‘霞多丽’葡萄果实中黄烷-3-醇合成酶活性及其相关基因表达特征
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宁夏大学

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宁夏自然科学基金项目(2020AAC03087)。


Activity of flavane-3-ol Synthetase and expression characteristics of related genes in "Chardonnay" Grape Fruit
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Project of Ningxia Natural Science Foundation (2020AAC03087).

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    摘要:

    为探究酿酒葡萄果实中黄烷-3-醇积累规律与合成相关酶、关键基因表达的关系,以贺兰山东麓主栽酿酒白葡萄品种‘霞多丽’为材料,采用比色法、高效液相色谱(HPLC)和荧光定量PCR技术,研究了葡萄果实发育过程中黄烷-3-醇单体物质的积累规律及其生物合成过程中相关酶活性、相关基因表达模式的变化。结果表明,(1)‘霞多丽’果实整个发育期,黄烷-3-醇主要单体物质没食子儿茶素(GC)、表儿茶素没食子酸酯含量(ECG)远高于儿茶素(CAT)、表儿茶素(EC)含量。ECG、EC 含量随着果实生育期均呈先升后降变化趋势,在花后50 d(7月13日)达最大值,分别为47.69和42.73 mg/g,而GC、CAT的含量则呈先降低后升高,后随果实成熟逐渐降低的变化趋势。(2)黄烷-3-醇生物合成相关酶PAL和ANS活性呈先升高后降低的变化趋势,F3H、F3"H、CHI活性均在花后20 d(6月13)出现峰值,而后活性迅速降低,DFR活性先升高后降低,花后50 d(7月13)再次升高,后随果实成熟逐渐降低。(3)黄烷-3-醇生物合成相关结构基因PAL、ANS 的表达水平随着果实生育期呈先上调后下调的趋势,与ECG、EC 含量变化趋势一致,F3"H、F3H、CHI 的表达水平却呈先下调后上调的趋势,DFR 的表达水平先上调,而后下调,随果实成熟再次上调。(4)相关性分析表明,F3H、F3"H和ANS酶活性与GC含量为显著正相关;F3H、F3"H 基因表达与GC、CAT的含量呈正相关,ANS、PAL 基因表达与ECG、GC的含量呈正相关。研究发现,黄烷-3-醇生物合成相关酶和相关基因表达影响了葡萄果实中黄烷-3-醇类化合物含量的积累,从而进一步影响葡萄酒的感官品质。

    Abstract:

    In order to explore the relationship between the accumulation of flavane-3-ol and the expression of synthesis-related enzymes and key genes in wine grape fruit, the white grape variety Chardonnay grown in Donglu of Helan Mountain was used as material by colorimetric method,high performance liquid chromatography (HPLC) and fluorescence quantitative PCR (FQ-PCR). The accumulation rule of flavane-3-ol monomer and the changes of related enzyme activity and related gene expression pattern during biosynthesis of grape fruit were studied.The results showed that: (1) During the whole fruit development period, the contents of Gallic catechin (GC) and epicatechin gallate (ECG) of flavane-3-ol were much higher than those of catechin (CAT) and epicatechin (EC). The contents of ECG and EC increased at first and then decreased with the fruit growth period, and reached the maximum at 50 days after anthesis, which were 47.69 mg/g and 42.73 mg/g, respectively, while the contents of GC and CAT decreased at first and then increased, and then decreased gradually with fruit ripening. (2) The activities of PAL and ANS related to flavane-3-ol biosynthesis increased at first and then decreased. The activities of F3H, F3"H and CHI peaked at 20 days after anthesis (June 13), and then decreased rapidly, and the activity of DFR increased at first and then decreased, increased again at 50 days after anthesis, and then decreased gradually with fruit ripening.(3)The expression level of structural genes PAL and ANS related to flavane-3-ol biosynthesis was up-regulated at first and then down-regulated with fruit growth period, which was consistent with the change trend of ECG and EC content, while the expression level of F3H, F3"H and CHI was down-regulated at first and then up-regulated. The expression level of DFR was up-regulated at first, then down-regulated, and then up-regulated again with fruit ripening. (4) Correlation analysis showed that the activities of F3H, F3"H and ANS were positively correlated with the content of GC.The expression of F3H and F3"H gene was positively correlated with the content of GC and CAT, and the expression of ANS and PAL gene was positively correlated with the content of ECG and GC. It was found that the synthase and gene expression related to flavane-3-ol biosynthesis affected the accumulation of flavane-3-ol compounds in grape fruit, which further affected the sensory quality of grape wine.

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  • 收稿日期:2021-09-08
  • 最后修改日期:2022-01-13
  • 录用日期:2022-01-14
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