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Atropa Belladonna

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Birgit Dräger – One of the best experts on this subject based on the ideXlab platform.

  • Overexpression of tropinone reductases alters alkaloid composition in Atropa Belladonna root cultures
    , 2013
    Co-Authors: Ute Richter, Grit Rothe, Anne-katrin Fabian, Bettina Rahfeld, Birgit Dräger

    Abstract:

    composition in Atropa Belladonna root culture

  • Alkaloids in plants and root cultures of Atropa Belladonna overexpressing putrescine N‐methyltransferase
    Journal of Experimental Botany, 2003
    Co-Authors: Grit Rothe, Takashi Hashimoto, Yasuyuki Yamada, Akira Hachiya, Birgit Dräger

    Abstract:

    Putrescine N-methyltransferase (PMT) is the first alkaloid-specific enzyme for nicotine and tropane alkaloid formation. The pmt gene from Nicotiana tabacum was fused to the CaMV 35S promoter and integrated into the Atropa Belladonna genome. Transgenic plants and derived root cultures were analysed for gene expression and for levels of alkaloids and their precursors. Scopolamine, hyoscyamine, tropine, pseudotropine, tropinone, and calystegines were found unaltered or somewhat decreased in pmt-overexpressing lines compared to controls. When root cultures were treated with 5% sucrose, calystegine levels were elevated in control roots, but were not affected in pmt-overexpressing roots. 1 microM auxin reduced calystegine levels in control roots, while in pmt-overexpressing roots all alkaloids remained unaltered. Expression level of pmt alone is apparently not limiting for tropane alkaloid formation in A. Belladonna.

  • alkaloids in plants and root cultures of Atropa Belladonna overexpressing putrescine n methyltransferase
    Journal of Experimental Botany, 2003
    Co-Authors: Grit Rothe, Takashi Hashimoto, Yasuyuki Yamada, Akira Hachiya, Birgit Dräger

    Abstract:

    Putrescine N-methyltransferase (PMT) is the first alkaloid-specific enzyme for nicotine and tropane alkaloid formation. The pmt gene from Nicotiana tabacum was fused to the CaMV 35S promoter and integrated into the Atropa Belladonna genome. Transgenic plants and derived root cultures were analysed for gene expression and for levels of alkaloids and their precursors. Scopolamine, hyoscyamine, tropine, pseudotropine, tropinone, and calystegines were found unaltered or somewhat decreased in pmt-overexpressing lines compared to controls. When root cultures were treated with 5% sucrose, calystegine levels were elevated in control roots, but were not affected in pmt-overexpressing roots. 1 microM auxin reduced calystegine levels in control roots, while in pmt-overexpressing roots all alkaloids remained unaltered. Expression level of pmt alone is apparently not limiting for tropane alkaloid formation in A. Belladonna.

Yasuyuki Yamada – One of the best experts on this subject based on the ideXlab platform.

  • Alkaloids in plants and root cultures of Atropa Belladonna overexpressing putrescine N‐methyltransferase
    Journal of Experimental Botany, 2003
    Co-Authors: Grit Rothe, Takashi Hashimoto, Yasuyuki Yamada, Akira Hachiya, Birgit Dräger

    Abstract:

    Putrescine N-methyltransferase (PMT) is the first alkaloid-specific enzyme for nicotine and tropane alkaloid formation. The pmt gene from Nicotiana tabacum was fused to the CaMV 35S promoter and integrated into the Atropa Belladonna genome. Transgenic plants and derived root cultures were analysed for gene expression and for levels of alkaloids and their precursors. Scopolamine, hyoscyamine, tropine, pseudotropine, tropinone, and calystegines were found unaltered or somewhat decreased in pmt-overexpressing lines compared to controls. When root cultures were treated with 5% sucrose, calystegine levels were elevated in control roots, but were not affected in pmt-overexpressing roots. 1 microM auxin reduced calystegine levels in control roots, while in pmt-overexpressing roots all alkaloids remained unaltered. Expression level of pmt alone is apparently not limiting for tropane alkaloid formation in A. Belladonna.

  • alkaloids in plants and root cultures of Atropa Belladonna overexpressing putrescine n methyltransferase
    Journal of Experimental Botany, 2003
    Co-Authors: Grit Rothe, Takashi Hashimoto, Yasuyuki Yamada, Akira Hachiya, Birgit Dräger

    Abstract:

    Putrescine N-methyltransferase (PMT) is the first alkaloid-specific enzyme for nicotine and tropane alkaloid formation. The pmt gene from Nicotiana tabacum was fused to the CaMV 35S promoter and integrated into the Atropa Belladonna genome. Transgenic plants and derived root cultures were analysed for gene expression and for levels of alkaloids and their precursors. Scopolamine, hyoscyamine, tropine, pseudotropine, tropinone, and calystegines were found unaltered or somewhat decreased in pmt-overexpressing lines compared to controls. When root cultures were treated with 5% sucrose, calystegine levels were elevated in control roots, but were not affected in pmt-overexpressing roots. 1 microM auxin reduced calystegine levels in control roots, while in pmt-overexpressing roots all alkaloids remained unaltered. Expression level of pmt alone is apparently not limiting for tropane alkaloid formation in A. Belladonna.

