G1Tn0-NB1
The transposon mutant library was reactivated following the same procedure as other samples. Bacterial cells were diluted to a concentration of 2  10 CFU/mL and infiltrated into tobacco leaves using a 1-mL sterile syringe. The inoculated plants were maintained at 28 C for 24 hours. Subsequently, the leaves were fragmented into 23 mm pieces using a blender and transferred to a glass dish containing ddHO. The mixture was subjected to ultrasonic treatment in an ultrasonic water bath for 15 minutes to lyse the apoplastic cells. The resulting homogenate was clarified either by centrifugation or filtration through a 40-m filter to reduce plant debris. Bacterial cells were pelleted by centrifugation at 5,000  g for 10 minutes.