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bodhisatta 
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Registered: 04/30/13
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afternoon reading on the topic of monocultures strains isolates varities hybrids 1
#21698708 - 05/19/15 02:54 PM (8 years, 8 months ago) |
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I understand that its possible to get two "varieties" to come together to make a "hybrid". They may end up showing similar features. But what defines a strain?
there's three kind of mycelium. primary secondary and tertiary.
secondary is what you think of as mycelium
tertiary is the mushroom itself, secondary differentiates into unique purpose built structures like the mushroom and becomes tertiary mycelium. most mushrooms can be cloned and the tertiary will turn back to secondary on a agar plate for instance
primary mycelium is what comes out of a spore
two primaries when they meet up if compatible will make a strain
https://www.shroomery.org/forums/showflat.php/Number/21283546#21283546 here's a post of mine explaining more about how those primary mycelia meet up and form a secondary mycelium
we call primary cubensis mycelium monokaryotic (one nucleus) and they meet up and form a Dikaryon which is secondary mycelium and we call that dikaryotic mycelium
dikaryotic mycelium is where our mushrooms come from. it takes that sharing of genetic information which in our fungi cubensis in this case they do it with a clamp connection https://www.shroomery.org/forums/showflat.php/Number/21432310 and https://www.shroomery.org/forums/showflat.php/Number/20947447#20947447
Like livestock I assume certain features are desired. Fast colonization, Large Fruits, heavy pinset, and of course potency. I also assume certain varieties react differently to what it is colonizing on. I'm sure some like grains where others like straw or w/e. But maybe cubensis psilocybe are for the most part the same?
all the same. no variety prefers any substrate over another but the whole species of cubensis may perffer some substrates over others but with all the ones everyone uses a clear pattern of superiority has yet to emerge and people still argue about what's the best substrate years and years later. with decades of trial and error if there was a best substrate out of all the common ones we use everyone would know about it by now. people base their baseless correlations on these things all the time. I could say substrate xyz sucks because the first time I tried to grow I failed the second time I had successes I used substrate ABC, so xyz sucks and ABC is great even though plenty of people other than me have great results with xyz it sucks because my ego is too big to let go that other people have opinions. or something like that is how the whole shroomery works.
Thinking in the future. What is the best way to isolate good mycelium? So far I've seen agar being used as a good isolation tool.
pretty much, there's a few ways to take a clone without agar but if you want to get into any real culture work you need to use a 2 dimensional media and agar is really the only one that's reliable.
Sorry if I asked too many questions. I overall really enjoy this and love learning about it. I just don't want to make people angry with stupid questions that I should be able to find on my own.
the only people that will yell at you saying you could go find it in the search engine are the people who also don't know the answer. they would have to go look it up too and post it and pretend like they knew it all along or they just say use the fucking search engine. sometimes UTFSE is warranted if the question is really dumb like, what's the PF tek ratio, since you should have the PF tek right in front of you already if you're making cakes. but even then I still post the answer as well as saying UTFSE. don't worry about those people just come here to learn and grow mushrooms and you'll have a way better time, people will tell you to calm down too but you'll just be chilling knowing they're the ones boiling over.

