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Way
The


Registered: 01/14/23
Posts: 4,336
Loc: A long way away
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Hey Stipe, I started with an understanding of strains, sectoring, and monocultures from Stro's post. I've since read some more of your explanations on various posts and obviously have come to realize that it's not entirely correct.
So with my new understanding of "monocultures" and "isolates", I still am left wondering about "sectors". You've explained that they aren't individual strains showing in agar, and I can get down with that. Makes complete sense to me.
What exactly are they though? Why does mycelium "sector" out like that and is there any benefit to paying attention to the "sectors"? Besides trying to get a more uniform culture of course.
I don't generally pay any attention to them besides just looking for the fastest, densest, most organized part of the culture, and that has been working great so far.
Just wondering if you can push my understanding of this forward a bit. Thanks for all the information you've provided to everyone, I really appreciate it.
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That's the way she goes, boys. Sometimes she goes, sometimes she doesn't, cause that's the fuckin way she goes.
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Re: Why Agar? [Re: Way] 5
#28583919 - 12/15/23 12:55 PM (1 month, 13 days ago) |
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Hyphae extend radially from a central point of inoculation. This central point consists of many aggregated strains which have branched and fused dependent upon mating type compatibility.
 
When starting from multispore, individual monokaryotic hypha branch out and immediately fuse with a suitable monokaryon, producing a dikaryotic single strain, until that dikaryotic strain finds further suitable monokaryons, then mate.
  
The dikaryons can anastomos with compatible monokaryons through what as know as "Bullers phenomenon". Dikaryons cannot fuse with other dikaryons, only compatible monokaryons, and monokaryons with other compatible monokaryons. This monokaryotic stage is short lived due to the high compatibility of mating types. This is taking place at microscopic scale, very tight quarters, which is why any attempt to tease out individual strains is impossible without the aid of serial dilution, microscopy, and specialized tools for manipulation.
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Hyphal aggregation may fulfil migratory or connective roles in the case of rhizomorphs and mycelial cords which, being corporate structures, are often capable of far more rapid extension than individual hyphae, whilst exhibiting parallel patterns of branching and anastomosis"
-Versatility and degeneracy p.25
Sectoring is the result of differences in growth speed, possible mutation, environmental factors, and possible differences in genetic grouping.
By the time your culture is visible to the naked eye it has become completely dikaryotic. Hyphal branching and fusing has jumbled all of strains together into an intricate mycelial matt which is rhizoid in morphology under a microscope:

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It is also arguable that without the ability to alternate exploratory and exploitative phases, mycelial margins would be unable, as they do, to maintain a constant rate of extension in more than one dimension without becoming progressively more sparse. The latter would follow from the inability of lateral branches to infill between leader hyphae that are already at maximum extension rate.
- The Growing Fungus
Sectors will contain many strains, how many? I really don't know, I'm not sure if anyone does, tbh. The likelihood of seriously distinct lines of genetic heritage being present on one side of the plate or the other, or between neighboring "sectors" seems to be extremely unlikely due to the tiny central location where the entirety of the genetic material originated from.
Further transfers from x plate will not significantly reduce the genetic diversity found on any given plate, perhaps slightly, but not significantly, but most likely not at all, tbh. The branching hyphae have so completely filled the microscopic gaps and fused where possible that most of the plate culture will be more or less identical, but not like an isolate in the sense that there's only one distinct set of genetics, there's just too much going on to separate those differences on agar, at the macroscopic level.
Genetic diversity remains on the plate, and will be expressed in your grows, but cannot be separated without serious intervention. The only way to separate the parental haplonts would be via chemical dedikaryotization, not scalpel transfers.
A simple thought experiment by argumentum ad absurdum: If heterozygosity was significantly reduced via scalpel/plate transfers, it would be possible to reduce any culture to a pure single strain isolate, eventually (which is what has been claimed). If this were true, we would have broken almost every cultivator multiple times over, making cultivation of demosticated lines either impossible, or incredibly difficult without the constant introduction of new gene flow from wild sources.
This is how I understand the literature, there are surely to be gaps in my understanding, but this is less incorrect than the previous understanding regarding sectors, etc.
I hope that helps
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Way
The


Registered: 01/14/23
Posts: 4,336
Loc: A long way away
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Hey thanks for taking the time to go through all of that. I'm very grateful to you.
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Stipe-n Cap said: -Versatility and degeneracy p.25
Sectoring is the result of differences in growth speed, possible mutation, environmental factors, and possible differences in genetic grouping.
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Stipe-n Cap said: Further transfers from x plate will not significantly reduce the genetic diversity found on any given plate, perhaps slightly, but not significantly, but most likely not at all, tbh.
I find these points in particular to be very interesting. I suspected both of these but having you confirm it really solidifies my understanding of how it all works. There really is a large misunderstanding around all of this on here.
Thanks again!
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That's the way she goes, boys. Sometimes she goes, sometimes she doesn't, cause that's the fuckin way she goes.
Edited by Way (12/15/23 01:33 PM)
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Re: Why Agar? [Re: Way] 3
#28583967 - 12/15/23 01:40 PM (1 month, 13 days ago) |
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Fungis is very counterintuitive. There's so much left unknown, and what is "known" will likely change in the future. Science is all about being able to change your mind when new information is presented.
I'll keep digging, I'm sure I'll be changing my mind about the above sooner or later, but it's the best I have for the time being. Maybe someone else with a deeper understanding will help fill in some of the blanks, contributing to the evolution of our understanding.
Time will tell. But until then, this is at least closer to the truth than what was previously available.
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tryptkaloids
Learner



