How to keep the mushroom mycelium long?
The mushroom mycelium wearing a synthetic medium (PDA, etc.) and other methods,
In reality, most affordable, ways that can keep for a long time to time mushroom mycelium and facilities?
There are several ways to keep your strains long, it depends on your facilities.
Best way: To keep them in liquid N2. You can take small plugs of 5 mm diameter form the edge of a colonie growth on PDA or MEA, and put for instance, five plugs into a 1.5 ml cryogenic tube and them fill the tube with 0.750 ml of a 10% glycerol solution. Then, you must inmerse the tubes in a N2 tank. If you do not have liquid N2 you can use a -80ºC or a -70ºC freezer. Once you want to recover your strains, it is enough to defreeze them by inmersion in hot water, afterwards you should empty the tube content over a plate amended with the medium chosen and remove the glycerol with a pipette. Finally: incubation.
Can you please specify, how long do you want to store mushroom mycelium? And what species?
We do need details of the mushroom you want to store and the duration. Further, what facilities you have - liquid nitrogen, - 80 degrees C freezer
Dear. Dr. Maria
My goal is mushroom (mycelium, spawn etc.)!
Thanks for your interests.
Sincerely Yours
2015.03.17.TUE. 09:04 (KOR)
By the method I explained, you can keep your strains at least two years, but even longer. Anyway I recommend to recover them every two years.
Dear Woo-Sik,
I asked that questions because as suggested low-temperature techniques are well in most cases, there can be some problems with several species and strains.
I faced problems with regrowth of Agaricus strains, especially homokaryotic after freezing and some others.
Anyway, some mushrooms can form dormnant structures such as arthrospores or sclerotia under certain conditions and can be successfully stored at 4ºC for about a year and even a little bit more in some cases.
Have a look at Hu et al. A new method for the preservation of axenic fungal cultures. DOI: 10.1016/j.mimet.2014.02.009 - the authors stored various fungi frozen for >10 years without loss of viability. Two caveats, based on how new the method is: 1) the authors tested the method with fast-growing r-strategist fungi. I have tried it with slow-growing fungi and it works, but I haven't tested that over the long periods the authors used. 2) We don't yet know if it affects fungal gene expression, etc - although the same can be said of many storage methods.
Dear. Jaime, Maria, Vikineswary, Ved, Sarah
The photo shows the mushroom attached occur at low temperatures.
On the same host in January 2015, occurred in December 2012. I'm here from January 2011 to January 2015 were observed four years occurred in the same period a year (winter time). This area is up more than 35 in the summer is winter, very cold areas, the minimum temperature falls to 30 degrees below zero.
In natural conditions the fungus in such bad condition will be preserved.
So What 's in the laboratory Otherwise, why?
My opinion: I think the biggest difference is that the host!
(natural: wood / laboratory: a synthetic medium).
Sincerely Yours
2015.03.19.THU. 18:30
http://gba.go.kr/page/page.htm?mode=view&&mcode=1363768370&mv_data=aWR4PTI5NTY4JnN0YXJ0UGFnZT0wJmxpc3RObz0zNTYyJnRhYmxlPWNzX2Jic19kYXRhJmNvZGU9Y29uc3VsdGluZyZzZWFyY2hfaXRlbT0mc2VhcmNoX29yZGVyPSZvcmRlcl9saXN0PSZsaXN0X3NjYWxlPQ==||
I'd suggest the main difference is cellular damage by ice crystals. The lab preservation techniques minimise this in 3 ways: 1) drying the culture before freezing; 2) very rapid freezing, which creates smaller ice crystals; 3) avoiding repeated freeze-thaw cycles.
All suggested techniques provide freezing rapid enough to prevent water crystallization. But in some cases there can be some problems apart from ice crystals.
Drying in mushrooms usually lead to decrease of its viability
Thank you. Sir fellow scientists [Especially Sarah and Maria]
"This poor vitality fungus in dry conditions," the idea is the same with me. Just a stream flows in the vicinity of the attached picture. One year to always maintain a relatively high humidity. However, the location of mushroom occurrence appears to be a change In time ever. The tree appears to slow the degradation of nutrients than synthetic medium.
2015.03.21. SAT. 00:05 [KOR]
Sincerely Yours
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