烟草—TMV相互作用中质膜NADPH氧化酶的组装  被引量:4

Assembly of Plasma Membrane NADPH Oxidase in Tobacco TMV Interaction

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作  者:蔡以滢[1] 唐慧 张宏明[1] 陈珈[1] 

机构地区:[1]中国农业大学生物学院,北京100094

出  处:《中国生物化学与分子生物学报》1999年第4期600-600,共1页Chinese Journal of Biochemistry and Molecular Biology

基  金:国家自然科学基金;植物病虫害生物学国家重点实验室基金

摘  要:比较 N N 烟草(与烟草花叶病毒( T M V)发生非亲和相互作用)和普通烟草 3002 品种(与 T M V 发生亲和相互作用)在烟草— T M V 的相互作用中质膜 N A D P H 氧化酶的组装激活、产生活性氧的差异.用两相法制备密闭的正向型质膜( P M)囊泡,以 S O D 敏感的 N A D P H 依赖的 Cyt c 的还原表示 N A D P H 氧化酶的活性,用人类噬中性白细胞 N A D P H 氧化酶亚基 p47phox 的抗体对烟草叶片蛋白进行免疫学检测.结果显示在两种烟草叶片胞质中均存在与 p47phox 亚基的抗体发生免疫交叉反应的相同分子量的蛋白,该蛋白在 T M V 侵染 N N 基因烟草后可向质膜发生转移,且伴随有氧化酶活性的升高.而对于普通烟草则无氧化酶膜组分和酶活性的明显变化.以上结果表明,烟草叶片质膜上存在与哺乳动物 N A D P H 氧化酶相类似的氧化酶,它的组装和激活可能是烟草— T M V 非亲和相互作用早期活性氧的主要来源.Assembly and activity of the plasma membrane NADPH oxidase and its role in the production of reactive oxygen species (ROS) in tobacco TMV (tobacco mosaic virus) interaction were examined by using tobacco cv, Samsun NN (incompatible with TMV, containing the N gene for resistance to TMV) and tobacco cv. 3002(compatible with TMV) as test materials. Plasma membrane (PM) vesicles were isolated from leaves of tobacco by two phase aqueous system. The oxidase activity was assayed by the rate of SOD sensitive Cyt c reduction in PM system. The antibody raised against human neutrophil p47 phox was found to cross react with a 47 kD cytosolic protein from both tobacco leaves. Furthermore, the amount of p47 phox in the PM and its oxidase activity increased in cv. Samsun NN leaves after infection, but not in cv. 3002 leaves, indicating that TMV could promote the translocation of this cytosolic component to the PM of tobacco cells carrying the resistance gene. These results provide evidence for involvement of the protein related to the neutrophil oxidase complex in the tobacco TMV incompatible interaction.

关 键 词:NADPH氧化酶 超氧阴离子 烟草 TMV 质膜 

分 类 号:Q948.122.4[生物学—植物学] Q946.33

 

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