Pathogenicity Assay of Penicillium expansum on Apple Fruits

Materials and Reagents

1. 90 x 15 mm Petri dish (any brand will suffice)
   
2. Plastic film (polyurethane material, any department store)
   
3. 1,000 µl pipette tips (Corning, Axygen^®, catalog number: TF-1000-R-S )
   
4. 200 µl pipette tips (Corning, Axygen^®, catalog number: TF-200-R-S )
   
5. 10 µl pipette tips (Corning, Axygen^®, catalog number: TF-300-R-S )
   
6. Polyester filter cloth cut into 8 x 8 cm squares (any fabric store)
   
7. Penicillium expansum T01: was isolated by our laboratory and whole-genome sequenced (Li et al., 2015)
   
8. Freshly harvested red Fuji apples
   
9. Glycerol (AMRESCO, catalog number: M152 )
   
10. Tween 20 (Sigma-Aldrich, catalog number: T2700 )
    
11. Sodium hypochlorite (Sigma-Aldrich, catalog number: 239305 )
    
12. Sterile distilled water
    
13. Potato
    
14. Dextrose (Macklin, catalog number: D823520 )
    
15. Agar (HUAAOBIO, catalog number: HA0552 )
    
16. 2% sodium hypochlorite solution (see Recipes)
    
17. PDA medium (see Recipes)
    

Equipment

1. Clean bench (Beijing Donglian Har Instrument Manufacture, model: SCB-1520 )
   
2. Sterile nail (approximately 3 mm in diameter, manual polishing)
   
3. Constant temperature incubator (TAICANG, model: THZ-C )
   
4. Hemacytometer (QIUJING, catalog number: XB-K-25 )
   
5. Vortexer (Select BioProducts, model: SBS100-2 )
   
6. 100 µl-1,000 µl pipette (Eppendorf, catalog number: 3120000267 )
   
7. 10 µl-1,00 µl pipette (Eppendorf, catalog number: 3120000240 )
   
8. 0.5µl-1,0 µl pipette (Eppendorf, catalog number: 3120000224 )
   
9. Optical microscope (CHONGQING OPTEC Instrument, model: B203LED )
   
10. 40 x 30 x 10 cm plastic basket (any brand will suffice)
    
11. Hand held sprayer (any brand will suffice)
    
12. 40 x 40 x 30 cm containers (any brand will suffice)
    
13. Hygrothermograph (Fisher Scientific, catalog number: 11-661-20 )
    

Software

1. SPSS version 13.0

Procedure