银杏雄株叶片类黄酮糖基转移酶活性分析  

Study on flavonoid glycosyltransferases activity in leaves of Ginkgo' male plants

作　　者：沈丹红[1] 陈鹏[1] 蒋菲[1] 马江黎 王金艳[1] 戴燕[1] 

机构地区：[1]扬州大学园艺与植物保护学院,扬州225009

出　　处：《果树学报》2014年第1期78-84,共7页Journal of Fruit Science

基　　金：国家农业综合开发项目(GH32311002);江苏省林业三项工程项目(lxsx[2011]16);江苏省林业三新工程重大项目(lysx[2013]05)

摘　　要：【目的】为银杏黄酮糖苷的生物合成代谢研究及其叶用资源评价和优选提供新的技术。【方法】根据UFGT催化黄酮苷元槲皮素与UDPG反应生成槲皮素糖苷和UDP的原理．建立经试验筛选得到的最佳pH、温度、底物浓度和反应时间组成的优化体系，采用反相离子对色谱法测定银杏雄株不同生长时期叶片的UFGT酶活性。【结果】反应产物UDP线性为12．5—300mg·L^-1（r=O．9994），回收率为98．76％，RSD为1.3％（n=5）；建立的UFGT活性测定优化体系为：pH=8．0、反应温度30℃、槲皮素浓度400μmol·L^-1、反应时间4h：银杏雄株叶片5—10月间UFGT活性变化表现为由5月份的0．2μmol·L^-1·min^-1逐渐上升，到9月份达到最大值，为0．62μmol·L^-1。【结论】该技术简便、准确、重复性好。可推荐供银杏黄酮糖基转移酶活性测定参考应用．银杏叶用雄株9月份采叶可获得较高的黄酮糖苷含量。[Objective]This research was conducted to provide a new technique for the research on biosyn- thetic metabolism of flavonoid glycoside and evaluation and selection of leaf-purpose resources in Ginkgo biloba L. [Method ]Based on the principle that UFGT catalyses the reaction of quereetin with UDP-glueose which produces quercetin glycoside and uridine diphosphate, the optimized system consisted of the suit- able pH, temperature, substrate concentration and reaction time. UFGT activities in leaves of male plants during different growth periods were determinated by reversed phase ion pair chromatography method. [Result]The results showed that UDP, a reactive product,had a significant linear relationship within 12.5-300 0μmol·L^-1（r=0.9994）, recovery rate of 98.76% and RSD = 1.3%（n=5）; the optimized system for determination of UFGT activity was： pH =8.0, reaction temperature 30 ℃, quercetin concentration 400μm and reaction time 4 hours; the UFGT activities in leaves of male plants gradually increased from 0.2 μmol·L^-1·min^-1 in May to 0.62μmol·L^-1·min^-1 in September when the peak appeared. [Conclusion]The above method is simple, accurate and reproducible, and could be for the detection of UFGT activity. Sam- piing in September could get a higher content of flavonoid glycoside in leaves of male plants of Ginkgo.

关 键 词：银杏雄株 叶片 UFGT活性 

分 类 号：S664.3[农业科学—果树学]