Expression analysis of the VTC2 and VTC5 genes encoding GDP-L-galactose phosphorylase, an enzyme involved in ascorbate biosynthesis, in Arabidopsis thaliana

Biosci Biotechnol Biochem. 2011;75(9):1783-8. doi: 10.1271/bbb.110320. Epub 2011 Sep 7.

Abstract

Arabidopsis thaliana contains two GDP-L-galactose phosphorylase genes, VTC2 and VTC5, which are critical for ascorbate (AsA) biosynthesis. We investigated the expression levels of both VTC2 and VTC5 genes in wild-type A. thaliana and the AsA deficient mutants during early seedling growth. Ascorbate accumulated to an equal extent in all genotypes up to 5 d post-germination (DPG). The transcript level of VTC2 was dominant, and increased in parallel with AsA accumulation in the wild type. On the other hand, the expression of VTC5 compensated for the reduced VTC2 transcription levels in the AsA deficient mutant vtc2-1 in young seedlings. A luciferase activity assay indicated that the VTC5 promoter was more active in young (2 DPG) cotyledons and that the VTC2 and VTC5 promoters drove a day-to-night variation in expression. The present work provides clues to the precise roles of VTC2 and VTC5 in AsA biosynthesis in A. thaliana at the young seedling stage.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / enzymology*
  • Arabidopsis / genetics
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism*
  • Ascorbic Acid / biosynthesis*
  • Ascorbic Acid / genetics
  • Cotyledon / enzymology
  • Cotyledon / genetics
  • Galactose / metabolism
  • Gene Expression Regulation, Plant*
  • Genes, Reporter
  • Germination
  • Guanosine Diphosphate / metabolism
  • Isoenzymes / genetics
  • Isoenzymes / metabolism*
  • Light
  • Luciferases / analysis
  • Mutation
  • Phosphorylases / genetics
  • Phosphorylases / metabolism*
  • Plants, Genetically Modified / enzymology*
  • Plants, Genetically Modified / genetics
  • Promoter Regions, Genetic
  • RNA, Messenger / analysis
  • RNA, Messenger / biosynthesis
  • Seedlings / enzymology
  • Seedlings / genetics
  • Time Factors
  • Transformation, Genetic

Substances

  • Arabidopsis Proteins
  • Isoenzymes
  • RNA, Messenger
  • Guanosine Diphosphate
  • Luciferases
  • Phosphorylases
  • Ascorbic Acid
  • Galactose