oh, and back to this
I understand that its possible to get two "varieties" to come together to make a "hybrid". They may end up showing similar features. But what defines a strain?
let's try to understand a "variety" first
a variety is a group of spores from a linage of your species(cubensis) that's been made more homozygous.
zygousness is just like having more tricks up your sleeve
wild cubensis is going to have be very heterozygous. that is there's lots of genetic things going on lots of different things IE hetero
when you start to print only the occasional mushroom that pops up and is white that's how you get something like AA+ a leuistic(not true albino) say you do 10MS grows see one white mushroom you print it. take that print grow 10MS grows see 2 white mushrooms. print both of those forget about the other one for now. do 10ms grows from one of those new syringes find 5 white mushrooms. take one of those and do 10 more and find 10 white musrhooms and so on. 7 generations down the line you're going to get nearly every single mushroom that you grow to be white. each time you're washing out genes for other things basically. you're loosing the tricks up your sleeve
Quote:
Workman said:
Now consider a wild collection of Psilocybe cubensis with a high heterozygosity. This basically means that most or all of each pair of genes in the mushroom are different from each other. Its the same gene location with the same basic function, but different versions. For example, if there is a single gene for height, you might have a version that gives short mushrooms and a version that gives tall mushrooms. If heterozygosity is high, you have one of each which may result in medium mushrooms unless one of the height genes is dominant.
Now, when you do multispore from a single mushroom you randomly get a mix of all the genes. Sticking to our height gene example, you could get two short copies, two tall copies or one of each. Obviously the strains with two short copies will be short and the ones with two tall copies will be tall.
Lets say we liked the short mushrooms so we saved that one and took a spore print for later. In this example the tall version of the height gene is lost to later generations. There is a net loss of heterozygosity. Over the entire genome the loss is about 50% per generation.
So mathematically we can figure out how many sequential multispore generations we need until the heterozygosity is reduced to an insignificant level and the strain is stable even from multispore.
Starting with a presumably high (~100%) heterozygosity from a wild collection. In reality, the heterozygosity is probably lower than 100%, but its an easy number to start with.
100% wild print 50% 1st generation from wild print 25% 2nd generation from 1st generation print 12.5% 3rd generation..... 6.25% 4th generation..... 3.12% 5th generation..... 1.56% 6th generation..... 0.78% 7th generation.....
You can see that the heterozygosity drops off quickly in the first few generations and is less than 1% after the 6th generation. This highlights the importance of choosing the best traits early on when there are more to choose from. Attempting to isolate traits in well established strains results in only minimal improvements unless spontaneous mutations increase the heterozygosity in a positive way (rare).
In summary:
Popular classic strains in circulation have all been grown well beyond 6 generations and are relatively stable from multispore with little need for isolation.
New strains, from wild material or cross breeding between different strains of the same species, can be stabilized fairly quickly with 6 or 7 generations of sequential multispore grows.
Selection is most important early in the process and if good genes are bred out, they are gone forever. Archiving original or early generation prints is recommended for preserving heterozygosity for later selective breeding. Continuous isolation of a bad strain with hopes of significant improvement is futile.
Does that help?
which this is a little bit better example of than what I just said.
a lot of people miss this post too https://www.shroomery.org/forums/showflat.php/Number/14525776#14525776
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wowimflabbergasted
supercalifragilistic



Registered: 07/16/12
Posts: 18,918
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: bodhisatta]
#21698876 - 05/19/15 03:40 PM (8 years, 8 months ago) |
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Ghatti
Totally not a Federal Agent

Registered: 06/23/14
Posts: 1,733
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: wowimflabbergasted]
#21699398 - 05/19/15 05:53 PM (8 years, 8 months ago) |
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No one has still given me a clear answer to "what does it look like on a microscopic level when 2 dikaryons of different species meet such as cubes and PS.cyans?"
Do they attempt to clamp but cannot exchange nuclei?
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: Ghatti]
#21699504 - 05/19/15 06:23 PM (8 years, 8 months ago) |
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they're completely different species I'm sure the receptors on their membranes react much like if you did an organ transplant with someone incomparable with you, there wouldn't be any attempt to get along. you have to induce a cross between species like that by dissolving membranes
they're not like oh hi happy to see you let's clamp and then realize oh shit you've got the wrong nucleus. the oh shit you're wrong happens sooner than that.
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Ghatti
Totally not a Federal Agent

Registered: 06/23/14
Posts: 1,733
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: bodhisatta]
#21699570 - 05/19/15 06:41 PM (8 years, 8 months ago) |
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But at what point? Likely have to start at when cubensis reach out to clamp. Do they reach out for one another or is there contact first to determine "who you're humping" cellularaly?
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: Ghatti]
#21699613 - 05/19/15 06:51 PM (8 years, 8 months ago) |
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what initiates the clamp connection? you would have to assume they try to fuck anything they touch with a clamp or assume that pan and cube have the same molecules and signals to imitate but down the road have a fail safe so they don't accidentally cross with a different species.
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Ghatti
Totally not a Federal Agent

Registered: 06/23/14
Posts: 1,733
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: bodhisatta]
#21699659 - 05/19/15 07:01 PM (8 years, 8 months ago) |
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So we did not really know what initiates the clamp? And on a cellular level what would happen if we actually just manually swapped or injected a nucleus?
I wanna know if they actually try to clamp but they both have "Y" hands and it just won't work.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: Ghatti]
#21699665 - 05/19/15 07:02 PM (8 years, 8 months ago) |
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i highly doubt they try to talk to each other at all most cells are pretty damn good at recognizing their own.
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Ghatti
Totally not a Federal Agent

Registered: 06/23/14
Posts: 1,733
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: bodhisatta]
#21699686 - 05/19/15 07:06 PM (8 years, 8 months ago) |
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I need a powerful scope
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Ghatti
Totally not a Federal Agent

