Isolation of Pythium from creeping bentgrass and annual bluegrass putting green plugs.
Symptoms on annual bluegrass tissues
Look for symptoms and mycelium on infected annual bluegrass tissues.
Selection of infected tissues
Selection of 3/4 leaves that show symptoms and mycelium.
Collection of infected tissues
Collect the infected tissues in a small Petri dish with a drop of D.I. water.
Surface sterilization
Prepare four small Petri dishes and fill three of them with sterile D.I. water.
Surface sterilization
Fill the Petri dish with the infected tissues with bleach 10%
Surface sterilization
Stir the tissues in bleach for one minute.
Surface sterilization
Transfer the tissues from bleach to the first dish with water and wash them for 30 seconds.
Surface sterilization
Transfer the tissues in the second dish with water and wash for 30 seconds.
Surface sterilization
After washing the tissues in the third dish filled with water for 30 seconds, transfer the tissues on a sterile filter paper and let it dry.
Surface sterilization
Transfer the dried tissues on a Petri dish with AWA and seal it.
