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Background  The  Arabidopsis thaliana-Pseudomonas syringae  model pathosystem is one of the most widely used systems to understand the mechanisms of microbial pathogenesis and plant innate immunity. Several inoculation methods have been used to study plant-pathogen interactions in this model system. However, none of the methods reported to date are similar to those occurring in nature and amicable to large-scale mutant screens.    Results  In this study, we developed a rapid and reliable seedling flood-inoculation method based on young  Arabidopsis  seedlings grown on MS medium. This method has several advantages over conventional soil-grown plant inoculation assays, including a shorter growth and incubation period, ease of inoculation and handling, uniform infection and disease development, requires less growth chamber space and is suitable for high-throughput screens. In this study we demonstrated the efficacy of the  Arabidopsis  seedling assay to study 1) the virulence factors of  P. syringae  pv.  tomato  DC3000, including type III protein secretion system (TTSS) and phytotoxin coronatine (COR); 2) the effector-triggered immunity; and 3)  Arabidopsis  mutants affected in salicylic acid (SA)- and pathogen-associated molecular pattern (PAMPs)-mediated pathways. Furthermore, we applied this technique to study nonhost resistance (NHR) responses in  Arabidopsis  using nonhost pathogens, such as  P. syringae  pv.  tabaci , pv.  glycinea  and pv.  tomato  T1, and confirmed the functional role of FLAGELLIN-SENSING 2 (FLS2) in NHR.    Conclusions  The  Arabidopsis  seedling flood-inoculation assay provides a rapid, efficient and economical method for studying  Arabidopsis-Pseudomonas  interactions with minimal growth chamber space and time. This assay could also provide an excellent system for investigating the virulence mechanisms of  P. syringae . Using this method, we demonstrated that FLS2 plays a critical role in conferring NHR against nonhost pathovars of  P. syringae , but not to  Xanthomonas campestris  pv.  vesicatoria . This method is potentially ideal for high-throughput screening of both  Arabidopsis  and pathogen mutants.

Arabidopsis seedling flood-inoculation technique: a rapid and reliable assay for studying plant-bacterial interactions