【作者】 冯真珍； 曹翠平； 邱海洪； 吴小锋； 鲁兴萌；

【Author】 FENG Zhen-Zhen CAO Cui-Ping QIU Hai-Hong WU Xiao-Feng LU Xing-Meng* (College of Animal Science of Zhejiang University,Hangzhou Zhejiang310029,China)

【机构】 浙江大学动物科学学院；

【摘要】 家蚕微粒子病是一种全身性寄生的蚕慢性疾病,其病原家蚕微孢子虫(Nosema bombycis,Nb)可通过胚种进行传染。随着昆虫细胞培养技术的发展,利用不同培养细胞研究病原微生物的侵染性成为研究病原侵染特异性的手段之一。本试验研究了Nb孢子经氢氧化钾发芽预处理,接种于鳞翅目昆虫—粉纹夜蛾(Trichoplusia ni,Tn)细胞系后,用电镜观察Nb孢子和Tn培养细胞的各种变化。结果发现Nb孢子经过氢氧化钾发芽预处理接种Tn培养细胞后,虽然可见Nb孢子在Tn培养细胞液中发芽,但并未观察到其孢原质进入Tn培养细胞。当Nb孢子的接种浓度达到2.0×105mL-1以上时,可以观察到Nb孢子通过内吞作用进入Tn培养细胞,但在Tn培养细胞内未见处于增殖阶段的Nb孢子。随着接种时间的延长和进入Tn培养细胞的Nb孢子数量的增加,可引起Tn培养细胞的超微结构发生了一些变化,细胞内有大空泡并出现较多溶酶体,线粒体和内质网结构减少,最终导致了Tn培养细胞的裂解。接种前后,Nb孢子数量也未有明显变化。即虽未见Nb以发芽形式感染Tn细胞系,但是能以吞噬途径进入Tn细胞系,并破坏其结构。据此推测Nb孢原质在进入不同寄主细胞(如Bm和Tn)的机制上不是简单的机械式注入,或其进入细胞后需要某些特定的因子和条件才能形成感染关系。更多还原

【Abstract】 Pebrine is one of chronic disease which parasitics in whole body of silkworm.its pathogen is Nosema bombycis,which can infect silkworm by semina.With the development of the technique of insect cells culture,people used different cultured cell lines to study and research the infected specificity of many pathogenic microorganism.In this test,we used electron microscope to study the changes of Nb spores and Tn cultured cells (belongs to Lepidoptera insect) after inoculating Nb spores which dealed with KOH pre-disposal.As the result,the process of germination of Nb spores had been observed in Tn cultured cells medium with the method of KOH pre-disposal,but the sporoplasm had not been observed in Tn cultured cell line.Nb spores had invaded into Tn cultured cells when inoculated concentration was more than 2.0×105 mL-1,but the process of propagation of Nb spores had not been observed in Tn cultured cells.As time went on and the quant of Nb spores which in Tn cultured cells were increased,the ultrastructure of Tn cultured cells were changed,there are big vacuoles and many lysosomes in the cytoplasm,and mitochondria and endocytoplasmic reticulum are decreased,these caused the disruption of Tn cultured cells at last.In addition,the quant of Nb spores had no remarkab rise.In a word,Nb spores can not infect the Tn cultured cells by the way of sprouting,but can be phagocytosed by Tn cultured cells and destroyed the cytoarchitectonics.So we presumed that the mechanism which Nb sporoplasm invade to host cells is not the simple injection,or Nb sporoplasm need special factors and conditions to infect host cells.更多还原