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InvisibleSWFL
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Spores on Agar Just Won't Take Off. Why?
    #28682434 - 03/01/24 10:54 AM (10 months, 11 days ago)

In the past I never had problems getting spores to take off on agar but my last 3 attempts have been total failures. It's almost like the spores are just dead for some reason. These are Pan cyan and cubensis spores that I've had for a few years but I didn't think they ever really expired.

I'm putting them on pre-poured (sterilized after pouring) malt extract agar in shallow mason jars made with a pretty standard recipe, inside of a still air box. I'm either getting nothing at all or contam that starts out looking kind of like mycellium but eventually turning gray and revealing its identity.

Is it possible that my spores are all dead for some reason or am I just screwing it up somehow that I'm not realizing?


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OfflineMycoplex
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Re: Spores on Agar Just Won't Take Off. Why? [Re: SWFL]
    #28682447 - 03/01/24 11:05 AM (10 months, 11 days ago)

Quote:

SWFL said:
In the past I never had problems getting spores to take off on agar but my last 3 attempts have been total failures. It's almost like the spores are just dead for some reason. These are Pan cyan and cubensis spores that I've had for a few years but I didn't think they ever really expired.

I'm putting them on pre-poured (sterilized after pouring) malt extract agar in shallow mason jars made with a pretty standard recipe, inside of a still air box. I'm either getting nothing at all or contam that starts out looking kind of like mycellium but eventually turning gray and revealing its identity.

Is it possible that my spores are all dead for some reason or am I just screwing it up somehow that I'm not realizing?




Have you tried switching up the agar recipe to see if the result is different?  Like using potato flake agar instead of MEA.

With older panaeolus spores, I've had both experiences:  germination and lack of after a couple weeks.    I don't think it's something specific about pans but I might be wrong, there are other growers here that have way more experience with them.

If your agar plates are contaminating eventually, try to narrow in on why that is happening.  If your plates are sterilized correctly and your SAB is working as intended they should remain sterile.

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OfflineYahra
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Re: Spores on Agar Just Won't Take Off. Why? [Re: Mycoplex] * 1
    #28682457 - 03/01/24 11:10 AM (10 months, 11 days ago)

Try to rehydrate the spores by making a syringe before you put them on agar.

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InvisibleSWFL
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Re: Spores on Agar Just Won't Take Off. Why? [Re: Mycoplex]
    #28682467 - 03/01/24 11:20 AM (10 months, 11 days ago)

I haven't tried potato agar recently but did use it successfully in the past so maybe I'll give it another shot. I've always been under the impression that the agar recipe wasn't really important as long as you've got a pretty reasonable balance of nutrition and proper viscosity.

I just looked at 8 non-inoculated agar plates I prepped a few months ago and 2 of them were contaminated, 6 were clean. And the two plates that are noc'd with spores are uncontaminated and have no growth at all after many weeks. I don't get it.

Maybe my agar is too dry? It's pretty rubbery. I think I'll go back to pouring plates with potato agar and try again unless somebody with a lot of pan cyan experience tells me otherwise. I really doubt that all my spore prints are duds even though they are a few years old. They've been folded in foil and tucked away in a ziplock bag so they should be fine.


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InvisibleSWFL
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Re: Spores on Agar Just Won't Take Off. Why? [Re: SWFL]
    #28682469 - 03/01/24 11:21 AM (10 months, 11 days ago)

I thought about this but don't really know how.

How does one hydrate spores without introducing contamination?


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OfflineRockinRobot
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Re: Spores on Agar Just Won't Take Off. Why? [Re: SWFL] * 1
    #28682473 - 03/01/24 11:24 AM (10 months, 11 days ago)

I use 1% LME 1.5% agar for germ plates.

You can hydrate spores by sterilizing a syringe of water in the PC and add 1 drop to the plate and streak the spores through it.

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OfflineYahra
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Re: Spores on Agar Just Won't Take Off. Why? [Re: SWFL]
    #28682475 - 03/01/24 11:26 AM (10 months, 11 days ago)

See Bod's So Easy A Caveman Could Do Itβ„’ Spore Syringe Making TEK

Let it sit a couple of days before you use it to inoculate the agar.

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InvisibleSWFL
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Re: Spores on Agar Just Won't Take Off. Why? [Re: RockinRobot]
    #28682492 - 03/01/24 11:41 AM (10 months, 11 days ago)

Quote:

RockinRobot said:
I use 1% LME 1.5% agar for germ plates.

You can hydrate spores by sterilizing a syringe of water in the PC and add 1 drop to the plate and streak the spores through it.




1% and 1.5% by weight of water? So like 1 c. water (240 grams) with 2.4 grams LME and 3.6 grams agar powder? Sorry, it's been a while and there are so many recipes here these days that I end up confused after searching the forum, lol.


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InvisibleFrank969
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Re: Spores on Agar Just Won't Take Off. Why? [Re: SWFL]
    #28682523 - 03/01/24 12:17 PM (10 months, 11 days ago)

Damn I had some spores of Orissa India that were some FIRE a couple years back and they all dried up and won't germinate. Gunna try to rehydrate them today. Will post results.

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OfflineRockinRobot
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Re: Spores on Agar Just Won't Take Off. Why? [Re: SWFL]
    #28682526 - 03/01/24 12:19 PM (10 months, 11 days ago)

Quote:

SWFL said:
Quote:

RockinRobot said:
I use 1% LME 1.5% agar for germ plates.

You can hydrate spores by sterilizing a syringe of water in the PC and add 1 drop to the plate and streak the spores through it.




1% and 1.5% by weight of water? So like 1 c. water (240 grams) with 2.4 grams LME and 3.6 grams agar powder? Sorry, it's been a while and there are so many recipes here these days that I end up confused after searching the forum, lol.




Your math is perfect there... and 240g of water is 240ml

Edited by RockinRobot (03/01/24 12:21 PM)

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InvisibleSWFL
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Re: Spores on Agar Just Won't Take Off. Why? [Re: RockinRobot]
    #28682593 - 03/01/24 01:12 PM (10 months, 11 days ago)

Thinking back now the spore prints are probably more like 8 years old. I'm guessing they're probably just too old and dead. Maybe I'll try to hydrate a print just for kicks anyway to see if they can be brought back to life but after some reading here it sounds like my chances aren't great.

Rainy season is not too far off now so worst case scenario I can go find a fresh specimen in a few months and start fresh...


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OfflineRockinRobot
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Re: Spores on Agar Just Won't Take Off. Why? [Re: SWFL]
    #28682659 - 03/01/24 02:12 PM (10 months, 11 days ago)

Quote:

SWFL said:
Thinking back now the spore prints are probably more like 8 years old. I'm guessing they're probably just too old and dead. Maybe I'll try to hydrate a print just for kicks anyway to see if they can be brought back to life but after some reading here it sounds like my chances aren't great.

Rainy season is not too far off now so worst case scenario I can go find a fresh specimen in a few months and start fresh...





Let me know if you don't have any luck. Be glad to send you some prints.

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