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OfflineDr.Ivan
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Please advise on mycelium on agar
    #26157111 - 08/28/19 12:52 PM (4 years, 4 months ago)

I have been working with agar for almost a year now and am seeing very mixed results, never really reaching the desired typical and beautiful rhizomorphic strains as shown here in the forum.

I have some plates to show you here with specific questions and would appreciated very much, if some of the pros here would chip in and help me understand what's going on. Please comment on the individual pics.

Here we go:

1. This is a plate with two mushroom pieces to clone. It looks strange how those strand are growing in a somewhat chaotic manner, not from inside towards the outside like it is normally seen.


2. With the Golden Teacher variety I always get plates with mycelium growing cloud like all over the place, not from the center outwards. This only happens for my golden teacher plates. All my plates come from the same spore prints, but different spore inoculations. Is this typical or is something wrong?




3. This A+ plate also shows this veiny structure, which I havent seen on any pics here in the forum. Is this because the plate is old already with end of may?


4. This is a grain spawn placed on agar and also shows this veiny structure, but its not as old as the previous one. Its from beginning of Jul. Any idea?


5. Does this look good at least? Does this need to be transferred still more times?


6. And is this good too?


7. This is also from may and looks like clouds somehow. Why is that?


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InvisibleMycolorado
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Re: Please advise on mycelium on agar [Re: Dr.Ivan]
    #26157121 - 08/28/19 01:01 PM (4 years, 4 months ago)

Those plates appear to be old.  Make transfers sooner, before they’re completely overgrown.  Start taking xfers when the culture is the size of a dime....look for stronger growth and make more xfers....repeat until you have uniform and strong growth.  Don’t just look for rhizo growth, but uniform and strong.


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OfflineDr.Ivan
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Re: Please advise on mycelium on agar [Re: Mycolorado]
    #26157125 - 08/28/19 01:05 PM (4 years, 4 months ago)

Quote:

Mycolorado said:
Those plates appear to be old.  Make transfers sooner, before they’re completely overgrown.  Start taking xfers when the culture is the size of a dime....look for stronger growth and make more xfers....repeat until you have uniform and strong growth.  Don’t just look for rhizo growth, but uniform and strong.



what should I do with those Golden Teacher cloudlike plates? How do you decide what to transfer and why do they continue to grow all over the place even after several transfers instead of from center out?


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OfflineJohnRainy
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Re: Please advise on mycelium on agar [Re: Dr.Ivan]
    #26157126 - 08/28/19 01:08 PM (4 years, 4 months ago)

Change your agar recipe is all I can suggest.

There may be other issues, or not, I am not a microbiology guy.

I have seen my transfers of strains go from rhyzomorphic to thin tomentose and back again...(I try to use a different nutrient in agar for each transfer because I read somewhere that is a good thing to do.)

Innoculations from those plates into grain always produced the same results, regardless of the plate myc's appearance.

Made me conclude that mycelium grows differently on different nutrient agars.


Edited by JohnRainy (08/28/19 01:21 PM)


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Offlinebadsponge
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Re: Please advise on mycelium on agar [Re: JohnRainy]
    #26157157 - 08/28/19 01:21 PM (4 years, 4 months ago)

Quote:

JohnRainy said:
Change your agar recipe is all I can say.
(I try to use a different nutrient in agar for each transfer because I read somewhere that is a good thing to do.)




I read that somewhere, too. But then later I read it only matters after several generations of transfers. I stick with MEA agar because it's clear, don't like the cloudiness of PDA. I only get up to 3-4 transfers before I use it all, anyway, and my growth is consistently strong.


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OfflineJohnRainy
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Re: Please advise on mycelium on agar [Re: Dr.Ivan]
    #26157167 - 08/28/19 01:24 PM (4 years, 4 months ago)

Quote:

why do they continue to grow all over the place even after several transfers instead of from center out?




Ive seen that too.  I don't like it.

