Heller-Dohmen Marion, Göpfert Jens C, Hammerschmidt Ray, Spring Otmar
Institute for Botany, University of Hohenheim, 70593 Stuttgart, Germany.
Mol Plant Pathol. 2008 Nov;9(6):777-86. doi: 10.1111/j.1364-3703.2008.00499.x.
Eight pathotypes of Plasmopara halstedii were screened to investigate the occurrence of virions and the potential viral influence on the pathogenicity of the sunflower downy mildew pathogen. In 23 of 26 P. halstedii isolates derived from eight countries in Europe, North America and South America, virions were detected by transmission electron microscopy. By contrast, there were no ultrastructural indications of virus-like particles in eight other related Oomycetes. The virions of representative P. halstedii isolates were morphologically and biochemically characterized and compared among each other. Regardless of their host's pathotypes, the geographical origin of the isolate and the sensitivity towards the fungicide metalaxyl, the viral characters obtained were uniform. The virions were isometric and measured approximately 37 nm in diameter. One polypeptide of c. 36 kDa and two segments of single-stranded RNA (3.0 and 1.6 kb) were detected. Both viral RNA segments were detected by capillary electrophoresis in the three remaining P. halstedii isolates where virions were undetectable by transmission electron microscopy. Virus-specific primers for the 1.6 kb-segment were synthesized and used to determine and compare a partial sequence of the viral coat protein among virions of different P. halstedii pathotypes. In all tested isolates, fragments of 0.7 kb were amplified which were directly sequenced. Sequence variation was insignificant. As both less aggressive and more aggressive P. halstedii isolates contained virions, the presence or absence of virions could not explain the diverse aggressiveness of the downy mildew pathogen towards sunflower. Moreover, the results indicated that pathogenicity of P. halstedii was not related to variation in morphological or biochemical characters of the virions.
对向日葵霜霉病菌(Plasmopara halstedii)的8个致病型进行了筛选,以研究病毒粒子的存在情况以及病毒对向日葵霜霉病病原菌致病性的潜在影响。在来自欧洲、北美和南美8个国家的26株向日葵霜霉病菌分离物中,有23株通过透射电子显微镜检测到了病毒粒子。相比之下,其他8种相关卵菌中没有病毒样颗粒的超微结构迹象。对代表性向日葵霜霉病菌分离物的病毒粒子进行了形态学和生化特征分析,并相互比较。无论其宿主致病型、分离物的地理来源以及对杀菌剂甲霜灵的敏感性如何,所获得的病毒特征都是一致的。病毒粒子呈等轴对称,直径约为37纳米。检测到一条约36 kDa的多肽和两段单链RNA(3.0和1.6 kb)。在另外3株向日葵霜霉病菌分离物中,通过毛细管电泳检测到了这两段病毒RNA,而通过透射电子显微镜未检测到病毒粒子。合成了针对1.6 kb片段的病毒特异性引物,并用于测定和比较不同向日葵霜霉病菌致病型病毒粒子中病毒外壳蛋白的部分序列。在所有测试的分离物中,扩增出了0.7 kb的片段并直接进行了测序。序列变异不显著。由于致病性较弱和较强的向日葵霜霉病菌分离物中都含有病毒粒子,因此病毒粒子的存在与否无法解释霜霉病病原菌对向日葵不同的致病力。此外,结果表明向日葵霜霉病菌的致病性与病毒粒子的形态或生化特征变异无关。