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Offlinegnirut0
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Registered: 02/18/20
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Hairy agar: is it a contam?
    #27015137 - 11/01/20 05:39 AM (4 years, 2 months ago)

Dear shroomerites,

After a few failed monotube attempts (got a contamination before pinning, I was told the contam was probably in the grain jars even if I couldn't see them) I decided to try agar again.

I had an old (~1 year) agar plate in the fridge and I did a transfer to two new agar plates. During one week, nothing happened, and during the second week, the mycelium started to grow. However, it looks a bit "hairy" and I'm a bit worried, I'd like an advice before transferring the agar to a grain jar.
I did a search on the forum for "hairy agar" and people told that it might be coweb, however here it is not growing fast, it took about a week to colonize the agar plate.

Here are my plates. I used red coloring on the agar, it did not cool very horizontally so on one side the agar is very thick, on the other side it is very thin. The agar is a bit messy at the center (there is a hole), because the mycelium was sticking to the scalpel and I had a hard time putting it on the plate...
First plate:


Second plate:


Here is a photo of the "hair":


What do you think about them?
Thanks!

Edited by gnirut0 (11/01/20 06:30 AM)

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InvisibleMateja
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Re: Hairy agar: is it a contam? [Re: gnirut0]
    #27015151 - 11/01/20 06:01 AM (4 years, 2 months ago)

What agar recipe is that?


--------------------
Cakes inside Water Tub

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Offlinegnirut0
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Re: Hairy agar: is it a contam? [Re: Mateja]
    #27015169 - 11/01/20 06:29 AM (4 years, 2 months ago)

I used 100g water, 2g agar, 4g liquid light malt extract, and some food coloring (didn't measure it).

I did a "no-pour" agar: I put 1 tablespoon of the mix in each petri dish, and sterilized it directly in the pressure cooker.

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OfflinePBJ710
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Re: Hairy agar: is it a contam? [Re: gnirut0]
    #27015282 - 11/01/20 08:18 AM (4 years, 2 months ago)

From what I can see it looks like its just tomatose growth which isn't necessarily a bad thing.  If you reduce the nutrients to 1.5-2% (vs. the current 4%) you should see more rhizomorphic growth out of them.  No obvious signs of contamination that I can see in those pics.

If you can take a picture of the plate with a light source behind it or at least white paper in the background, it may make it easier for us to see whats going on in there.

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Offlinegnirut0
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Re: Hairy agar: is it a contam? [Re: PBJ710]
    #27015764 - 11/01/20 01:30 PM (4 years, 2 months ago)

Indeed, I think I doubled the LME dose because since it's liquid I assumed that it would be less nutritious. I'll try to use 2% next time.

I didn't know about tomatose vs rhizomorphic! It seems it's rarely mentioned in TEKs for beginners.
From what I understood, rhizomorphic happen when you try to do strain isolation? Since I didn't do a lot of agar transfer (I did spore -> agar -> agar), it's probably normal to have tomatose growth.

Here are some photos with a light in the back:


Do you think I should try to inoculate a grain jar with one of the plates?

Thanks!

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OfflinePBJ710
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Re: Hairy agar: is it a contam? [Re: gnirut0]
    #27015875 - 11/01/20 02:43 PM (4 years, 2 months ago)

Yeah, it looks clean - just tomatose.  It's not a guaranteed thing by any means, but mycelium growing on really nutritious agar tends to be denser and fluffy like that since it has plenty of food where it already is living.  If it has to chase after the food it tends to grow outward more to find new food sources - IME using lower nute levels helps make the agressive growth show itself to help identify the best places to transfer from.

You might need a few more transfers as well just to thin down the genetic pairings in that culture.

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Offlinegnirut0
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Re: Hairy agar: is it a contam? [Re: gnirut0]
    #27015963 - 11/01/20 03:59 PM (4 years, 2 months ago)

Okay, thanks!

I think I'll try to inoculate a grain jar with on of the plates and try to fruit it, in the meantime I'll do some strain selection.
If I understood correctly, doing some strain selection can be dangerous since we can select a strain that fruits less easily or that is less potent?
If I fruit a mycelium with a lot of strains, I guess that there is a better chance that some strains give fruits?

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OfflinePBJ710
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Re: Hairy agar: is it a contam? [Re: gnirut0]
    #27016335 - 11/01/20 08:09 PM (4 years, 2 months ago)

It can, but it doesn't seem to occur very often.  Honestly I suspect most that don't fruit are actually environmental/culture cleanliness issues maninfesting as not fruiting, but that's still an unproven theory.  Maybe I just need to increase my sample size to see it occur?

Basically yeah, but I'll let people more adept in discussing isolation handle that subject :wink:

https://www.shroomery.org/forums/showflat.php?Number=27001555&page=0

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Offlinegnirut0
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Re: Hairy agar: is it a contam? [Re: PBJ710]
    #27016663 - 11/02/20 02:05 AM (4 years, 2 months ago)

Okay, thank you so much!

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