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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
Article

Summary

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 

Abbreviations

MS

Murashige-Skoog (Murashige and Skoog 1962)

1/2 MS

half strength MS

HF

phytohormone-free

NAA

1-naphthaleneacetic acid

ELISA

enzyme-linked immunosorbent assay

HPLC

high performance liquid chromatography

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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

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