Establishment of Rhizobium-mediated transformation of Coptis japonica and molecular analyses of transgenic plants
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概要
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Coptis japonica is a perennial medicinal plant grown in Asia, and its rhizome is used as crude drug, in which berberine is highly accumulated as the main alkaloid. In this study, a transformation method for C. japonica using Rhizobium has been established. Young petioles were infected with Rhizobium radiobactor Gv3101 (pMP90) harboring a plasmid with β-glucuronidase (gus) driven by cauliflower mosaic virus 35S promoter and hygromycin phosphotransferase (hpt) gene as the selection marker. GUS assay showed that 46% of the calli grown on hygromycin plates clearly expressed the gus gene. As an application of the transformation method, an endogenous cDNA encoding an ABC protein, CjMDR1, which is involved in berberine transport, was introduced in sense orientation. Cjmdr1 transgenic plant (Sense-2) regenerated from hygromycin resistant callus line had a single copy of integrated hpt gene in its genomic DNA. Northern analyses of the Sense-2 plant showed that Cjmdr1 mRNA levels were lower in all organs examined than those of wild-type plant, which suggested that co-suppression of Cjmdr1 expression occurred. Indeed, the berberine content in Sense-2 was also suppressed. This is the first report of stable transformation of Coptis spp. and also of the alteration of a secondary metabolite by transport engineering.
- 日本植物細胞分子生物学会の論文
- 2005-06-01
著者
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Yazaki Kazufumi
Laboratory of Plant Gene Expression, Research Institute for Sustainable Humanosphere, Kyoto Universi
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Yazaki Kazufumi
Kyoto Univ. Kyoto Jpn
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Sato Fumihiko
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University
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Yazaki Kazufumi
Lab. Of Plant Gene Expression Res. Inst. For Sustainable Humanosphere Kyoto Univ.
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Yazaki Kazufumi
Laboratory Of Gene Expression Wood Research Institute Kyoto University
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KIUCHI Fumiyuki
Research Center for Medicinal Plant Resources National Institute of Biomedical Innovation
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SHITAN Nobukazu
Laboratory of Plant Gene Expression, Research Institute for Sustainable Humanosphere, Kyoto Universi
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YOSHIMATSU Kayo
Research Center for Medicinal Plant Resources, National Institute of Biomedical Innovation
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Kiuchi F
Graduate School Of Pharmaceutical Sciences Kyoto University
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Kiuchi Fumiyuki
国立医薬品衛生研究所
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Kikuchi F
Faculty Of Pharmaceutical Sciences Kanazawa University:(present Address)graduate School Of Pharmaceu
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Kikuchi F
Faculty Of Pharmaceutical Sciences Kanazawa University
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Kiuchi Fumiyuki
National Inst. Biomedical Innovation Ibaraki Jpn
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Kiuchi Fumiyuki
Graduate School Of Pharmaceutical Sciences Kyoto University
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Sato Fumihiko
Division Of Integrated Life Science Graduate School Of Biostudies Kyoto University
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Sato Fumihiko
Laboratory Of Molecular And Cellular Biology Of Totipotency Division Of Integrated Life Science Grad
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Yoshimatsu K
National Inst. Biomedical Innovation Ibaraki Jpn
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Yoshimatsu K
National Inst. Health Sci. Ibaraki Jpn
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Yoshimatsu Kayo
Research Center For Medicinal Plant Resources National Institute Of Biomedical Innovation
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Shitan Nobukazu
Laboratory Of Molecular And Cellular Biology Of Totipotency Division Of Integrated Life Science Grad
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Sato Fumihiko
Division Of Applied Life Sciences Graduate School Of Agriculture Kyoto University
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Kiuchi Fumiyuki
Faculty Of Pharmaceutical Sciences Kanazawa University:(present Address)graduate School Of Pharmaceu
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Shitan Nobukazu
Department Of Natural Medicinal Chemistry Kobe Pharmaceutical University
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