Vilches S, Castillo A
Departamento de Ciencias Biológicas, Facultad de Química y Biologia, Universidad de Santiago de Chile, Santiago, Chile.
FEMS Microbiol Lett. 1997 Oct 1;155(1):125-30. doi: 10.1111/j.1574-6968.1997.tb12696.x.
In wild-type Botrytis cinerea CVg25 strain we have detected the presence of extrachromosomal genetic elements corresponding to double-stranded RNA molecules. These genetic elements have been designated L, M1 and M2 with molecular sizes of 8.3, 2.0 and 1.4 kb, respectively. The visualization by electron microscopy of mycelium ultrathin sections from B. cinerea CVg25 showed the presence of isometric virus-like particles of about 40 nm in diameter. Linear sucrose gradient centrifugation of mycelium-free extracts was done to determine if the double-stranded RNAs were associated with virus-like particles. The gradient profile obtained at 260 and 280 nm revealed a major peak that was analyzed by both agarose-gel electrophoresis and electron microscopy. It was observed that only the L-double-stranded RNA molecule copurified with isometric virus-like particles. These virus-like particles had a similar morphology and size as those detected by electron microscopy in the mycelium sections. These results suggest that only the L-double-stranded RNA would be encapsidated.
在野生型灰葡萄孢CVg25菌株中,我们检测到了对应于双链RNA分子的染色体外遗传元件的存在。这些遗传元件分别被命名为L、M1和M2,分子大小分别为8.3、2.0和1.4 kb。对灰葡萄孢CVg25的菌丝体超薄切片进行电子显微镜观察,结果显示存在直径约40 nm的等轴病毒样颗粒。对不含菌丝体的提取物进行线性蔗糖梯度离心,以确定双链RNA是否与病毒样颗粒相关联。在260和280 nm处获得的梯度图谱显示有一个主峰,通过琼脂糖凝胶电泳和电子显微镜对其进行了分析。观察到只有L双链RNA分子与等轴病毒样颗粒共纯化。这些病毒样颗粒的形态和大小与在菌丝体切片中通过电子显微镜检测到的颗粒相似。这些结果表明只有L双链RNA会被衣壳化。