Surface sterilization of plant material

An elaborated example of surface sterilization
  • Cut the plant material to an appropriate size to fit the container
    which will be used during the sterilsation procedure
  • Rinse plant material under running tap water 
  • Shake for a few seconds in alcohol
  • HgCl2 (0.1 - 1%) + Teepol 2 drops/100ml: 3-5 min
  • Rinse in autoclaved distilled water
  • Commercial bleach 7-15% + Teepol 2 drops/100ml: 10-30 min 
  • Rinse several times in autoclaved distilled water

 

 
sterilization of orchid flower stalks

 

Notes: 
  • Tap water: to remove large debris, detergent can be used to shrub the surface
  • Ethanol: 70% is most effective to denaturate proteins. Sometimes 95% is used to dissolve the surface wax layer
  • NaOCl (commercial bleach) or Ca(OCl)2 : activity is based on both
    Cl- and other oxidation reactions
  • HgCl2* activity based on Cl- and heavy metal denaturation of proteins.
  • wetting agent (e.g. “Teepol”) can be added to the forementioned agents

* Hg is toxic for the environment, therefore recuperate the Hg-solution after use and collect in a large container. Hg can be precipitated by adding ammonia to the solution, and siphoning the supernatant