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Offlinecatterpillar
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Difficulty getting spores to germinate
    #21335431 - 02/26/15 08:44 PM (9 years, 10 months ago)

So I have a few spore prints which I'm having a lot of trouble getting anything to grow from. I don't have any reason to believe that they are all dead. The prints haven't been subjected to any heat or chemicals or anything. I've been trying to grow them on agar. One is a panaeolus cyanescens, one is a psilocybe cyanescens and the other is an Agaricas Augustus. I used an agar recipe with 9 grams of agar, 10 grams of malt extract and 500mL of water. The Augustus I've had trouble with on other batches of agar too.

What might have gone wrong? If it's the prints is there anything I can do to make them more likely to germinate?

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OfflineMushroom_J
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Re: Difficulty getting spores to germinate [Re: catterpillar]
    #21335461 - 02/26/15 08:49 PM (9 years, 10 months ago)

try 6-7 grams agar.
spores need more liquid to germinate. 9 grams to 500ml of water is pretty stiff. Other than that... i have no idea.


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Offlinecatterpillar
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Re: Difficulty getting spores to germinate [Re: Mushroom_J]
    #21335557 - 02/26/15 09:10 PM (9 years, 10 months ago)

Thanks! I'll try that.

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InvisibleGrey
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Re: Difficulty getting spores to germinate [Re: catterpillar]
    #21335594 - 02/26/15 09:17 PM (9 years, 10 months ago)

How long have they been on agar?


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InvisibleSpitballJedi
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Re: Difficulty getting spores to germinate [Re: Grey]
    #21335641 - 02/26/15 09:25 PM (9 years, 10 months ago)

Sometimes I just make a small grain jar and inoculate that with spores. Then transfer a single colonized grain to agar.


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Offlinecatterpillar
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Re: Difficulty getting spores to germinate [Re: SpitballJedi]
    #21335659 - 02/26/15 09:30 PM (9 years, 10 months ago)

Quote:

9 grams to 500ml of water is pretty stiff. Other than that... i have no idea.




Now that I think about it I'm pretty sure the agar recipe was supposed to be 5 grams of agar and 6 grams of malt extract.... Which would also explain why the cube spores that germinated are growing really slowly.

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Offlinemushpunx
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Re: Difficulty getting spores to germinate [Re: SpitballJedi]
    #21335700 - 02/26/15 09:37 PM (9 years, 10 months ago)

Quote:

SpitballJedi said:
Sometimes I just make a small grain jar and inoculate that with spores. Then transfer a single colonized grain to agar.





Im with SBJ on this one, I took this advice from him before. Now whem I have trouble getting germination on agar its my go to

Pan Cyan man thats the species that gives me trouble getting germinatiom. Even on grain I could only get a few grains to take :shrug: its from syringe I have better luck swiping prints


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Invisibled0urd3n
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Re: Difficulty getting spores to germinate [Re: mushpunx]
    #21335723 - 02/26/15 09:42 PM (9 years, 10 months ago)

Do you make a spore syringe or just wipe the spores from the loop onto the grain?


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Offlinecatterpillar
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Re: Difficulty getting spores to germinate [Re: catterpillar]
    #21335755 - 02/26/15 09:48 PM (9 years, 10 months ago)

I usually transfer the spores onto the agar using an inoculating loop. The spores have been on the agar a couple of weeks... Though the Agaricus I've previously left on there for much longer.

But the more I think about it the more I think I used a completely different recipe and just had a brain fail when mixing this one up. I think maybe I used 6 grams of agar and 5 grams of malt extract... Does that sound right? Most recipies on the net say 10 grams of agar though. But I never used to have this problem before.

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OfflineJunior Fungus
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Re: Difficulty getting spores to germinate [Re: SpitballJedi]
    #21335902 - 02/26/15 10:22 PM (9 years, 10 months ago)

Quote:

SpitballJedi said:
Sometimes I just make a small grain jar and inoculate that with spores. Then transfer a single colonized grain to agar.




