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Plant Molecular Biology

, Volume 48, Issue 4, pp 401–411 | Cite as

Two flavonoid glucosyltransferases from Petunia hybrida: molecular cloning, biochemical properties and developmentally regulated expression

  • Mami Yamazaki
  • Emiko Yamagishi
  • Zhizhong Gong
  • Masako Fukuchi-Mizutani
  • Yuko Fukui
  • Yoshikazu Tanaka
  • Takaaki Kusumi
  • Masaatsu Yamaguchi
  • Kazuki Saito
Article

Abstract

Two flavonoid glucosyltransferases, UDP-glucose:flavonoid 3-O-glucosyltransferase (3-GT) and UDP-glucose: anthocyanin 5-O-glucosyltransferase (5-GT), are responsible for the glucosylation of anthocyani(di)ns to produce stable molecules in the anthocyanin biosynthetic pathway. The cDNAs encoding 3-GT and 5-GT were isolated from Petunia hybrida by hybridization screening with heterologous probes. The cDNA clones of 3-GT, PGT8, and 5-GT, PH1, encode putative polypeptides of 448 and 468 amino acids, respectively. A phylogenetic tree based on amino acid sequences of the family of glycosyltransferases from various plants shows that PGT8 belongs to the 3-GT subfamily and PH1 belongs to the 5-GT subfamily. The function of isolated cDNAs was identified by the catalytic activities for 3-GT and 5-GT exhibited by the recombinant proteins produced in yeast. The recombinant PGT8 protein could convert not only anthocyanidins but also flavonols into the corresponding 3-O-glucosides. In contrast, the recombinant PH1 protein exhibited a strict substrate specificity towards anthocyanidin 3-acylrutinoside, comparing with other 5-GTs from Perilla frutescens and Verbena hybrida, which showed broad substrate specificities towards several anthocyanidin 3-glucosides. The mRNA expression of both 3-GT and 5-GT increased in the early developmental stages of P. hybrida flower, reaching the maximum at the stage before flower opening. Southern blotting analysis of genomic DNA indicates that both 3-GT and 5-GT genes exist in two copies in P. hybrida, respectively. The results are discussed in relation to the molecular evolution of flavonoid glycosyltransferases.

anthocyanin flavonoid glucosyltransferase Petunia hybrida 

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Copyright information

© Kluwer Academic Publishers 2002

Authors and Affiliations

  • Mami Yamazaki
    • 1
  • Emiko Yamagishi
    • 1
  • Zhizhong Gong
    • 1
  • Masako Fukuchi-Mizutani
    • 2
  • Yuko Fukui
    • 2
  • Yoshikazu Tanaka
    • 2
  • Takaaki Kusumi
    • 2
  • Masaatsu Yamaguchi
    • 3
  • Kazuki Saito
    • 1
  1. 1.Laboratory of Molecular Biology and Biotechnology, Graduate School of Pharmaceutical SciencesChiba UniversityInage-kuJapan
  2. 2.Institute for Fundamental Research, Suntory Ltd.MishimaJapan
  3. 3.Faculty of HorticultureMinami Kyushu UniversityTakanabeJapan

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