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Analysis of 6-methoxy podophyllotoxin and podophyllotoxin in hairy root cultures of Linum album Kotschy ex Boiss

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Abstract

Linum album is a herbaceous medicinal plant that contains some lignans with antiviral and anticancer properties such as podophyllotoxin (PTOX) and 6-methoxy podophyllotoxin (MPTOX). In this research, hairy root cultures of L. album were established by transformation with Agrobacterium rhizogenes strains LBA9402, A4, AR15834 and Agrobacterium tumefaciens strain C58C1 (pRiA4). The presence of PTOX and MPTOX in the hairy roots was verified by ESI/MS in positive ion mode. MPTOX was confirmed and its enantiomer determined by nuclear magnetic resonance spectroscopy and circular dichroism spectroscopy, respectively. PTOX and MPTOX production was determined by HPLC, in different lines of hairy roots. The results showed that all obtained hairy root lines produced higher yield of lignans than mother plant roots. In addition, the lignan content in the roots derived from A. rhizogenes strain LBA9402 was higher than in those obtained from A. tumefaciens strain C58C1.

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Abbreviations

Ags :

Agropine synthase

CD:

Circular dichroism

LC–ESI–MS/MS:

Liquid chromatography–electrospray–mass spectrometry/mass spectrometry

MPTOX:

6-methoxy podophyllotoxin

NMR:

Nuclear magnetic resonance

PTOX:

Podophyllotoxin

References

  • Bais HP, Sudha G, George J, Ravishankar GA (2001) Influence of exogenous hormones on growth and secondary metabolite production in hairy root cultures of Cichorium intybus L. Cv. Lucknow local. In Vitro Cell Dev Biol Plant 37:293–299

    CAS  Google Scholar 

  • Botta B, Delle Monache G, Misiti D, Vitali A, Zappia G (2001) Aryltetralin lignans: chemistry, pharmacology and biotransformation. Curr Med Chem 8:1363–1381

    Article  PubMed  CAS  Google Scholar 

  • Chattopadhyay S, Srivastava AK, Bisaria VS (2002) Optimization of culture parameters for production of podophyllotoxin in suspension culture of Podophyllum hexandrum. Appl Biochem Biotechnol 102:381–393

    Article  PubMed  Google Scholar 

  • Chattopadhyay S, Bisaria VS, Srivastava AK (2003a) Enhanced production of podophyllotoxin by Podophyllum hexandrum using in situ cell retention bioreactor. Biotechnol Prog 19:1026–1028

    Article  PubMed  CAS  Google Scholar 

  • Chattopadhyay S, Bisaria VS, Bhojwani SS, Srivastava AK (2003b) Enhanced production of podophyllotoxin by fed-batch cultivation of Podophyllum hexandrum. Can J Chem Eng 81:1–8

    Article  Google Scholar 

  • Chattopadhyay S, Farkya S, Srivastava AK, Bisaria VS (2003c) Bioprocess considerations for production of secondary metabolites by plant cell suspension cultures. Biotechnol Bioprocess Eng 7:138–149

    Article  Google Scholar 

  • Dewik PM (2002) Medicinal natural products, a biosynthetic approach. Wiley, Chichester, pp 121–136

    Google Scholar 

  • Eberhardt TL, Bernards MA, He L, Davin LB, Wooten JB, Lewis NG (1993) Lignification in cell suspension cultures of Pinus taeda: in situ characterization of a Gymnosperm lignin. J Biol Chem 268:21088–21096

    PubMed  CAS  Google Scholar 

  • Farkya S, Bisaria VS (2008) Exogenous hormones affecting morphology and biosynthetic potential of hairy root line (LYR2i) of Linum album. J Biosci Bioeng 105:140–146

    Article  PubMed  CAS  Google Scholar 

  • Farkya S, Bisaria VS, Srivastava AK (2004) Biotechnological aspects of the production of the anticancer drug podophyllotoxin. Appl Microbiol Biotechnol 65:504–519

    Article  PubMed  CAS  Google Scholar 

  • Giri A, Narasu ML (2000) Transgenic hairy roots: recent trends and applications. Biotechnol Adv 18:1–22

    Article  PubMed  CAS  Google Scholar 

  • Giri A, Giri CC, Dhingra V, Narasu ML (2001) Enhanced podophyllotoxin production from Agrobacterium rhizogenes transformed cultures of Podophyllum hexandrum. Nat Prod Lett 15:229–235

    Article  PubMed  CAS  Google Scholar 

  • Gordaliza M, Garc PA, del Corral M, Castro MA, Gomez-Zurita MA (2004) Podophyllotoxin: distribution, sources, applications and new cytotoxic derivatives. Toxicon 44:441–459

    Article  PubMed  CAS  Google Scholar 

  • Khan S, Irfan Qureshi M, Kamaluddin Alam T, Abdin MZ (2007) Protocol for isolation of genomic DNA from dry and fresh roots of medicinal plants suitable for RAPD and restriction digestion. Afr J Biotechnol 6:175–178

    CAS  Google Scholar 

  • Kuhlmann S, Kranz K, Lücking B, Alfermann AW, Petersen M (2002) Aspects of cytotoxic lignan biosynthesis in suspension cultures of Linum nodiflorum. Phytochem Rev 1:37–43

