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Enhanced morphinan alkaloid production in hairy root cultures of Papaver bracteatum by over-expression of salutaridinol 7-o-acetyltransferase gene via Agrobacterium rhizogenes mediated transformation

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Abstract

Papaver bracteatum is an important medicinal plant valued for its high content of thebaine and an alternative to P. somniferum for benzylisoquinoline alkaloid production. Salutaridinol 7-o-acetyltransferase (SalAT) is a key gene in morphinan alkaloids biosynthesis pathway. Over expression of SalAT gene was used for metabolic engineering in P. bracteatum hairy root cultures. Transcript level of the salutaridinol 7-o-acetyltransferase gene in transgenic hairy root lines increased up to 154 and 128 % in comparison with hairy roots without SalAT over expression and wild type roots, respectively. High performance liquid chromatography analysis showed that the transgenic hairy roots relatively improved levels of thebaine (1.28 % dry weight), codeine (0.02 % dry weight) and morphine (0.03 % dry weight) compared to those hairy roots without SalAT over expression. This suggests that P. bracteatum hairy roots expressing the SalAT gene could be potentially used for the production of valuable morphinan alkaloids.

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References

  • Allen RS, Millgate AG, Chitty JA, Thisleton J, Miller JA, Fist AJ, Gerlach WL, Larkin PJ (2004) RNAi-mediated replacement of morphine with the nonnarcotic alkaloid reticuline in opium poppy. Nat Biotechnol 22:1559–1566

    Article  CAS  Google Scholar 

  • Allen RS, Miller JAC, Chitty JA, Fist A, Gerlach WL, Larkin PJ (2008) Metabolite engineering of morphinan alkaloids by over expression and RNAi suppression of Salutaridinol 7-O-acetyltransferase in opium poppy. Plant Biotechnol J 6:22–30

    CAS  Google Scholar 

  • Azadi P, Chin DP, Kuroda K, Khan RS, Mii M (2010) Macro elements in inoculation and co-cultivation medium strongly affect the efficiency of Agrobacterium-mediated transformation in Lilium. Plant Cell Tiss Organ Cult 101:201–209

    Article  CAS  Google Scholar 

  • Cao D, Hou W, Liu W, Yao W, Wu C, Liu X, Han T (2011) Overexpression of TaNHX2 enhances salt tolerance of composite and whole transgenic soybean plants. Plant Cell Tiss Organ Cult 107:541–552

    Article  CAS  Google Scholar 

  • Carolan JC, Hook ILI, Walsh JJ, Hodkinson TR (2002) Using AFLP markers for species differentiation and assessment of genetic variability of in vitro cultured Papaver bracteatum (section oxytona). In Vitro Cell Dev Biol Plant 39:300–307

    Google Scholar 

  • Chattopadhyay T, Roy S, Mitra A, Maiti MK (2011) Development of a transgenic hairy root system in jute (Corchorus capsularis L.) with gusA reporter gene through Agrobacterium rhizogenes mediated co-transformation. Plant Cell Rep 30:485–493

    Article  CAS  Google Scholar 

  • Dehghan E, Hosseini B, Naghdi Badi H, Shariari Ahmadi F (2010) Application of conventional and new biotechnological approaches for improving of morphinane alkaloids production. J Med Plants 9:33–50

    CAS  Google Scholar 

  • Desgagne-Penix I, Farrow SC, Cram D, Nowak J, Facchini PJ (2012) Integration of deep transcript and targeted metabolite profiles for eight cultivars of opium poppy. Plant Mol Biol 79:295–313

    Article  CAS  Google Scholar 

  • Geng L, Niu L, Gresshoff PM, Shu C, Song F, Huang D, Zhang J (2012) Efficient production of Agrobacterium rhizogenes-transformed roots and composite plants in peanut (Arachis hypogaea L.). Plant Cell Tiss Organ Cult 109:491–500

    Article  CAS  Google Scholar 

  • Grothe T, Lenz R, Kutchan TM (2001) Molecular characterization of the salutaridinol 7-O-acetyltransferase involved in morphine biosynthesis in opium poppy (Papaver somniferum). J Biol Chem 276:30717–30723

    Article  CAS  Google Scholar 

  • Hosseini Khalifani B (2007) Genetic manipulation of Papaver somniferum for increasing morphinan alkaloids. PhD Thesis, Ferdowsi University of Mashhad, Iran

  • Kempe K, Higashi Y, Frick S, Sabarna K, Kutchan TM (2009) RNAi suppression of the morphinan biosynthetic gene SalAT and evidence of association of pathway enzymes. Phytochemistry 70:579–589

    Article  CAS  Google Scholar 

  • Larkin PJ, Miller JA, Allen RS, Chitty JA, Gerlach WL, Frick S, Kutchan TM, Fist AJ (2007) Increasing morphinan alkaloid production by over-expressing codeinone reductase in transgenic Papaver somniferum. Plant Biotechnol J 5:26–37

