Macho Alberto P, Rufián José S, Ruiz-Albert Javier, Beuzón Carmen R
Shanghai Center for Plant Stress Biology, Chinese Academy of Sciences, Shanghai, China.
Dpto. Biología Celular, Genética y Fisiología, Instituto de Hortofruticultura Subtropical y Mediterránea, Universidad de Málaga-Consejo Superior de Investigaciones Científicas (IHSM-UMA-CSIC), Campus de Teatinos, Avda. Cervantes 2, Málaga, E-29071, Spain.
Methods Mol Biol. 2016;1363:209-17. doi: 10.1007/978-1-4939-3115-6_17.
When studying bacterial plant pathogens, the genetic analysis of the contribution of virulence factors to the infection process has traditionally been hindered by their high degree of functional redundancy. In recent years, it has become clear that the use of competitive index in mixed infections provides an accurate and sensitive manner of establishing virulence phenotypes for mutants for which other assays have failed. Such increases in sensitivity and accuracy are due to the direct comparison between the respective growths of the co-inoculated strains within the same infection, each strain replicating as they would in individual infections. Interferences between the co-inoculated strains must be therefore avoided using the appropriate experimental settings. In this chapter, we will present the optimal experimental conditions to achieve maximum sensitivity on virulence assays using the phytopathogenic bacterium Pseudomonas syringae, as well as some additional considerations to ensure the correct interpretations of the results.
在研究细菌性植物病原体时，毒力因子对感染过程的贡献的遗传分析传统上一直受到其高度功能冗余的阻碍。近年来，很明显，在混合感染中使用竞争指数为其他检测方法失败的突变体建立毒力表型提供了一种准确且灵敏的方式。这种灵敏度和准确性的提高是由于在同一感染中共同接种的菌株各自生长之间的直接比较，每个菌株都像在单独感染中一样进行复制。因此，必须使用适当的实验设置避免共同接种的菌株之间的干扰。在本章中，我们将介绍使用植物致病细菌丁香假单胞菌在毒力检测中实现最大灵敏度的最佳实验条件，以及一些确保正确解释结果的其他注意事项。
