Plant Cell Reports

, Volume 11, Issue 3, pp 132–136 | Cite as

Transformation of opium poppy (Papaver somniferum L.) with Agrobacterium rhizogenes MAFF 03-01724

  • Kayo Yoshimatsu
  • Koichiro Shimomura


Transformed cultures of opium poppy (Papaver somniferum L.) were established by infecting hypocotyl segments with Agrobacterium rhizogenes MAFF 03-01724. Undifferentiated calli formed on the infected site grew satisfactorily on phytohormone-free solid medium in the dark and produced opine, mikimopine, which could not be detected in a normal culture. Numerous adventitious shoots developed from transformed calli during subculture. The transformed shoots separated individually were cultured on phytohormone-free MS solid medium at 22 ° C under 14 h/day light. They displayed wider leaves and longer internodes than shoots established from seeds or non-transformed root culture. The content of morphinan alkaloids in the cultures and regenerated shoots were quantitatively analyzed by enzyme-linked immunosorbent assay and high performance liquid chromatography. HPLC analysis revealed that non-transformed shoots contained much more codeine (1310 gmg/g dry wt.) than morphine (50 μg/g dry wt.), while the transformed shoot cultures did not contain morphine, although the level of morphinan alkaloids in the transformed shoots (213 μg morphine equivalents/g fr. wt.) was comparable to that in non-transformed shoots (182 μg morphine equivalents/g fr. wt.) by ELISA.

Key words

Transformation Palaver somniferum Agrobacterium rhizogenes Morphinan alkaloids ELISA HPLC 



Murashige-Skoog (Murashige and Skoog 1962)

1/2 MS

half strength MS




1-naphthaleneacetic acid


enzyme-linked immunosorbent assay


high performance liquid chromatography


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Furuya T, Ikuta A and Syôno K (1972) Phytochem. 11: 3041–3044Google Scholar
  2. Ikeda K, Teshima D, Aoyama T, Satake M and Shimomura K (1988) Plant cell Rep. 7: 288–291Google Scholar
  3. Ikuta A, Syôno K and Furuya T (1974) Phytochem. 13: 2175–2179Google Scholar
  4. Isogai A, Fukuchi N, Hayashi M, Kamada H, Harada H and Suzuki A (1988) Agric. Biol. Chem. 52: 3235–3237Google Scholar
  5. Isogai A, Fukuchi N, Hayashi M, Kamada H, Harada H and Suzuki A (1990) Phytochem. 29: 3131–3134Google Scholar
  6. Kamo KK, Kimoto W, Hsu A-F, Mahlberg PG and Bills DD (1982) Phytochem. 21: 219–222Google Scholar
  7. Mano Y, Nabeshima S., Matsui C and Ohkawa H (1986) Agric. Biol. Chem. 50: 2715–2722Google Scholar
  8. Murashige T and Skoog F (1962) Physiol. Plant. 15: 473–497Google Scholar
  9. Nessler CL (1990) In: Handbook of Plant Cell Culture vol. 5, Poppy, Ammirato PV, Evans DR, Sharp WR and Bajaj YPS (ed), pp 693–715, McGraw-Hill Publishing Co., New YorkGoogle Scholar
  10. Petit A, David C Dahl GA, Ellis JG, Guyon P, Casse-Delbart F and Tempe J (1983) Mol. Gen. Gent. 190: 204–214Google Scholar
  11. Roberts MF (1988) In: Cell Culture and Somatic Cell Genetics of Plants vol. 5, Isoquinolines (Papaver Alkaloids), Constabel F and Vasil IK (ed), pp 315–334, Academic Press Inc., San DiegoGoogle Scholar
  12. Sawada J, Janejai N, Nagamatsu K and Terao T (1988) Molecular Immunology 9: 937–943Google Scholar
  13. Schuchmann R and Wellmann E (1983) Plant Cell Rep. 2: 88–91Google Scholar
  14. Songstad DD, Giles KL, Park J, Novakovski D, Epp D, Friesen L and Roewer I (1989) Plant Cell Rep. 8: 463–466Google Scholar
  15. Staba EJ, Zito S and Amin M (1982) J. Nat. Prod. 45: 256–262Google Scholar
  16. Trease GE and Evans WC (1989) In: Pharmacognosy 13th edition, p 582–591, Baillière Tindall, LondonGoogle Scholar
  17. Tyler RT, Eilert U, Rijnders COM, Roewer IA and Kurz WGW (1988) Plant Cell Rep. 7: 410–413Google Scholar
  18. Yoshikawa T. and Furuya T. (1985) Planta Med. 110–113Google Scholar
  19. Yoshimatsu K, Satake M, Shimomura K, Sawada J and Terao T (1990) J. Nat. Prod. 53: 1498–1502Google Scholar

Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • Kayo Yoshimatsu
    • 1
  • Koichiro Shimomura
    • 1
  1. 1.Tsukuba Medicinal Plant Research Station, National Institute of Hygienic SciencesIbarakiJapan

Personalised recommendations