漆酶诱导北美沙地葡萄抗毒素Res合成及代谢研究  

Biosynthesis and Metabolism of Phytoalexin Resveratrol Induced by Laccase in Vitis rupestris Cell of North American

作　　者：常小丽[1] 龚国淑[1] 廖建霞[1] 戴浩[1] 郑晓琴[2] 崔永亮[2] 张翠苹[1] 

机构地区：[1]四川农业大学农学院,成都611130 [2]四川省自然资源科学研究院,成都610015

出　　处：《西北农业学报》2015年第4期105-113,共9页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：四川农业大学优秀引进人才启动金项目;四川省教育厅项目(13ZB0278)

摘　　要：为明确真菌漆酶(laccase)对北美沙地葡萄Vitis rupestris抗毒素白藜芦醇(Resveratrol)合成及代谢的影响,采用高效液相色谱(HPLC)法,对白腐菌云芝漆酶与白藜芦醇的体外催化反应条件及产物进行研究;并通过病原菌harpin蛋白预诱导抗病沙地葡萄细胞悬浮系后,采用漆酶处理或非处理,检测白藜芦醇合成及代谢情况。试验结果显示,漆酶在醋酸缓冲液pH 4.5、25℃、150r·min-1、黑暗条件下,与白藜芦醇催化反应2h,可将白藜芦醇氧化形成二聚体葡萄素(δ-viniferin)。漆酶作为一种真菌致病因子能诱导Vitis rupestris细胞合成反式白藜芦醇(trans-resveratrol),并迅速将其氧化为抗性更强的δ-viniferin。同时,漆酶能与病原菌致病蛋白harpin协同诱导葡萄白藜芦醇合成及代谢,以提高沙地葡萄抗性。For studying the influence of laccase fromTrametes versicolor on the biosynthesis and metabolism of the phytoalxin resveratrol in cell cultures of North American Vitis rupestris,high performance liquid chromatography（HPLC）was used to study the condition of catalysis reaction and products of laccase.Resveratrol were investigated in vitro,and accumulation of resveratrol and its metabolic derivatives in cell culture of resistant Vitis rupestris were examined under treatment of pretreated with harpin,and then supplemented with or without laccase,or harpin,or harpin followed by laccase in vivo.We found that laccase was capable of catalyzing resveratrol into its dimmer-viniferin in acetate buffer（pH 4.5）at 25℃,150r·min-1 on a rotatory shaker in the dark in vitro.As a class of fungical virulence factor,application of laccase alone induced accumulation of trans-resveratrol and its toxic oxidized dimmer in Vitis rupestris cell.Meanwhile,there was a synergistic effect of laccase and the bacterial harpin protein on activating the resistance level of Vitis rupestris in terms of the phytoalexin resveratrol and its metabolism.This is very significant for further uncovering the role of laccase in the grapevine-pathogen interaction mechanism.

关 键 词：漆酶 沙地葡萄 白藜芦醇 HPLC 葡萄素 HARPIN蛋白 

分 类 号：S663.1[农业科学—果树学]