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Re: AGAR ENVY! (Anything and All things agar!) [Re: LewDoja] * 3
    #29552835 -

In 25 years I have never used food coloring in media. It is 100% not needed and can be detrimental.


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Question everything, it's very responsive.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Nichrome] * 1
    #29552900 -

I got a T1 plate where I took 4 transfers from a germ plate of semperviva/subtropicalis and put them in a hole in the agar.

From what I can see there is several sectors in the growth, the top right wedge doesnt look to good to me so I would avoid taking transfers from this one. The leftmost growth seems to be the most vigorous to me and this is what I would normally transfer.

When it comes to the 'mixed' sectors, from what I gathered from treefrog above, is that when two dikaryons meet and they 'find out' that there is a better mating partner then they will form a new dikaryon with this nuclei combination. My question is then, would I want to rather take my transfers from the areas in between the wedges where it looks to be a new sector? In my head this will give a better mating combination than the wedges by themselves.

Input appreciated :biggrin:


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Re: AGAR ENVY! (Anything and All things agar!) [Re: DiscoDave]
    #29552919 -

Second question for another plate.

I recently streaked a 0.5% LME plate with TTBVI spores using grab and drag. When looking at the plate under the microscope (20x occular and 10x objective lenses) from the backside of the plate. I saw a lot of small threadlike grows, but I also saw what I think is the TTBVI spores next to them. It looks very similar in shape to the microscopy photos treefrog posted in her thread. My question is then, since the threads are much smaller than the spores, is it safe to assume that I messed up and introduced some contaminants? There are also thicker strands in the photo on the right side below the streak mark which I would think to be the TTBVI mycelium.

Input welcome :lol:



I really need to get a better microscope :rofl:

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Re: AGAR ENVY! (Anything and All things agar!) [Re: DiscoDave] * 1
    #29552925 -

You still want to select for the most vigorous growth.

The last couple of days I've been diving into this rabbit hole pretty deeply, and my current understanding is that dikaryons occasionally break down into monos.  And that's likely when the crossing happens.  Two dikaryons don't otherwise mate.

So it gives you more choices, but the best choice is likely still vigorous growth.

Anyway, I would give that plate a few more days.  Run the two most aggressive cultures and also store them both.  Then keep the best one for future projects.

As far as storage, stuffing a few wedges in a baggie (like a smash wedge style but not smashed) and storing them in jars in the fridge so they don't dry out is working pretty well for short-term storage.  Then when I revive them I just put them in a cabin to make sure I'm not transferring bacteria.

This was a revived pan cross that came from that technique.  It performed well so is back on AIO and also a master for summer grows.



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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]
    #29552934 -

Thank you for your wisdom O' great treefrog :biggrin:

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Re: AGAR ENVY! (Anything and All things agar!) [Re: DiscoDave] * 1
    #29552937 -

I wouldn't assume it's contamination yet.  It could be artifacts from the glass or alcohol etc.  Unless it's visible without a microscope.

The threads in the lower right look like mycelium.  But with a microscope I rely on clamps to tell mushroom producing mycelium from other fungi.

If you're trying to snag monos, I pull them early and scope them after they've grown out enough to tell if they're mold without a microscope (usually speed, growth patterns, and sporulation signify mold).

Then I use the scope to check for clamps by taking a wedge to a slide (I don't use a cover slip for mycelium anymore).  If I don't find any clamps I use it for breeding.


--------------------
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: DiscoDave] * 2
    #29552939 -

DiscoDave said:
Thank you for your wisdom O' great treefrog :biggrin:


Well I hope it's as accurate as it goes.  Some of the papers I was reading are only a couple of years old, and the exact mechanisms are both complicated and aren't settled science.


--------------------
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] * 4
    #29552977 -

Some pretty treasure coast mycelium. Too bad for that little bacterial colony up top. Oh well, opportunity for a nice picture :shrug::mushroom2:



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Island clean up
Pasteurizing Straw with Hydrated Lime
Swiss Cheese Fruiting Chamber

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Re: AGAR ENVY! (Anything and All things agar!) [Re: tree frog] * 1
    #29552987 -

tree frog said:
I wouldn't assume it's contamination yet.  It could be artifacts from the glass or alcohol etc.  Unless it's visible without a microscope.

The threads in the lower right look like mycelium.  But with a microscope I rely on clamps to tell mushroom producing mycelium from other fungi.

If you're trying to snag monos, I pull them early and scope them after they've grown out enough to tell if they're mold without a microscope (usually speed, growth patterns, and sporulation signify mold).

Then I use the scope to check for clamps by taking a wedge to a slide (I don't use a cover slip for mycelium anymore).  If I don't find any clamps I use it for breeding.


