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Elicitor enhanced production of protoberberine alkaloids from in vitro cell suspension cultures of Tinospora cordifolia (Willd.) Miers ex Hook. F. & Thoms

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Abstract

The present research investigates the effect of Piriformospora indica, an endophytic fungus, on production of protoberberine alkaloids in in vitro cell suspension cultures of Tinospora cordifolia. Although T. cordifolia produces a number of protoberberine alkaloids, the simultaneous production of jatrorrhizine and palmatine in cell suspension cultures of T. cordifolia was observed for the first time with the use of P. indica as biotic elicitor. The cells in suspension cultures were elicitated with P. indica on 14th day of culture initiation and the production of the alkaloids on 16th day was monitored. The autoclaved as well as filter sterilized cultures of P. indica were used in addition to the use of fungal cell extract. The elicitor effect of P. indica was analyzed and compared with other abiotic elicitor (methyl jasmonate) and biotic elicitors (chitin and chitosan). The culture filtrate of P. indica in the filter sterilized (5.0% v/v) form gave better response with enhanced 4.2-fold production of jatrorrhizine (10.72 mg/g DW) and 4.0-fold production of palmatine (4.39 mg/g DW). The production of these compounds was at par with that achieved in methyl jasmonate (at 250 µM) treated cell suspension cultures.

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References

  • Adya AK, Gautam A, Zhang L, Varma A (2013) Characterization of Piriformospora indica culture filtrate. In: Varma A, Kost G, Oelmüller R (eds) Piriformospora indica, soil biology, vol 33. Springer, Berlin, pp 345–375

    Chapter  Google Scholar 

  • Ahmed AB, Kim SK (2011) Chitin, chitosan derivatives induce the production of secondary metabolites and plant development through in vitro and in vivo techniques. In: Kim SK (ed) Chitin, chitosan, oligosaccharides and their derivatives: biological activities and applications. CRC Press, Florida, pp 589–603

    Google Scholar 

  • Bagde US, Prasad R, Varma A (2013) Impact of culture filtrate of Piriformospora indica on biomass and biosynthesis of active ingredient aristolochic acid in Aristolochia elegans Mart. Int J Biol 6:29–37

    Article  Google Scholar 

  • Bajaj R, Agarwal A, Rajpal K, Asthana S, Kumar R, Prasad R, Kharkwal AC, Sherameti I, Oelmüller R, Varma A (2014) Co-cultivation of Curcuma longa with Piriformospora indica enhances the yield and active ingredients. Am J Curr Microbiol 2:6–17

    Google Scholar 

  • Baldi A, Farkya S, Jain A, Gupta N, Mehra R, Datta V, Srivastava AK, Bisaria VS (2010) Enhanced production of podophyllotoxins by co-culture of transformed Linum album cells with plant growth-promoting fungi. Pure Appl Chem 82:227–241

    Article  CAS  Google Scholar 

  • Breuling M, Alfermann A, Reinhard E (1985) Cultivation of cell cultures of Berberis wilsonae in 20-L airlift bioreactors. Plant Cell Rep 4:220–223

    Article  CAS  PubMed  Google Scholar 

  • Cai Z, Kastell A, Mewis I, Knorr D, Smetanska I (2012) Polysaccharide elicitors enhance anthocyanin and phenolic acid accumulation in cell suspension cultures of Vitis vinifera. Plant Cell Tissue Organ Cult 108:401–409

    Article  CAS  Google Scholar 

  • Chintalwar G, Gupta S, Roja G, Bapat V (2003) Protoberberine alkaloids from callus and cell suspension cultures of Tinospora cordifolia. Pharm Biol 41:81–86

    Article  CAS  Google Scholar 

  • Cho HY, Son SY, Rhee HS, Yoon SY, Lee-Parsons CW, 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  CAS  PubMed  Google Scholar 

  • Franken P (2012) The plant strengthening root endophyte Piriformospora indica: potential application and the biology behind. Appl Microbiol Biotechnol 96:1455–1464

