【作者】 余兴；

【导师】 高丽萍；

【作者基本信息】 安徽农业大学 ， 果树学， 2005， 硕士

【摘要】 白藜芦醇是葡萄在抵御真菌侵染、机械和紫外线等伤害时产生的植保素次生代谢产物,对人类健康特别是预防心血管疾病等方面也具有显著功效。 本试验以酿酒葡萄品种(系)梅鹿特(Merlot)、巴柯(Bacol Noir)、烟 73(yan73)、蛇龙珠(Carbernet Gernischet)、白羽(Rkatsiteli)为材料,系统研究了生长发育过程中白藜芦醇及其糖苷的变化与相关酶活性间的关系。同时探讨了紫外线处理对果实的白藜芦醇及其相关酶活性的影响。主要研究结果如下: 1. 利用高效液相色谱(HPLC)建立了葡萄中白藜芦醇及其糖苷的测定方法。等度洗脱条件 I 适合于反式白藜芦醇的测定,而等度洗脱条件 II 和梯度洗脱适合于顺、反式白藜芦醇及其糖苷的分析。同时建立了顺式白藜芦醇及其糖苷的测定方法。 2. 通过正交试验,确定了白藜芦醇样品的最佳提取条件,即 60%甲醇提取 6 h,SPE 小柱除杂质,50%甲醇溶解样品,试验结果表明该方法回收率高。 3. 生长发育期间,不同品种果实的白藜芦醇及其糖苷含量的变化趋势不同,其中蛇龙珠果实的白藜芦醇及其糖苷含量变化呈双峰形态;白羽的白藜芦醇苷随果实生长而逐渐降低,而白藜芦醇缓慢增加。 4. 首次研究了葡萄果实生长发育过程中苯丙氨酸解氨酶(PAL),β-葡萄糖苷酶与白藜芦醇及其糖苷之间的关系,结果表明果实生长发育过程中白羽的 PAL 活性较低,且随着果实的生长发育而逐渐降低,而蛇龙珠的 PAL 活性呈现出明显高峰。PAL 和果实中的白藜芦醇的形成存在一定的相关性。蛇龙珠、白羽果实中β-葡萄糖苷酶的活性高峰均出现在生长发育初期,并随果实的生长发育而迅速下降。β-葡萄糖苷酶可能与白藜芦醇苷水解转变成白藜芦醇有关。果实生长发育过程中未检测到过氧化物酶(POD)的活性。 5. 研究了紫外线(UV)辐射对葡萄中白藜芦醇合成的诱导作用,研究表明 UV能诱导果实中 PAL、POD 酶的产生,UV 的适宜辐射剂量为 200 s,UV 辐射后在 30℃下贮存 48 h,可使白藜芦醇含量提高 7 倍。UV 处理后未检测到反式白藜芦醇苷和β-葡萄糖苷酶活性。更多还原

【Abstract】 Resveratrol (trans-3, 4’, 5-trihydoystilbene), as a phytoalexin, exists in berry peelof most grapes cultivars and its synthesis was stimulated by the UV ray, fungal infectionand mechanical injury. Recently, more and more researches proved that resveratrol hadmany beneficial effects on human health, such as reducing the risk of coronary heartdisease, inhibiting platelets aggregation and low-density lipoproteins oxidation. The relationships between the changes of resveratrol, piceid and relevant enzymaticactivities during the development of grape were studied, taking the fruits of Merlot, BacolNoir, Yan73, Carbernet Gernisch Rkatsiteli grapes as experimental material. The effect ofUV treatment on the changes of resveratrol and piceid and relevant enzymatic activities inpost-harvest grape was discourvered. The main results gained in this study were asfollows: 1. The testing methods of resveratrol and piceid in grape were determined by meansof high performance liquid chromatography(HPLC). The isocratic elution condition Iwas suitable to trans-resveratrol test, and both isocratic elution condition II and gradientelution condition were fit methods for trans-resveratrol test, cis-resveratrol and its piceid.The testing method of cis-resveratrol, trans-resveratrol and piceid was determined too. 2. The optimum extraction condition of resveratrol in grape with a high recovery wasdetermined by orthogonal design, i.e. sample was extracted with 60% methanol for 6 h,purified through a solid-phase extraction column, then dissolved in the solution of 50%methanol. 3. The trends of resveratrol and piceid chages in different grape cultivars weredifferent during the development of fruit. The chages of resveratrol and its piceid contentin Carbernet Gernischet was a curve of double peaks. The content of resveratrol inRkatsiteli slowly decreased, whereas its piceid gradually increased during thedevelopment of fruit. 4. It was frist time that the relationships among resveratrol and its piceid andphenylalanineammonia-lyase(PAL), and β-glucosidase, had been studied during thegrowth and development of grape. The PAL activity in Rkatsiteli was lower, anddecreased gradually as the fruit grew, which in Carbernet Gernischet increased andappeared an obvious peak in the chromatogram. There was a positive correlation betweenPAL activity and the formation of resveratrol. The β-glucosidase activity in CarbernetGernischet and Rkatsiteli was at high level in the early stage of the fruit development andthen decreased rapidly. It was postulated that this kind of chage of β-glucosidase probablyattributed to the enzymatic hydrolysis of piceid indican into resveratrol. The Peroxidase(POD) activity hadn’t been tested during the fruit development of grape. 5. The enzymatic activities of PAL and POD could be stimulated by UV radiation ingrape. In the treatment of Carbernet Gernischet by UV radiation for 200 s and thenstoring for 48 h at 30℃, the maximum concentrations of resveratrol could be increase upto seven times of that in the contrast. But β-glucosidase activity and piceid had not beenchecked in grape after the treatment of UV radiation.更多还原