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Invisiblehamloaf
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Re: Cloning from dried mushroom tissue techniques? [Re: fastfred]
    #18977365 - 10/14/13 04:20 PM (10 years, 4 months ago)

Haha.


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OfflineRogerRabbitM
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Re: Cloning from dried mushroom tissue techniques? [Re: hidyn]
    #18978579 - 10/14/13 08:43 PM (10 years, 4 months ago)

Quote:

hidyn said:
Quote:

RogerRabbit said:
If the grower was worth his salt he already expanded his spawn as far as practicable before inoculating the bulk substrate to grow those mushrooms.  If you create spawn from them, you start out with geriatric cell lines before you do your first grain to grain tranfer.
RR




Could you please expand on this?

Are you saying that the grower would just keep dividing their spawn until they had a pile of spawn bags full of mycelium that just didn't want to colonize anymore, then fruited them?

If a mycelial network can in fact 'die of old age', do you have any sorts of information as to how long this can take? How do you reset the clock? Can you just germinate spores from the fruiting bodies? If so, is there enough genetic material in the spores to start anew? Is letting multiple spores germinate and mesh together enough, or would you eventually need to cross your strains with someone else's?




Senescence is a real problem.  My best shiitake strain is over 30 years old and still performs very well.  My oyster strain is one of the best in the world.  I cloned it from a wild fruiting in 2004.

However, I'll take out a refrigerated slant to inoculate agar with the culture, and then inoculate a grain master.  After that, I'll do up to 5 or so grain to grain transfers.  I'm using 7 pounds of grains in each bag.  This gives the possibility for that tiny piece from my master slant to expand to over 70,000 pounds of spawn.  That spawn is then used to inoculate ten pound sawdust blocks.  Trust me, by the time that all colonizes and fruits, you don't want to clone a mushroom and start over.

Chronological age counts less than the number of cell divisions, but being a life form both are important and both contribute to the decline of a culture.
RR


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Offlinerev0kadavur
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Re: Cloning from dried mushroom tissue techniques? [Re: RogerRabbit]
    #18999341 - 10/19/13 11:34 AM (10 years, 4 months ago)

Quote:

RogerRabbit said:
I have a 150 year old mushroom growing book and in it they describe saving dried spawn for years.  They hadn't yet learned sterile technique in those days so they would dig up some mycelium from the mushroom beds, dry it, and then use it to spawn new beds later, often much later.
RR





I would love to find this book....


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OfflineKiya_Star427
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Registered: 10/14/13
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Re: Cloning from dried mushroom tissue techniques? [Re: RogerRabbit]
    #19001162 - 10/19/13 07:41 PM (10 years, 4 months ago)

Quote:

RogerRabbit said:
Peroxide is just going to damage the mycelium right when you want it to grow.

Use distilled water with no nutrients.  Wash a dozen or so small chunks about 2mm in diameter in a jar of distilled water.  Strain off and repeat the wash with fresh, sterilized distilled water. Repeat a few times and then leave the pieces floating after the last rinse in a jar with proper filter.

Wait for a piece or two to fuzz up, and then transfer to antibiotic agar.

Transfer as necessary until you have a clean culture.
RR




:like:


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Invisiblewildlifesc
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Re: Cloning from dried mushroom tissue techniques? [Re: RogerRabbit]
    #27690232 - 03/10/22 08:54 PM (1 year, 11 months ago)

So does this mean you should not clone from the fruiting body of a sawdust spawn or fruiting block that you buy?

I thought when you expand from the fruiting body it's considered a "mother culture".


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Offlinesedulous
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Re: Cloning from dried mushroom tissue techniques? [Re: wildlifesc]
    #27713131 - 03/29/22 04:33 PM (1 year, 10 months ago)

Cloning? Yes, but I've also "reanimated" dried fungal matter two ways:

1.) Not actually cloning but works: In the laminar flow, into poured LME/agar I carefully chose a specimen and removed the veil and scraped the gills into the agar. This worked and took several passes before I had a solid culture. It was tomentose for quite awhile. I believe that so many spores were mating at the same time. Eventually I was able to achieve a nice culture and began to work it towards a mono culture.

2.) In the laminar flow, into poured LME/agar as above I carefully chose specimens and proceeded to ONE AT A TIME hydrate sample A.) with sterile water from a pressure cooked jar. B.) hydrogen peroxide. Within 20-30 seconds of initial hydration, I was able to use my sterile tools to remove a section of tissue from inside the stipe and place upon agar.

Each specimen grew within seven days, albeit slow at first, it recovered, and grow for several generations. I was surprised as anyone would have been growing zombies and it's evidence of the unstoppable nature of the mycelium! RESILIENCE! Nothing abnormal to report otherwise.


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