月季花瓣中类黄酮和类胡萝卜素的提取与测定

Extraction and Determination of Flavonoids and Carotenoids in Petals of Roses (Rosa spp.)

月季(Rosa spp.)是一种具有重要价值的观赏植物,花色是其最重要的观赏性状之一。为了解析月季花色形成的分子机理,本研究以‘云蒸霞蔚’和‘太阳城’的杂交后代为试验材料,探索类黄酮和类胡萝卜素的提取与测定方法,并且对‘梅朗口红’花瓣中类黄酮和类胡萝卜素进行定性定量分析。结果表明:(1)类黄酮提取的最佳浸提剂为甲醇-水-甲酸-三氟乙酸(70∶27∶2∶1, V/V),采用超声波辅助提取,流动相为0.5%甲酸水溶液(A)和乙腈(B),洗脱梯度为0 min 5%B,5 min 10%B,30 min 19%B,50 min 40%B,50.01~60 min 5%B;(2)类胡萝卜素提取的最佳浸提剂为甲醇(含0.1%BHT),需要皂化,最佳复溶溶剂为甲醇-甲基叔丁基醚(1∶1,V/V,含0.1%BHT);(3)‘梅朗口红’花瓣中总共检测到1种花青苷、8种黄酮醇和6种类胡萝卜素,其中花青苷含量最高,黄酮醇次之,类胡萝卜素最低,开花过程中色素含量变化显著,矢车菊3,5-双氧葡萄糖苷的含量先升高后降低,在S3时期,最大值为1 705.90μg/g FW,槲皮素3-O-鼠李糖苷、槲皮素总含量以及黄酮醇总含量先升高后降低,在S2时期达到最大值,槲皮素7-O-鼠李糖苷和山奈酚总含量逐渐下降;(4)相关分析显示,明度L*与花青苷含量极显著负相关,色相a*与花青苷含量显著正相关。本研究不仅为月季花色的分子育种提供帮助,同时也为月季花瓣中功能物质的开发提供借鉴。

Rose is one of the most economically significant ornamental crops, and flower color is one of the most important trait. In order to illuminate the molecular mechanism of the formation of rose flower color, progenies from ’Yunzheng Xiawei’ and ’Sun City’ were used to explore the methods of extraction and determination of flavonoids and carotenoids. The composition of flavonoids and carotenoids were further determined in petals of red modern rose cultivar ’Rouge Meilland’. Results showed that:(1) The optimal solvent for the flavonoids extraction from rose petals was methanol:water:formic acid:trifluoroacetic acid(70∶27∶2∶1;V/V) and ultrasonic assisted extraction was more effective. The mobile phases comprised 0.5% aqueous formic acid(A) and acetonitrile(B) with the following gradient: 0 min, 5% B, 5 min, 10% B, 30 min, 19% B, 50 min, 40% B, and 50.01~60 min, 5% B.(2)The optimal solvent for the carotenoids extraction from rose petals was methanol(0.1% BHT). Saponification displayed better separation of carotenoids. The best solvent of carotenoid dry powder was methanol:methyl tertiary butyl ether(1∶1, V/V, with 0.1% BHT).(3) In total, 1 anthocyanin, 8 flavonols and 6 carotenoids were detected and identified. With regard to the concentration, anthocyanin was the highest, flavonol was next and carotenoid was the lowest. Signifficant changes were observed in their proffiles during seven blooming stages. The anthocyanin contents increased firstly and then decreased, and was highest at S3, which was 1 705.90 μg/g FW. The content of quercetin 3-O-rhamnoside, total quercetins and total flavonols increased firstly and then decreased, and was highest at S2. The content of quercetin 7-O-rhamnoside and total kaempferols showed a downward trend.(4)The results of correlation analysis showed that, anthocyanin content had great significantly negative correlation with L*, and had significantly positive correlation with a*. The results of this study not only can be helpful for the breeding of flower color of rose, but also will provide referential for the development of functional compounds in rose petals.