    Article  CAS  Google Scholar 

  • Le Flem-Bonhomme V, Laurain-Matter D, Fliniavx MA (2004) Hairy root induction of Papaver somniferum var. album, a difficult-to-transform plant, by A. rhizogenes LBA 9402. Planta 218:890–893

    Article  Google Scholar 

  • Lenz R, Zenk MH (1994) Closure of the oxide bridge in morphine biosynthesis. Tetrahedron Lett 35:3897–3900

    Article  CAS  Google Scholar 

  • Leonard E, Runguphan W, Connor SO, Prather KJ (2009) Opportunity in metabolic engineering to facilitate scalable alkaloid production. Nat Chem Biol 5:292–300

    Article  CAS  Google Scholar 

  • Liscombe DK, Facchini PJ (2008) Evolutionary and cellular webs in benzylisoquinoline alkaloid biosynthesis. Curr Opin Biotech 19:173–180

    Article  CAS  Google Scholar 

  • Ono NN, Tain L (2011) The multiplicity of hairy root cultures: prolific possibilities. Plant Sci 180:439–446

    Article  CAS  Google Scholar 

  • Ono NN, Bandaranayake PCG, Tain L (2012) Establishment of pomegranate (Punica granatum) hairy root cultures for genetic integration of hydrolysable tannin biosynthetic pathway. Planta 236:931–941

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Park SU, Yu M, Facchini PJ (2002) Antisense RNA-mediated suppression of benzophenanthridine alkaloid biosynthesis in transgenic cell cultures of California poppy. Plant Physiol 128:696–706

    Article  CAS  Google Scholar 

  • Pienky S, Brandt W, Schmidt J, Karmell R, Zigler J (2009) Functional characterization of involvement in papaverine biosynthesis in opium poppy (Papaver somniferum). Plant J 60:56–67

    Article  Google Scholar 

  • Sharafi A, Hashemi Sohi H, Mousavi A, Azadi P, Hosseini Khalifani B, Razavi K (2013a) Metabolic engineering of morphinan alkaloids by over expression of codeinone reductase in transgenic hairy root of Papaver bracteatum. Biotechnol Lett 35:445–453

    Article  CAS  Google Scholar 

  • Sharafi A, Hashemi Sohi H, Mousavi A, Azadi P, Razavi K, Ntui VO (2013b) A reliable and efficient protocol for inducing hairy roots in Papaver bracteatum. Plant Cell Tiss Organ Cult 113:1–9

    Article  CAS  Google Scholar 

  • Tisserat B, Berhow M (2009) Production of pharmaceutical from Papaver cultivars. In vitro Eng Life Sci 3:190–196

    Article  Google Scholar 

  • United Nations Document. Scientific Research on Papaver bracteatum. Report of the Fourth Working Group. ST/SOA/SER.J/23; 1976:6

  • Zhang HC, Liu JM, Lu HY, Gao SL (2009) Enhanced flavonoid production in hairy root cultures of Glycyrrhiza uralensis Fisch by combining the over-expression of chalcone isomerase gene with the elicitation treatment. Plant Cell Rep 28:1205–1213

    Article  CAS  Google Scholar 

  • Ziegler J, Voigtländer S, Schmidt J, Kramell R, Miersch O, Ammer C, Gesell A, Kutchan TM (2006) Comparative transcript and alkaloid profiling in Papaver species identifies a short chain dehydrogenase/reductase involved in morphine biosynthesis. Plant J 48:177–192

    Article  CAS  Google Scholar 

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Acknowledgments

The authors thank to Dr. Houshang Ghasemi and Mr. Saeed Dini (TEMAD Co.) for providing HPLC services.

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Authors and Affiliations

  1. National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran

    Ali Sharafi, Haleh Hashemi Sohi, Amir Mousavi & Bahareh Dehsara

  2. Tissue Culture and Gene Transformation Department, Agricultural Biotechnology Research Institute of Iran (ABRII), P. O. Box 31535-1897, Karaj, Iran

    Pejman Azadi

  3. Faculty of Horticulture, College of Agriculture and Natural Resources, Urmia University, Urmia, Iran

    Bahman Hosseini Khalifani

  4. Biotechnology Incubator of NIGEB, Novin Giti Gene R&D Co., Tehran, Iran

    Ali Sharafi

Authors
  1. Ali Sharafi
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  2. Haleh Hashemi Sohi
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  3. Amir Mousavi
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  4. Pejman Azadi
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  5. Bahareh Dehsara
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  6. Bahman Hosseini Khalifani
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Correspondence to Haleh Hashemi Sohi or Amir Mousavi.

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Sharafi, A., Hashemi Sohi, H., Mousavi, A. et al. Enhanced morphinan alkaloid production in hairy root cultures of Papaver bracteatum by over-expression of salutaridinol 7-o-acetyltransferase gene via Agrobacterium rhizogenes mediated transformation. World J Microbiol Biotechnol 29, 2125–2131 (2013). https://doi.org/10.1007/s11274-013-1377-2

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  • DOI: https://doi.org/10.1007/s11274-013-1377-2

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