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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Re: AGAR ENVY! (Anything and All things agar!) [Re: tree frog] * 1
    #29516478 -

That makes more sense post germination, and it’s badass.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: tree frog]
    #29516581 -

I just read that page and it's at the very edge of my comprehension but

'
The basis of a compatibility test carried out in the laboratory is that small pieces of the strains that are to be tested are placed side by side on the surface of an agar medium, and hyphal interactions in these ‘confrontations’ usually show phenotypes that imply self/nonself recognition. When the confrontations are incubated, leading hyphae may mingle, and hyphal anastomoses occur between their branches. If the confronting strains are compatible the heterokaryon may proliferate so that the whole mycelium becomes heterokaryotic
"

I'm interested in agar plate wizardry and strain isolation, and how to get into mating and selecting and whatnot. The stuff in that page wasn't really a method or TEK and I want to learn how pick and choose the way you do.

Do I need to get a microscope or will using magnifying glass work?


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Edited by Archespore (03/15/26 12:09 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Archespore] * 1
    #29516589 -

To isolate monos and verify them, you need a microscope.

But if you want to isolate possible monos and possible two nuceli strains...

I suggest learning to do grab and drags.  And then just observing how they behave together on a plate.

https://www.shroomery.org/forums/showflat.php/Number/29505739


--------------------
And if I only could
I'd make a deal with God
And I'd get him to swap our places
I'd be running up that road
Be running up that hill
With no problems

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Re: AGAR ENVY! (Anything and All things agar!) [Re: tree frog]
    #29516611 -

I think I am ignorant of some fundamental concepts, I thought the paper was talking about two strains merging and becoming one, somehow. I'll do some more reading thanks boss


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Re: AGAR ENVY! (Anything and All things agar!) [Re: tree frog]
    #29516650 -

tree frog said:


And this plate has one promising looking culture on it and two the don't look great.  Because the cultures are right next to each other, it makes observing bacteria and other sus growth easier.





Just to be on the same page on the first photo in my quote, is what you're after the section where they have mated? And if so is this because it shows their compatibility?

And in the 2nd image its the left culture that looks the best? just to verify my guess :lol:

Is this also a good way to get a good culture from a print even if im not doing the grab and drag for monos and mating?? Just to get a bit more genetic diversity competing early?

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Re: AGAR ENVY! (Anything and All things agar!) [Re: DiscoDave]
    #29516655 -

So in that photo what happened likely was a parasexual cross.  Which is a whole other complicated topic.

Mostly what I was looking for was clean growth. And ensuring that they're dikaryons.  If they were monos or not well suited for fruiting, they likely would have crossed earlier.  And then I would have selected the cross.

I don't remember which culture I ended up running from that plate.  Probably the parasexual cross.

And yes, the left culture is the healthy looking one in the other photo.

And yes, doing this with a regular germination helps you observe how they behave when they are competing with each other.  If they're crossing early they're likely hooking up with more compatible partners.  If they're growing out for awhile and avoiding each other, they're likely happy with their current mate.

If they start knotting because they sense competition and ran out of room to colonize, you know you have a strain that can fruit.

RogerRabbit said:
BPJohnny said:
The Mushroom Cultivator

"Two dikaryotik mycelial networks can also gro together, exchange genetic material and form a new strain. Such an encounter, where two hyphal systems fuse, is known as anastomosis. When two incompatible colonies of mycelia meet, a zone of inhibited growth frequently forms. On agar media, this zone of incompatibility is visible to the unaided eye."

Now correct me if I'm wrong but in ANY multi spore inoculation

there are many different strains so this process will happen

regardless if you mix different syringes or just use one.


To a certain extent, yes.  They may fuse, and they may remain separate and fruit separate.  When unlike mycelium are placed together on a petri dish, this can be observed.  As Paul said, a line of inhibited growth forms between the two strains.  This can be observed below.  Both of these growths originated from the same sporeprint, but were isolated down to single sector isolated strains.  These two isolates are not compatible, therefore a line of separation has formed between them.


Other times, after the zone of isolation has formed, a distinct third growth will emerge from it. This third growth as shown below emerging from the 12 o'clock position is a hybrid of the two mycelium, and is usually more vigorous than either parent.


When spores germinate and then pair up with compatable hyphae to form multiple strains that then fuse by anastomosis, the results can be seen in the picture below.  The single sector at the bottom of the growth is steadly becoming more of the total as it assimilates the other strains into its network.  If allowed to grow, it might have consolidated the entire dish into one strain or it may have ran up against an incompatible strain as seen in the pics above and stopped.

Holding a petri dish up to light and looking at it from the agar side is a good way to see the line of inihibition between the strains.
RR


Fuck or Fight isn't a new tek :lol:


--------------------
And if I only could
I'd make a deal with God
And I'd get him to swap our places
I'd be running up that road
Be running up that hill
With no problems

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Re: AGAR ENVY! (Anything and All things agar!) [Re: tree frog]
    #29516662 -

Thank you for the clarification sensei treefrog, i am gonna try to put multiple transfers onto T1 plates next time I germinate. Will also post the results here when I get to it :biggrin:

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Re: AGAR ENVY! (Anything and All things agar!) [Re: DiscoDave]
    #29516669 -

It does likely work better if you isolate first.  The grab and drag allows for early isolation.  Which also helps with getting clean growth early. It's a simple tek I hope more folks try it, and aren't intimidated by the fact that it allows folks with microscopes to get and verify monos easily.

