Baturo-Ciesniewska Anna, Groves Carol L, Albrecht Kenneth A, Grau Craig R, Willis David K, Smith Damon L
Department of Phytopathology and Molecular Mycology, UTP University of Science and Technology, Kordeckiego 20, 85-225 Bydgoszcz, Poland.
Department of Plant Pathology.
Plant Dis. 2017 Jan;101(1):192-199. doi: 10.1094/PDIS-06-16-0896-RE. Epub 2016 Oct 11.
Symptoms of clover rot caused by Sclerotinia trifoliorum or S. sclerotiorum are identical, making differentiation and identification of the causal species difficult and time consuming. Polymerase chain reaction (PCR) amplification and nucleotide sequencing were used to examine 40 isolates of S. trifoliorum (29 from Poland, 11 from the United States) and 55 isolates of S. sclerotiorum (26 from Poland, 29 from the United States). We determined that amplification of the β-tubulin and calmodulin genes with TU1/TU2/TU3 and SscadF1/SscadR1 PCR primers and the presence of introns and single-nucleotide polymorphisms (SNP) within the ribosomal DNA (rDNA) as detected with NS1/NS8 and internal transcribed spacer (ITS)1/ITS4 PCR primers are effective for rapidly and accurately differentiating between the two species of Sclerotinia. In addition, our research revealed a lack of intraspecies variation within S. sclerotiorum isolates from the United States and Poland using these same molecular markers. We detected a relatively high degree of intraspecies variability among isolates of S. trifoliorum from the United States and Poland using the presence of introns and SNP within the rDNA. SNP and nuclear small-subunit rDNA analyses revealed distinct groups of S. trifoliorum among the isolates used in this study. The results of this study provide useful information for clover breeders and pathologists looking to develop clover varieties with durable resistance.
由三叶草核盘菌或核盘菌引起的三叶草腐烂病症状相同，因此区分和鉴定致病菌种既困难又耗时。采用聚合酶链反应（PCR）扩增和核苷酸测序技术，对40株三叶草核盘菌（29株来自波兰，11株来自美国）和55株核盘菌（26株来自波兰，29株来自美国）进行了检测。我们确定，使用TU1/TU2/TU3和SscadF1/SscadR1 PCR引物扩增β-微管蛋白和钙调蛋白基因，以及使用NS1/NS8和内转录间隔区（ITS）1/ITS4 PCR引物检测核糖体DNA（rDNA）中的内含子和单核苷酸多态性（SNP），对于快速准确地区分这两种核盘菌是有效的。此外，我们的研究表明，使用这些相同的分子标记，美国和波兰的核盘菌分离株之间缺乏种内变异。利用rDNA中的内含子和SNP的存在，我们在美国和波兰的三叶草核盘菌分离株中检测到了相对较高程度的种内变异性。SNP和核小亚基rDNA分析揭示了本研究中使用的分离株中三叶草核盘菌的不同组群。这项研究的结果为希望培育具有持久抗性的三叶草品种的育种者和病理学家提供了有用的信息。
