Expression of the grape dihydroflavonol reductase gene and analysis of its promoter region.
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.
Gollop
Rachel
R
Department of Fruit Tree Breeding and Molecular Genetics, ARO, The Volcani Center, PO Box 6, Bet-Dagan 50250, Israel. rachel_gollop@yahoo.com
Even
Sylvie
S
Colova-Tsolova
Violeta
V
Perl
Avihai
A
D000429:Alcohol Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D000872:Anthocyanins / Q000096:biosynthesis; D001483:Base Sequence; D002118:Calcium / Q000494:pharmacology; D002478:Cells, Cultured; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000528:radiation effects; D018506:Gene Expression Regulation, Plant / Q000187:drug effects / Q000528:radiation effects; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D008027:Light; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified; D044945:Proanthocyanidins; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D015398:Signal Transduction; D013395:Sucrose / Q000494:pharmacology; D027843:Vitis / Q000201:enzymology / Q000235:genetics*
D000872:Anthocyanins; D018744:DNA, Plant; D044945:Proanthocyanidins; D011993:Recombinant Fusion Proteins; C013221:proanthocyanidin; D013395:Sucrose; D000429:Alcohol Oxidoreductases; C056570:dihydroflavanol 4-reductase; D005966:Glucuronidase; D002118:Calcium