I have some Pleurotus eryngii on Petri dishes with alternative solid culture medium and need to get fresh mycelium weight without the medium weight influence, which are the best and efficient techs to make clean separation?
Hi Samuel: there is a very simple recipe: place the agar on a fine methallic mesh or filter and pour boiling water in in until the whole agar desapears... you will get the clean mycelium. Then you can recover it by inverting the mesh on a filter paper, pouring water again to release the mycelium from the mesh. You need to know the dry weight of the fitler paper sheet beforehand so you can let it dry with the mycelium. Finally, you weight the filter paper with the fungus and substract the weight of the paper. Maybe you can get the fresh weight before driing, or carefully separating the mycelium from the mesh and weighting it... but I don't think that the fresh weight of an agarized culture means anything.
Instead of growing on solid medium, grow in broth. Then the mycelium can be harvested using cheesecloth, or muslin, as a filter (sterilised of course- by autoclaving). When growing in broth, the medium has to be shaken (gently for fungi- not the same as bacteria). We use Erlenmyer flasks, and a rotating shaking plate. Most common mycological media for plant pathogens is available in broth form.
Dear Samuel,
Simply you could cultivate the fungus on a PDA plate covered with a sheet of sterile cellophane. The cellophane separate mycelium from medium, which make the recovery of fungal mycelium is easy. However, one disadvantage of this method is that some fungal species are capable to degrade the cellophane. Good luck!
Best regards
Afra
Thanks for the answers friends i gone take these informations and put on my projects. Afra Khiralla I don't understand the principle and how this can separate the mycelium, you can explain more clear?
Hi Samuel,
Before you culture the fungus on petri dish, put a sterile sheet of cellophane on the selected medium surface, in order to seprate the mycelium from the medium. Then culture your fungus on the cellophane. Don't worry about that, because the transability of celluphane is good, so fungus can grow normally. Once you got a full growth, you could scratch fungal mycelium easily using steril spatula. I have used this technic to collect the mycelium for DNA extraction. Please find out this links:
DOI: 10.1093/femsle/fnw089
DOI: 10.1016/S0168-1656(99)00154-6
Good luck
Afra
1. Just add the desired solvent directly on Petridishdish, for example, methanol, slightly scrap with a sterile spatula.
2. Hot water treatment (suggested by Francisco Kuhar ) is also an good option but it will deteriorate the heat heat-sensitive metabolites.
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