芜菁花叶病毒外壳蛋白在寄主植物叶绿体中的积累及其对光系统II活性的影响  

Accumulation of Coat Protein of Turnip mosaic virus in Host Chloro-plasts and Its Effect on PS II Activity

作　　者：付东亚[1] 洪健[2] 陈集双[1] 吴建祥[2] 

机构地区：[1]浙江大学生命科学学院,杭州310029 [2]浙江大学生物技术研究所,杭州310029

出　　处：《植物生理与分子生物学学报》2004年第1期34-40,共7页Journal Of Plant Physiology and Molecular Biology

基　　金：国家自然科学基金项目(No.39970067)资助。~~

摘　　要：分别以接种感染芜菁花叶病毒(TuMV)和健康对照的青菜苏州青品种(Brassica chinensis L. cv.Suzhou)和芥菜温州芥菜品种(B.juncea L. cv.Wenzhou)叶片为材料提取完整叶绿体,用胰蛋白酶消除其表面蛋白后,抽提总蛋白,经SDS-PAGE电泳和Western blot检测,发现TuMV的外壳蛋白(CP)存在于感病寄主的叶绿体中。免疫金标记电镜实验显示TuMV-CP定位在感病青菜和芥菜的细胞质和叶绿体中。对两种寄主植物叶片的叶绿素荧光动力学参数测定结果显示,青菜、芥菜的Fv/Fo、Fv/Fm、φPSII、qp值都有不同程度的降低,qN值增大。实验结果表明TuMV侵染后在寄主细胞叶绿体中积累的CP,抑制了光系统II(PS II)的光化学活性,这可能是影响寄主植物光合作用的一个重要原因。Intact chloroplasts were isolated fromChinese cabbage (Brassica chinensis L. cv.Suzhou) and mustard (B. juncea L. cv. Wenzhou)leaves inoculated with Turnip mosaic virus(TuMV). The proteins attached to the surface ofchloroplasts were digested, then the total pro-teins of chloroplasts were extracted. By analysisof the SDS-PAGE pattern, one extra protein bandwith the same mobility and molecular weight asTuMV-coat protein was found in the chloroplastsof infected leaves (Fig.1). Results of Westernblotting showed that this band was highly reac-tive to the antiserum against TuMV-CP (Fig.2).The detection of immunogold-particles alsoshowed the presence of TuMV-CP in chloroplastsof both host plant species as compared with thehealthy control plant (Fig.4-14). Electron mi-croscopic photograph showed some virus par-ticles and cylindrical inclusions in the cytoplasmof host plants(Fig.3-1, 2). Some chloroplastsshowed abnormalities, including inhibited lamel-lar development, swollen and rounded, mem-brane vesiculation and disintegration (Fig.3-3, 4).On the tenth day after TuMV inoculation, Fv /Fo, Fv/ Fm, 諴SII and qP values all became lower,but the qN values were higher, as compared withthe healthy control plants(Table 1). The resultsprovided the evidence for the presence of TuMV-CP in the diseased chloroplasts. It suggests thatthe accumulation of TuMV-CP inside chloro-plasts affect photosynthesis in virus-infectedplants by inhibiting the PS II activity.

关 键 词：芜菁花叶病毒 外壳蛋白 叶绿体 光系统Ⅱ 

分 类 号：S432.41[农业科学—植物病理学]