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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Spore revival from desiccated fruits * 13
    #28789802 -

Spore revival from fruits dried at 160F for longer than 24 hours:

 

  The following will be a departure from most of my cult posts which tend to be a tad long-winded, this is short and sweet. As many are aware I have been on an extended hiatus from cultivation, however, I still like to find ways to contribute to the knowledge base of the forum without the requirement for the production of fruit bodies.

I have often read arguments over the years concerning dried fruits producing growth on agar. The argument tends toward the assumption that both mycelium and spores die at or below the temperature used to desiccate fruits, without providing any definite answers for either argument. At the time I didn't own any live cultures or spore stock so, I approached a friend who happened to have some old fruits I had grown about three years ago stored in a Mason jar in some junk drawer in his spare room. These fruits were dried at 160F in my Cabela's 160L dehydrator for 36 hours; Why 36 hours? because a 160L dehydrator packed to the titts with dense PE takes a while to dehydrate properly.:lol:



The Process:

Pretty standard inoculation stuff here, sort of.:lol:

I took some cap material from the storage jar (3yo), then scrapped the gills onto 1.5% MEA using a disposable inoculation loop:



I then used a dash of sterile distilled H2O from a syringe to aid in spreading the spore-laden gill fragments evenly around the surface of the plate using a disposable cell spreader. I made 6 plates and then placed them on a shelf at room temp to do their thing. Roughly 4-5 days later I saw my first signs of growth:





One down, 5 to go:



The next to fall would be from mold and another to bacteria:



Then, about 8 days in:



It was too early to be certain but, it was looking promising. A few days later:



then another plate:



2 out of six were good to go, so I snagged a transfer from both, however, one of the 2 produced scalloped edges, which I trashed:



Leaving me with one viable culture:





The transfer expressed itself with half rhizo/tomentose, so I snagged a nice piece of rhizo:



It would appear that spores can be revived from old desiccated fruits at the temperature commonly used to dry fruits. It stands to reason that spores are hardier than mycelium, it also stands to reason that the above growth displayed morphology typical of spore germination and not that of revived tertiary mycelium.

Check out my Spore cleaning without a centrifuge or agar thread, which evolved from this topic.

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Re: Spore revival from desiccated fruits [Re: Stipe-n Cap] * 1
    #28789820 -

Cool!

Coincidentally yesterday I started a test with sprinkling grinded and dried fruits from capsule on agar. To see what happens.

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Re: Spore revival from desiccated fruits [Re: cybork] * 1
    #28789842 -

Great write up. I've always been under the impression they can be revived so long as cell walls stay intact. How far they can be pushed before failing has always been a question. I'm liking the answer.
:solidnod:

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Re: Spore revival from desiccated fruits [Re: the_chosen_one]
    #28789886 -

This is good stuff to know.


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          you can't learn anything.

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Re: Spore revival from desiccated fruits [Re: morningWood]
    #28790064 -

This is a very small sample size but proves the concept. Results may vary but, the odds are in your favor (for anyone attempting) considering this worked on the first attempt with 3yo gill fragments dried for longer than the average 24.

More work needs to be done to verify whether or not dried mushroom tissue is truely dead at those temps, however, it seems reasonable to assume that it is. I'm sure someone out there will read this and test to verify, hint, hint.

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Re: Spore revival from desiccated fruits [Re: Stipe-n Cap] * 3
    #28790092 -

I’ve always kinda figured tissue revival isn’t what was going on. It might be interesting to test some internal tissue that is dehydrated on its own, but then again there’s so many spores in my dehydrator it would be difficult to know if you were getting just tissue.

But I do think people “resurrecting” desiccated fruits are just putting spores to agar.


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Re: Spore revival from desiccated fruits [Re: Stipe-n Cap] * 3
    #28790097 -

I like this, I can’t even count how many cube varieties I collected using this method. When I started out it was a great way to sample mushrooms before I decided I wanted to grow them…

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Re: Spore revival from desiccated fruits [Re: Smellyhobbit] * 1
    #28790126 -

Internal tissue should be fee of environmental spore load, if done carefully, but I doubt it survives; if it had grown as a result of revived tissue I'd expect rhizomorphic growth from the gills off the hop, instead of floccose.

And to insert some uncertainty....here's some rhizomorphs emerging from the otherwise standard floccose growth:





I'm willing to bet that the spores germinated first, with revival of some gill fragments happening a week or so later, which would make sense.....maybe.


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Re: Spore revival from desiccated fruits [Re: Stipe-n Cap] * 2
    #29010952 -

Stipe-n Cap said:


I'm willing to bet that the spores germinated first, with revival of some gill fragments happening a week or so later, which would make sense.....maybe.




Hey can I get her number?  Just asking for a friend...

I'd be real interested to know if tissue is being revived, but like smelly I assume it's just spores?

