PDF(Pubmed)
Abstract：
Fusarium crown rot (FCR), caused by Fusarium spp., is a devastating disease in wheat growing areas. Previous studies have shown that FCR is caused by co-infection of F. graminearum, F. pseudograminearum, F. proliferatum and F. verticillioides in Hubei Province, China. In this study, a method was developed to simultaneously detected DNAs of F. graminearum, F. pseudograminearum, F. proliferatum and F. verticillioides that can efficiently differentiate them. Whole genome sequence comparison of these four Fusarium spp. was performed and a 20 bp sequence was designed as an universal upstream primer. Specific downstream primers of each pathogen was also designed, which resulted in a 206, 482, 680, and 963 bp amplicon for each pathogen, respectively. Multiplex PCR specifically identified F. graminearum, F. pseudograminearum, F. proliferatum and F. verticillioides but not from other 46 pathogens, and the detection limit of target pathogens is about 100 pg/μl. Moreover, we accurately determined the FCR pathogen species in wheat samples using the optimized multiplex PCR method. These results demonstrate that the multiplex PCR method established in this study can efficiently and rapidly identify F. graminearum, F. pseudograminearum, F. proliferatum, and F. verticillioides, which should provide technical support for timely and targeted prevention and control of FCR.
摘要：
镰刀菌冠腐病(FCR),由镰刀菌引起。,是小麦种植区的毁灭性疾病。以前的研究表明，FCR是由同时感染禾谷镰刀菌引起的，F.假赤霉，湖北省F.proliferatum和F.writicillioides，中国。在这项研究中,开发了一种同时检测F.graminearumDNA的方法，F.假赤霉，可以有效区分它们的F.proliferatum和F.roticillioides。这四个镰刀菌的全基因组序列比较。进行，并设计了20bp序列作为通用上游引物。还设计了每种病原体的特异性下游引物，每个病原体产生206、482、680和963bp的扩增子，分别。多重PCR专门鉴定了禾谷镰刀菌，F.假赤霉，F.增殖和轮虫，但不来自其他46种病原体，目标病原体的检测限约为100pg/μl。此外，我们使用优化的多重PCR方法准确地确定了小麦样品中的FCR病原体种类。这些结果表明，本研究建立的多重PCR方法可以高效、快速地鉴定禾谷菌株。F.假赤霉，F.增殖，和F.verticillioides，为及时、有针对性地预防和控制FCR提供技术支持。
