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Hairy root induction of Papaver somniferum var. album, a difficult-to-transform plant, by A. rhizogenes LBA 9402

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Abstract

Two strains of Agrobacterium rhizogenes (15834, LBA 9402) and one Agrobacterium tumefaciens strain [GV 3101 (PMP90RK, p35SGUS-2)] and four culture media were tested and compared for their ability to induce hairy root formation on wounded Papaver somniferum L. hypocotyls. Five weeks after the infection with A. rhizogenes LBA 9402, hairy roots appeared on 80% of the hypocotyls maintained in the hormone-free liquid medium. Six hairy-root cultures were established. Transformation was confirmed by polymerase chain reaction analysis. One clone was analysed for its alkaloid production. The total alkaloid content was higher in the transformed roots (0.46±0.06% DW) than in the untransformed roots (0.32±0.05% DW). The transformed roots accumulated three times more codeine (0.18±0.02% DW) than intact roots (0.05±0% DW). Moreover, morphine (0.255±0.03% DW) and sanguinarine (0.014±0% DW) were found in the liquid culture medium.

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Abbreviations

2,4-D :

2,4-Dichlorophenoxyacetic acid

LS :

Linsmaier and Skoog

References

  • Arellano J, Vasquez F, Villegas T, Hernandez G (1996) Establishment of transformed root cultures of Perezia cuernavacana producing the sesquiterpene quinone perezone. Plant Cell Rep 15:455–458

    Article  CAS  Google Scholar 

  • Bonhomme V, Laurain-Mattar D, Lacoux J, Fliniaux MA, Jacquin-Dubreuil A (2000a) Tropane alkaloid production by hairy roots of Atropa belladonna obtained after transformation with Agrobacterium rhizogenes 15834 and Agrobacterium tumefaciens containing rol A, B, C genes only. J Biotechnol 81:151–158

    Article  CAS  Google Scholar 

  • Bonhomme V, Laurain-Mattar D, Fliniaux MA (2000b) Effects of the rol C gene on hairy roots: induction development and tropane alkaloid production by Atropa belladonna. J Nat Prod 63:1249–1252

    Article  CAS  PubMed  Google Scholar 

  • Caboni E, Lauri P, Tonelli M, Falasca G, Damiano C (1996) Root induction by Agrobacterium rhizogenes in walnut. Plant Sci 118:203–208

    Article  CAS  Google Scholar 

  • Cline SD, Coscia CJ (1988) Stimulation of sanguinarine production by combined fungal elicitation and hormonal deprivation in cell suspension cultures of Papaver bracteatum. Plant Physiol 86:161–165

    CAS  Google Scholar 

  • Eilert U, Kurz WGW, Constatel F (1985) Stimulation of sanguinarine accumulation in Papaver somniferum cell cultures by fungal elicitors. J Plant Physiol 119:65–76

    CAS  Google Scholar 

  • Hooykaas PJJ, Shilperoort RA (1992) Agrobacterium and plant genetic engineering. Plant Mol Biol 19:15–38

    CAS  PubMed  Google Scholar 

  • Jouhikainen K, Lindgren L, Jokelainen T, Hiltumen R, Teeri TH (1999) Enhancement of scopolamine production in Hyoscyamus muticus L. hairy root cultures by genetic engineering. Planta 208:545–551

    Article  CAS  Google Scholar 

  • Kamo KK, Kimoto W, Hsu AF, Mahlberg PG, Bills DD (1982) Morphinan alkaloids in cultured tissues and redifferentiated organs of Papaver somniferum. Phytochemistry 21:219–222

    Article  CAS  Google Scholar 

  • Kifle S, Shao M, Jung C, Cai D (1999) An improved transformation protocol for studying gene expression in hairy roots of sugar beet (Beta vulgaris L.). Plant Cell Rep 18:514–519

    CAS  Google Scholar 

  • Kutchan TM, Rush MD, Coscia CJ (1986) Subcellular localization of alkaloids and dopamine in different vacuolar compartments of Papaver bracteatum. Plant Physiol 81:161–166

    CAS  Google Scholar 

  • Laurain-Mattar D, Gillet-Manceau F, Buchon L, Nabha S, Fliniaux M-A, Jacquin-Dubreuil A (1999) Somatic embryogenesis and rhizogenesis of tissue cultures of two genotypes of Papaver somniferum: relationships to alkaloid production. Planta Med 65:167–170