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Godoy-Hernández G, Vázquez-Flota FA (2012) Growth measurements: estimation of cell division and cell expansion. In: Loyola-Vargas VM, Ochoa-Alejo N (eds) Plant cell culture protocols, methods in molecular biology, vol 877. Humana Press, New York, pp 41–48

    Chapter  Google Scholar 

  • Hara M, Kitamura T, Fukui H, Tabata M (1993) Induction of berberine biosynthesis by cytokinins in Thalictrum minus cell suspension cultures. Plant Cell Rep 12:70–73

    Article  CAS  PubMed  Google Scholar 

  • Käfer E (1977) Meiotic and mitotic recombination in Aspergillus and its chromosomal aberrations. Adv Genet 19:33–131

    PubMed  Google Scholar 

  • Kumar V, Sahai V, Bisaria VS (2013) Effect of Piriformospora indica on enhanced biosynthesis of anticancer drug, podophyllotoxin, in plant cell cultures of Linum album. In: Varma A, Kost G, Oelmüller R (eds) Piriformospora indica, soil biology, vol 33. Springer, Berlin, pp 119–137

    Chapter  Google Scholar 

  • Kumar P, Chaturvedi R, Sundar D, Bisaria VS (2016) Piriformospora indica enhances the production of pentacyclic triterpenoids in Lantana camara L. suspension cultures. Plant Cell Tissue Organ Cult 125:23–29

    Article  CAS  Google Scholar 

  • Nair AJ, Sudhakaran PR, Rao JM, Ramakrishna SV (1992) Berberine synthesis by callus and cell suspension cultures of Coscinium fenestratum. Plant Cell Tissue Organ Cult 29:7–10

    Article  CAS  Google Scholar 

  • Patel MB, Mishra S (2011) Hypoglycemic activity of alkaloidal fraction of Tinospora cordifolia. Phytomedicine 18:1045–1052

    Article  CAS  PubMed  Google Scholar 

  • Patil D, Gautam M, Mishra S, Kulkarni P, Suresh K, Gairola S, Jadhav S, Patwardhan B (2010) Quantitative determination of protoberberinealkaloids in Tinospora cordifolia by RP-LC-DAD. Chromatographia 71:341–345

    Article  CAS  Google Scholar 

  • Philogene BJ, Arnason JT, Towers GH, Abramowski Z, Campos F, Champagne D, McLachlan D (1984) Berberine: a naturally occurring phototoxic alkaloid. J Chem Ecol 10:115–123

    Article  CAS  PubMed  Google Scholar 

  • Rai M, Varma A, Pandey A (2004) Antifungal potential of Spilanthes calva after inoculation of Piriformospora indica. Mycoses 47:479–481

    Article  CAS  PubMed  Google Scholar 

  • Rao BR, Kumar DV, Amrutha RN, Jalaja N, Vaidyanath K, Rao AM, Rao S, Polavarapu R, Kishor P (2008) Effect of growth regulators, carbon source and cell aggregate size on Berberine production from cell cultures of Tinospora cordifolia Miers. Curr Trends Biotechnol Pharm 2:269–276

    CAS  Google Scholar 

  • Schäfer P, Pfiffi S, Voll LM, Zajic D, Chandler PM, Waller F, Scholz U, Pons-Kühnemann J, Sonnewald S, Sonnewald U, Kogel KH (2009) Manipulation of plant innate immunity and gibberellin as factor of compatibility in the mutualistic association of barley roots with Piriformospora indica. Plant J 59:461–474

    Article  PubMed  Google Scholar 

  • Sharma G, Agrawal V (2013) Marked enhancement in the artemisinin content and biomass productivity in Artemisia annua L. shoots co-cultivated with Piriformospora indica. World J Microbiol Biotechnol 29:1133–1138

    Article  CAS  PubMed  Google Scholar 

  • Shields VD, Smith KP, Arnold NS, Gordon IM, Shaw TE, Waranch D (2008) The effect of varying alkaloid concentrations on the feeding behavior of gypsy moth larvae, Lymantria dispar (L.) (Lepidoptera: Lymantriidae). Arthropod-plant Interact 2:101–107

