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InvisibleInocuole
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Anastomosis
    #23572992 - 08/24/16 02:24 PM (7 years, 5 months ago)

Too many non-advanced topics in Advanced myco. Let's talk about something slightly more advanced than "gargantuous monotubs" or "is this contaminated"?

What I'd really like is to see some pictures, and discussion of, anastomosis. For those who don't know, or don't like using google, anastomosis is the process by which the same, or different hyphae, fuse together. When this happens the individual stands begin to share genetic information with one another. This is an important concept when breeding to select desired traits, because if you are able to identify where anastomosis has occurred, you will more easily be able to select growth which is likely to share traits from its constituent spores/hyphae. ( https://en.wikipedia.org/wiki/Anastomosis )

Problem is, not many people do this kind of work, and even fewer do it publicly. As such, there are basically no pictures of anastomosis occurring with psilocybe mycelium. I don't know how many of you are working on strain-crossing projects currently, but if anyone has any pictures of anastomosis occurring on agar, I would be VERY interested to see them posted here. The more awareness surrounding this process, the less likely that incredible rare genetics will slip by people unnoticed, or be thrown away in unused agar plates.

I want to see this hobby take another huge step forward, and I think getting some more accessible information on this phenomenon in particular will help pave the way. If nobody has any pictures I'll keep trying until I get something and share my own pictures. Either way, let's get some discussion going and figure this shit out.


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InvisibleinskiM
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Re: Anastomosis [Re: Inocuole]
    #23574584 - 08/24/16 11:43 PM (7 years, 5 months ago)

It's a very good topic Inocuole and I too am getting tired of the off topic threads in this forum.

I wonder if it would be possible to stain one or both of the cultures to make it easy to differentiate between the two and then do time lapse photography through the microscope in order to observe anastomosis take place.


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InvisibleInocuole
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Re: Anastomosis [Re: inski]
    #23574592 - 08/24/16 11:45 PM (7 years, 5 months ago)

That's a really good idea...  problem I can see maybe encountering is that the process of staining the mycelium with something it can't quickly metabolize may slow it down.  Last time I used really dark food-colored agar, the mycelium took on that hue, and was a bit slower than usual.


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Re: Anastomosis [Re: Inocuole]
    #23574612 - 08/24/16 11:55 PM (7 years, 5 months ago)

Yes, i think the type of stain would be very important and some research and experimentation would be needed to find the right ones.

Regarding food colouring in agar, I purchased some black food colouring and found that it contained two preservatives, (211) and (202).
211 is Sodium benzoate.
202 is Potassium sorbate.

These two are the likely cause of the slow growth in your cultures on agar containing food colouring.


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OfflineGreg
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Re: Anastomosis [Re: inski]
    #23581794 - 08/26/16 09:08 PM (7 years, 5 months ago)

I have nice time-lapse gear and a 40x-2500x microscope.
I also have congo red stain which seems to dye cube myc nicely, not sure how it affects growth though.

I've been considering doing some timelapses of spores germinating, but I've had trouble with tiny vibrations making the captured images blurry.

If someone could capture this process (anastomosis) happening though that would be very interesting.


Edited by Greg (08/26/16 09:22 PM)


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Invisiblec10h12n2o
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Re: Anastomosis [Re: Greg]
    #23601830 - 09/01/16 04:52 PM (7 years, 4 months ago)

Wow, killer topic buddy, thanks for the great info as usual

I'm glad you put a word to this concept for me, I've been trying to learn more about this kind of thing and knowing the right terms will be tremendously useful while I am digging through academic databases :smile:

Is anastomosis what's going on here? I was throwing out some old plates and saw something crazy on one, never seen growth like this. My assumption was a reaction to some kind of bacteria or contam myc, but the thought of strains converging/exchanging crossed my mind



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InvisibleInocuole
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Re: Anastomosis [Re: c10h12n2o]
    #23601841 - 09/01/16 04:55 PM (7 years, 4 months ago)

That might be contaminated, but yes, anastomosis IS going on there.  I just can't say for sure that it's not the contam that's experiencing the anastomosis.


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OfflineTerpfreak
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Re: Anastomosis [Re: Inocuole]
    #23608304 - 09/03/16 02:53 PM (7 years, 4 months ago)

I'm having trouble understanding where should I look for more info?


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InvisibleInocuole
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Re: Anastomosis [Re: Terpfreak]
    #23608375 - 09/03/16 03:13 PM (7 years, 4 months ago)

Well, that's kind of the point of the thread.  I don't see much info on it, would like people with experience to offer some input.  If before that happens, I happen to be able to discern some things, then I'll certainly share those results.


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Re: Anastomosis [Re: Terpfreak]
    #23608449 - 09/03/16 03:44 PM (7 years, 4 months ago)



Edited by Psilosopherr (09/03/16 03:57 PM)


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OfflineTerpfreak
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Re: Anastomosis [Re: Psilosopherr]
    #23608531 - 09/03/16 04:10 PM (7 years, 4 months ago)

Real question here; why focus on michorizae if most fungi does this?


