Centre of Competence for the Innovation in the Agro-environmental Sector-AGROINNOVA, University of Turin, via Paolo Braccini 2, I-10095 Grugliasco, TO, Italy.
Department of Agricultural, Forestry and Food Sciences, University of Torino, via Paolo Braccini 2,I-10095 Grugliasco, TO, Italy.
Plant Dis. 2019 Sep;103(9):2305-2314. doi: 10.1094/PDIS-01-19-0035-RE. Epub 2019 Jul 12.
and are two causal agents of brown rot, one of the most important diseases in stone fruit. Two species cause blight on blossoms and twigs and brown rot on fruit in pre- and postharvest. Both species are distributed worldwide in North and South America, Australia, and Japan. In Europe, is endemic, while was introduced in 2001 and it is now widespread in several countries. Currently, both species coexist in European stone fruit orchards. spp. overwinter in cankers and mummified fruit. Mummy monitoring during winter permits growers to understand which species of will be prevalent in an orchard during the following season, permitting planning of an appropriate crop protection. Traditionally, the identification has been carried out using morphological features and even with polymerase chain reaction (PCR)-based assays that requires time and well-equipped laboratories. In this study, two isothermal-based methods were designed to identify these pathogens in a faster way than using traditional methods. The loop-mediated amplification (LAMP) assays were validated on some isolates of spp. coming from the mummy monitoring according to the international European and Mediterranean Plant Protection Organization standard (PM7/98), taking into account specificity, sensitivity, repeatability, and reproducibility. The sensitivity of both assays was checked by monitoring (at different time points) two nectarine varieties artificially inoculated and stored at two different temperatures. The reliability of both LAMP assays against the quantification of the inoculum was compared with previously published quantitative PCR assays. Both LAMP methods were able to detect a low number of cells. These LAMP methods could be a useful tool for monitoring brown rot causal agents in the field and during postharvest.
和 是桃褐腐病的两种病原体,这是核果类水果最重要的病害之一。这两种病原菌均可引起花果和嫩枝凋萎病以及采后果实的褐腐病。这两种病原菌在南北美洲、澳大利亚和日本的世界各地都有分布。在欧洲, 是地方性的,而 于 2001 年引入,现已在多个国家广泛传播。目前,这两个物种同时存在于欧洲核果果园中。 spp. 在溃疡和木乃伊果实中越冬。冬季对木乃伊的监测使种植者能够了解在下一个季节果园中哪种 物种将占优势,从而可以计划进行适当的作物保护。传统上,鉴定是使用形态特征进行的,甚至使用聚合酶链反应(PCR)为基础的检测方法,这需要时间和设备齐全的实验室。在这项研究中,设计了两种基于等温的方法,以比传统方法更快地识别这些病原体。环介导的扩增(LAMP)检测根据国际欧洲和地中海植物保护组织标准(PM7/98)对来自木乃伊监测的一些 spp. 分离物进行了验证,考虑了特异性、敏感性、重复性和再现性。通过监测(在不同时间点)两种油桃品种的人工接种和在两种不同温度下储存,检查了两种检测方法的敏感性。两种 LAMP 检测方法与先前发表的定量 PCR 检测方法相比,对接种物数量的可靠性进行了比较。两种 LAMP 方法都能够检测到少量细胞。这些 LAMP 方法可能是田间和采后监测桃褐腐病病原体的有用工具。