Registered: 02/08/15
Posts: 12,641
Loc: Exact Center
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Stipe, what's your background if you don't mind me asking?
I hope to be as smart as you one day
-------------------- "Remember, kids, the difference between science and screwing around is writing it down" -adam savage Flowchart for Recommended plan of action. Learn the tried and true way to grow mushrooms Use the Damn search engine After you know what you're doing, take a break Pick a book, Make some chips! Josex said:Don't take the site seriously bro, ain't worth it.
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Just a guy on the internet who enjoys engaging in these kinds of topics.
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Bajazly
Stranger... Than You Think



Registered: 09/02/22
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Loc: BC, Mexico
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Stipe, who are these guys? The gardeners?
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Re: Why Agar? [Re: Bajazly]
#28584267 - 12/15/23 05:48 PM (1 month, 12 days ago) |
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Lol, probs your friendly neighborhood bacteria 
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HappinessStan
Fungivore



Registered: 10/10/12
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Loc: Worcester, UK
Last seen: 1 hour, 50 minutes
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Quote:
Stipe-n Cap said: Just a guy on the internet who enjoys engaging in these kinds of topics.
Never ending learning. Thanks for the detailed explanation. I'm half way there but there's always still so much more to learn. Organisms are just fantastic, such complex lifeforms that never cease to amaze us. What a beautiful world we live in.
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Limitless curiosity and some critical thinking goes a long way, you can teach yourself anything.
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Shroomish8
Stranger

Registered: 12/25/18
Posts: 14
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#28583919
Wow. That could be its own post, but it's just an amazing reply to an amazing post. Great explanation. Thank you for the clear, detailed write-up on agar AND hyphae interactions! Definitely cleared up some misconceptions and confusion I've had for a while.
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c01h
Aspiring Psychonaut



Registered: 11/24/23
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Last seen: 8 days, 3 hours
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Quote:
Stipe-n Cap said: Limitless curiosity and some critical thinking goes a long way, you can teach yourself anything.
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Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
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Re: Why Agar? [Re: c01h] 1
#28586450 - 12/17/23 07:25 AM (1 month, 11 days ago) |
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The quickest way to reduce heterozygosity in a culture (other than with serial dilution/microscopy/dedikaryotization) is through successive generations of spore printing. Not by plate transfers:
Generation zero - wild specimen heterozygosity 100%
F1 multispore of wild specimen 50% F2 multispore of F1 specimen 25% F3 ....12.5% F4 ....6.25% F5 ....3.12% F6 ....1.56%
and so on.
After the F6 generation there is such little heterozygosity (variability) left that all the mushrooms generated by multispore are virtually identical and the strain is considered stable.
The immediate descendants of the initial parents are the first filial generation (F1), much of the cultivars in circulation are well past f10, some may be close to f100+, which is why I believe some cultivars are beginning to, dare I say, "break", like golden teacher, for instance; Mutations abound.
Any of the successive generations of descent from an original parental generation contain less genetic material than the previous generation, which means one should be very careful when selecting specimens for printing.
Haphazard printing of just any ol fruitbody is reducing genetic variance along a specific trajectory, whether the cultivator realizes it or not (most very new growers probably don't consider it). Some trajectories are better than others, and once genes have been removed from a given culture, they're gone.
When selecting genetic material for your stock, the source of those spores will be a very important variable, because you'll need to trust that the source materials have been carefully handled/preserved intentionally, otherwise you may be receiving or producing less than desirable traits for future generations.
There are far more haphazardly taken prints in circulation than not, syringes are no different as they're the end product of spore production. With the influx of interest and new growers, people are swapping spores like cousins swap spit in Appalachia.
Food for thought.
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Thankful
Seeker

Registered: 01/17/24
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Last seen: 42 minutes, 30 seconds
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Stipe - this is an awesome post full of great educational value. Thank you so much!
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HILLBILLY OUTLAW
Above And Beyond!



Registered: 04/21/22
Posts: 2,854
Loc: Luckenbach Texas
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 Awesome write up. Your knowledge is and ability to share it is a vital part of our community!!!
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 🅃 🄴 🄰 🄼 🄲 🄻 🄸 🄽 🄶 🅆 🅁 🄰 🄿 TEAM SPREAD THE LOVE! Smellyhobbit said: Embarrassment and bashfulness are leeches on your ability to learn.
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