Journal Article

Expression of the grape dihydroflavonol reductase gene and analysis of its promoter region

Perl, Avihai, Even, Sylvie, Gollop, Rachel, Colova‐Tsolova, Violeta

2002

Overview

Dihydroflavonol reductase (DFR) is a key enzyme involved in anthocyanin biosynthesis and proanthocyanidin synthesis in grape. DFR catalyses the reduction of dihydroflavonols to leucoanthocyanidins in the anthocyanin pathway. The DFR products, the leucoanthocyanidins, are substrates for the next step in the anthocyanin pathway and are also the substrates for the proanthocyanidin pathway. In the present study the promoter of the grape dfr gene was cloned. Analysis of the dfr promoter sequence revealed the existence of several putative DNA binding motifs. The dfr promoter was fused to the uidA gene and the control of this fusion and the endogenous dfr gene expression, was studied in transformed plants and in red cell suspension originated from fruits. The dfr promoter–uidA gene fusion was expressed in leaves, roots and stems. Deletions of the dfr promoter influenced the specificity of the expression of the GUS gene fusion in plantlet roots and the level of expression in plants and in the red cell suspension originated from fruits. The deletion analysis of the dfr promoter suggests that a specific sequence located between −725 to −233 might be involved in expression of the dfr gene in fruits. Light, calcium and sucrose induced the dfr gene expression. In the transformed suspension cultures, expression of both the endogenous dfr gene and the dfr promoter–uidA gene fusions was induced by white light. The induction by both light and calcium suggests the possible involvement of a UV receptors signal transduction pathway in the induction of the dfr gene. The induction of the dfr gene and the dfr promoter–uidA gene fusions by light and sucrose indicates a close interaction between sucrose and light signalling pathways.

• Publisher: Oxford University Press,OXFORD UNIVERSITY PRESS,Oxford Publishing Limited (England)
• Subject: Agronomy. Soil science and plant productions / Alcohol Oxidoreductases / Alcohol Oxidoreductases - genetics / Alcohol Oxidoreductases - metabolism / amino acid sequences / anthocyanins / Anthocyanins - biosynthesis / Base Sequence / beta-glucuronidase / Binding sites / biochemical pathways / Biological and medical sciences / Biosynthesis / Calcium / Calcium - pharmacology / Cell and Molecular Biology, Biochemistry and Molecular Physiology / Cell physiology / cell suspension culture / Cells, Cultured / chemistry / Developmental expression / dfr / DNA, Plant / DNA, Plant - chemistry / DNA, Plant - genetics / DNA-binding domains / drug effects / Economic plant physiology / enzymology / Fruits / Fundamental and applied biological sciences. Psychology / Gene expression / Gene Expression Regulation, Enzymologic / Gene Expression Regulation, Enzymologic - drug effects / Gene Expression Regulation, Enzymologic - radiation effects / Gene Expression Regulation, Plant / Gene Expression Regulation, Plant - drug effects / Gene Expression Regulation, Plant - radiation effects / Gene induction / Genes / genetic transformation / genetics / Glucuronidase / Glucuronidase - genetics / Glucuronidase - metabolism / grape / Light / light signalling / Metabolism / Metabolism. Physicochemical requirements / Molecular and cellular biology / Molecular Sequence Data / Nitrogen metabolism and other ones (excepting carbon metabolism) / Nutrition. Photosynthesis. Respiration. Metabolism / oxidoreductases / pharmacology / Plant cells / Plant physiology and development / Plant roots / Plantlets / Plants / Plants, Genetically Modified / Proanthocyanidins / promoter regions / Promoter Regions, Genetic / Promoter Regions, Genetic - genetics / radiation effects / receptors / Recombinant Fusion Proteins / Recombinant Fusion Proteins - genetics / Recombinant Fusion Proteins - metabolism / reporter genes / RNA / Roots / Sequence Analysis, DNA / Signal Transduction / Sucrose / Sucrose - pharmacology / sucrose signalling / ultraviolet radiation / Vitis / Vitis - enzymology / Vitis - genetics / Vitis vinifera