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OfflineRogerRabbitM
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Re: clone from dried caps [Re: vinsue]
    #2976099 - 08/07/04 07:11 PM (14 years, 8 months ago)

Glad you guys got the bugs worked out. This is a great way to store cultures.


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Re: clone from dried caps [Re: RogerRabbit]
    #2976312 - 08/07/04 08:22 PM (14 years, 8 months ago)

I wouldn't go that far... I still think that spores are germinating from the caps or that they are incompletely dried.

It wouldn't surprise me too much to find out that the mycelium can withstand drying with out dying though. It would be an excellent survival technique.

I highly doubt that a vacuum desicated or freeze-dried mushroom could be regenerated though. I also doubt that normaly dried mushrooms are viable for very long.


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Offlinephucknut68
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Re: clone from dried caps [Re: fastfred]
    #2976502 - 08/07/04 09:46 PM (14 years, 8 months ago)

take a look at that cluster of shrooms they all look the same he cloned it.

pinback: did the pinset/flush look and perform like a clone??


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Offlinescatmanrav
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Re: clone from dried caps [Re: phucknut68]
    #2978449 - 08/08/04 04:08 PM (14 years, 8 months ago)

Well I was doing a petri yesterday and did one using a dried shroom thats at least 2 years old as thats when I bought it. Cracker dry and has remained that way in an airtight jar with silica gel. I cut a peice of the gill, a peice from inside the cap and a peice from the base of the stem, the soft spongy part and threw them all on a plate. I dont really care if its a spore or the shroom that goes if it goes, which I doubt completly, but took only a second to try. I just would like to be able to produce these again, not sure what they were but they were very large, and very potent. Well I'll be posting if it ever shows anything.


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Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
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Turn your closet into a fruiting chamber
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OfflineRogerRabbitM
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Re: clone from dried caps [Re: vinsue]
    #2979280 - 08/08/04 10:24 PM (14 years, 8 months ago)

Totally, completely dried mushroom mycelia will grow again. It's a fact and not to be argued. It happens every year in the wild. Look at oyster mushrooms. In my area, there is NO rain from about mid May until early October. The forest fire danger is very high, and all the dead wood in the forest is cracker dry. If you cut into a log, as I did last week, you will see the mushroom myc, and it's as dry as a bone and looks dead. However, come October when the rains start, there will be oyster mushrooms by the truckload popping up from nearly every fallen tree, all over the forest. It happens every year. Culture banks store dried mycelia long term as a way of preserving isolated strains of edibles. The dried mycelia grows again when re-hydrated. Those aren't spores germinating, it's mycelium. Besides, I challenge anyone to grow oyster or shiitaki mushrooms from spores.


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InvisiblePinback
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Re: clone from dried caps [Re: phucknut68]
    #2979791 - 08/09/04 01:18 AM (14 years, 8 months ago)

Quote:

phucknut68 said:
take a look at that cluster of shrooms they all look the same he cloned it.

pinback: did the pinset/flush look and perform like a clone??




I don't think it's any unequivocal evidence either for or against:



The thing that makes me believe it's not spores is simply because it was an agar wedge I used. There is of course a (IMO small) possibility that cubensis spores in the air snuck into the petri dish where the wedges were drying, and landed on them. What speaks against this is that there are lots of other spores and bacteria in the air that could just as likely have landed on the wedges, but, there were no contaminations.

Nothing dried at room temperature will be "completely dry", there's always a small amount of water left, but I don't really get that argument. The wedge (dried for 55 days) was very hard, and actually split into two pieces when I speared it with a scalpell. I don't think it will get much drier than that at RT.

From what I gather, only four of the posters in this thread has actually tried this. I would be great if more did, especially the more skeptic ones.


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Re: clone from dried caps [Re: Pinback]
    #2980098 - 08/09/04 02:54 AM (14 years, 8 months ago)

A dried agar wedge?
How did you dry and store it ?

I wonder how long the dried mycelium would be viable ...
Would be a new method for long-term storage ...

Maybe one could make sure that no spores are germinating by using peroxidated agar ?


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Re: clone from dried caps [Re: ragadinks]
    #2980366 - 08/09/04 04:28 AM (14 years, 8 months ago)

I cut a wedge from a colonized plate, put it in a petri dish (the kind that doesn't make a perfect seal, I don't know the english word for it) and put it on a shelf away from direct sunlight, but still not dark. It was simply kept there for the entire time. I'm guessing the temperature was between 20 and 24?C.

It would really be interesting to see if it even survives on peroxidated agar. The reason vegetative mycelium survives, but not spores, is that the mycelium produces catalase, an enzyme that breaks down hydrogen peroxide. It is possible that the catalase might be denatured and no longer able to protect the cells when the mycelium is dried, but the only way of finding out is trying. Too bad it takes such a long time...


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Offlineragadinks
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Re: clone from dried caps [Re: Pinback]
    #2980375 - 08/09/04 04:31 AM (14 years, 8 months ago)

Hmmh, I guess the catalase won't survive the drying process, but as you mentioned: You only can be sure when you try it out.


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Re: clone from dried caps [Re: ragadinks]
    #2980606 - 08/09/04 07:56 AM (14 years, 8 months ago)

>Totally, completely dried mushroom mycelia will grow again. It's a fact and not to be argued.