Transferring a wedge off of a regular germ plate, you are likely to end up with multiple cultures already that will sector, mate, compete, etc.

They'll just grow out from the single wedge rather than multiple different ones.  Which can make it harder to observe what is what.


--------------------
And if I only could
I'd make a deal with God
And I'd get him to swap our places
I'd be running up that road
Be running up that hill
With no problems

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Re: AGAR ENVY! (Anything and All things agar!) [Re: tree frog] * 2
    #29516820 -

Paying taxes, just made



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Beef’s The Hitchikers Guide to Shroomery
Stipe’s Clean Spawn Checklist
Gummies by milkboy
Eat’s Easy Oats Prep
DV8DUG’s Recognizing Contamination in Grain Jars

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Headass] * 4
    #29517124 -

Ha! Finally got some spores to germinate! :rockon:

Special thank you to Phaethon for the prints! You're a life saver! :cheers: :hug:

Made T1 transfers today. Yeah boi!! :awegroove:     

 


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Re: AGAR ENVY! (Anything and All things agar!) [Re: Tormato] * 4
    #29517168 -

I made my first attempt at agar the other day. :dancer:

I used Bod's agar tek found via the HHG.

10 plates, about 25ml per plate using MEA recipe.

Poured inside SAB and then wrapped in cling wrap (figured it wouldn't hurt right?).

I'm going to keep them closed up for a couple of weeks to see how clean they stay. This is day 3 and so far so good!





All the plates look clean close up. Any other artifacts in the pics are just photo shenanigans. Pretty happy!!

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Tormato] * 1
    #29517261 -

:rockon:


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Re: AGAR ENVY! (Anything and All things agar!) [Re: unhinge9315]
    #29517265 -

Quote:
unhinge9315 said:.

I'm going to keep them closed up for a couple of weeks to see how clean they stay. This is day 3 and so far so good!!!


Maybe nocc up a few in the meantime? Couldn’t hurt. That’s what agar’s for. Maybe lick one also.

Congrats on your cake germinations!

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Re: AGAR ENVY! (Anything and All things agar!) [Re: ModularMind] * 1
    #29517307 -

ModularMind said:
Maybe nocc up a few in the meantime? Couldn’t hurt. That’s what agar’s for. Maybe lick one also.

Congrats on your cake germinations!


Thanks! Plan is to watch them for a few more days to make sure I haven't stuffed it up, then inoculate some of them with the LC syringes I have. One GT that I used for the PF cakes and another B+ that I've got in reserve.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: unhinge9315]
    #29517413 -

unhinge9315 said:
I made my first attempt at agar the other day. :dancer:

I used Bod's agar tek found via the HHG.

10 plates, about 25ml per plate using MEA recipe.

Poured inside SAB and then wrapped in cling wrap (figured it wouldn't hurt right?).

I'm going to keep them closed up for a couple of weeks to see how clean they stay. This is day 3 and so far so good!





All the plates look clean close up. Any other artifacts in the pics are just photo shenanigans. Pretty happy!!


I think couple of weeks is not necessary. Mold will grow in 3-5 days, bacteria in 7-10 max.


--------------------
.
"If a person has arrived, at whatever age, at views and beliefs that later make him unhappy, then he can also, through effort of mind and sharpness of reasoning, learn to free himself from these ideas and replace them with ones that correspond more closely to reality."
-Lucien Augen

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Blizzloo]
    #29517415 -

Also,
Does anyone know what's going on here? It's a mush tissue


--------------------
.
"If a person has arrived, at whatever age, at views and beliefs that later make him unhappy, then he can also, through effort of mind and sharpness of reasoning, learn to free himself from these ideas and replace them with ones that correspond more closely to reality."
-Lucien Augen

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Blizzloo] * 1
    #29517430 -

Blizzloo said:
Also,
Does anyone know what's going on here? It's a mush tissue



I'm not sure what you mean by "going on" but if this is a tissue culture, it looks to me like you've got some reasonable mycillium growing there. 😎


--------------------
-Noku
Eskawata kaya waikiki
HHG
"Shake it like you would a newborn baby" - BeefExtreme

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Tormato]
    #29517431 -

Tormato said:
Ha! Finally got some spores to germinate! :rockon:

Special thank you to Phaethon for the prints! You're a life saver! :cheers: :hug:

Made T1 transfers today. Yeah boi!! :awegroove:     

 




You haven't had any issues using red food coloring in your agar? I wanted to use some but found out the red dye has antifungal characteristics so I never tried it


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Re: AGAR ENVY! (Anything and All things agar!) [Re: Blizzloo] * 1
    #29517434 -

If you mean the little tendrils it's just horny for food bro


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Re: AGAR ENVY! (Anything and All things agar!) [Re: Nokumana]
    #29517442 -

Nokumana said:
Blizzloo said:
Also,
Does anyone know what's going on here? It's a mush tissue



I'm not sure what you mean by "going on" but if this is a tissue culture, it looks to me like you've got some reasonable mycillium growing there. 😎


I don't know exactly how to describe it, but it reminds me of wet fur, or ice crystals on a sunny day. It's Iceberg with a history of reverting, so I also assume the mycelium itself is already starting to look a little different.

But I asked the question because I was also wondering if such an unusual appearance could indicate some kind of contamination.


--------------------
.
"If a person has arrived, at whatever age, at views and beliefs that later make him unhappy, then he can also, through effort of mind and sharpness of reasoning, learn to free himself from these ideas and replace them with ones that correspond more closely to reality."
-Lucien Augen

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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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