General protocol for completing Koch's Postulates to prove that a pathogen is causing a specific disease.
Collection of samples
Collection of the samples from the field with a zoysia plugger.
Plants in pots
Collected samples from the field are planted in pots filled with green sand in the greenhouse.
Pure culture of the isolates on PDA
Preparation of the plugs from pure culture on PDA for inoculations of the plants.
Inoculation
Inoculations of the plants with plugs from pure culture on PDA. The plug is introduced in the canopy close to the soil.
Spritz plants with D.I. water
Spritzing the inoculated plants with D.I. water before incubate them.
Preparation of plants for incubation
Sealing the plants with a plastic bag and a rubber band before the incubation in the growth chamber.
Pot in Petri dish with water
Each plant is set in a Petri Dish filled with water.
Inoculated plants in the growth chamber
Plants arranged in the growth chambers previously set up on 25°C with a photoperiod of 12 hours night/day.