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OfflineTerpfreak
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Re: Anastomosis [Re: Terpfreak]
    #23608549 - 09/03/16 04:15 PM (7 years, 4 months ago)

Inocuole have you heard of carmine for dye? Its a bug thats powdered and is extremely powerful for coloring . Its only red though .


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Invisiblec10h12n2o
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Re: Anastomosis [Re: Inocuole]
    #23608569 - 09/03/16 04:20 PM (7 years, 4 months ago)

Quote:

Terpfreak said:
I'm having trouble understanding where should I look for more info?



Quote:

Inocuole said:
Well, that's kind of the point of the thread.  I don't see much info on it, would like people with experience to offer some input.  If before that happens, I happen to be able to discern some things, then I'll certainly share those results.




there are several thousand articles published in academic databases on the subject, though most of it pertains to cancer, and other versions of the phenomenon that dont really apply to our hobby

then there is stuff like this that has useful info, but is very dense and a totally different type of fungus, but it shows the type of diversity that can be seen within cultures even very similar cultures

Highly polymorphic in silico-derived microsatellite loci in the potato-infecting fungal pathogen Rhizoctonia solani anastomosis group 3 from the Colombian Andes.
Authors:
FERRUCHO, R. L.
ZALA, M.
ZHANG, Z.
CUBETA, M. A.
GARCIA-DOMINGUEZ, C.
CERESINI, P. C.
Source:
Molecular Ecology Resources. May2009, Vol. 9 Issue 3, p1013-1016. 4p. 1 Chart.

thanks again for sharing this post inocuole, very interesting stuff. i will also share if i find anything interesting

btw i did transfer some of that crazy crackle growth to see if it persists in subsequent plates

Quote:

Terpfreak said:
Real question here; why focus on michorizae if most fungi does this?




thats a valid question, although i am not sure if most fungi does this.

its so bizarre the transformations myc can go through, rhizo, tomentose, linear, and back again. and then this stuff! lots of dynamics to consider. makes me wonder what we might be missing out on by focusing on aggressive root-like growth


Edited by c10h12n2o (09/03/16 04:24 PM)


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InvisibleInocuole
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Re: Anastomosis [Re: Terpfreak]
    #23608590 - 09/03/16 04:26 PM (7 years, 4 months ago)

Quote:

Terpfreak said:
Inocuole have you heard of carmine for dye? Its a bug thats powdered and is extremely powerful for coloring . Its only red though .




I haven't, but one thing I'd be worried about is that the mycelial network doesn't really seem to work like veins with dye.  I've colored some agar black and paid close attention to the absorption of the color into the mycelium and it seemed to spread rather... indiscriminately, I guess I want to say?  That's not quite the same though, so maybe if the dye was very very concentrated and only placed in one tiny insertion point, it could be useful for highlighting what we're looking for.  I can't help but presume that it would just spread out as pink in all directions though.


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OfflineGreg
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Re: Anastomosis [Re: Inocuole] * 1
    #23608616 - 09/03/16 04:33 PM (7 years, 4 months ago)

That's why dyes like congo red that specifically bind to hyphae are so useful.


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Invisiblec10h12n2o
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Re: Anastomosis [Re: Greg]
    #23608674 - 09/03/16 04:55 PM (7 years, 4 months ago)

i had no idea!!! thats killer info greg :smile:

is congo red a good dye for working with fungi in general?


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OfflineTerpfreak
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Re: Anastomosis [Re: Greg]
    #23608689 - 09/03/16 05:00 PM (7 years, 4 months ago)

Im not sure what most of this means but here take this- sorry if Im de-railing please let me know.


Quote:

Carmine can be used as a staining agent in histology, as a Best's carmine to stain glycogen, mucicarmine to stain acidic mucopolysaccharides, and carmalum to stain cell nuclei. In these applications, it is applied together with a mordant, usually an Al(III) salt.


-wiki


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Re: Anastomosis [Re: Terpfreak]
    #23609375 - 09/03/16 08:57 PM (7 years, 4 months ago)

Kinda unable to post now but I will be back. . .


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Re: Anastomosis [Re: Pastywhyte]
    #23614485 - 09/05/16 11:26 AM (7 years, 4 months ago)

Cool thread.

I remember reading something about adding rattlesnake venom to agar encouraging anastomosis.

Stamets also talks about serially diluting spore solution so that you can germinate single spores and isolate the cultures before they fuse with other nearby single strains. So if I remember what he wrote correctly, you can put 2 cultures from single, unfused, strains on the same plate. IIRC this is basically the sexual reproduction method of mushrooms, and normally when you streak some spores on a plate this happens when nearby spores germinate and converge. But, if the spores are germinated at a sufficient distance from each other that you can take a wedge before it meets another culture, this is a good way to create hybrids.


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InvisibleInocuole
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Re: Anastomosis [Re: NumeroEno]
    #23614848 - 09/05/16 01:39 PM (7 years, 4 months ago)

:standingby:


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