No matter what thanks for posting this up as proof of concept :thumbup:


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New to cultivation or the shroomery?  Observe the hitchhiker's guide

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Smellyhobbit said:
"Wiping a needle with isopropyl after flame sterilization is like shitting after you wipe your ass."

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Re: Spore revival from desiccated fruits [Re: Mr Piggy]
    #29011886 -

Give it a shot. I'm currently unable to investigate further but would love to see someone else give it a go. If anyone happens to take up the baton, please post here so I can see the results.

I just noticed that the bacterial plate pic in the OP is a double :kingcrankey:

Well, Y'all get the point. Imagine the pictured plate has different bacterial growth than the first. :lol:

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Re: Spore revival from desiccated fruits [Re: Stipe-n Cap] * 1
    #29011992 -

You've accidentally tested something I've been wondering recently: whether heating spores will reduce the contaminant load appreciably, especially for the white scalloping molds. Apparently not. Glad I read this on impulse.


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The fool who persists in his folly will become wise.

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Re: Spore revival from desiccated fruits [Re: phenyl] * 1
    #29012099 -

Once all the cervid hunting seasons are over and I stop going crazy I might try to revive using internal stipe tissue :shrug:

See if we can reduce or eliminate the possibility of spores, fuck around and find out.


--------------------
🅃🄴🄰🄼 🄸🄷🄾🄿


New to cultivation or the shroomery?  Observe the hitchhiker's guide

The Hitchhiker's Guide to the Shroomery



Smellyhobbit said:
"Wiping a needle with isopropyl after flame sterilization is like shitting after you wipe your ass."

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Re: Spore revival from desiccated fruits [Re: Mr Piggy] * 1
    #29012106 -

Most def.

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Re: Spore revival from desiccated fruits [Re: Mr Piggy] * 5
    #29012115 -

Next time I Fire up the dehydrator I'll toss an actual print in there lol.
As for the tissue ... 160 is basically pasturizing Temps, be easy enough to toss a non dehydrated fresh fruit  with no cap into water at those Temps for an hour then try to clone , this would at least prove or disprove the temp parts but be easier than trying to sample from dry fruits that likely have spores all over them


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Re: Spore revival from desiccated fruits [Re: cronicr]
    #29012133 -

:popcorn:

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Re: Spore revival from desiccated fruits [Re: Stipe-n Cap] * 1
    #29012463 -

I had some Panaeolus cinctulus from Sept 2019 that were sun-dried and put into a ziplock baggie. That baggie found its way under a car seat and sat there through summer temps and winter temps for over 2 years before I found the baggie. They were then thrown into a box with other prints/specimens and forgotten about for another 2-3 years.

Dried gills were crushed and dumped onto 2 agar plates.



Clean transfers were obtained after 2 transfers from the original plate(s).



Still trying to fruit the 3 isolates I obtained but luck has been shitty for the past year of attempts.


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Re: Spore revival from desiccated fruits [Re: BlimeyGrimey]
    #29127396 -

Damn, just noticing this now. Pretty cool. :rockon:

Also bumping to remind cron of his reply above...

:standingby:

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Re: Spore revival from desiccated fruits [Re: Stipe-n Cap] * 5
    #29130710 -

So, I just had an idea...

Old gill fragments are obviously pretty likely to be carrying bacteria, as pictured in the OP. Now, if spores can survive 36 hours at 160F, that means spores can survive pasteurization...

If spores survive pasteurization temps, that means dirty syringes can be submerged  in a pasteurization bath to neutralize most pesky contaminants, thereby making them more reliable as inoculum.

Infected prints can be loaded into syringes or vials to be pasteurized in the same fashion.

Spore cleaning without a centrifuge or agar :rockon:

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Re: Spore revival from desiccated fruits [Re: Stipe-n Cap] * 1
    #29131111 -

Stipe-n Cap said:
So, I just had an idea...

Old gill fragments are obviously pretty likely to be carrying bacteria, as pictured in the OP. Now, if spores can survive 36 hours at 160F, that means spores can survive pasteurization...

If spores survive pasteurization temps, that means dirty syringes can be submerged  in a pasteurization bath to neutralize most pesky contaminants, thereby making them more reliable as inoculum.

Infected prints can be loaded into syringes or vials to be pasteurized in the same fashion.

Stipes spore cleaning prior to agar tek :rockon:


I have experimented with this by sous vide immersion of prints as I alluded to above, as well as other things to decrease load on spores -> decrease undetected molds in culture. Results were quite mixed. Some molds were quite shockingly thermo tolerant in experimental conditions to various... treatments.


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The fool who persists in his folly will become wise.

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Re: Spore revival from desiccated fruits [Re: phenyl] * 1
    #29131132 -

Perhaps running the spores at 160°F while suspended in vial or syringe will help. Let's continue this new topic in the new thread.

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