I like to tell myself that maybe some cells of mycelium got spread around via a water droplet as I was parafilming the plate...


Edited by JohnRainy (08/28/19 01:25 PM)


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OfflineDr.Ivan
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Re: Please advise on mycelium on agar [Re: badsponge]
    #26157224 - 08/28/19 02:01 PM (4 years, 4 months ago)

I use MEA with 20gr agar and 15gr ME per l. I used do 20/20, but had the impression, that it makes the mycelium a bit lazier. Maybe I shoudl try different agar and ME brands and see if this changes anything. Still, it would surprise me, because with the same brands and recipe I had seen a few good plates too.

With regard to the Golden Teacher appearance, I think I can guarantee that there was no water droplet rolling around. It just magically seems to grow all over the place, so that I wonder, if that's even mycelium...


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InvisibleMycolorado
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Re: Please advise on mycelium on agar [Re: Dr.Ivan]
    #26157242 - 08/28/19 02:08 PM (4 years, 4 months ago)

Your media recipe sounds good.  What agar are you using?  That gt does look sketch.  I’d start fresh and make early xfers.  How are you noc’ing the plates with the spores and what’s the origin of the spores?


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OfflineDr.Ivan
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Re: Please advise on mycelium on agar [Re: Mycolorado]
    #26157255 - 08/28/19 02:25 PM (4 years, 4 months ago)

Quote:

Mycolorado said:
Your media recipe sounds good.  What agar are you using?  That gt does look sketch.  I’d start fresh and make early xfers.  How are you noc’ing the plates with the spores and what’s the origin of the spores?




I am using this agar here:
https://www.amazon.com/Hoosier-Hill-Farm-Agar-powder/dp/B0093DB9QO/ref=cm_cr_arp_d_product_top?ie=UTF8

I did already start fresh from spores, but have only 5 prints, each of different varieties, so had to use the same print. I tried to replicate this inoculation tek where you swipe 3 different overlapping sectors of the plate at the edges, one after another with loop sterilization in between. can't find it in the forum and forgot, if this has a special name, but I think t was supposed to be the professional way used in labs.

I am starting to question the spore quality, when I see some pics here with such different looking mycelium (like here:https://www.shroomery.org/forums/showflat.php/Number/18430998/fpart/all/vc/1). The spores are from an Austrian vendor and I have no idea, how old they are and how they have been stored and handled. I am actually unhappy with all 5 varities so far. Either I have a systematic problem or the spores a really bad. I am based in EU and probably should try another source.


Edited by Dr.Ivan (08/28/19 02:28 PM)


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OfflineDr.Ivan
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Re: Please advise on mycelium on agar [Re: Dr.Ivan]
    #26158802 - 08/29/19 10:29 AM (4 years, 4 months ago)

I only now realized that my vendor is a sponsor here and seems well known. That's my source for spores. They are regarded as a very good supplier it seems, so I would exclude that the spore quality has issues. I will now change my ME and agar brands and see, if this changes anything.

Thanks!


Edited by Dr.Ivan (08/29/19 10:57 AM)


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InvisibleMycolorado
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Re: Please advise on mycelium on agar [Re: Dr.Ivan]
    #26158822 - 08/29/19 10:46 AM (4 years, 4 months ago)

:rules: You need to edit the vendor/sponsor name out of your post as it’s against forum rules.  Vendor spores are for microscopy purposes only.


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OfflineDr.Ivan
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Re: Please advise on mycelium on agar [Re: Mycolorado]
    #26158844 - 08/29/19 10:58 AM (4 years, 4 months ago)

Quote:

Mycolorado said:
:rules: You need to edit the vendor/sponsor name out of your post as it’s against forum rules.  Vendor spores are for microscopy purposes only.



sorry, wasn't aware, thanks for reminding me


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InvisibleMycolorado
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Re: Please advise on mycelium on agar [Re: Dr.Ivan]
    #26158846 - 08/29/19 10:59 AM (4 years, 4 months ago)

:thumbup:


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