Photo description of Sclerotinia homoeocarpa fungicide resistance protocol.
Prepare PDA Media
Prepare potato dextrose agar media by adding 39 g of PDA powder to 1 L of distilled water. Heat while mixing.
Serial dilution preparation
While the media is in the autoclave, prepare desired serial dilutions in sterile water
Example using Boscalid—Emerald(0.70g a.i/ 1g product)
Serial dilutions were created using Boscalid to final concentrations of 1000, 100,10, 1 ug a.i/mL
1000 ug stock solution
Stock solution was 1000 ug a.i/mL—(7.14 g of product mixed with 10 mL sterile water)
10-fold serial dilution
1 mL of 1000 ug solution to 9 mL sterile water to create 100 ug solution
10-fold serial dilution
1 mL of 100 ug solution to 9 mL sterile water to create 10 ug solution
10-fold serial dilution
1 mL of 100 ug solution to 9 mL sterile water to create 1 ug solution
10-fold serial dilution
Final Concentrations: 1000, 100, 10, 1 ug a.i/mL
Allow media to cool
Allow molten PDA to cool to 50C in a water bath or on the lab bench
Amend media
Amend cooled media with 1 mL of your prepared fungicide concentration and mix using the magnetic stir bar for 10 seconds
Amend media
Amend cooled media with 1 mL of your prepared fungicide concentration and mix using the magnetic stir bar for 10 seconds
Pour plates
Pour approximately 20 mL of amended media into sterile petri plates and allow plates to cool
