(1.江西农业大学 食品科学与工程学院/江西省天然产物与功能食品重点实验室, 江西 南昌 330045;2.广东省农业科学院 蚕业与农产品加工研究所/农业农村部功能食品重点实验室/ 广东省农产品加工重点实验室, 广东 广州 510610)
(1.College of Food Science and Engineering/Jiangxi Key Laboratory of Natural Products and Functional Foods, Jiangxi Agricultural University, Nanchang 330045, China;2. Sericultural & Agri-Food Research Institute, Guangdong Academy of Agricultural Sciences/ Key Laboratory of Functional Foods, Ministry of Agriculture and Rural Affairs/Guangdong Key Laboratory of Agricultural Products Processing, Guangzhou 510610, China)
显齿蛇葡萄叶富含多酚类化合物,是一种具有很高开发利用价值的新资源食品原料。以显齿蛇葡萄幼叶为外植体诱导出了2种典型的愈伤组织,通过激素优化和连续筛选获得了性状优良的愈伤组织,在此基础上建立并优化了显齿蛇葡萄叶细胞悬浮培养体系,对悬浮培养细胞中的主要多酚类物质进行了鉴定,并评价了多酚的抗氧化活性。采用超高压液相色谱飞行时间质谱 (UPLC-Q-TOF MS/MS)和高效液相色谱 (HLPC)检测技术,分别从愈伤组织和悬浮培养细胞中鉴定出了7种和10种多酚类化合物,其中原花青素B1、(+)-儿茶素、表儿茶素没食子酸酯为愈伤组织和悬浮培养细胞中共有的物质,且含量较高,其中以表儿茶素没食子酸酯含量最高,在愈伤组织和悬浮培养细胞中分别达到了5020.965μg/g和1044.725μg/g。抗氧化活性评价结果显示,悬浮培养细胞多酚提取物具有很强的抗氧化活性,对2,2-联氮-二 (3-乙基-苯并噻唑-6-磺酸)二铵盐 (ABTS+)和2,2-联苯基-1-苦基肼基 (DPPH)自由基清除的IC50分别为30.681μg/mL和3.685μg/mL,相同质量浓度下对DPPH自由基的清除和总还原能力均强于维生素C,其抗氧化能力主要源于表儿茶素没食子酸酯和(+)-儿茶素。希望研究可为显齿蛇葡萄细胞培养及其多酚类次生代谢产物的合成调控提供参考。
Ampelopsis grossedentata leaf is rich in polyphenols, and now is regarded as a new food material with high value for utilization. Two typical kinds of calluses were induced from Ampelopsis grossedentata young leaves. The excellent calluses were successfully obtained by hormone optimization and successive screening, which were further used to develop the cell suspension culture system of Ampelopsis grossedentata leaf. The main polyphenols in the suspensively cultured cells were identified, and their antioxidant activities were evaluated in the meantime. Seven polyphenols were identified in the extracts of calluses by the UPLC-Q-TOF MS/MS and HLPC detection, while ten polyphenols were identified in the suspensively cultured cells. Three polyphenols including proanthocyanidin B1, (+)-catechin, epigetechin gallate were found both in calluses and cells with high concentration, among which epicatechin gallate was the most abundant compound, whose contents reached 5020.965μg/g and 1044.725μg/g in calluses and cells, respectively. In addition, total polyphenols extracts from the suspensively cultured cells displayed strong antioxidant activities, with IC 50values of 30 681 μg/mL and 3.685μg/mL respectively in the ABTS+ and DPPH free radical scavenging tests. What's more, both the DPPH radical scavenging and total reduction ability of the polyphenol extracts were significantly stronger than vitamin C under the same mass concentration, which were mainly attributed to epicatechin gallate and (+)-catechin. It was hoped that this study would provide a reference for the culture of Ampelopsis grossedentata leaf cells and the regulation of its polyphenolic secondary metabolite synthesis.