Cloning and molecular analysis of structural genes involved in flavonoid and stilbene biosynthesis in grape (Vitis vinifera L.) |
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Authors: | Francesca Sparvoli Cathie Martin Attilio Scienza Giuseppe Gavazzi Chiara Tonelli |
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Institution: | (1) Dipartmento di Genetica e di Biologia dei Microorganismi, Università degli Studi di Milano, Via Celoria 26, 20133 Milano, Italia;(2) Istituto di Coltivazioni Arboree, Università degli Studi di Milano, Via Celoria 2, 20133 Milano, Italia;(3) Department of Genetics, John Innes Institute, Colney Lane, NR4 7HU Norwich, UK |
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Abstract: | Genes involved in flavonoid and stilbene biosynthesis were isolated from grape (Vitis vinifera L.). Clones coding for phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydoxylase (F3H), dihydroflavonol 4-reductase (DFR), leucoanthocyanidin dioxygenase (LDOX) and UDP glucose:flavonoid 3-O-glucosyl transferase (UFGT), were isolated by screening a cDNA library, obtained from mRNA from seedlings grown in light for 48 h using snapdragon (Antirrhinum majus) and maize heterologous probes. A cDNA clone coding for stilbene synthase (StSy) was isolated by probing the library with a specific oligonucleotide. These clones were sequenced and when the putative products were compared to the published amino acid sequence for corresponding enzymes, the percentages of similarity ranged from 65% (UFGT) to 90% (CHS and PAL). The analysis of the genomic organization and expression of these genes in response to light shows that PAL and StSy genes belong to large multigene families, while the others are present in one to four copies per haploid genome. The steady-state level of mRNAs encoded by the flavonoid biosynthetic genes as determined in young seedlings is coordinately induced by light, except for PAL and StSy, which appear to be constitutively expressed. |
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Keywords: | anthocyanins cDNA cloning flavonoids gene expression genomic organization stilbenes Vitis vinifera L |
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