    Article  PubMed  PubMed Central  Google Scholar 

  • Singh S, Pandey S, Srivastava S, Gupta V, Patro B, Ghosh A (2003) Chemistry and medicinal properties of Tinospora cordifolia (Guduchi). Indian J Pharmacol 35:83–91

    CAS  Google Scholar 

  • Singh A, Rajpal K, Singh M, Kharkwal AC, Arora M, Varma A (2013) Mass cultivation of Piriformospora indica and Sebacina species. In: Varma A, Kost G, Oelmüller R (eds) Piriformospora indica, soil biology, vol 33. Springer, Heidelberg, pp 377–392

    Chapter  Google Scholar 

  • Sirrenberg A, Göbel C, Grond S, Czempinski N, Ratzinger A, Karlovsky P, Santos P, Feussner I, Pawlowski K (2007) Piriformospora indica affects plant growth by auxin production. Plant Physiol 131:581–589

    Article  CAS  Google Scholar 

  • van der Plas LH, Eijkelboom C, Hagendoorn MJ (1995) Relation between primary and secondary metabolism in plant cell suspensions. Plant Cell Tissue Organ Cult 43:111–116

    Article  Google Scholar 

  • Varma A, Verma S, Sahay N, Bütehorn B, Franken P (1999) Piriformospora indica, a cultivable plant-growth-promoting root endophyte. Appl Environ Microbiol 65:2741–2744

    CAS  PubMed  PubMed Central  Google Scholar 

  • Vennerstrom JL, Klayman DL (1988) Protoberberine alkaloids as antimalarials. J Med Chem 31:1084–1087

    Article  CAS  PubMed  Google Scholar 

  • Wang X, Yao X, Zhu ZA, Tang T, Dai K, Sadovskaya I, Flahaut S, Jabbouri S (2009) Effect of berberine on Staphylococcus epidermidis biofilm formation. Int J Antimicrob Agents 34:60–66

    Article  PubMed  Google Scholar 

  • Wiktorowska E, Długosz M, Janiszowska W (2010) Significant enhancement of oleanolic acid accumulation by biotic elicitors in cell suspension cultures of Calendula officinalis L. Enzyme Microb Technol 46:14–20

    Article  CAS  Google Scholar 

  • Yu HH, Kim KJ, Cha JD, Kim HK, Lee YE, Choi NY, You YO (2005) Antimicrobial activity of berberine alone and in combination with ampicillin or oxacillin against methicillin-resistant Staphylococcus aureus. J Med Food 8:454–461

    Article  CAS  PubMed  Google Scholar 

  • Zhao J, Zhu W-H, Hu Q (2001a) Enhanced catharanthine production in Catharanthus roseus cell cultures by combined elicitor treatment in shake flasks and bioreactors. Enzyme Microb Technol 28:673–681

    Article  CAS  PubMed  Google Scholar 

  • Zhao J, Zhu W-H, Hu Q (2001b) Selection of fungal elicitors to increase indole alkaloid accumulation in Catharanthus roseus suspension cell culture. Enzyme Microb Technol 28:666–672

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

The authors are thankful to Mr Sumit Singh, Department of Chemical Engineering, Indian Institute of Technology Delhi, New Delhi for assisting in MS data analysis and to Department of Biotechnology, Ministry of Science and Technology, Govt. of India for partially supporting the work.

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Correspondence to V. S. Bisaria.

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Communicated by Wagner Campos Otoni.

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Kumar, P., Srivastava, V., Chaturvedi, R. et al. Elicitor enhanced production of protoberberine alkaloids from in vitro cell suspension cultures of Tinospora cordifolia (Willd.) Miers ex Hook. F. & Thoms. Plant Cell Tiss Organ Cult 130, 417–426 (2017). https://doi.org/10.1007/s11240-017-1237-0

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