Recovery of C. graminicola from Root Infected Plants

Once a set of root inoculated plants reaches three weeks of age, transfer the plants to a 0.5L pot and fill using the green house soil.  (The green house soil is a mixture of a of 1:2 store soil and metromix soil)

  1. Six weeks after the plants are transferred, each plant needs to be surfaced sterilized by washing roots and stalk for 40 seconds in 10% bleach and then washing in dd H2O. Leaves should be washed for 30 seconds in 10% bleach and then washed in dd H2O.
  2. Plate the plant parts in Acid ½ PDA with Hygromycin and Ampicillin. 
    1. Add 19.5g of PDA to 1L of dd H2O
    2. Autoclave
    3. Once the media reaches 55°C, add in this order 130ml of Hygromycin, 1ml Ampicillin and 1.47 ml of sterile lactic acid.
  3. Two days after plating, begin observing the plates for fungal growth.  Make slides of these and identify under a microscope.

 

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