Tropane Alkaloid Production and Shoot Regeneration in Hairy and Adventitious Root Cultures of Duboisia myoporoides-D. leichhardtii Hybrid(Pharmacognosy)

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概要

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• Co-culture conditions for Duboisia myoporoides-D. leichhardtii hybrid hairy root induction were investigated using leaf explants and Agrobacterium rhizogenes ATCC 15834. The bacteria density and duration of coculture greatly affected the induction rate ; the highest rate of 50% was obtained when the leaf explants were cocultured for 2 d with 106 bacteria. One hairy root clone that showed the fastest root growth was selected and used for comparison study with adventitious roots cultured with 0.5 mg/l indole-3-acetic acid (IAA). The hairy roots cultured in Murashige and Skoog (MS) liquid medium grew well and yielded much more tropane alkaloids (35 mg/l scopolamine and 17 mg/l hyoscyamine) than adventitious roots cultured in 0.5 mg/l IAA after 6 weeks of culture at 25 ℃ in the dark. The hairy and adventitious roots (2.5 cm) grown in liquid media were divided into 5 parts (each 0.5 cm) along the root axis. Distribution of scopolamine and IAA was then determined by enzyme-linked immunosorbent assay (ELISA). Inverse relationship between contents of scopolamine and IAA was observed in the hairy roots ; increase of scopolamine and decrease of IAA were proportional to the distance from the root meristem. In contrast, the contents of scopolamine and IAA were relatively constant in the adventitious roots. In shoot regeneration experiments, the hairy and adventitious root segments (1 cm) were placed onto 1/2 MS solid medium containing various concentrations of IAA and BA cultured at 25 ℃ under 16 h light. In adventitious roots, the shoots regenerated on media containing 6-benzyladenine (BA) (0.5 to 5 mg/l), and 100% regeneration was observed in medium with 0.1 mg/l IAA and 2 mg/l BA. On the other hand, shoot regeneration was only observed in 33% of hairy roots cultured on medium containing 5 mg/l BA.

• 社団法人日本薬学会の論文
• 2004-08-01

著者

• Sawada Jun-ichi

  Department Of Immunobiology Faculty Of Applied Biological Science Hiroshima University

• Sawada Jun-ichi

  Division Of Functional Biochemistry And Genomics National Institute Of Health Sciences

• SHIMOMURA Koichiro

  Tsukuba Medicinal Plant Research Station, National Institute of Health Sciences

• YOSHIMATSU KAYO

  Tsukuba Medicinal Plant Research Station, National Institute of Health Sciences

• KAMADA Hiroshi

  Institute of Biological Sciences, University of Tsukuba

• Sawada Jun-ichi

  Division Of Biochemistry And Immunochemistry National Institute Of Health Sciences

• Sawada Jun-ichi

  Division Of Biochemistry And Immunochemistry Natinal Institute Of Health Sciences

• SUDO Hiroshi

  Institute of Biological Sciences, University of Tsukuba

• KIUCHI Fumiyuki

  Tsukuba Medicinal Plant Research Station, National Institute of Health Sciences

• KIKUCHI Yutaka

  Division of Biochemistry and Immunochemistry, National Institute of Health Sciences

• KIUCHI Fumiyuki

  Research Center for Medicinal Plant Resources National Institute of Biomedical Innovation

• Kiuchi F

  Graduate School Of Pharmaceutical Sciences Kyoto University

• Kiuchi Fumiyuki

  国立医薬品衛生研究所

• Kikuchi F

  Faculty Of Pharmaceutical Sciences Kanazawa University:(present Address)graduate School Of Pharmaceu

• Kikuchi F

  Faculty Of Pharmaceutical Sciences Kanazawa University

• Kiuchi Fumiyuki

  National Inst. Biomedical Innovation Ibaraki Jpn

• SHIMOMURA Koichiro

  Research Center for Medicinal Plant Resources, National Institute of Biomedical Innovation

• Kiuchi Fumiyuki

  Graduate School Of Pharmaceutical Sciences Kyoto University

• Kiuchi Fumiyuki

  Tsukuba Medicinal Plant Research Station National Institute Of Health Sciences

• Shimomura Koichiro

  Tsukuba Medicinal Plant Research Station National Institute Of Health Sciences

• Shimomura Koichiro

  Tsukuba Medicinal Plant Research Station National Institute Of Health Sciences:faculty Of Life Scien

• Kamada Hiroshi

  Institute Of Biological Sciences Univ.

• Sudo H

  Department Of Molecular Biology And Biotechnology Graduate School Of Pharmaceutical Sciences Chiba U

• Yoshimatsu K

  National Inst. Biomedical Innovation Ibaraki Jpn

• Yoshimatsu K

  National Inst. Health Sci. Ibaraki Jpn

• Yoshimatsu Kayo

  Tsukuba Division Research Center For Medicinal Plant Resources National Instituite Of Biomedical Inn

• Kiuchi Fumiyuki

  Faculty Of Pharmaceutical Sciences Kanazawa University:(present Address)graduate School Of Pharmaceu

• Kikuchi Yutaka

  Division Of Biochemistry And Immunochemistry National Institute Of Health Sciences:division Of Micro

• Shimomura Koichiro

  Tsukuba Medicinal Plant Research Station National Institute Of Health Science

• Kamada Hiroshi

  Univ. Tsukuba

• Kamada Hiroshi

  Institute Of Biological Sciences Tsukuba University

• Kiuchi Fumiyuki

  Tsukuba Div. Res. Center For Medicinal Plant Resources National Inst. Of Biomedical Innovation

• Yoshimatsu Kayo

  Tsukuba Div. Res. Center For Medicinal Plant Resources National Inst. Of Biomedical Innovation

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