| 摘要: |
| 运用透射电镜对异常死亡文蛤的细胞病理变化及感染的病毒粒子进行了超微结构观察,并通过差速离心、酒石酸钾超速离心和氯化铯密度梯度离心结合等方法对病毒粒子分离纯化。超微结构观察结果显示,患病个体的鳃、消化盲囊、消化道和外套膜均大量存在一种病毒颗粒,该病毒呈球形,无囊膜,直径45～55nm,核心与核衣壳间无明显界限,未发现病毒包涵体。切片结果显示该病毒存在于细胞间质及被破坏的细胞质中,经纯化后负染显示,该病毒粒子呈球形,二十面体对称,无囊膜,直径在60～75nm,表面结构粗糙,壳粒结构较明显。超薄切片观察表明,由该病毒粒子的感染引起的细胞病理变化主要表现为：细胞核染色质集聚,溶酶体增多,线粒体肿涨,粗面内质网膨胀,肌原纤维排列混乱,细胞结构变得无规则直至解体。比较了蔗糖、酒石酸钾-甘油与氯化铯三种介质密度梯度离心的方法,结果表明氯化铯适合该病毒粒子的提纯。由称量法得到该病毒粒子在氯化铯中的浮密度为1.554~1.582g/mL 。本文还从形态学特征等方面讨论了该病毒的初步定位。 |
| 关键词: 文蛤 球形病毒 分离纯化 细胞病理 |
| DOI：10.11759/hykx20141209001 |
| 分类号:S944 |
| 基金项目:国家科技支撑计划课题(2012BAC07B03)；海洋行业公益专项项目(2013418007） |
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| Purification and ultrastructure of a spherical virus in cultured hard clam Meretrix meretrix Linnaeus |
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SHEN Hui,WAN Xi-he,HE Pei-min,YAO Guo-xing,CHEN Ai-hua
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1.Jiangsu institute of Marine Fisheries;2.Shanghai Ocean University,Shanghai
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| Abstract: |
| Since last decade, the disease out breaking in cultured hard clams along Nantong coastal line has caused great deal of death in Meretrix meretrix Linnaeus. Electron microscopic examinations of ultrathin sections revealed the presence of spherical virus particles in the intercellular substance and cytoplasm of the gill, digestive diverticula, mantle, and digestive tract of the moribund hard clam Meretrix meretrix Linnaeus in 2013. And the virions are approximately 45 to 55 nm in diameter , having no envelope,and there are no ambit between the core and nucleocapsid. In the infected cells with the virus, the cellular organelles presented obvious pathological changes. For example, the endoplasmic reticulum swelling and mostly turning into vesicles, the nuclear chromatin condensed, the mitochondria swelled and dissolved , the muscle fiber was disarray, the number of lysosome raised , and so on .The method adapts for purifying the virus from liquid of organism homogenization by differential centrifugation, potassium tartrate cushion ultracentrifugation, CsCl density gradient centrifugation, compared the sucrose or potassium tartrate-glycerol density gradient ultracentrifugation. With the negative staining, the virions are isometric symmetry, and approximately 60 to 75 nm in diameter. And there are many obvious capsomers on the surface of the virions. The buoyant density of virions is 1.554 to 1.582g/mL in the isopycnic CsCl gradient. In this paper, we also discussed the systematic status of the virus by the morphological characters. |
| Key words: Meretrix meretrix Linnaeus spherical virus purification cytopathology |