Materials and Reagents

1. Petri dish, triple vent 94 x 15mm (Greiner Bio One, catalog number: 633 185 )
   
2. Filter paper (Whatman, catalog number: 1001-090 )
   
3. Parafilm (Parafilm, catalog number: PM992 )
   
4. Plant container pots, 3 L (National Agrochemical Distributors, catalog number: POTS34 )
   
5. John Innes compost No. 2 (Westland Horticulture)
   
6. Square Petri dish 100 x 100 x 20 mm (SARSTEDT, catalog number: 82.9923.422 )
   
7. Glass Pasteur pipette (VWR, catalog number: 14673-010 )
   
8. Fusarium graminearum strain GZ3639 (Proctor et al., 1995)
   
9. Plant agar (Duchefa, catalog number: P1001.1000 )
   
10. Benzimidazole (Stock solution at 500 mM in ethanol stored in aliquots at -20 °C) (Sigma-Aldrich, catalog number: 194123 )
    
11. Ethanol (Sigma-Aldrich, catalog number: E7023 )
    
12. Tween-20 (Sigma-Aldrich, catalog number: P2287 )
    

Equipment

1. Incubator (20 °C)
   
2. Glasshouse (Cambridge HOK production glasshouse) (20-22 °C with a 16 h light/8 h dark photoperiod at 300 μmol m^-2 s^-1 and 70% relative humidity)
   
3. Plant growth room (20 °C with a 16 h light/8 h dark photoperiod at 200 μmol m^-2 s^-1 and 70% relative humidity)
   
4. Digital camera (Nikon, model: COOLPIX P500 with a NIKKOR 36X wide optical ZOOM ED VR, 4.0 - 144 mm, 1:3.4 - 5.7)
   
5. Copy stand (RPS Studio RS-CS920 copy stand)
   
6. Lights (cold-light fluorescent lighting system)
   

Software

1. Fiji (http://fiji.sc/)
   Note: Fiji is an open source software using the same functionality as ImageJ with many bundled plugins. For that reason the image analysis presented here could be executed with ImageJ.

Procedure