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Enhanced accumulation of atropine in Atropa belladonna transformed by Rac GTPase gene isolated from Scoparia dulcis

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Abstract

Leaf tissues of Atropa belladonna were transformed by Sdrac2, a Rac GTPase gene, that is isolated from Scoparia dulcis, and the change in atropine concentration of the transformants was examined. Re-differentiated A. belladonna overexpressing Sdrac2 accumulated considerable concentration of atropine in the leaf tissues, whereas the leaves of plants transformed by an empty vector accumulated only a very low concentration of the compound. A. belladonna transformed by CASdrac2, a modified Sdrac2 of which translate was expected to bind guanosine triphosphate (GTP) permanently, accumulated very high concentrations of atropine (approximately 2.4-fold excess to those found in the wild-type plant in its natural habitat). In sharp contrast, the atropine concentration in transformed A. belladonna prepared with negatively modified Sdrac2, DNSdrac2, expected to bind guanosine diphosphate instead of GTP, was very low. These results suggested that Rac GTPases play an important role in the regulation of secondary metabolism in plant cells and that overexpression of the gene(s) may be capable of enhancing the production of natural products accumulated in higher plant cells.

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Abbreviations

GFP:

Green fluorescent protein

HPLC:

High performance liquid chromatography

MJ:

Methyl jasmonate

RACE:

Rapid amplification of cDNA end

PCR:

Polymerase chain reaction

RT:

Reverse transcription

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Acknowledgments

This work was supported, in part, by research grants from the Ministry of Education, Culture, Sports, Science & Technology in Japan, and from Cosmetology Research Foundation.

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Authors and Affiliations

  1. Laboratory of Medicinal Bio-resources, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, Sugitani 2630, Toyama, 930-0194, Japan

    Kyouhei Asano, Jung-Bum Lee, Yoshimi Yamamura & Fumiya Kurosaki

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  1. Kyouhei Asano
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  2. Jung-Bum Lee
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  3. Yoshimi Yamamura
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  4. Fumiya Kurosaki
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Correspondence to Fumiya Kurosaki.

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Asano, K., Lee, JB., Yamamura, Y. et al. Enhanced accumulation of atropine in Atropa belladonna transformed by Rac GTPase gene isolated from Scoparia dulcis . Transgenic Res 22, 1249–1255 (2013). https://doi.org/10.1007/s11248-013-9733-4

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  • DOI: https://doi.org/10.1007/s11248-013-9733-4

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