structures of new triterpene glycosides, gymnemasides C, D, E and F from the leaves of Gymnema sylvestre R. Br.: influence of Gymnema glycosides on glucose uptake in rat small intestinal fragments. Chem Pharm Bull 45:2034–2038

    Google Scholar 

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

    Article  CAS  Google Scholar 

  • Nadkarni AK, Nadkarni KM (1976) Indian materia medica, vol 1. Popular Prakashan, Bombay, p 569

    Google Scholar 

  • Nadakarni AK (1992) Indian Materia Medica. In: Gymnema sylvestre Popular Prakashan: Bombay, vol 1, pp 596

  • Okabayashi Y, Tani S, Fujisawa T, Koide M, Hasegawa H, Nakamura T, Fujii M, Otsuki M (1990) Effect of Gymnema sylvestre R.Br. on glucose homeostasis in rats. Diabetes Res Clin Pract 9:143–148

    Article  PubMed  CAS  Google Scholar 

  • Toshihiro Y, Kenzi M, Kenzo O, Osamu, T (1994) Nippon Shokuhin Kagaku Kaishi. 41:202–205 (In Japanese)

    Google Scholar 

  • Persaud SJ, Al-Majed H, Raman A, Jones PM (1999) Gymnema sylvestre stimulates insulin releases in vitro by increased membrane permeability. J Endocrinol 163:207–212

    Article  PubMed  CAS  Google Scholar 

  • Rachh RR, Rachh NR, Ghadiya DC, Modi DC, Modi KP, Patel NM, Rupareliya MT (2010) Antihyperlipidemic activity of Gymnema sylvestre R. Br. leaf extract on rats fed with high cholesterol diet. Int J Pharmacol 6(2):138–141

    Article  Google Scholar 

  • Rafiullah MRM, Siddiqui AW, Mir SR, Ali M, Pillai KK, Singh S (2006) Antidiabetic activity of some Indian medicinal plants. Pharm Biol 44:95–99

    Article  Google Scholar 

  • Rana AC, Avadhoot Y (1992) Experimental evaluation of hepato-protective activity of Gymnema sylvestre and Curcuma zedoaria. Fitoterapia 63:60–62

    Google Scholar 

  • Rao GS, Sinsheimer JE (1968) Structure of gymnemagenin. Chem Commun 1681–1682

  • Rao GS, Sinsheimer JE (1971) Constituents from Gymnema sylvestre leaves VIII: isolation, chemistry and derivatives of gymnemagenin and gymnestrgenin. J Pharma Sci 60:190–193

    Article  CAS  Google Scholar 

  • Ren YY, West CA (1992) Elicitation of diterpene biosynthesis in rice (Oryza sativa L.) by chitin. Plant Physiol 99:1169–1178

    Article  PubMed  CAS  Google Scholar 

  • Rhee HS, Cho H-Y, Son SY, Yoon S-YH, Park JM (2010) Enhanced accumulation of decursin and decursinol angelate in root cultures and intact roots of Angelica gigas Nakai following elicitation. Plant Cell Tiss Organ Cult 101:295–302

    Article  CAS  Google Scholar 

  • Sahu NP, Mahato SB, Sarkar SK, Poddar G (1996) Triterpenoid saponins from Gymnema sylvestre. Phytochemistry 41:1181–1185

    Article  PubMed  CAS  Google Scholar 

  • Selvanayagam ZE, Gnanavendhan SG, Chandrashekhran P, Balakrishna K, Rao RB (1995) Plants with anti-snake venom activity–a review on pharmacological and clinical studies. Fitoterapia 65:99–111

    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 

  • Shanmugasundaram ERB, Gopinath KL, Shanmugasundaram KR, Rajendran VM (1990a) Possible regeneration of the islets of Langerhans in streptozotocin-diabetic rats given Gymnema sylvestre leaf extracts. J Ethnopharmacol 30:265–279

    Article  PubMed  CAS  Google Scholar 

  • Shanmugasundaram ERB, Rajeswari G, Baskaran K, Kumar RBR, Shanmugasundaram KR, Ahmath BK (1990b) Use of Gymnema sylvestre leaf extract in the control of blood glucose in insulin-dependent diabetes mellitus. J Ethnopharmacol 30:281–294

    Article  PubMed  CAS  Google Scholar 

  • Sharan M, Taguchi G, Gonda K, Jouke T, Shimosaka M, Hayashida N, Okazaki M (1998) Effects of methyl jasmonate and elicitor on the activation of phenylalanine ammonialyase and the accumulation of scopoletin and scopolin in tobacco cell cultures. Plant Sci 132:13–19

    Article  CAS  Google Scholar 

  • Shigematsu N, Asano R, Shimosaka M, Okazaki M (2001) Effect of administration with the extract of Gymnema sylvestre R. Br leaves on lipid metabolism in rats. Biol Pharm Bull 24:713–717