    Article  CAS  Google Scholar 

  • Li Q, Yan G, Ge T (2007) A fragmentation study of podophyllotoxin and its 40-demethyl-4b-substituted derivatives by electrospray ionization ion-trap time-of-flight tandem mass spectrometry. Rapid Commun Mass Spectrom 21:2843–2852

    Article  PubMed  CAS  Google Scholar 

  • Lin H, Kwok KH, Doran PM (2003) Development of Linum flavum hairy root cultures for production of Coniferin. Biotechnol Lett 25:521–525

    Article  PubMed  CAS  Google Scholar 

  • Lin MC, Lin JH, Chen SK, Cheng YW, Cheng HW (2008) Simultaneous determination of podophyllotoxin, quercetin and kaempferol in podophyllin by liquid chromatography tandem mass Spectrometry. J Food Drug Anal 16:29–40

    Google Scholar 

  • Mishra J, Bhandari H, Singh M, Rawat S, Agnihotri RK, Mishra S, Purohit S (2011) Hairy root culture of Picrorhiza kurroa Royle ex Benth.: a promising approach for the production of picrotin and picrotoxinin. Acta Physiol Plant 33(5):1841–1846

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Nikolay V, Ionkova I (2005) Lignan production by cell cultures of Linum setaceum and Linum campanulatum. Pharm Biol 43:509–511

    Article  Google Scholar 

  • Park S, Facchini P (2000) Agrobacterium rhizogenes-mediated transformation of opium poppy, Papaver somniferum L., and California poppy, Escholizia californica cham., root cultures. J Exp Bot 51:1005–1016

    Article  PubMed  CAS  Google Scholar 

  • Ramachandra RS, Ravishankar GA (2002) Plant cell cultures: chemical factories of secondary metabolites. Biotechnol Adv 20:101–153

    Article  Google Scholar 

  • Smollny T, Wichers H, Kalenberg S, Shahsavari A, Petersen M, Alfermann WA (1998) Accumulation of podophyllotoxin and related lignans in cell suspension cultures of Linum album. Phytochemistry 48:975–979

    Article  CAS  Google Scholar 

  • Van Uden W, Pras N, Visser JF, Malingre TM (1989) Detection and identification of podophyllotoxin produced by cell cultures derived from Podophyllum hexandrum Royle. Plant Cell Rep 8:165–168

    Article  Google Scholar 

  • Verma PC, Rahman L, Negi AS, Jain DC, Khanuja SPS, Banerjee S (2007) Agrobacterium rhizogenes mediated transformation of Picrorhiza kurroa Royle ex Benth.: establishment and selection of superior hairy root clone. Plant Biotechnol Rep 1:169–174

    Article  Google Scholar 

  • Wang B, Zhang G, Zhu L, Chen L, Zhang Y (2006) Genetic transformation of Echinacea purpurea with Agrobacterium rhizogenes and bioactive ingredient analysis in transformed cultures. Colloid Surface B 53:101–104

    Article  CAS  Google Scholar 

  • Yousefzadi M, Sharifi M, Behmanesh M, Ghasempour A, Moyano E, Palazon J (2010a) Salicylic acid improves podophyllotoxin production in cell cultures of Linum album by increasing the expression of genes related with its biosynthesis. Biotechnol Lett 32:1739–1743

    Article  PubMed  CAS  Google Scholar 

  • Yousefzadi M, Sharifi M, Ahmadian Chashmi N, Behmanesh M, Ghasempour A (2010b) Optimization of podophyllotoxin extraction method from Linum album cell cultures. Pharm Biol 48:1421–1425

    Article  PubMed  CAS  Google Scholar 

  • Zhong JJ (2002) Plant cell culture for production of paclitaxel and other taxanes. J Biosci Bioeng 94:591–599

    PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Plant Biology, Faculty of Biological Sciences, Tarbiat Modares University, P.O. Box: 14115-154, Tehran, Iran

    Najmeh Ahmadian Chashmi & Mozafar Sharifi

  2. Department of Biology, Faculty of Sciences, Hormozgan University, Bandar Abbas, Iran

    Morteza Yousefzadi

  3. Department of Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran

    Mehrdad Behmanesh

  4. Department of Phytochemistry, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, Tehran, Iran

    Hassan Rezadoost

  5. Catedra de Microbiología Industrial y Biotecnología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina

    Alejandra Cardillo

  6. Laboratori de Fisiologia Vegetal, Facultat de Farmacia, Universitat de Barcelona, Barcelona, Spain

    Javier Palazon

Authors
  1. Najmeh Ahmadian Chashmi
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  2. Mozafar Sharifi
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  3. Morteza Yousefzadi
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  4. Mehrdad Behmanesh
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  5. Hassan Rezadoost
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  6. Alejandra Cardillo
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  7. Javier Palazon
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Correspondence to Mozafar Sharifi.

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Chashmi, N.A., Sharifi, M., Yousefzadi, M. et al. Analysis of 6-methoxy podophyllotoxin and podophyllotoxin in hairy root cultures of Linum album Kotschy ex Boiss. Med Chem Res 22, 745–752 (2013). https://doi.org/10.1007/s00044-012-0067-1

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  • DOI: https://doi.org/10.1007/s00044-012-0067-1

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