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葡萄(Vitis vinifera L. cv. 赤霞珠)发育过程中花、幼果和果皮中无色花青素还原酶和花青素还原酶的基因表达及活性

Leucoanthocyanidin reductase and anthocyanidin reductase gene expression and activity in flowers, young berries and skins of Vitis vinifera L. cv. Cabernet-Sauvignon during development.

作者信息

Gagné Séverine, Lacampagne Soizic, Claisse Olivier, Gény Laurence

机构信息

Université de Bordeaux, UMR 1219 Oenologie, INRA, ISVV, 210 Chemin de Leysotte, CS 58008, 33 140 Villenave D'Ornon, France.

出版信息

Plant Physiol Biochem. 2009 Apr;47(4):282-90. doi: 10.1016/j.plaphy.2008.12.004. Epub 2008 Dec 16.

Abstract

Proanthocyanidins, or condensed tannins, are crucial polyphenolic compounds for grape and wine quality. Recently, significant advances were achieved in understanding the biosynthesis of their main subunits: (+)-catechin and (-)-epicatechin, produced by catalysis of leucoanthocyanidin reductase (LAR) and anthocyanidin reductase (ANR), respectively. Expression studies had been published but no data were available on enzyme activity. In our work, we devised assays to measure LAR and ANR activity and determine their development throughout the growth of flowers, young berries, and skins of Vitis vinifera L. cv. Cabernet-Sauvignon. We also investigated the accumulation of compounds in these tissues and focused on the expression of both the structural genes and the transcription factors involved in regulating them: VvMYB5a and VvMYBPA1. Biosynthetic genes were expressed early and LAR and ANR were already active during flowering and at the beginning of berry growth, as well as during colour-change in skins. The profiles we determined correlated with total tannin, catechin, and epicatechin concentrations. The involvement of VvMYB5a and VvMYBPA1 was confirmed and specific expression patterns were also established for VvLAR transcripts.

摘要

原花青素,即缩合单宁,是影响葡萄和葡萄酒品质的关键多酚类化合物。最近,在了解其主要亚基(+)-儿茶素和(-)-表儿茶素的生物合成方面取得了重大进展,它们分别由无色花青素还原酶(LAR)和花青素还原酶(ANR)催化产生。此前已有关于表达研究的报道,但尚无关于酶活性的数据。在我们的研究中,我们设计了测定方法来测量LAR和ANR的活性,并确定它们在酿酒葡萄品种赤霞珠花朵、幼果和果皮生长过程中的变化情况。我们还研究了这些组织中化合物的积累,并重点关注了调控它们的结构基因和转录因子:VvMYB5a和VvMYBPA1的表达。生物合成基因在早期表达,LAR和ANR在开花期、果实生长初期以及果皮变色期均已具有活性。我们测定的图谱与总单宁、儿茶素和表儿茶素的浓度相关。证实了VvMYB5a和VvMYBPA1的作用,并确定了VvLAR转录本的特定表达模式。

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