This plate had a bunch of condensation on it, and the grooves in the agar was a prime spot for the water to settle down in and diffuse into the agar a little. So there is some visible patterns without a microscope but I mostly think this is just water being deposited on the surface and becoming a part of the agar (it reflects light the same way as the agar next to it, but with a raised surface). But I guess if its mold it will take over quite quickly so I will be patient. Thanks again :biggrin:

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Re: AGAR ENVY! (Anything and All things agar!) [Re: RustBucketz]
    #29553025 -

RustBucketz said:
Found a Syzygy plate from a few years ago…





The plate itself is from years ago or the picture?

I dig the siggy pics, when did you grow those?


--------------------
- My trade list
- New here? Me too! Pop by and say hi c:

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Archespore]
    #29553120 -

Archespore said:
RustBucketz said:
Found a Syzygy plate from a few years ago…





The plate itself is from years ago or the picture?

I dig the siggy pics, when did you grow those?


Everything in the sig is from 3/4 years ago and the plate is just as old, I would say late 21’ - early 22’


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bucket full of rust

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Re: AGAR ENVY! (Anything and All things agar!) [Re: RustBucketz]
    #29553585 -

Saw a user (maybe Asura?) mention in a thread about agar transfer that they do not transfer beyond t2 and start over if it has to go beyond that. Is this standard practice? Is my agar going to be not as good after the 2nd transfer?

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Dj_karma_magnet] * 2
    #29553696 -




Getting back into it after a couple years and very excited. Bought some pre-made plates and made up some no-pours which I am very happy with.

I put a couple spore varieties down but haven't gotten any growth even after two weeks (no contamination though :cool:). The spores are old but I will keep waiting. Have some different spores coming so will try those. Lots of fun to come home and check on these haha. :grin:


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Re: AGAR ENVY! (Anything and All things agar!) [Re: Dj_karma_magnet]
    #29554046 -

Why are you transferring? For contams or for uniform growth, rhizo vs tomentose?

I like to try transferring to different recipes to encourage growth change if that's what you mean. Lme, PDA, adjust nutrient levels, water agar.

Today I poured reg strength PDA, low nute PDA, grain water agar, water agar. Got to test out my new stir magnet thingy. Was able to pour 6 bottles of agar without anything clumping, maybe one or two plates. I lost far more last time due to temperature gradients. I'm happy with the purchase



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- My trade list
- New here? Me too! Pop by and say hi c:

Edited by Archespore (04/20/26 05:48 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Archespore]
    #29554292 -

Transferring for contam. I like your idea about different recipes though

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Dj_karma_magnet] * 1
    #29554510 -

Dj_karma_magnet said:
Saw a user (maybe Asura?) mention in a thread about agar transfer that they do not transfer beyond t2 and start over if it has to go beyond that. Is this standard practice? Is my agar going to be not as good after the 2nd transfer?


Not at all, maybe in some exotics it's beneficial to keep diversity high.
Take as many as needed for clean uniform growth.

Don't forget about the cabin sequest.... Will speed up the process with problematic cultures

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Re: AGAR ENVY! (Anything and All things agar!) [Re: LewDoja] * 2
    #29554636 -

@TreeFrog

Suppose there's a big fat mono sitting there and on one side of it another mono slides in and that is dikaryon clamp history. But what if at the same time another swinger, from another mother, monokaryon jumps in and clamps up too on the other side? More interestingly what happens when the duality of the relationship meets in the middle?

Is this close to your B + E example from a page or two back?

This is the trikaryon? Double clamp connections possible?

Am I understanding this correctly as a real possibility(but not likely)?


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Agar Plates and Removing Hidden Contaminants - A Visual Notebook
Recognizing Contamination in Grain Jars - A Visual Notebook
Contaminated Condiment Cup Cleanup v1.0

Green Cap Pheno Isolation

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Re: AGAR ENVY! (Anything and All things agar!) [Re: DV8DUG] * 3
    #29554646 -



How's she looking so far? Is that 8 o'clock a potential problem area? I feel like there's an indent there vs the rest of the growth.

Thanks as always 🤗

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Re: AGAR ENVY! (Anything and All things agar!) [Re: DV8DUG] * 1
    #29554654 -

DV8DUG said:
@TreeFrog

Suppose there's a big fat mono sitting there and on one side of it another mono slides in and that is dikaryon clamp history. But what if at the same time another swinger, from another mother, monokaryon jumps in and clamps up too on the other side? More interestingly what happens when the duality of the relationship meets in the middle?

Is this close to your B + E example from a page or two back?

This is the trikaryon? Double clamp connections possible?

Am I understanding this correctly as a real possibility(but not likely)?


I like how your mind works :rofl:



--------------------
“All Fungi are edible.

Some fungi are only edible once.”
― Terry Pratchett


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Re: AGAR ENVY! (Anything and All things agar!) [Re: auroradawn]
    #29554681 -

Wow, that's gonna be beautiful. Looks like gecko fingers.

I can't wait to see what it looks like once it grows a bit more, please share. Looks great!


--------------------
- My trade list
- New here? Me too! Pop by and say hi c:

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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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