    CAS  Google Scholar 

  • Linsmaier EM, Skoog F (1965) Organic growth factor requirement of tobacco tissue cultures. Physiol Plant 18:100–127

    CAS  Google Scholar 

  • Nessler CL, Allen RD, Galewsky S (1985) Identification and characterization of latex-specific proteins in opium poppy. Plant Physiol 79:499–504

    CAS  Google Scholar 

  • Palazon J, Cusido RM, Gonzalio J, Bonfill M, Morales C, Pinol T (1998) Relation between the amount of rol C gene product and indole alkaloid accumulation in Catharanthus roseus transformed root cultures. J Plant Physiol 153:712–718

    CAS  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • Roberts MF, Carthy DMc, Kutchan TM, Coscia CJ (1983) Localization of enzymes and alkaloidal metabolites in Papaver latex. Arch. Biochem Biophys 222:599–609

    CAS  PubMed  Google Scholar 

  • Schuchmann R, Wellmann E (1983) Somatic embryogenesis of tissue of Papaver somniferum and Papaver orientale and its relationship to alkaloid and lipid metabolism. Plant Cell Rep 2:88–91

    CAS  Google Scholar 

  • Tepfer D (1984) Transformation of several species of higher plants by Agrobacterium rhizogenes: sexual transmission of the transferred genotype and phenotype. Cell 37:959–967

    PubMed  Google Scholar 

  • Trick HN, Finer JJ (1997) SAAT: sonication-assisted Agrobacterium-mediated transformation. Transgenic Res 6:329–336

    CAS  Google Scholar 

  • Vanhala L, Hiltumen R, Oksman-Caldentey KM (1995) Virulence of different Agrobacterium strains on hairy root formation of Hyoscyamus muticus. Plant Cell Rep 14:236–240

    CAS  Google Scholar 

  • Vervliet G, Holsters M, Teuchy H, Van Montagu M, Schell J (1975) Characterisation of different plaqueforming and defective temperate phages in Agrobacterium strains. J Gen Virol 26:33–48

    CAS  PubMed  Google Scholar 

  • Yoshikawa T, Furuya T (1985) Morphinan alkaloid production by tissue cultures differentiated from cultured cells of Papaver somniferum. Planta Med 2:110–113

    Google Scholar 

  • Yoshimatsu K, Shimomura K (1992) Transformation of opium poppy (Papaver somniferum L.) with Agrobacterium rhizogenes MAFF 03-01724. Plant Cell Rep 11:132–136

    CAS  Google Scholar 

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Acknowledgements

We are grateful to Mr. Pierrel, Botanical Garden of Nancy and Mr. Tepfer, INRA Versailles for the generous gifts, respectively, of P. somniferum (Psa) seeds and A. rhizogenes LBA 9402. We thank Dr. E. Lainé (Laboratoire de Biotechnologie et Physiologie Végétales, Amiens, France) for his assistance with the molecular biology. The authors express their thanks to Prof. Annie Jacquin-Dubreuil for helpful discussions and comments on the manuscript. This research was supported by grant 97-5-11657 from the French Ministry for Education and Research (MENRT).

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  1. V. Le Flem-Bonhomme

    Present address: Immunopathologie et Infections, UPRES EA-2128, UFR de Médecine, CHU Côte de Nacre, 14032, Caen Cedex, France

Authors and Affiliations

  1. Laboratoire de Phytotechnologie, Faculté de Pharmacie, Université de Picardie Jules Verne, 1 rue des Louvels, 80037, Amiens Cedex 1, France

    V. Le Flem-Bonhomme, D. Laurain-Mattar & M. A. Fliniaux.

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  1. V. Le Flem-Bonhomme
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  2. D. Laurain-Mattar
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  3. M. A. Fliniaux.
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Correspondence to V. Le Flem-Bonhomme.

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Le Flem-Bonhomme, V., Laurain-Mattar, D. & Fliniaux., M.A. Hairy root induction of Papaver somniferum var. album, a difficult-to-transform plant, by A. rhizogenes LBA 9402. Planta 218, 890–893 (2004). https://doi.org/10.1007/s00425-003-1196-z

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