    Article  PubMed  CAS  Google Scholar 

  • Shimizu K, Iino A, Nakajima J, Tanaka K, Nakajyo S, Urakawa N, Atsuchi M, Wada T, Yamashita C (1997) Suppression of glucose absorption by some fractions extracted from Gymnema sylvestre leaves. J Vet Med Sci 59:245–251

    Article  PubMed  CAS  Google Scholar 

  • Sinsheimer JE, Rao GS (1970) Constituents from Gymnema sylvestre leaves VI: acylated genins of the gymnemic acids-isolation and preliminary characterization. J Pharma Sci 59:629–632

    Article  CAS  Google Scholar 

  • Sinsheimer JE, Rao GS, McIlhenny HM (1970) Constituents from Gymnema sylvestre leaves V: isolation and preliminary characterization of the gymnemic acids. J Pharma Sci 59:622–628

    Article  CAS  Google Scholar 

  • Stöcklin W (1967) Gymnemagenin, vermutliche Struktur. Helvetica Chimica Acta 50:491–503

    Article  Google Scholar 

  • Subathra Devi C, Murugesh S, Mohana VS (2006) Gymnemic acid production in suspension cell cultures of Gymnema sylvestre. J App Sci 6(10):2263–2268

    Article  Google Scholar 

  • Sugihara Y, Nojima H, Matsuda H, Murakami T, Yoshikawa M, Kimura I (2000) Antihyperglycemic effects of gymnemic acid IV, a compound derived from Gymnema sylvestre leaves in streptozotocin-diabetic mice. J Asian Nat Prod Res 2:321–327

    Article  PubMed  CAS  Google Scholar 

  • Tsuda Y, Kiuchi F, Liu HM (1989) Establishment of the structure of gymnemagenin by x-ray analysis and the structure of deacylgymnemic acid. Tetrahedron Lett 30(3):361–362

    Article  CAS  Google Scholar 

  • Uniyal MR (1993) Some popular and traditional ayurvedic herbs useful in family planning. Sachiitra Ayurved 45:665–668

    Google Scholar 

  • Walker TS, Bais HP, Vivanco JM (2002) Jasmonic acid-induced hypericin production in cell suspension cultures of Hypericum perforatum L. (St. John’s Wort). Phytochemistry 60:289–293

    Article  PubMed  CAS  Google Scholar 

  • Walling LL (2000) The myriad plant responses to herbivores. J Plant Growth Regul 19:195–216

    PubMed  CAS  Google Scholar 

  • Wang LF, Luo H, Miyoshi M, Imoto T, Hiji Y, Sasaki T (1998) Inhibitory effect of gymnemic acid on intestinal absorption of oleic acid in rats. Can J Physiol Pharmacol 76:1017–1023

    Article  PubMed  CAS  Google Scholar 

  • Yazaki K, Takeda K, Tabata M (1997) Effects of Methyl Jasmonate on Shikonin and Dihydroechinofuran production in Lithospermum Cell Cultures. Plant Cell Physiol 38(7):776–782

    CAS  Google Scholar 

  • Ye WC, Zhang QW, Che CT, Zhao SX (2000) Oleanane saponins from Gymnema sylvestre. Phytochemistry 53:893–899

    Article  PubMed  CAS  Google Scholar 

  • Ye WC, Liu X, Zhao SX, Che CT (2001) Triterpenes from Gymnema sylvestre growing in China. Biochem Sys Eco 29:93–1195

    Google Scholar 

  • Yoon JH, Kim KH, Ma JC, Huh H (2000) Induced accumulation of triterpenoids in Scutellaria baicalensis suspension cultures using a yeast elicitor. Biotechnol Lett 22:1071–1075

    Article  CAS  Google Scholar 

  • Yoshikawa K, Arihara S, Matsuura K (1991) A new type of anti-sweet principles occurring in Gymnema sylvestre. Tetrahedron Lett 32:789–792

    Google Scholar 

  • Yoshikawa K, Arihara S, Matsuura K, Miyase T (1992) Dammarane saponins from Gymnema sylvestre. Phytochemistry 31: 237–241

    Google Scholar 

  • Yoshikawa K, Kondo Y, Arihara S, Matsuura K (1993) Anti-sweet natural products. IX structures of gymnemic acids XV-XVIII from Gymnema sylvestre R. Br. Chem Pharma Bull 40:1779–1782

    Google Scholar 

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

    Article  CAS  Google Scholar 

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Acknowledgments

We thank Dr. Manjappa and Dr. Suresh, Department of Chemistry, Bapuji Institute of Engineering and Technology, Davangere, for the help in HPLC analysis.

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Veerashree, V., Anuradha, C.M. & Kumar, V. Elicitor-enhanced production of gymnemic acid in cell suspension cultures of Gymnema sylvestre R. Br.. Plant Cell Tiss Organ Cult 108, 27–35 (2012). https://doi.org/10.1007/s11240-011-0008-6

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