Vinopal Dominik, Bryan Anthony, Hodson David P, Saunders Diane G O
John Innes Centre, Norwich Research Park, Norwich, UK.
International Maize and Wheat Improvement Center (CIMMYT), Texcoco, Mexico.
Methods Mol Biol. 2025;2898:93-111. doi: 10.1007/978-1-0716-4378-5_6.
Plant pathogens are a serious constraint to crop production worldwide, being responsible for considerable economic loss annually. Rapid detection of disease incidences can be critical for enacting timely control measures and thereby limiting the severity of costly disease outbreaks. With recent technological advances in genome and gene sequencing technologies and parallel reductions in cost, DNA-based plant disease diagnostics methods are now becoming embedded in surveillance systems for many critical crop disease threats. In this chapter, we consider one such diagnostic technique, termed Mobile And Real-time PLant disEase (MARPLE) diagnostics. The MARPLE diagnostics platform is a point-of-care (POC) platform that utilizes amplicon re-sequencing on the portable MinION sequencer to type individual strains of the wheat yellow rust (Puccinia striiformis f. sp. tritici; Pst) or wheat stem rust (Puccinia graminis f. sp. tritici; Pgt) pathogens. The system is designed to amplify genes highly variable between strains of the Pst or Pgt pathogens, and following amplicon re-sequencing, the sequences of these genes can be used for phylogenetic analysis, comparing "new" samples to a set of reference isolates from around the world. The MARPLE diagnostics method is also very simple, with very little training required and equipment needed, making the method highly suited for use in resource-poor environments. The following chapter contains everything you need to know to set up and execute MARPLE diagnostics.
植物病原体是全球作物生产的严重制约因素，每年造成相当大的经济损失。快速检测疾病发病率对于及时采取控制措施从而限制代价高昂的疾病爆发的严重程度至关重要。随着基因组和基因测序技术的最新技术进步以及成本的同步降低，基于DNA的植物病害诊断方法现在正被纳入许多关键作物病害威胁的监测系统中。在本章中，我们将介绍一种这样的诊断技术，称为移动实时植物病害（MARPLE）诊断。MARPLE诊断平台是一种即时检测（POC）平台，它利用便携式MinION测序仪上的扩增子重测序来鉴定小麦条锈病（条形柄锈菌小麦专化型；Pst）或小麦秆锈病（禾柄锈菌小麦专化型；Pgt）病原体的各个菌株。该系统旨在扩增Pst或Pgt病原体菌株之间高度可变的基因，在扩增子重测序之后，这些基因的序列可用于系统发育分析，将“新”样本与来自世界各地的一组参考分离株进行比较。MARPLE诊断方法也非常简单，所需的培训和设备很少，这使得该方法非常适合在资源匮乏的环境中使用。以下章节包含设置和执行MARPLE诊断所需了解的一切内容。
