Colour variation in red grapevines (Vitis vinifera L.): genomic organisation, expression of flavonoid 3'-hydroxylase, flavonoid 3',5'-hydroxylase genes and related metabolite profiling of red cyanidin-/blue delphinidin-based anthocyanins in berry skin.
Colour variation in red grapevines (Vitis vinifera L.): genomic organisation, expression of flavonoid 3'-hydroxylase, flavonoid 3',5'-hydroxylase genes and related metabolite profiling of red cyanidin-/blue delphinidin-based anthocyanins in berry skin.
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红色葡萄藤（Vitis Vinifera L.）的颜色变化：基因组组织，类黄酮3'-羟化酶的表达，类黄酮3'，5'-羟化酶基因以及相关的红色氰化物/蓝色Delphinidin arthocyin atthocyin in Berry sigry simmiken。
DOI：
10.1186/1471-2164-7-12
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发表时间：
2006-01-24
期刊：
影响因子：
4.4
通讯作者：
Testolin, Raffaele
中科院分区：
文献类型：
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作者：
Castellarin, Simone D;Di Gaspero, Gabriele;Marconi, Raffaella;Nonis, Alberto;Peterlunger, Enrico;Paillard, Sophie;Adam-Blondon, Anne-Francoise;Testolin, Raffaele
编码类黄酮3 '-和3'，5 '-羟化酶（F3' H和F3 '5'H）的苯丙素途径的结构基因长期以来被用来解释在所谓的葡萄藤红色品种中基于花青素和飞燕草素的花青素色素的生物合成。这两种类型的花青素的相对比例在很大程度上受遗传控制，并决定了红/紫/蓝浆果葡萄品种及其相应葡萄酒之间的颜色变化。根据植物同源基因设计简并引物，从葡萄属葡萄品种赤霞珠（Cabernet Sauvignon）中分离到VvF 3 'H和VvF 3' 5 'H基因片段，分别翻译成313和239个氨基酸的蛋白质片段，与植物细胞色素P450单加氧酶CYP 75的同源性分别高达76%和82%。根据序列同源性、表达谱及其与十个不同成熟阶段代谢产物积累的相关性，对推定的功能进行了分配。在颜色转变开始时，VvF 3 'H和VvF 3' 5 'H的转录诱导在时间上与花青素苷生物合成的开始相协调，在红色浆果中的表达分别比绿色浆果高2倍和50倍。VvF 3 '5'H表达的峰值在两周后观察到，伴随着飞燕草素/矢车菊素衍生物的比率的增加。结构基因组学分析表明，两个拷贝的VvF 3 'H物理连接在第17号连锁群上，几个拷贝的VvF 3' 5 'H紧密聚集并嵌入到第6号连锁群上的节段重复中，揭示了与迄今为止已知的其他植物类黄酮羟化酶基因（主要在植物中）相比的高度复杂性。我们已经证明，编码类黄酮3 '-和3'，5 '-羟化酶的基因在积累类黄酮的葡萄植物的任何组织中表达，特别是在主要合成花青素的成熟红色浆果的皮肤中。转录谱与基于红色/花青素和蓝色/飞燕草素的花青素的积累动力学之间的相关性表明，VvF 3 'H和VvF 3' 5 'H表达与成熟束的颜色演变一致。当地的物理地图周围的VvF 3 'H和VvF 3' 5 'H基因座的构建应有助于促进每个异构体的监管元件的识别和未来的葡萄藤和葡萄酒的颜色通过农艺学，环境和生物技术工具的操作。
Structural genes of the phenyl-propanoid pathway which encode flavonoid 3'- and 3',5'-hydroxylases (F3'H and F3'5'H) have long been invoked to explain the biosynthesis of cyanidin- and delphinidin-based anthocyanin pigments in the so-called red cultivars of grapevine. The relative proportion of the two types of anthocyanins is largely under genetic control and determines the colour variation among red/purple/blue berry grape varieties and their corresponding wines. Gene fragments of VvF3'H and VvF3'5'H, that were isolated from Vitis vinifera 'Cabernet Sauvignon' using degenerate primers designed on plant homologous genes, translated into 313 and 239 amino acid protein fragments, respectively, with up to 76% and 82% identity to plant CYP75 cytochrome P450 monooxygenases. Putative function was assigned on the basis of sequence homology, expression profiling and its correlation with metabolite accumulation at ten different ripening stages. At the onset of colour transition, transcriptional induction of VvF3'H and VvF3'5'H was temporally coordinated with the beginning of anthocyanin biosynthesis, the expression being 2-fold and 50-fold higher, respectively, in red berries versus green berries. The peak of VvF3'5'H expression was observed two weeks later concomitantly with the increase of the ratio of delphinidin-/cyanidin-derivatives. The analysis of structural genomics revealed that two copies of VvF3'H are physically linked on linkage group no. 17 and several copies of VvF3'5'H are tightly clustered and embedded into a segmental duplication on linkage group no. 6, unveiling a high complexity when compared to other plant flavonoid hydroxylase genes known so far, mostly in ornamentals. We have shown that genes encoding flavonoid 3'- and 3',5'-hydroxylases are expressed in any tissues of the grape plant that accumulate flavonoids and, particularly, in skin of ripening red berries that synthesise mostly anthocyanins. The correlation between transcript profiles and the kinetics of accumulation of red/cyanidin- and blue/delphinidin-based anthocyanins indicated that VvF3'H and VvF3'5'H expression is consistent with the chromatic evolution of ripening bunches. Local physical maps constructed around the VvF3'H and VvF3'5'H loci should help facilitate the identification of the regulatory elements of each isoform and the future manipulation of grapevine and wine colour through agronomical, environmental and biotechnological tools.