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Characterization and structural features of a chalcone synthase mutation in a white-flowering line of Matthiola incanaR. Br. (Brassicaceae)

Plant Molecular Biology volume 55, pages 455–465 (2004)Cite this article

Abstract

For Matthiola incana (Brassicaceae), used as a model system to study biochemical and genetical aspects of anthocyanin biosynthesis, several nearly isogenic colored wild type lines and white-flowering mutant lines are available, each with a specific defect in the genes responsible for anthocyanin production (genes e, f, and g). For gene f supposed to code for chalcone synthase (CHS; EC 2.3.1.74), the key enzyme of the flavonoid/anthocyanin biosynthesis pathway belonging to the group of type III polyketide synthases (PKS), the wild type genomic sequence of M. incana line 04 was determined in comparison to the white-flowering CHS mutant line 18. The type of mutation in the chs gene was characterized as a single nucleotide substitution in a triplet AGG coding for an evolutionary conserved arginine into AGT coding for serine (R72S). Northern blots and RT-PCR demonstrated that the mutated gene is expressed in flower petals. Heterologous expression of the wild type and mutated CHS cDNA in E. Scherichia coli, verified by Western blotting and enzyme assays with various starter molecules, revealed that the mutant protein had no detectable activity, indicating that the strictly conserved arginine residue is essential for the enzymatic reaction. This mutation, which previously was not detected by mutagenic screening, is discussed in the light of structural and functional information on alfalfa CHS and related type III PKS enzymes.

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Affiliations

  1. Department of General Genetics, Center of Plant Molecular Biology (ZMBP), University of Tübingen, Auf der Morgenstelle 28, Germany

    Vera Hemleben, Angela Dressel & Bernhard Epping

  2. Institute for Pharmaceutical Biology, Philipps-University Marburg, Deutschhausstr, 17A, Germany

    Richard Lukačin & Stefan Martens

  3. Structural Biology Laboratory – Noel group, The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, CA, USA

    Michael Austin

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  1. Vera Hemleben
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  2. Angela Dressel
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  3. Bernhard Epping
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  4. Richard Lukačin
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  5. Stefan Martens
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  6. Michael Austin
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Hemleben, V., Dressel, A., Epping, B. et al. Characterization and structural features of a chalcone synthase mutation in a white-flowering line of Matthiola incanaR. Br. (Brassicaceae). Plant Mol Biol 55, 455–465 (2004). https://doi.org/10.1007/s11103-004-1125-y

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  • DOI: https://doi.org/10.1007/s11103-004-1125-y

  • anthocyanin
  • evolution
  • flavonoid biosynthesis
  • Medicago sativa
  • polyketide synthase
  • protein structure