title: control_2
design: This protocol is modified from Ishiga et al., 2011. The procedure for the growth and inoculation of Arabidopsis seedlings and pathogen preparation and inoculation begins with the sterilization and stratification of Arabidopsis seeds. These seeds are then sown on 1/2 MS square agar plates and grown vertically in a plant tissue culture Percival at 22C under long-day conditions. Six days post-sowing, normal-sized seedlings are transferred to new round plates of 1/2 MS for continued growth until the twelfth day after sowing, maintaining uniform inoculation. At this stage, the plates are placed horizontally. It is crucial to perform all these steps under sterile conditions. For the pathogen preparation, fresh pathogen is scraped from the plates 24-48 hours post-inoculation using pipette tips and resuspended in distilled water containing 10 mM MgCl2. The OD600 is adjusted to 0.1 for testing Avr effector-triggered ETI. However, for other purposes, the OD can be varied. Silwet L-77 is added to this preparation just before use to achieve a final concentration of 0.028% (w/v). The pathogen inoculation involves the gentle pouring of the pathogen suspension onto plates to fully immerse the seedlings. After a brief immersion period of 2-3 minutes, the liquid is carefully drained off. The plates are then covered with lids and incubated at 22C in a Percival for observation.
library_name: control_2