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Molecular identification of 1-Cys peroxiredoxin and anthocyanidin/flavonol 3-O-galactosyltransferase from proanthocyanidin-rich young fruits of persimmon (Diospyros kaki Thunb.)

机译:柿子富含原花青素的年轻果实中的1-Cys过氧化物酶和花色素苷/黄酮醇3-O-半乳糖基转移酶的分子鉴定(Diospyros kaki Thunb。)

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摘要

Fruits of persimmon (Diospyros kaki Thunb.) accumulate large amounts of proanthocyanidins (PAs) in the early stages of development. Astringent (A)-type fruits remain rich in soluble PAs even after they reach full-mature stage, whereas non-astringent (NA)-type fruits lose these compounds before full maturation. As a first step to elucidate the mechanism of PA accumulation in this non-model species, we used suppression subtractive hybridization to identify transcripts accumulating differently in young fruits of A- and NA-type. Interestingly, only a few clones involved in PA biosynthesis were identified in A–NA libraries. Represented by multiple clones were those encoding a novel 1-Cys peroxiredoxin and a new member of family 1 glycosyltransferases. Quantitative RT-PCR analyses confirmed correlation of the amount of PAs and accumulation of transcripts encoding these proteins in young persimmon fruits. Furthermore, the new family 1 glycosyltransferase was produced in Escherichia coli and shown to efficiently catalyze galactosylation at 3-hydroxyl groups of several anthocyanidins and flavonols. These findings suggest a complex mechanism of PA accumulation in persimmon fruits.Electronic supplementary materialThe online version of this article (doi:10.1007/s00425-009-0989-0) contains supplementary material, which is available to authorized users.
机译:柿果(Diospyros kaki Thunb。)的果实在发育的早期阶段会积累大量原花青素(PAs)。收敛性(A)型水果即使达到成熟阶段后仍然富含可溶性PA,而非收敛性(NA)型水果在完全成熟之前会丢失这些化合物。为了阐明该非模式物种中PA积累的机制的第一步,我们使用抑制消减杂交来鉴定在A型和NA型幼果中积累不同的转录本。有趣的是,在A–NA文库中仅鉴定出少数参与PA生物合成的克隆。由多个克隆代表的是那些编码新的1-Cys过氧化物酶和家族1糖基转移酶的新成员的克隆。定量RT-PCR分析证实了柿子幼果中PA含量与编码这些蛋白的转录本积累之间的相关性。此外,新的家族1糖基转移酶在大肠杆菌中产生,并且显示出可有效催化几种花色素和黄酮醇的3-羟基处的半乳糖基化。这些发现表明柿子果实中PA积累的复杂机制。电子补充材料本文的在线版本(doi:10.1007 / s00425-009-0989-0)包含补充材料,授权用户可以使用。

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