Development of Cell Suspension Culture System for Ampelopsis grossedentata Leaf and Its Production of Polyphenolic Compounds(显齿蛇葡萄叶细胞悬浮培养体系的建立及其产多酚类物质研究)
摘要
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<sup>+</sup> 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.(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