Dear colleagues. Recently I'm trying to grow fungal endophytes from liverwort Aneura on potato dextrose agar medium. I've tried to sterilise samples with 70% and 96% ethanol for 3-5 minutes. Unfortunately I can find optimal way of doing it, my results are: none growth on medium or only bacteria growth. Can anyone suggest better way of sterilisation?
I worked with Lunularia and other liverworths, even if my intent was to obtain axenic coulture of plants. In my experience, I had better results using sodium hypochloride solution, since some plant tissues sesm to be more sensitive to ethanol. My suggestion is to shortly sterilize the surface with a 5% sodium hypochloride (2-5 min), washing 4-5 times in sterile distilled water, and axenically cut the tissue; then put the tissue sections on agar plates (better if contaning a bacterial inhibitor, i.e. chloramphenicol).
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I agree with Francesca, you use a shorthly time for sterilize the surface with a 5% sodium hypochloride
Dear Piotr Wawrzyniak hi
I sugest you sterilize with 70% ethanol for 60-79 second and use a fungicide for 7 min+sodium hypochloride or Pbcl2
I agree with all my colleagues above.
A different methodology (if any of those listed above works for you... ) , is "flambé" the plant material ( this works very well for me when I have to isolate endophytic from some tissues, such as seeds and fruits , and some types of leavesr) .
Just immerse in 70% ethanol, put in the flame and take enough attention to the flame, killing the fire as soon as you you realize that the ethanol has dried (can not let it burn !).
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