感染HaSNPV棉铃虫幼虫的组织病理和免疫组化研究  

Pathogenic and Immunohistochemicai Studies of the HaSNPV Infected Helicoverpa armigera Larvae

作　　者：欧阳志荃[1,2] 孙修炼[1,2] 邓菲[1,2] 袁丽[1,2] 胡志红[1,2] 

机构地区：[1]中国科学院武汉病毒研究所分子病毒学重点实验室,湖北武汉430071 [2]中国科学院武汉病毒研究所无脊椎动物病毒学联合开放实验室,湖北武汉430071

出　　处：《中国病毒学》2003年第6期603-606,T003,T004,共6页Virologica Sinica

基　　金：国家杰出青年基金(3002503);中国科学院创新工程重要方向性项目(kscx21-02;kscx-sw-301-09);863项目(2001AA214031:101-06-10-01);国家973计划(2003CB114202)

摘　　要：本文报道了棉铃虫单核衣壳核多角体病毒(Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus,HaSNPV)感染棉铃虫幼虫的病理时相及HaSNPV多角体蛋白在幼虫组织中表达的免疫组化研究.以5×103PFU的HaSNPV出芽病毒粒子(BV)注射4龄初的棉铃虫幼虫,石蜡切片的H.E染色表明,在感染后24h,病理变化不明显;48h后脂肪体、气管组织的细胞核开始肿大,细胞开始变形;72h后脂肪体、气管、真皮细胞核肿大十分明显,组织结构松散;但中肠和肌肉组织未见明显病变;96h后,脂肪体、气管、真皮组织结构完全被破坏,肌肉组织变疏松.免疫组化结果表明,感染后24h,未能检测到多角体蛋白在幼虫组织中的表达;感染后48h,多角体蛋白可在部分脂肪体、气管组织、血细胞和真皮组织的细胞核中表达;72h后,被感染的脂肪体、气管组织、和真皮组织的细胞数比48h多; 96h后,多角体蛋白可以在脂肪体、气管、真皮组织中大量表达,48h到96h间,被感染的血细胞数目基本不变,中肠组织和肌肉都未检测到多角体蛋白的表达;幼虫死亡后,可在中肠上皮细胞的基底膜间隙中检测到多角体蛋白的表达,肌肉组织中未见表达信号,其它组织全部被感染并且组织结构被破坏, H.E的结果与免疫组化的结果基本相符.Here we report the pathogensis process of Helicoverpa armigera larvae infected with Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HaSNPV) using H.E staining and immunohistochemistry with antibody against polyhedrin. The fourth instar Helicoverpa armigera larvae were injected with 5×103 PFU HaSNPV per larva. H.E staining showed that no apparent pathogensis could be detected in all tissues at 24 hours post infection (hpi). But at 48 hpi, the nucleus of the fat body and trachea epidermis became swollen, and the shape of the cells in these tissues were changed. At 72 h p i, significant hispathogensis were observed in fat body, trachea epidermis and the epidermis, while no changes were found in midgut and muscles. At 96 hpi, the constitution of the fat body, trachea and epidermis were destroyed. The immunohistochemistry results indicated the polyhedrin protein could not be detected at 24 hpi. At 48 hpi, it was found partially expressed in the tissues of fat body, trachea epidermis and epidermis. From 72 to 96 hpi, the polyhedrin was found heavily expressed in fat body, trachea epidermis and epidermis. No polyhedrin was found expressed in midgut untill larvae dead and the muscule was not be infected all the time.

关 键 词：棉铃虫核多角体病毒 病理时相 多角体蛋白 免疫组化 

分 类 号：S476[农业科学—农业昆虫与害虫防治]