Transformation of opium poppy (Papaver somniferum L.) with Agrobacterium rhizogenes (MAFF 03-01724)
1992
Yoshimatsu, K. | Shimomura, K.
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 degrees 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 micrograms/g dry wt.) than morphine (50 micrograms/g dry wt.), while the transformed shoot cultures did not contain morphine, although the level of morphinan alkaloids in the transformed shoots (213 micrograms morphine equivalents/g fr. wt.) was comparable to that in non-transformed shoots (182 micrograms morphine equivalents/g fr. wt.) by ELISA.
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