Skip to main content
SpringerLink
Log in

Characterization of transgenic plants derived from hairy roots ofHyoscyamus muticus

  • Published:
Plant Cell Reports Aims and scope Submit manuscript

Abstract

Mature plants were regenerated via protoplasts fromAgrobacterium rhizogenes-transformed root cultures ofHyoscyamus muticus L., and chemical analyses were performed on 34 individual plants. The regenerated plants showed strong phenotypic differences from clone to clone as well as from the control plants. Polymerase chain reaction studies revealed that the plants exhibiting the strongest phenotypic alterations contained therol (A, B and C) genes, whereas the plants with fewer alterations had lost them. The plants produced hyoscyamine, scopolamine and a range of different calystegins, and considerable somaclonal variation was observed. Alkaloid production in the plants transgenic for therol genes was clearly reduced. The pattern of calystegins was similar within all the regenerated plants lackingrol genes. Among the plants withrol genes, the calystegin B1 was not detectable. It seems clear that the presence ofrol genes is detrimental to the alkaloid accumulation in the transgenic plants in contrast to hairy root cultures.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

PCR :

Polymerase chain reaction

References

  • Altvorst AC van, Bino RJ, Dijk AJ van, Lamers AMJ, Lindhout WH, Mark F van der, Dons JJM (1992) Effects of the introduction ofAgrobacterium rhizogenes rol genes on tomato plant and flower development. Plant Sci 83:77–85

    Google Scholar 

  • Arroo RRJ, Develi A, Meijers H, Van de Westerlo E, Kemp AK, Croes AF, Wullems GJ (1995) Effects of exogenous auxin on root morphology and secondary metabolism inTagetes patula hairy root cultures. Physiol Plant 93:233–240

    Google Scholar 

  • Atkinson RG, Gardner RC (1991)Agrobacterium-mediated transformation of pepino and regeneration of transgenic plants. Plant Cell Rep 10:208–212

    Google Scholar 

  • Bilang J, Sturm A (1995) Cloning and characterization of a glutathione S-transferree that can be photolabeled with 5-azido-indole-3-acetic acid. Plant Physiol 109:253–260

    Google Scholar 

  • Burtin D, Martin-Tanguy J, Tepfer D (1991) Alpha-dl-difluorometh-ylornithine, a specific, irreversible inhibitor of putrescine biosynthesis, induces a phenotype in tobacco similar to that ascribed to the root-inducing, left-hand transferred DNA ofAgrobacterium rhizogenes. Plant Physiol 95:461–468

    Google Scholar 

  • Cardarelli M, Mariotti D, Pomponi M, Spanò L, Capone I, Constantino P (1987)Agrobacterium rhizogenes T-DNA genes capable of inducing hairy root phenotype. Mol Gen Genet 209:475–480

    Google Scholar 

  • Chilton M-D, Tepfer DA, Petit A, David C, Casse-Delbart F, Tempé J (1982)Agrobacterium rhizogenes inserts T-DNA into the genomes of the host plant root cells. Nature 295:432–434

    Google Scholar 

  • Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Mol Biol Rep 1: 19–21

    Google Scholar 

  • Dräger B (1995) Identification and quantification of calystegines, polyhydroxyl nortropane alkaloids. Phytochem Anal 6:31–37

    Google Scholar 

  • Dräger B, Funck C, Höhler A, Mrachatz G, Nahrstedt A, Portsteffen A, Schaal A, Schmidt R (1994) Calystegines as a new group of tropane alkaloids in Solanaceae. Plant Cell Tissue Organ Cult 38:235–240

    Google Scholar 

  • Fladung M (1990) Transformation of diploid and tetraploid potato clones with therol C genes ofAgrobacterium rhizogenes and characterization of transgenic plants. Plant Breed 104:295–304

    Google Scholar 

  • Goldmann A, Milat M-L, Ducrot P-H, Lallemand J-Y, Maille M, Lepingle A, Charpin I, Tepfer D (1990) Tropane derivatives fromCalystegia sepium. Phyotochemistry 29: 2125–2127

    Google Scholar 

  • Hamill JD, Rounsley S, Spencer A, Todd G, Rhodes MJC (1991) The use of the polymerase chain reaction in plant transformation studies. Plant Cell Rep 10:221–225

    Google Scholar 

  • Hänisch ten Cate CH, Loonen AEHM, Ottaviani MP, Ennik L, Eldik G van, Stiekema WJ (1990) Frequent spontaneous deletions of Ri T-DNA inAgrobacterium rhizogenes transformed potato roots and regenerated plants. Plant Mol Biol 14:735–741

    Google Scholar 

  • Kawata M, Harada S, Antonio B, Oono K (1992) Protoclonal variation of plant regeneration in rice. Plant Cell Tissue Organ Cult 28:1–10

    Google Scholar 

  • Larkin PJ, Scowcroft WR (1981) Somaclonal variation — a novel source of variability from cell cultures for plant improvement. Theor Appl Genet 60:197–214