  • Expression of Atropa Belladonna putrescine N-methyltransferase gene in root pericycle.
    Plant and Cell Physiology, 1999
    Co-Authors: Ken-ichi Suzuki, Yasuyuki Yamada, Takashi Hashimoto

    Abstract:

    The cDNAs encoding putrescine N-methyltransferase (PMT), which catalyzes the S-adenosylmethionine-dependent N-methylation of putrescine at the first committed step in the biosynthetic pathways of tropane alkaloids, were isolated from Atropa Belladonna and Hyoscyamus niger. These PMTs, however, lacked the N-terminal tandem repeat arrays previously found in Nicotiana PMTs. AbPMT1 RNA was much more abundant in the root of A. Belladonna than was AbPMT2 RNA. The 5′-flanking region of the AbPMT1 gene was fused to the beta-glucuronidase (GUS) reporter gene and transferred to A. Belladonna. Histochemical analysis showed that GUS is expressed specifically in root pericycle cells and that the 0.3-kb 5′-upstream region was sufficient for pericycle-specific expression. Treatment of A. Belladonna roots with methyl jasmonate did not up-regulate the expression of GUS or endogenous AbPMT genes. The regulation of tropane alkaloid biosynthesis is discussed and compared with that of nicotine biosynthesis.

Koichiro Shimomura – One of the best experts on this subject based on the ideXlab platform.

  • Salicylic acid carboxyl methyltransferase induced in hairy root cultures of Atropa Belladonna after treatment with exogeneously added salicylic acid.
    Plant and Cell Physiology, 2002
    Co-Authors: Hiroyuki Fukami, Koichiro Shimomura, Hiroshi Hirano, Tomiko Asakura, Keiko Abe, Takashi Yamakawa

    Abstract:

    ;In Atropa Belladonna hairy roots, exogeneously added salicylic acid (SA) is converted to methyl salicylate (MSA) through the reaction, which might be catalysed by Sadenosyl-L-methionine: salicylic acid carboxyl methyltransferase (SAMT). Here we cloned a cDNA for A. Belladonna SAMT (AbSAMT1), which consisted of 357 aa residues. It was expressed in E. coli, and the recombinant AbSAMT1 showed SAMT activity. When A. Belladonna hairy roots were exposed to a high concentration of SA, AbSAMT1 mRNA begins to be expressed 12 h after the exposure, and steady expression continued over 144 h.

  • Responses of Transformed Root Culture of Atropa Belladonna to Salicylic Acid Stress
    Journal of Bioscience and Bioengineering, 2002
    Co-Authors: Kung-ta Lee, Hiroshi Hirano, Takashi Yamakawa, Tohru Kodama, Yasuo Igarashi, Koichiro Shimomura

    Abstract:

    The effect of salicylic acid (SA) on tropane alkaloid production and the responses to SA stress of transformed root cultures of Atropa Belladonna (Belladonna) were investigated. Treatment of A. Belladonna transformed roots with 0.2 mM SA did not have any effect on tropane alkaloid production, but two compounds were produced in the medium. These were identified as the SA derivatives methylsalicylate and methyl-o-methoxybenzoate by high-resolution mass spectrometry and UV spectrometry. In contrast, treatment with 2 mM salicylic acid stimulated tropane alkaloid release from the transformed roots into the medium by up to 35% of the total alkaloids after 24 h, and the SA derivatives were not observed in the medium. These results revealed that transformed root of A. Belladonna exhibits distinct by different responses to SA stress depending on the SA concentration.

  • Genetic Transformation of Atropa Belladonna
    Transgenic Medicinal Plants, 1999
    Co-Authors: Mondher El Jaziri, Kayo Yoshimatsu, Koichiro Shimomura

    Abstract:

    Leaves of Atropa Belladonna L. (Solanaceae) are mainly used as a source for the tropane alkaloids, hyoscyamine and scopolamine, which are not easily produced by chemical synthesis. Thus because of its high morphogenetic potential, the in vitro culture of A. Belladonna has been actively investigated as an alternative to field cultivation and as a possible source for tropane alkaloid production (for review see Bajaj and Simola 1991) Several studies have confirmed that the product levels in unorganized A. Belladonna callus and suspension cultures are generally very low and that root morphology is required for synthesis of tropane alkaloid in vitro. However, molecular genetic technology is now available to achieve transformation and regeneration of transgenic A. Belladonna plants in which morphological traits and/or particular metabolic step(s) are artificially modified. The hairy root system is demonstrated to be much more genetically stable than dedifferentiated normal (callus) or transformed (crown gall) plant cells. In this chapter some important recent studies related to the genetic transformation of A. Belladonna are discussed.