Seems like a great way to do it. Have you ever gone from print to grain jar directly? I guess I would use my inoculation loop and just rub spores onto the grain... Any advice on doing that?


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OfflinePsyCLown89
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Re: Difficulty getting spores to germinate [Re: Junior Fungus]
    #21337631 - 02/27/15 10:23 AM (9 years, 10 months ago)

Hmm, that is a lot of agar.

I have found using little again is graet for getting spores to germinate - have had amazing success since reading about this and giving it a try. I think it was quoted by RR somewhere, I took it and adjusted it to what works for me.


I use 200ml water, 1.5g agar and 1.5g potato flakes. The agar is fairly "soupy" and works very well for getting spores to germinate I have found.
Try it and adjust a bit.


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InvisibleSpitballJedi
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Re: Difficulty getting spores to germinate [Re: PsyCLown89]
    #21337763 - 02/27/15 10:50 AM (9 years, 10 months ago)

When I go print to grain, I have a long tweezer and just grab a grain and dab it on the print. I flame sterilize the tweezer and I use half pint wide-mouth jars. It's a small amount of grain, maybe like a 1/4 cup.


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Invisibled0urd3n
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Re: Difficulty getting spores to germinate [Re: SpitballJedi]
    #21337793 - 02/27/15 10:54 AM (9 years, 10 months ago)

:takingnotes:

Great idea. Haven't heard of this as a method. :thumbup:

Might work well with grass seed for hard to germ Panaeolus species.


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InvisibleSpitballJedi
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Re: Difficulty getting spores to germinate [Re: d0urd3n]
    #21337876 - 02/27/15 11:09 AM (9 years, 10 months ago)

Once some of the grain colonizes, I just grab one and put it on agar and continue from there as usual.


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Invisibled0urd3n
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Re: Difficulty getting spores to germinate [Re: SpitballJedi]
    #21337894 - 02/27/15 11:12 AM (9 years, 10 months ago)

Yeah makes sense. Thanks SBJ. :thumbup:


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InvisibleSpitballJedi
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Re: Difficulty getting spores to germinate [Re: d0urd3n]
    #21337924 - 02/27/15 11:19 AM (9 years, 10 months ago)

:thumbup:


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Offlinestreamer

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Re: Difficulty getting spores to germinate [Re: SpitballJedi]
    #21337964 - 02/27/15 11:32 AM (9 years, 10 months ago)

Quote:

SpitballJedi said:
When I go print to grain, I have a long tweezer and just grab a grain and dab it on the print. I flame sterilize the tweezer and I use half pint wide-mouth jars. It's a small amount of grain, maybe like a 1/4 cup.




Great tip!! After many months of reading, I'm always amazed to still learn something almost every day!

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OfflinePsyCLown89
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Re: Difficulty getting spores to germinate [Re: streamer]
    #21337970 - 02/27/15 11:35 AM (9 years, 10 months ago)

Hmm, that is really interesting. I think I should try print to grain like that sometime.

Thanks SBJ!


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OfflineKizzle
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Re: Difficulty getting spores to germinate [Re: PsyCLown89]
    #21339019 - 02/27/15 03:35 PM (9 years, 10 months ago)

As long as you're not seeing any bacteria or anything I would suggest trying more transfers. You could even do multiple streak on a single plate to save on agar. Got to be some viable spores left somewhere on those prints.


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Offlinecatterpillar
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Re: Difficulty getting spores to germinate [Re: Kizzle]
    #21341255 - 02/28/15 01:28 AM (9 years, 10 months ago)

Thanks everyone for the advice. This discussion has been very helpful!

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Offlinetourrat
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Re: Difficulty getting spores to germinate [Re: catterpillar]
    #21341841 - 02/28/15 08:03 AM (9 years, 10 months ago)

Thank you for the grain idea...I've got some azures and cyans that I'm having a hell of a time with. I'm gonna give this a try.


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