Hi!
I would like to try to grow some wood decay fungus into petri dishes, but unluckily I know nothing about which is the method I should follow if I want to take them from some dead tree in the forest.
Is there any easy method that I can follow?
Thanks a lot!
Dear Aitor,
The most easy way for macromycetes is to establish a tissue axenic culture by isolation from a surface-sterilized fruitbody. To obtain a culture from mycelia can be a challenge due to numerous microorganisms adjacent to it in dead wood substrata. If isolation from fruitbody is suitable for your purposes I can add some references for technique, but actually this method is similar for most macromycete species.
With best regards,
Elena
Hi. thanks a lot for your fast answer. I'm not sure if that would be suitable, while I would like to use it afterward to try to decay some wood. If that works I think it could be possible to use the fruitbody.
Thanks a lot
Dear Aitor,
I believe that you can work with the technique of cultivation and isolation by peal. It usually works.
Hugs
I would suggest cutting several +/- 3x3cm pieces of wood including the limit between decayed and symptomless areas. Strong surface sterilization by 1% sodium hypochloride for 3 minutes, followed by 3 washings with sterile water and wrapping in sterile filter or absorbent paper.
Under aseptic condition carefully split the piece of wood with a sterile chisel or cutter and take aseptically several +/- 2x2x2 mm pieces of tissues across the section untuched by the splitting tool, starting from the symptomless side.
Transfer to Petri dishes containing water agar or a 1/10 strenght malt or PDA medium, keeping in mind that the agar concentration need to be kept high (15-20g/l) for consistency of the medium.
Incubate at room temperature and regularly check for mycelium growth and sporulation on the samples. Under the stereomicroscope check for absence of bacterial contamination and take hyphal tips or conidia for subcultures on fresh PDA or other general medium in Petri dishes or tubes. Check for absence of contamination.
In case of tenacious bacterial contamination the samples could be transferred to streptomycin amended media or dusted with a scalpel tip of streptomycin powder.
77
Hi Aitor
You can read this paper "A Biomolecular Method for the Detection of Wood Decay Fungi"
https://joa.isa-arbor.com/request.asp?JournalID=1&ArticleID=3080&Type=2
Also "Detection and Identification of Decay Fungi in Spruce Wood"
https://aem.asm.org/content/66/11/4725.full
And "Early-diverging wood-decaying fungi detected using three"
https://www.sciencedirect.com/science/article/pii/S1055790316000312
And "Control of growth of wood decay Basidiomycetes by Trichoderma spp"
https://www.fpl.fs.fed.us/documnts/pdf1991/bruce91a.pdf
Also "DIFFEEENTIATION OF WOOD-DECAYING FUNGI"
https://naldc.nal.usda.gov/naldc/download.xhtml?id=IND43969196&content=PDF
Also "Isolation of wood rot fungi from Maruthamalai hills Western Ghats"
https://www.ijcmas.com/vol-2-9/M.Meenakshisundaramand%20C.Bharathiraja.pdf
And "Wood decay under the microscope"
http://www.trepleieforum.no/foredrag/Francis_Schwarze_Wood_decay_under_the_microscope.pdf
Aseptically inoculate wood decay fungi into potato dextrose agar plate
Hello, Aitor Barbero-López
You can take a parts from infected wood (fruiting bodies, spores or directly from the infected pars). Inoculate on petri dishes containing the following composition: 1.0 g KH2PO4, 0.2 g MgSO4. 7H2O, 0.1g CaSO4, 0.01g Fe2(SO4)3, 10.0 g glucose, tap water to 1 liter; 20.0 g agar. Fungal hyphae of wood decay fungi will grow the whole surface of the dish so that you can purify a culture and identify the fungi.
Good luck
I also go inline with what the Prof. Jawad Abood A. Said but any of Potato dextrose agar or sabouroud dextrose agar can also be used for the culture
You can take a piece from surface-sterilized fruitbody (from the inside fruitbody, because we suppose that it is the "cleanest" part) by sterile scalpel and transfer it to the Petri dish with agar (you can try isolation on PDA or Malt agar). You have to work aseptically. When I preparing agar, I put hot agar into dishes and close them immediately due to keep moisture in the dishes. I mean that conditions of cultivation depend on taxonomic categories (e.g. genus). If you want to work with basidiocarps, you should use fresh fruiting bodies (only a few days after collection from nature). It´s not easy. Good Luck!
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