Centre for Biotechnology, Brock University, St. Catharines, ON L2S 3A1, Canada.
Agriculture and Agri-Food Canada, Vineland Station, ON L0R 2E0, Canada.
Viruses. 2019 Oct 1;11(10):910. doi: 10.3390/v11100910.
is a globally devastating pathogen of apple, pear, and other Rosaceous plants. The use of lytic bacteriophages for disease management continues to garner attention as a possible supplement or alternative to antibiotics. A quantitative productive host range was established for 10 phages using 106 wild type global isolates of , and the closely related , to investigate the potential regional efficacy of these phages within a biopesticide. Each host was individually infected with each of the 10 phages and phage production after 8 h incubation was measured using quantitative real time PCR (qPCR) in conjunction with a standardized plasmid. PCR amplicons for all phages used in the study were incorporated into a single plasmid, allowing standardized quantification of the phage genome copy number after the infection process. Nine of the tested phages exhibited a broad host range, replicating their genomes by at least one log in over 88% of tested hosts. Also, every Amygdaloideae infecting host was able to increase at least one phage by three logs. Bacterial hosts isolated in western North America were less susceptible to most phages, as the mean genomic titre produced dropped by nearly two logs, and this phenomenon was strongly correlated to the amount of exopolysaccharide produced by the host. This method of host range analysis is faster and requires less effort than traditional plaque assay techniques, and the resulting quantitative data highlight subtle differences in phage host preference not observable with typical plaque-based host range assays. These quantitative host range data will be useful to determine which phages should be incorporated into a phage-mediated biocontrol formulation to be tested for regional and universal control of .
是一种全球性的苹果、梨和其他蔷薇科植物病原体。溶菌噬菌体作为抗生素的补充或替代品,用于疾病管理的应用不断受到关注。使用 106 种野生型全球 分离株和密切相关的 ,建立了 10 种噬菌体的定量生产宿主范围,以调查这些噬菌体在生物农药中的潜在区域疗效。将每个宿主分别用 10 种噬菌体中的每一种感染,并在 8 小时孵育后使用定量实时 PCR(qPCR)结合标准化质粒测量噬菌体的产量。研究中使用的所有噬菌体的 PCR 扩增子都被整合到一个质粒中,允许在感染过程后标准化定量噬菌体基因组拷贝数。测试的 9 种噬菌体表现出广泛的宿主范围,在 88%以上的测试宿主中至少复制了一个对数的基因组。此外,感染的每个 宿主都能够使至少一种噬菌体增加三个对数。来自北美西部的细菌宿主对大多数噬菌体的敏感性较低,因为产生的基因组滴度平均下降了近两个对数,这种现象与宿主产生的胞外多糖量强烈相关。这种宿主范围分析方法比传统的噬菌斑测定技术更快,所需工作量更少,并且产生的定量数据突出了噬菌体宿主偏好的细微差异,这些差异在典型的基于噬菌斑的宿主范围测定中是无法观察到的。这些定量宿主范围数据将有助于确定应将哪些噬菌体纳入噬菌体介导的生物防治制剂中,以测试其对的区域和普遍控制。