Garcia M L, Grau O, Sarachu A N
Area de Química Biológica y Biología Molecular, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, Argentina.
Res Virol. 1991 Jul-Aug;142(4):303-11. doi: 10.1016/0923-2516(91)90017-w.
Isolate 90-1-1 Concordia (Argentina) of the citrus psorosis agent was graft-transmitted to citrus and mechanically transmitted to Chenopodium quinoa, which was used as a local lesion assay host. Infected citrus and C. quinoa plant lesions were used as starting materials for the purification of the psorosis-associated agent. In extracts partially purified by differential centrifugation, infectivity was abolished by RNase treatment, even in 0.3 M NaCl, indicating that ssRNA is required for biological activity. The total loss of infectivity produced by proteinase K treatment and the decline in infectivity caused by phenol extraction indicated that protein may be essential for infectivity. When partially purified extracts were subjected to sucrose density gradient centrifugation, infectivity on C. quinoa from certain 2-fraction combinations was higher than expected, compared to the infectivity of the individual fractions. Therefore, infectivity was not associated with a single component but with the combination of at least two components which were distinguishable on sedimentation. The infectious material was present in the top and bottom zones of a sucrose gradient, which on further purification by a second gradient and agarose gel electrophoresis, revealed the presence of a 50-kDa protein. This protein was absent in comparable gradient fractions from healthy plants, and therefore most likely represented the capsid protein of both the top and bottom sucrose gradient zone components. Taken together, these results led to the conclusion that the citrus-psorosis-associated virus (CPsAV) is a multipartite virus, containing ssRNA and a 50-kDa coat protein.(ABSTRACT TRUNCATED AT 250 WORDS)
柑橘鳞皮病病原体的分离物90 - 1 - 1康科迪亚(阿根廷)通过嫁接传播到柑橘上,并通过机械接种传播到藜麦上,藜麦用作局部病斑检测宿主。受感染的柑橘和藜麦植株病斑用作纯化与鳞皮病相关病原体的起始材料。在通过差速离心部分纯化的提取物中,即使在0.3 M NaCl中,核糖核酸酶处理也会使感染性丧失,这表明单链核糖核酸(ssRNA)是生物活性所必需的。蛋白酶K处理导致感染性完全丧失,苯酚提取导致感染性下降,这表明蛋白质可能是感染性所必需的。当部分纯化的提取物进行蔗糖密度梯度离心时,与单个组分的感染性相比,某些2组分组合对藜麦的感染性高于预期。因此,感染性与单一成分无关,而是与至少两种在沉降上可区分的成分的组合有关。感染性物质存在于蔗糖梯度的顶部和底部区域,通过第二次梯度和琼脂糖凝胶电泳进一步纯化后,发现存在一种50 kDa的蛋白质。在来自健康植株的可比梯度组分中不存在这种蛋白质,因此它很可能代表顶部和底部蔗糖梯度区组分的衣壳蛋白。综上所述,这些结果得出结论,柑橘鳞皮病相关病毒(CPsAV)是一种多分体病毒,含有单链核糖核酸和一种50 kDa的外壳蛋白。(摘要截短于250字)