8个葡萄品种的花色苷组分及含量分析

Analysis of anthocyanin compositions and contents in eight grape cultivars

【目的】了解不同葡萄品种的花色苷组分特征。【方法】采用高效液相色谱-质谱(HPLC-MS)法对8个特色葡萄品种成熟期果皮花色苷组分及含量进行测定分析。【结果】不同品种的果皮花色苷质量分数以‘摩尔多瓦’最高,为8 131.08 mg·kg^(-1),其次是‘蜜萄斯’‘阿考龙’‘福客’,为7 016.25~7 546.66 mg·kg^(-1),而‘天赐’‘赤霞珠’的花色苷较低,分别为2 608.59和2 442.87 mg·kg^(-1),且均由二甲花翠素类、甲基花翠素类、花翠素类,以及花青素类、甲基花青素类花色苷组成,但不同葡萄品种单体花色苷所占比例不同。二甲花翠素类花色苷是所测品种的主要花色苷,其所占比例最高,达80.76%。‘赤霞珠’与‘天赐’、‘蜜萄斯’与‘阿考龙’、‘早巨峰’与‘摩尔多瓦’的花色苷组成比例相似度较高,而‘贝克’及‘福客’花色苷组成与上述品种差异较大。不同品种的花色苷修饰程度以‘贝克’最高,为99.41%,其次是‘摩尔多瓦’及‘天赐’,分别为96.51%、94.60%,‘福客’修饰程度最低,为81.20%。‘赤霞珠’‘天赐’‘蜜萄斯’与‘阿考龙’不含双葡萄糖苷,属欧亚种葡萄,而‘早巨峰’‘摩尔多瓦’‘福客’及‘贝克’主要为双葡萄糖苷花色苷,不属于欧亚种葡萄。通过主成分及聚类分析,‘蜜萄斯’与‘阿考龙’、‘赤霞珠’与‘天赐’最为接近,‘早巨峰’‘贝克’‘摩尔多瓦’‘福客’花色苷组分关系则较远。【结论】不同葡萄品种单体花色苷所占比例有明显差异,二甲花翠素类花色苷是所测品种的主要花色苷,大部分欧亚种葡萄中含量次高的花色苷是甲基花青素类,而欧美杂交种葡萄中则是甲基花翠素类;‘摩尔多瓦’‘蜜萄斯’及‘阿考龙’花色苷含量及修饰程度均较高,具有染色品种的功能。

