Leaf Disc Stress Tolerance Assay for Tobacco

Materials and Reagents

1. Young tobacco plants (to be tested)
   
2. Distilled water
   
3. Acetone (Sigma-Aldrich, catalog number: I10010 )
   
4. Stress reagents (such as NaCl, Sigma-Aldrich, catalog number: S7653 ; for salinity stress)
   
5. Liquid nitrogen
   
6. Extraction buffer (see Recipes)
   

Equipment

1. Leaf puncher/ cork borer (Sigma-Aldrich, catalog number: Z165220 )
   
2. Petri plates (60 x 15 mm)
   
3. Microcentrifuge tube
   
4. Tissue paper
   
5. Graduated cylinder
   
6. Mortar and pestle (Sigma-Aldrich, catalog number: Z112496 )
   
7. Glass cuvette (1 ml) (Sigma-Aldrich, catalog number: C8550-1EA )
   
8. Electronic balance
   
9. Spectrophotometer
   
10. Benchtop refrigerated centrifuge (Eppendorf, catalog number: 5427 R )
    
11. Vortexer (Sigma-Aldrich, catalog number: Z755613-1EA )
    

Procedure