Phytopathology. 2001 Sep;91(9):905-12. doi: 10.1094/PHYTO.2001.91.9.905.
ABSTRACT Molecular phylogenetic analyses were performed on 40 isolates of Verticillium fungicola collected from various Pennsylvania mushroom farms in 1999 and 28 isolates of Verticillium spp. collected during the last 50 years from various geographic locations. Sequence analysis of internal transcribed spacers 1 and 2 (ITS1 and ITS2) and 5.8S regions of the nuclear ribosomal DNA (rDNA) transcriptional unit and analysis of random amplified polymorphic DNA (RAPD) data were performed for the 68 isolates of Verticillium spp. Identical rDNA sequences were obtained for all 40 Pennsylvania isolates collected during 1999, 13 North American isolates collected during the last 50 years, and the ex-type strain of V. fungicola var. aleophilum. Sequence analysis of European isolates revealed a close relationship to the ex-type strain V. fungicola var. fungicola. No European-like isolates of V. fungicola var. fungicola were detected in the collection of North American isolates examined. Results from six decamer RAPD primers strongly indicate the presence of a clonal population of V. fungicola among Pennsylvania isolates. In addition, RAPD data delineated a Korean isolate (DC130) and ex-type strain V. fungicola var. aleophilum from the North American group. Virulence assays, based on spore inoculation of mushroom pilei, revealed variation corresponding to each neighbor-joining and RAPD grouping. All isolates with rDNA sequence and RAPD grouping similarity to ex-type strains V. fungicola var. aleophilum and V. fungicola var. fungicola displayed the highest level of virulence. Based on rDNA sequence and RAPD analyses, isolates displaying reduced or no virulence were distantly related to these two varieties. All results obtained for the analyses of ex-type strain V. fungicola var. flavidum suggested that this fungal isolate should not be considered a variety of V. fungicola, but rather a distinct species.
摘要 1999 年，从宾夕法尼亚州的多个蘑菇农场采集了 40 株Verticillium fungicola 分离株，并从过去 50 年的不同地理位置采集了 28 株Verticillium spp.分离株，进行了分子系统发育分析。对核核糖体 DNA（rDNA）转录单位的内部转录间隔区 1 和 2（ITS1 和 ITS2）和 5.8S 区以及随机扩增多态性 DNA（RAPD）数据进行了序列分析，对 68 株Verticillium spp.分离株进行了分析。所有 40 株 1999 年采集的宾夕法尼亚分离株、过去 50 年采集的 13 株北美分离株和 V. fungicola var.aleophilum 的模式菌株均获得了相同的 rDNA 序列。欧洲分离株的序列分析显示与模式菌株 V. fungicola var. fungicola 密切相关。在检查的北美分离株中未检测到类似欧洲的 V. fungicola var. fungicola 分离株。六聚体 RAPD 引物的结果强烈表明宾夕法尼亚分离株中存在 V. fungicola 的无性繁殖种群。此外，RAPD 数据将韩国分离株（DC130）和模式菌株 V. fungicola var.aleophilum 与北美组分开。基于蘑菇堆中孢子接种的毒力测定，根据每个邻接和 RAPD 分组显示了变化。与 V. fungicola var. aleophilum 和 V. fungicola var. fungicola 的模式菌株的 rDNA 序列和 RAPD 分组具有相似性的所有分离株均显示出最高水平的毒力。基于 rDNA 序列和 RAPD 分析，显示出降低或无毒性的分离株与这两个品种的关系较远。对模式菌株 V. fungicola var. flavidum 的分析结果表明，该真菌分离株不应被视为 V. fungicola 的一个品种，而应被视为一个独特的物种。