Registered: 06/23/14
Posts: 1,733
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: Ghatti]
#21699736 - 05/19/15 07:18 PM (8 years, 8 months ago) |
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So bodhista sorry to stray off topic but tell me if this would be mono or dikaryotic mycelium clamping? I assume it is dikaryotic since it seems to be clamping on itself.
 I would assume monokaryotic mycelium would have 2 hyphae strands that clamp onto each other to exchange and create dikaryotic mycelium right?
Edited by Ghatti (05/19/15 07:22 PM)
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: Ghatti]
#21699818 - 05/19/15 07:35 PM (8 years, 8 months ago) |
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clamps are part of how the mycelium grows and colonizes dikaryotic mycelium makes them as it grows.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: bodhisatta]
#21699832 - 05/19/15 07:38 PM (8 years, 8 months ago) |
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Quote:
inski said:
 In (a) we have two Monokaryons coming together, note, Monokaryons have septa and look like Dikaryons without clamps, this is known as "Mating" (b) shows Fusion or "Cytogamy" (c) shows Nuclear Division or "Karyokinesis". (d) shows Clamp Connection formation. And (e) shows a stable Dikaryon.
thats how two monokaryotic make dikaryotic. the image you just posted from the clamp connection wiki is how a dikaryotic strain keeps going and going
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Ghatti
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: bodhisatta]
#21699850 - 05/19/15 07:42 PM (8 years, 8 months ago) |
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Ahh ty. Then what I would like to know is what happens when 2 monokaryotic separate strains meet. Do they attempt to mate. I was saying clamp but maybe the answer is the same.
So clamping is simply mycelium cell division correct?
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: Ghatti]
#21699866 - 05/19/15 07:44 PM (8 years, 8 months ago) |
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except when two monokaryons meet up they clamp to exchange and then each time the newly formed strain wants to grow out it elongates separates forms a clamp and that's how the mycelium grows out.
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,810
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: bodhisatta]
#21699910 - 05/19/15 07:54 PM (8 years, 8 months ago) |
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Nice thread. I love this kinda stuff. Directly relates to some of my current experiments. I really need to get a scope as I have many questions regarding genetics fruiting in concert, aggressive anastomosis, etc.
Nice write bodhi.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: Pastywhyte]
#21699916 - 05/19/15 07:56 PM (8 years, 8 months ago) |
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,810
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: bodhisatta]
#21700215 - 05/19/15 09:32 PM (8 years, 8 months ago) |
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Well its probably gonna end up being a birthday gift to myself so I will probably have a budget of $600 or so. I want something I gonna be happy with for some time
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MonkeyJesusFresco
am i suspended in agar?



Registered: 10/09/12
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: Pastywhyte]
#21703065 - 05/20/15 04:53 PM (8 years, 8 months ago) |
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Quote:
all the same. no variety prefers any substrate over another but the whole species of cubensis may perffer some substrates over others but with all the ones everyone uses a clear pattern of superiority has yet to emerge and people still argue about what's the best substrate years and years later. with decades of trial and error if there was a best substrate out of all the common ones we use everyone would know about it by now. people base their baseless correlations on these things all the time. I could say substrate xyz sucks because the first time I tried to grow I failed the second time I had successes I used substrate ABC, so xyz sucks and ABC is great even though plenty of people other than me have great results with xyz it sucks because my ego is too big to let go that other people have opinions. or something like that is how the whole shroomery works.
hahaha, I read that the other day right after I had whiped up a 5 gallon batch of MonkeyJesusFresco's pH Balanced Adjusted Hand-Mixed 100% Fair-Trade Organic Gourmet Bulk Mushroom Substrate⢠
why do I use and make it? because: as stated above
Edited by MonkeyJesusFresco (05/20/15 05:01 PM)
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: MonkeyJesusFresco]
#21703090 - 05/20/15 04:58 PM (8 years, 8 months ago) |
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to balance a pH you must first assume it's unbalanced.
pH adjusted
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MonkeyJesusFresco
am i suspended in agar?



Registered: 10/09/12
Posts: 3,306
Loc: South East USA
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: bodhisatta]
#21703115 - 05/20/15 05:03 PM (8 years, 8 months ago) |
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um...and a side note, I checked on my MonkeyJesusFresco's pH Adjusted Hand-Mixed 100% Fair-Trade Organic Gourmet Bulk Mushroom Substrate⢠and it's swarming with gnats, will standard pasturization temps suffice or should I up it to the 170s nevermind, I kid, I kid 
edit:
that is a bad ass picture of a broken spore, btw; you rock.
Edited by MonkeyJesusFresco (05/20/15 05:04 PM)
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: afternoon reading on the topic of monocultures strains isolates varities hybrids [Re: MonkeyJesusFresco]
#22063717 - 08/08/15 05:23 PM (8 years, 5 months ago) |
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afternoon bump
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