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Offlineouuwee
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Invisible looking ring around mycelium
    #26830564 - 07/18/20 10:53 AM (3 years, 6 months ago)

All my plates have this invisible looking ring around the mycelium and I'm not sure what it is

This is how it looks to the naked eye. Looks normal right?


This is how it looks if you angle it and have correct lighting. You can see this transparent looking ring around the piece I've transferred.


I've already made transfers from over 20 plates that have this issue and all the new plates have this invisible ring too! I found a picture (not mine) that looks exactly like the issue that I have. (Ignore the obvious contamination on the outer edge) What is it?? Is it contamination? If so, it's weird that it happened to all 40+ plates. I always flame sterilize on each transfer so it can't be cross contamination.



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InvisibleRoger Clemency
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Re: Invisible looking ring around mycelium [Re: ouuwee]
    #26830577 - 07/18/20 11:02 AM (3 years, 6 months ago)

I get the little exploratory outer ring on a lot of plates. I guess there are different situations though because mine have always been clean but I've seen other people saying they're always mold when they see them so IDK.

That bottom pic you posted looks suspect to me. When I get it it's not such a clean solid break between the thicker myc and thinner.

It's hard to tell anything though until they grow out some more.


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Offlineouuwee
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Re: Invisible looking ring around mycelium [Re: Roger Clemency]
    #26830765 - 07/18/20 01:21 PM (3 years, 6 months ago)

Ever since I got the invisible ring, growth has been slow. It's as if the ring is hindering the growth. I would have transfer plates sit for a week and a half with little to no growth.


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InvisibleLadysKnight
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Re: Invisible looking ring around mycelium [Re: ouuwee]
    #26830810 - 07/18/20 01:37 PM (3 years, 6 months ago)

Try just transferring a bit of the transparent ring to a new plate, it should show itself.


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InvisibleRoger Clemency
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Re: Invisible looking ring around mycelium [Re: LadysKnight]
    #26830844 - 07/18/20 02:01 PM (3 years, 6 months ago)

That sounds bad, 10 days after a transfer you should have a 3/4 full plate at least.


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Offlinecowcatcher
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Re: Invisible looking ring around mycelium [Re: Roger Clemency]
    #26831030 - 07/18/20 03:41 PM (3 years, 6 months ago)

I just started using Agar and I have the same thing on all of my plates. I'm using Bod's oat prep water agar recipe. I am hoping it isn't contamination as it appears on all of my transfers as well.


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:mushroom2:Micro dosing my blues away:mushroom2:


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OfflineItsaMeShroomio
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Re: Invisible looking ring around mycelium [Re: cowcatcher]
    #26831149 - 07/18/20 04:46 PM (3 years, 6 months ago)

If it's causing growth to slow/stop, then its probably a contam. I would try transferring to a different agar recipe and see if the myc can outrun it on that. I have found rye soak water agar to work well when I see a contam like that on standard LME agar. BRF agar seems to work too, but not as well. Weaker LME agar might also work, but I haven't tried that yet.


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OfflineA.k.aM
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Re: Invisible looking ring around mycelium [Re: ItsaMeShroomio]
    #26831158 - 07/18/20 04:52 PM (3 years, 6 months ago)

Mine do that and slow down and I’m pretty sure it slows because it’s started to grow through the agar instead of on top. Idk though.


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Offlinecowcatcher
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Re: Invisible looking ring around mycelium [Re: ItsaMeShroomio]
    #26831370 - 07/18/20 07:00 PM (3 years, 6 months ago)

It doesn't seem to slow mine down any, I have steady growth from start to finish. I have made a few small half pint jars of oats to test it out on before trying it on my larger oat jars. If it is contaminated it should show up quickly in the small jars. Depending on what I find I may try a different agar recipe.


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:mushroom2:Micro dosing my blues away:mushroom2:


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Invisibledfwerydfhg
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Re: Invisible looking ring around mycelium [Re: cowcatcher]
    #26834551 - 07/20/20 04:05 PM (3 years, 6 months ago)

I get this a ton, would love to definitively know what it is. It doesn't seem to have caused any problems, I've inoculated multiple batches of grain with plates that show it with no obvious contamination or problems fruiting. But I'm still a total rookie at this.


