I am currently working on tissue culture of orchids especially Dendrobium sps. and i am not able to overcome with the fungal contaminations. Can anyone suggest the surface sterilization technique to be followed for orchids?
@ Leeba Balan
My suggestions,
Along with the general surface sterilization procedure (after tween 20 or soap solution wash), 0.1 - 0.5 % of Bavistin (antifungal) can be used to wash the explants for 15 to 30 minutes to eradicate surface fungi.
If it is endophytic fungal contamination, there are some commercially available chemicals (from Sigma- I forgot the name!!!) which can be added in tissue culture media to control.
Best wishes.
I had the same issue with Brachypodium for a long time. I would suggest sterilization for a slighter longer time than you normally do and keep everything super clean, your bench, gloves, incubation cabinet etc. Make several batches of explants daily because sometimes you just cannot contain or get rid of the fungus once it spreads in one plate and then easily contaminates others. My best plate had got contaminated with fungus and I couldnt get rid of it no matter how hard I tried, I had to dispose of it. Adding antifungal to tissue culture media and ensuring your plates and explants remain totally dry before you incubate them are extremely essential. Overall better sterile technique during handling is imp. in this case prevention is better than cure.
Best of Luck!
To eradicate fungal contamination for orchids in plant tissue culture sterilization by washing with NaOCl, is recommended.
It is important to know that sometimes the contamination can be caused by your own mistakes during work or equipment pollution used or the failure of the laminar hood or autoclave. You should also consider endogenous contamination.
I have experience in sterilization of leaf and floral node stalk explants in Phalaenopsis orchids. n my experience, Silver nanoparticles is successful at 200 ppm concentration for 10 minutes or, this treatment [Sodium hypochlorite (20% with TWEEN® 20; 20 min)+ Alcohol (70%; 60 second)+ twice washing with sterille-distilled water (3 and 5 min, respectively)].
- Badges
- Science topic
- Similar topics
- Mathematics