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

Abstract

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.

Naturally Occurring Anthocyanin, Structure, Functions and Biosynthetic Pathway in Fruit Plants

“…There are numerous structures and anthocyanin profile has been reported in different plant species [11,12].…”

“…They are glycosides of polymethoxy and polyhdroxy derived of flavylium salts or 2-phenylbenzopyrylium [19]. So far 600 dissimilar entities of anthocyanins have been discovered from different species and this quantity is still rising with the recent advancements in research [11,12,15]. In the plant kingdom, it was documented that through methylation of the six ordinary anthocyanidins, approximately 90% of the anthocyanins were modified of three methylated anthocyanidins, malvidin, petunidin and peonidin, comprised of 20% of the overall described anthocyanidins [17].…”

“…Glycosyl moieties transformed anthocyanidins in various synthetic ways in a specific to family or species, and its color pattern significantly rely on hydroxyl groups on the B-ring; as the number of larger groups, the darkeners the blue color [22,31]. In grape berries anthocyanins is synthesized through the flavonoid path, which split the similar upstream pathway throughout proanthocyanidins, in anticipation of the development of anthocyanidins by the catalysis of anthocyanidin synthase (ANS), also known as leucoanthocyanidin dioxygenase (LDOX) [11,79].…”

A reliable protocol for the stable transformation of non-embryogenic cells cultures of grapevine (Vitis vinifera L.) and Taxus x media

“…Several groups have published reports on the transformation of Vitis [33][34][35][36][37][38][39][40]. To date, Gollop et al [41] have provided evidence for the stable transformation of non-embryogenic cultures (NEC). Nevertheless, the transformation success rate and long-term sustainability of the established transformed cell culture did not seem to be a crucial requirement in their study, and neither was reported.…”

A Medicago truncatula H+-pyrophosphatase gene, MtVP1, improves sucrose accumulation and anthocyanin biosynthesis in potato (Solanum tuberosum L.)

“…Many anthocyanin biosynthetic genes are induced by sugars (Gollop et al, 2001(Gollop et al, , 2002Martin et al, 2002;Hiratsu et al, 2003). Expression of the petunia (Petunia hybrida) chalcone synthase (CHS) gene in transgenic Arabidopsis leaves is induced by sugars.…”