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Offlinemeowjinx
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Re: Invisible looking ring around mycelium [Re: dfwerydfhg]
    #26834590 - 07/20/20 04:48 PM (3 years, 6 months ago)

Could just be the fungus exploring its environment. If you're gonna conquer a new land isn't the first thing you do is send surveyors to draw out its boundaries? It could be sending microscopic hyphal filaments to establish the boundaries of its substrate and plan its growth strategy accordingly.

If it's trapped in a tiny space then it may start diverting resources earlier into entering its reproductive phase

Or it could just be looking for where the nutrient source ends and then growing outwards back to the main mycelial body to "gather up" all of the resources it discovered along its way

I dunno, I'm totally just guessing. But fungi are eukaryotic organisms just like animals and plants and their behavior can be much more complex than most people may assume. Just because aesthetically WE want mycelium to grow evenly outward in a pretty little circle does not mean that that is what the mycelium "wants"


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Invisibledfwerydfhg
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Re: Invisible looking ring around mycelium [Re: meowjinx] * 1
    #26835696 - 07/21/20 08:52 AM (3 years, 6 months ago)

I get the feeling that there's a bit of "tea leaf reading" going on with agar. Half the time I see threads about this issue there's at least one person (usually with far more experience than me) insisting that the ring indicates mold. Without microscopy I guess the best confirmation is to grow it out and try to fruit.

Doesn't help that people usually post either their best rhizo plates, or obvious contams. Everything in the middle mostly doesn't show up so rookies like me don't get an idea of what's normal, except through experimentation. Which is the best way to learn anyway!

I might try varying my agar recipe, see if that alters the way this shows up.


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InvisibleLadysKnight
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Re: Invisible looking ring around mycelium [Re: dfwerydfhg]
    #26835774 - 07/21/20 09:53 AM (3 years, 6 months ago)

This is about the best pic (d3monic's) I've seen of the moldy halo. (@10:00)
Notice how the clear halo is only around the mold, on the left of the culture.



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Invisibledfwerydfhg
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Re: Invisible looking ring around mycelium [Re: LadysKnight]
    #26835825 - 07/21/20 10:42 AM (3 years, 6 months ago)

Is that mold though? Or just the interface between straggly tomentose mycelium, and a faster growing rhizomorphic part? A ton of my early transfer plates look like that, I've just never paid too much attention to the "halo" and whether it appears on the rhizo bit also.

If you took the bit from 7-8 (on D3monic's plate) and translated it all the way around in an image editor, you'd have what looks like several plates I've successfully taken to fruiting on grain, with no signs of contamination. It's just not rhizo. What if the halo is the way this particular kind of growth happens?

Here's an old pic of mine that shows the same thing. Obviously not as nice as D3monic's!

My usual practice is to take a transfer from the rhizo bit and think no more of it. But sometimes the new plate grows out looking like the bit with the ring. And if it's even, symmetrical growth, it (so far) hasn't caused me any problems.

Here's a similar plate that's grown out all the way: the suspect bit is knotting!


Is there any other experiment that might positively identify that stuff as mold? Next time I see it, I'll for sure grab a bit of the "halo" and put it to it's own plate and see what happens.


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OfflineAtmozFear
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Re: Invisible looking ring around mycelium [Re: dfwerydfhg]
    #26835861 - 07/21/20 10:55 AM (3 years, 6 months ago)

Quote:

dfwerydfhg said:
I get the feeling that there's a bit of "tea leaf reading" going on with agar. Half the time I see threads about this issue there's at least one person (usually with far more experience than me) insisting that the ring indicates mold. Without microscopy I guess the best confirmation is to grow it out and try to fruit.

Doesn't help that people usually post either their best rhizo plates, or obvious contams. Everything in the middle mostly doesn't show up so rookies like me don't get an idea of what's normal, except through experimentation. Which is the best way to learn anyway!

I might try varying my agar recipe, see if that alters the way this shows up.



:whathesaid: well said.