I remain skeptical...

>It happens every year in the wild.

Do you have a citation for that? Perhaps it happens in the desert with certain species, but I've never read in any life cycle description that it actually dries out completely.

Culture banks store dried mycelia long term[...]

I don't mean to be argumentative, but could you provide some more information on that? Culture banks usually freeze samples in liquid nitrogen or keep them on slants. I've heard of them freeze-drying specimins, but not with the hope of reanimating them.

Could somebody describe the media they are using to get these results? The only mention I see is PDY... I would suggest that this is not the media to use as the yeast could contribute to the reanimation effect.

A foaf tried a similar experiment with LB (admittedly a shitty media) and did not get any growth.

-FF


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Re: clone from dried caps [Re: fastfred]
    #2980783 - 08/09/04 10:10 AM (14 years, 8 months ago)

In an earlier post in this thread, there is a link to a document from a culture bank describing this. The title of the document is "Instructions for Reviving Freeze-Dried Fungal Cultures"! Sorry, I don't have any specific details about the process, but it shows that it is actually done by professionals.


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Re: clone from dried caps [Re: Pinback]
    #2980914 - 08/09/04 11:28 AM (14 years, 8 months ago)



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Offlinehyphae
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Re: clone from dried caps [Re: fastfred]
    #2981290 - 08/09/04 01:18 PM (14 years, 8 months ago)

The reason freeze drying works for long term is that metabolic activity is almost stopped. Eventually a specimen will die if something is not done to stop this activity and we in our limited knowledge as humans cannot revive anything that is dead (actually dead) :wink: We can preserve and revive without question. Dried specimens over 1-20yrs. has not to my knowledge been successful in reviving, I'm open to documentation although, as apparently even after 55 days a "dried" specimen still is active metabolically. Like I first said I do not see this as any type of long term solution although thats not what the thread started out as anyway. :wink: GL


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Offlinescatmanrav
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Re: clone from dried caps [Re: hyphae]
    #2992448 - 08/11/04 08:10 PM (14 years, 8 months ago)

Well I grew green. I could try to dip in H202 and try again but I have not the will nor patience for that so good luck to you all with it :smile:


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay


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OfflineAquaman
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Re: clone from dried caps [Re: vinsue]
    #2999989 - 08/13/04 11:59 AM (14 years, 8 months ago)

Coming in late on this, but FWIW...

An aquafriend of mine re-animated some shake, from a 3+ year old bag, in honeywater.  This shake had been stored in a ziplock, with dessicant, at room temperature the entire time. 

Marginal, but definite mycelium growth. (more than likely spore germination & not clone, but still a bit exciting).  This particular jar of honeywater was forgotten about for over a year until he recently cleaned his fridge.  That same jar had a rather large culture in it, quite a bit larger than originally stored. 

Instead of tossing it, he decided to see if it was still viable.  He let the jar warm in a cabinet for ~12 hrs, innoculated some sterilized rye, and has mycelial growth in just 2 days  :smile:
Tough stuff.


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Re: clone from dried caps [Re: Aquaman]
    #4770204 - 10/07/05 07:25 PM (13 years, 6 months ago)

Sorry to bump this old thread, but I have some new data.

A little over a year ago, I took small samples of colonized grain and bird seed and put it in sterilized test tubes. I plugged the opening with sterilized cotton, and put the tubes on a shelf where they resided in ambient light at room temperature. Note that only a small amount was used to ensure fast drying.


Some two weeks ago, I removed a few kernels and put them on an agar plate (standard PDYA plus 0.024% H2O2). This is what the plate looked like yesterday:


It does seem a bit fishy - at least on the left side. It looks better today though, the far left mycelium looks very similar to the right side on the picture. I have transfered a few wedges of the good looking side to a jar with grain to test it for fruiting.

To sum it up, the mycelium has survived after having been dried for >380 days, and could be revived on peroxidated agar.


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OfflineRogerRabbitM
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Re: clone from dried caps [Re: Pinback]
    #4770623 - 10/07/05 09:55 PM (13 years, 6 months ago)

Thanks for the update and picture. I've long believed that dried mycelium will outlast even spores. You culture above looks like it was from multispore with two distinct substrains saved. That's the difference you're seeing.
RR


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Re: clone from dried caps [Re: Pinback]
    #4770751 - 10/07/05 10:37 PM (13 years, 6 months ago)

Quote:

Pinback said:
Sorry to bump this old thread, but I have some new data.





Hey, thank you for giving it that bump.
I was just asking RR if he could find it,lol.


Doc

:cool: :thumbup:


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Re: clone from dried caps [Re: RogerRabbit]
    #4772149 - 10/08/05 09:12 AM (13 years, 6 months ago)

You are correct, I was almost certain that it was a clone, but when I checked my notes it was from multispore. :blush:


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OfflineTerantula
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Re: clone from dried caps [Re: Pinback]
    #4776105 - 10/09/05 10:09 AM (13 years, 6 months ago)

totally amazing...

I thought this property was exclusive only to sclerota producing myceliums? Cool.

Learn something RADICAL today... lol


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Mushrooms, Mycology and Psychedelics >> Advanced Mycology

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