Kong Yixuan, Jin Peiyuan, Jia Chenchen, Qiao Panpan, Zhang Huangwei, Dong Yinglu, Yang Zhimin, Zhou Yuxin, Jung Geunhwa, Hu Jian
College of Agro-Grassland Science, Nanjing Agricultural University, Nanjing, China.
Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing, China.
Pest Manag Sci. 2025 Jan;81(1):196-203. doi: 10.1002/ps.8422. Epub 2024 Sep 19.
Sparse leaf patch (SLP) is one of the most significant diseases affecting seashore paspalum (Paspalum vaginatum Sw.), caused by Microdochium paspali. Fast and accurate detection of this pathogen is crucial for effective disease management. However, conventional culture-based methods are time-consuming and often compromised by the presence of other saprophytic or endophytic fungi.
In this study, we developed a real-time fluorescent quantitative (q)PCR method based on the internal transcribed spacer (ITS) region of the ribosomal RNA gene to rapidly detect and quantify M. paspali. The qPCR assay demonstrated the ability to detect all 12 tested isolates of M. paspali, with no cross-reactions observed when tested against 30 isolates of other fungal pathogens from turfgrass samples. The detection limit of the qPCR method was as low as 3.65 × 10 copies μL of M. paspali genomic DNA, and the entire detection process could be completed within 1 h. The fluorescence signal was detectable in the leaf tissues of seashore paspalum without apparent disease symptoms as early as 24 h postinoculation with M. paspali. Moreover, the qPCR method successfully detected M. paspali in both asymptomatic and symptomatic turfgrass samples, including leaf, stem, root and rhizosphere soil, indicating that this assay can significantly enhance the detection of M. paspali.
The study developed a rapid real-time qPCR assay for the detection of M. paspali causing SLP on seashore paspalum and in environmental samples, which has important implications for early warning and management of SLP. © 2024 Society of Chemical Industry.
稀叶斑块病（SLP）是影响海滨雀稗（Paspalum vaginatum Sw.）的最严重病害之一，由李氏禾附球菌（Microdochium paspali）引起。快速准确地检测这种病原体对于有效的病害管理至关重要。然而，传统的基于培养的方法耗时且常因其他腐生或内生真菌的存在而受到影响。
在本研究中，我们基于核糖体RNA基因的内部转录间隔区（ITS）开发了一种实时荧光定量（q）PCR方法，用于快速检测和定量李氏禾附球菌。该qPCR检测方法能够检测所有12株受试李氏禾附球菌分离株，在与来自草坪草样品的30株其他真菌病原体进行检测时未观察到交叉反应。qPCR方法的检测限低至3.65×10个李氏禾附球菌基因组DNA拷贝/μL，整个检测过程可在1小时内完成。早在接种李氏禾附球菌后24小时，在没有明显病害症状的海滨雀稗叶片组织中就能检测到荧光信号。此外，qPCR方法成功地在无症状和有症状的草坪草样品中检测到李氏禾附球菌，包括叶片、茎、根和根际土壤，表明该检测方法可显著提高李氏禾附球菌的检测率。
本研究开发了一种快速实时qPCR检测方法，用于检测引起海滨雀稗稀叶斑块病的李氏禾附球菌以及环境样品中的该菌，这对稀叶斑块病的早期预警和管理具有重要意义。© 2024化学工业协会。
