Soybean Cyst Nematode, Heterodera glycines, Infection Assay Using Soybean Roots

Materials and Reagents

1. Plastic Pasteur pipettes (Corning, Falcon^®, catalog number: 357575 )
   
2. Nylon cloth at 30 micron pore
   
3. 90 mm Whatman circle filter size
   
4. White filter paper (GE Healthcare, catalog number: 10347009 )
   
5. Fleaker filtration unit (Cole Parmer fluid handling and analysis Item) (Cole-Parmer Instrument Compan, catalog number: EW-08917-50 )
   
6. Disposable Petri Dishes (Kord-Valmark Labware Products, catalog number: 2900 )
   
7. Compost soil
   
8. Soybean seeds (William 82 or Essex) (Glycine max) (ARS-USDA)
   
9. Sodium hypochlorite (Commercial Bleach-Clorox contains 8.3% Na-hyperchlorite)
   
10. Sucrose (Baker Analyzed Reagent, catalog number: 4072-1 )
    
11. Table sugar (Commercial sugar)
    
12. Ethyl alcohol (The Warner-Graham Company, catalog number: 64-17-5 )
    Note: Currently, it is “Sigma-Aldrich, catalog number: 64-17-5 ”.
    
13. Zinc sulfate heptahydrate (Sigma-Aldrich, catalog number: 91f-0135 )
    
14. Sucrose solution (see Recipes)
    
15. Sodium hypochlorite solution (see Recipes)
    
16. 3 mm Zinc sulfate heptahydrate (ZnSO₄) solution (see Recipes)
    

Equipment

1. Greenhouse with a sink to divert drain water to a chlorine holding tank to kill nematodes that go down the drain
   
2. 8-10 inch round Pots (Myers Lawn&Garden)
   
3. Sieves (Sieve sizes: #20 = 841 micron; #60 = 250 micron; #80 = 177 micron; #100 = 149 micron; #250 = 58 micron; #500 = 25 micron) (U.S Standard Sieve Series)
   
4. 1 liter glass cylinder (Pyrex USA)
   
5. Dissecting microscope (Spenser)
   
6. Gyrotory water bath Shaker (LabX, New Brunswick Scientific, model: G76 )
   
7. 1 liter glass beaker (Pyrex)
   
8. Platform Shaker (New Brunswick Scientific)
   
9. 10 ml pipette (Thermo Fisher Scientific, Fisherbrand)
   

Procedure