Kurera Shimosh, McCallum Brent, Brar Gurcharn Singh, Nilsen Kirby, Aboukhaddour Reem, Fetch Tom, Bakker Matthew, Walkowiak Sean
Canadian Grain Commission, Winnipeg, MB, Canada.
Department of Microbiology, University of Manitoba, Winnipeg, MB, Canada.
Methods Mol Biol. 2025;2898:129-137. doi: 10.1007/978-1-0716-4378-5_8.
Rust diseases are a major limitation to grain production in Canada and have caused devastating yield losses worldwide [1]. The identification and characterization of pathogenic fungi responsible for rust diseases are important for implementing effective disease management practices. Modern DNA-based methods of identification, such as quantitative polymerase chain reaction (PCR) and sequencing, are time-consuming, costly, and require well-trained laboratory personnel. Here, we describe a protocol to rapidly identify rust samples to the species level using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Biochemical profiles generated by MALDI-TOF MS are used for identification. The method is commonly used in clinical settings and can accurately identify hundreds of microbes in minutes at a low cost. The MALDI-TOF MS method described here is simple and can assess samples of rust spores and spore-mats originating from different cereal hosts and identify them to the species level with high accuracy, high throughput, and low cost. The method is not currently able to accurately distinguish among races, though this may be possible with additional research and sample fractionation.
锈病是加拿大谷物生产的主要限制因素，在全球范围内已造成毁灭性的产量损失[1]。鉴定和表征引起锈病的致病真菌对于实施有效的病害管理措施至关重要。现代基于DNA的鉴定方法，如定量聚合酶链反应（PCR）和测序，既耗时又昂贵，还需要训练有素的实验室人员。在此，我们描述了一种使用基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）将锈病样本快速鉴定到物种水平的方案。由MALDI-TOF MS生成的生化图谱用于鉴定。该方法常用于临床环境，能够在几分钟内以低成本准确鉴定数百种微生物。这里描述的MALDI-TOF MS方法简单，可以评估来自不同谷物宿主的锈孢子和孢子垫样本，并以高精度、高通量和低成本将它们鉴定到物种水平。尽管通过进一步研究和样本分级可能实现，但该方法目前尚无法准确区分生理小种。