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InvisibleLadysKnight
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Re: Invisible looking ring around mycelium [Re: LadysKnight]
    #26836017 - 07/21/20 12:30 PM (3 years, 6 months ago)

I never saw a halo til last year I got a new print and everything that came from it had this halo. I suspected it to be contamination because every plate took a month to grow out. Every jar of grain took a month or more to finish. Every shoebox took a month to colonize. When they fruited, it took SEVERAL weeks to go from tiny pin to full size. Sometimes the whole tub grew with the caps leaning upside down. I tried for 6 months to clean up the cultures, even clones had the halo, til finally I isolated the halo and saw it was an extremely slow white mold with a clear edge. So yes cultures would still look totally normal on plates other than the halo and the myc would still fruit.

I understand there are other types of halos, and other people have other experiences, but I will be avoiding halos.


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OfflineA.k.aM
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Re: Invisible looking ring around mycelium [Re: LadysKnight]
    #26836023 - 07/21/20 12:32 PM (3 years, 6 months ago)

How would a clone have the mold?


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Invisibledfwerydfhg
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Re: Invisible looking ring around mycelium [Re: LadysKnight]
    #26836117 - 07/21/20 01:36 PM (3 years, 6 months ago)

Quote:

LadysKnight said:
I never saw a halo til last year I got a new print and everything that came from it had this halo. I suspected it to be contamination because every plate took a month to grow out. Every jar of grain took a month or more to finish. Every shoebox took a month to colonize. When they fruited, it took SEVERAL weeks to go from tiny pin to full size. Sometimes the whole tub grew with the caps leaning upside down. I tried for 6 months to clean up the cultures, even clones had the halo, til finally I isolated the halo and saw it was an extremely slow white mold with a clear edge. So yes cultures would still look totally normal on plates other than the halo and the myc would still fruit.

I understand there are other types of halos, and other people have other experiences, but I will be avoiding halos.




Thanks for the background! To be clear, when you said you isolated the mold, do you mean you got a plate that was free of mycelium, only the mold? I'm interested if it grew out in a circle that was transparent the whole way, or did it change its morphology at some stage? Can you remember if it was associated only with wispy growth, or did you see it on the edge of strongly rhizo mycelium also?

By the sounds of it so far, your experience is very different to mine- my "halo" plates grew out at a normal rate. So we may be talking about different things that happen to look the same.

I spent a little time trawling through D3monic's photos, just because he is the most prolific photographer of agar that I could think of (and takes great photos). One good example is this one. You can see there's several examples of a plate being shared between rhizo mycelium and wispier stuff, and in most cases the rhizo has no halo but the wispy stuff does (to varying degrees). You can also see that the growth rate has been about the same on either side.


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InvisibleLadysKnight
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Re: Invisible looking ring around mycelium [Re: dfwerydfhg]
    #26836176 - 07/21/20 02:00 PM (3 years, 6 months ago)

Quote:

dfwerydfhg said:

Thanks for the background! To be clear, when you said you isolated the mold, do you mean you got a plate that was free of mycelium, only the mold? I'm interested if it grew out in a circle that was transparent the whole way, or did it change its morphology at some stage? Can you remember if it was associated only with wispy growth, or did you see it on the edge of strongly rhizo mycelium also?




The first isolation was accidental. It came from touching a transfer to a spot on the plate before placing it in the center. The transfer grew out normally (for that print), looking like rhizomorphic mycelium with the shiny, clear halo at the margins. The accidental spot grew a clear, shiny round culture that eventually whitened with age, from the center out. After that accidental isolation, I transferred some to a clean plate so it was the only thing on the plate. This plate grew the same as the accidental isolation and smelled distinctly of mold when none of the other plates nor jars did.


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OfflineMarxcelium
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Re: Invisible looking ring around mycelium [Re: dfwerydfhg]
    #26836205 - 07/21/20 02:17 PM (3 years, 6 months ago)



This is unfortunately the best picture I have of these cultures right now, but the Puerto Rican on the right (purple agar) has the halo 100% of the time on LME agar (way less noticable on BRF agar, but that could be because it tends to be more matte than shiny). It's also always tomentose and extremely fluffy. Can't get it to go rizo no matter what I do.  It's never showed contams on plates, in jars, or spawned to tubs in the two plus months I've been working with it, and it's just as vigorous if not more so than the other cubes I've got around.  The clear halo very obviously seems like the leading edge of the myc in this case, so it's not always mold, at least.


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