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,该研究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,分析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)在‘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(ECG)含量远高于儿茶素(CAT)、表儿茶素(EC)含量,且ECG、EC含量均随果实发育期进程呈先升后降变化趋势,并于花后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)葡萄果实中黄烷-3-醇生物合成相关酶PAL和ANS活性随果实发育期进程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,而DFR活性呈先升高后降低,并于花后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的表达水平随果实发育期进程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,并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含量呈显著正相关关系;F3H、F3′H基因表达与GC、CAT的含量呈正相关关系,ANS、PAL基因表达与ECG、GC的含量呈正相关关系。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。
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,this study analyzed the white grape variety Chardonnay grown in Donglu of Helan Mountain with 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 activities and related gene expression patterns 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 development of fruit,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 flavane-3-alcohol biosynthesis related enzymes PAL and ANS in grape fruits increased at first and then decreased with the development of fruit.The activities of F3H,F3′H and CHI peaked at 20 days after anthesis(June 13),and then decreased rapidly,while 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 of structural genes PAL and ANS related to flavane-3-ol biosynthesis was up-regulated at first and then down-regulated with the development of fruit,which was consistent with the change trend of ECG and EC contents,while the expression of F3H,F3′H and CHI was down-regulated at first and then up-regulated.The expression 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 genes was positively correlated with the contents of GC and CAT,and the expression of ANS and PAL genes was positively correlated with the contents 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.
作者
沙晓蓉
张萍
王云霞
卜虎柏
马莹
靳磊
SHA Xiaorong;ZHANG Ping;WANG Yunxia;BO Hubai;MA Ying;JIN Lei(Agricultural College of Ningxia University, Yinchuan 750000, China)
出处
《西北植物学报》
CAS
CSCD
北大核心
2022年第2期263-271,共9页
Acta Botanica Boreali-Occidentalia Sinica
基金
宁夏自然科学基金(2020AAC03087)。