    Google Scholar 

  • Lee HS, Kim SW, Lee K-W, Eriksson T, Liu JR (1995)Agrobacterium-mediated transformation of ginseng (Panax ginseng) and mitotic stability of the insertedβ-glucuronidase gene in regenerants from isolated protoplasts. Plant Cell Rep 14:545–549

    Google Scholar 

  • Martin-Tanguy J, Corbineau F, Burtin D, Ben-Hayyim G, Tepfer D (1993) Genetic transformation with a derivative ofrol C fromAgrobacterium rhizogenes and treatment with alpha-aminoisobutyric acid produce similar phenotypes and reduce ethylene production and the accumulation of water-insoluble polyamine-hydroxycinnamic acid conjugates in tobacco flowers. Plant Sci 93:63–76

    Google Scholar 

  • Molyneux RJ, Pan YT, Goldmann A, Tepfer DA, Elbein DA (1993) Calystegines: a novel class of alkaloid glycosidase inhibitors. Arch Biochem Biophys 304:81–88

    Google Scholar 

  • Oksman-Caldentey K-M, Hiltunen R (1996) Transgenic crops for improved pharmaceutical products. Field Crops Res 45:57–69

    Google Scholar 

  • Oksman-Caldentey K-M, Strauss A (1986) Somaclonal variation of scopolamine content in protoplast-derived cell culture clones ofHyoscyamus muticus. Planta Med 52:6–12

    Google Scholar 

  • Oksman-Caldentey K-M, Vuorela H, Strauss A, Hiltunen R (1987) Variation in the tropane alkaloid content ofHyoscyamus muticus plants and cell culture clones. Planta Med 53:349–354

    Google Scholar 

  • Oksman-Caldentey K-M, Kivelä O, Hiltunen R (1991) Spontaneous shoot organogenesis and plant regeneration from hairy root cultures ofHyoscyamus muticus. Plant Sci 78:129–136

    Google Scholar 

  • Saito K, Yamazaki M, Murakoshi I (1992) Transgenic medicinal plants:Agrobacterium-mediated foreign gene transfer and production of secondary metabolites. J Nat Prod 55:149–162

    Google Scholar 

  • Schmülling T, Schell J, Spéna A (1988) Single genes fromAgrobacterium rhizogenes influence plant development. EMBO J 7:2621–2629

    Google Scholar 

  • Sevón N, Hiltunen R, Oksman-Caldentey K-M (1992) Chitosan increases hyoscyamine content in hairy root cultures ofHyoscyamus muticus. Pharma Pharmacol Lett 2:96–99

    Google Scholar 

  • Sevón N, Oksman-Caldentey K-M, Hiltunen R (1995) Eficient plant regeneration from hairy root-derived protoplasts ofHyoscyamus muticus. Plant Cell Rep 14:738–742

    Google Scholar 

  • Shen WH, Petit A, Guern J, Tempo J (1988) Hairy roots are more sensitive to auxin than normal roots. Proc Natl Acad Sci USA 85:3417–3421

    Google Scholar 

  • Spéna A, Schmülling T, Koncz C, Schell J (1987) Independent and synergistic activity ofrolA, B and C loci in stimulating abnormal growth in plants. EMBO J 6:3891–3899

    Google Scholar 

  • Sree Ramulu K, Dijkhuis P, Hänisch ten Cate CH, De Groot B (1985) Patterns of DNA and chromosome variation during in vitro growth in various genotypes of potato. Plant Sci 41:69–78

    Google Scholar 

  • Tepfer D (1990) Genetic transformation usingAgrobacterium rhizogenes. Physiol Plant 79:140–146

    Google Scholar 

  • Trulson AJ, Simpson RB, Shahin EA (1986) Transformation of cucumber (Cucumis sativus L.) plants withAgrobacterium rhizogenes. Theor Appl Genet 73:11–15

    Google Scholar 

  • Yukimune Y, Hara Y, Yamada Y (1994) Tropane alkaloid production in root cultures ofDuboisia myoporoides obtained by repeated selection. Biosci Biotech Biochem 58:1443–1446

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Pharmacognosy, Department of Pharmacy, University of Helsinki, (Viikinkaari 5), PO Box 56, FIN-00014, Helsinki, Finland

    N. Sevón, R. Hiltunen & K. -M. Oksman-Caldentey

  2. Institut für Pharmazeutische Biologie der Martin-Luther-Universität, Hoher Weg 7, D-06108, Halle, Germany

    B. Dräger

Authors
  1. N. Sevón
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. B. Dräger
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. R. Hiltunen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. K. -M. Oksman-Caldentey
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to K. -M. Oksman-Caldentey.

Additional information

Communicated by W. Barz

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sevón, N., Dräger, B., Hiltunen, R. et al. Characterization of transgenic plants derived from hairy roots ofHyoscyamus muticus . Plant Cell Reports 16, 605–611 (1997). https://doi.org/10.1007/BF01275500

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01275500

Key words

Search

Navigation