[Objective] The content and composition of anthocyanins is an important determinant of the color and functional value of grape berries and wine, and thus is crucial for their market value. In order to de- termine the anthocyanin content and composition in different grape varieties for breeding table grape or wine grape varieties with rich anthocyanins, we introduced some varieties with deep-color skin and good resistance in recent years. [ Methods ]The test was carried out in a demonstration vineyard in Shandong Ag- ricultural University, where a warm temperate semi-humid continental monsoon climate dominates. The vineyard was established in 2011, the vines were planted with vertical trellis systems, in a north-south ori- entation, with 1.2 m between plants, 2.2 m between rows and 2 m in height. The training system was spur- pruned vertical shoot-positioned （VSP）, and all the grapes were under the same cultivation management. The tested materials included Vitis vinifera ＇ Cabernet Sauvignon＇, ＇ Manna＇ （a hybrid of ＇ Cabernet Sauvi- gnon＇）, ＇ Cabernet Mitos＇, ＇ Acolon＇, V. vinifera- V. labrusca ＇ Kyoho Early＇, ＇ Moldova＇ （＇ Guzal kara＇ × ＇ SV. 12-375 ＇）, ＇ Frontenac＇ （＇ Riparia 89＇ × ＇ Landot4511＇）, ＇ Becker＇ （an interspecific species with parents unknown, bred by Becker and presented by Hance）. For each variety, plants with uniform vigor and size were selected, and 3 plants per plot with replicates （totally 9 plants） were used for the study. Four ma- ture berry clusters born in the middle shoots were taken from every plot （12 clusters in total） and all the berries from the same plot were peeled, pooled and rapidly frozen in liquid nitrogen for measuring anthocy- anins. Extraction of anthocyanins was conducted using the method of Zhaoquan. The HPLC-MS analyses were carried out using a WATERS ACQUITY UPLC-PDA detector. Chromatographic separation was car- ried out using a 100 mm ×2.1 mm i.d. thermostated at 45℃. The mobile phase was a linear gradient of wa- ter/formic acid （98 ： 2） （solvent B） in acetonitrile （solvent A）, at a flow rate of 0.3 mL. min-1. The following gradient was used： 0-20 min, 6%-16% A; 20-28 rain, 16%-23% A; 28-35 min, 23%-50% A; 35-37 min, 50% A; 37-40 min, 50%-60% A; and MS analyses. MS operating parameters were： capillary voltage, 3 000 V; cone voltage, 30.0 V; cone gas flow, 10 L·h^-1; desolvation gas flow, 700 L·h^-1; ion source temperature, 100 ℃; desolvation temperature, 400 ℃. The instrument was operated in a positive ion mode scanning from m·z^-1 50 to 2 000. [ Results ]The total anthocyanin content in ＇Moldova＇ skin was the highest （8 131.08 mg·kg^-1）, followed by ＇Cabernet Mitos＇, ＇ Acolon＇ and ＇ Frontenac＇ in a range of 7 016.25- 7 546.66 mg·kg^-1. It was the lowest in ＇Manna＇ and ＇Cabernet Sauvignon＇, being 2 608.59 and 2 442.87 mg＂ kg^-1, respectively. Malvidin, petunidin, delphindin, cyanidin, peonidin, and their derivatives could be detected in all varieties tested, although their proportions were different among varieties. Malvidin and its derivative malvidins were the main components in all varieties, accounting for 80.76%. Anthocyanin com- position was similar between ＇Cabernet Sauvignon＇ and ＇Manna＇, ＇Cabernet Mitos＇ and ＇Acolon＇, ＇ Kyoho Early＇ and ＇ Moldova＇, while ＇ Becker＇ and ＇ Frontenac＇ had an anthocyanin composition which was different with the others. ＇Becker＇ had the highest modification degree of the anthocyanins, which reached 99.41%, followed by ＇ Moldova＇ and ＇Manna＇ with a modification degree of 96.51% and 94.60% respectively. ＇Frontenac＇ had the lowest modification degree of 81.20% only. The result also showed that V. vinifera varieties ＇ Cabernet Sauvignon＇, ＇ Manna＇, ＇ Cabernet Mitos＇ and ＇ Acolon＇ did not contain diglucosides, while ＇Kyoho Early＇, ＇Moldova＇ , ＇Frontenac＇ and ＇Becker＇ contained much diglucosides, indicating that they did not belong to V. vinifera. The principal component and cluster analyses indicated that ＇ Cabernet Mitos＇, ＇ Acolon＇, ＇ Cabernet Sauvignon＇ and ＇ Manna＇ were closer in relationship, but ＇Kyoho Early＇, ＇Moldova＇, ＇Frontenac＇ and ＇Becket＇ had a distant relationship based on the composition of anthocyanins. [Conclusion] Different varieties differ significantly in anthocyanin content and composition in their skin. Methylation is the main modification form of anthocyanins. ＇Moldova＇, ＇Cabernet Mitos＇, ＇ Acolon＇ and ＇ Frontenac＇ have a high content of total anthocyanins, but a high degree of modifi- cation of anthocyanins occurs in ＇Becker＇, ＇Moldova＇ and ＇Manna＇. Malvidins are the main components in all grape varieties, and peonidin and its derivatives （peonidins） are the second most abundant anthocy-anin components in most V. vinifera, while V. vinifera×V, labrusca＇s is dominant by petunidin and its derivatives （petunidins）. ＇Moldova＇, ＇Cabernet Mitos＇ and ＇Acolon＇ can be used as a source of dyes, as they have a higher anthocyanin content with a higher modification degree. Therefore they can be cultivated in large scale as important pigmented varieties. Anthocyanin composition analysis can be an effective method to classify grape cultivars.