Nanjing Agricultural University, Weigang 1, Nanjing, Jiangsu, China
Mycologia. 2012 Sep-Oct;104(5):1250-9. doi: 10.3852/11-365. Epub 2012 Apr 9.
In North America, one of the most important root diseases of Poa and Festuca turf is summer patch, caused by Magnaporthe poae. Detection and identification of M. poae in infected roots by conventional culture-based methods is difficult and time consuming, typically taking 3 wk or longer to accomplish. In this study, a culture-independent, TaqMan real-time PCR assay was developed for the detection of M. poae from the roots of fungicide treated and non-treated Kentucky bluegrass (Poa pratensis) turf. The assay was validated with the target pathogen, closely related fungal species and a number of other microorganisms that inhabit the same host and soil environment. This assay was more sensitive (could detect as little as 3.88 pg genomic DNA of M. poae), rapid and accurate compared to direct microscopic observation and isolation on a selective medium. The real-time PCR detection results corresponded closely to visual assessments of disease severity in the field. Utilization of this assay in diagnostic laboratories will enable turfgrass managers to more quickly and effectively detect and potentially reduce fungicide usage through early and accurate identification of the pathogen.
在北美,导致草地早熟禾和羊茅草坪最重要的根部病害之一是夏季斑枯病,该病由禾谷镰刀菌引起。传统的基于培养的方法很难且费时地检测和鉴定受感染根部中的禾谷镰刀菌,通常需要 3 周或更长时间才能完成。在这项研究中,开发了一种非培养的 TaqMan 实时 PCR 检测方法,用于检测经杀菌剂处理和未经处理的肯塔基蓝草(Poa pratensis)草坪根部中的禾谷镰刀菌。该检测方法使用目标病原体、密切相关的真菌种和其他一些栖息在同一宿主和土壤环境中的微生物进行了验证。与直接显微镜观察和选择性培养基上的分离相比,该检测方法更灵敏(可以检测到低至 3.88 pg 禾谷镰刀菌的基因组 DNA)、快速且准确。实时 PCR 检测结果与田间疾病严重程度的目视评估密切吻合。在诊断实验室中使用该检测方法将使草坪管理者能够更快、更有效地通过早期和准确鉴定病原体来检测并潜在地减少杀菌剂的使用。