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Offlinekingjames488
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perfecting multispore strains... hu? (i don't get it)
    #10242078 - 04/27/09 01:08 PM (11 years, 10 months ago)

ok...
I don't get this one bit.

I've been reading about strains lately and some mention that the strain has been perfected, or sub strains isolated, or something like that.

but i also hear everything with MS is COMPLETELY random...

so how can these people alter strains if its random, seems like the information is contradicting it's self.

can someone please clarify?


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Re: perfecting multispore strains... hu? (i don't get it) [Re: kingjames488]
    #10242103 - 04/27/09 01:14 PM (11 years, 10 months ago)

you work the spores on agar, and isolate a substrain (cut it from the rest of the growth) and grow it out more on agar. You can tell the difference between substrains and you go with the ones with the best qualities. It may take a few transfers, but if done correctly, you should end up with one set of genetics.


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OfflineChrisWho
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Re: perfecting multispore strains... hu? (i don't get it) [Re: iskinbash]
    #10242117 - 04/27/09 01:16 PM (11 years, 10 months ago)

Not to hi-jack the thread or anything, but I know about isolation and stuff, but after you grow out an isolated strain, are the spores from the strain also isolated? or do you have to make a live tissue transfer or something?


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Re: perfecting multispore strains... hu? (i don't get it) [Re: kingjames488]
    #10242127 - 04/27/09 01:18 PM (11 years, 10 months ago)

Not completely random- a multispore grow is going to have the genetics of the parent generations. In the same way that human children will tend to look like their parents. Two people with brown hair and brown eyes won't have blue-eyed children, right? Same principle.

Of course, it could very well be vendor BS also. Stick with sponsors.


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Re: perfecting multispore strains... hu? (i don't get it) [Re: ChrisWho]
    #10242129 - 04/27/09 01:18 PM (11 years, 10 months ago)

Heres a couple paragraphs from RRs mushroomvideos.com

Mushroom Strain Isolation

Most mushrooms produce spores. When a mushroom spore germinates, the mycelium that emerges from the spore generally has one nucleus per cell, and we refer to this as hyphae, or monkaryotic mycelium. When hyphae that emerged from two individual spores of the same species cross paths, if they’re compatible, they join together to exchange DNA between them. The resulting mycelium generally has two or more nuclei per cell, and is referred to as dikaryotic mycelium. This mycelium, formed from two separate hyphae, is the very definition of a strain.

Since it only takes two spores to make a strain, you can easily see how many strains can potentially differentiate when you use an inoculating loop to swipe thousands of spores from a sporeprint onto a Petri dish of agar media. The process of strain isolation involves transferring these individual growths, or sectors away from each other, so that each can be expanded and fruited to determine the best performing strains. Strains with the desired characteristics will then become part of your permanent collection, stored long-term in test tubes as master culture slants.


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Re: perfecting multispore strains... hu? (i don't get it) [Re: ChrisWho]
    #10242154 - 04/27/09 01:23 PM (11 years, 10 months ago)

Quote:

ChrisWho said:
Not to hi-jack the thread or anything, but I know about isolation and stuff, but after you grow out an isolated strain, are the spores from the strain also isolated? or do you have to make a live tissue transfer or something?




Spores from an isolate will have the same potential for random results as the original spore print. By selecting specimens and growing new generations, you can inbreed and select for certain traits- but it's slow and tedious work. Better to simply isolate a substrain you like and grow that out.


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OfflineChrisWho
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Re: perfecting multispore strains... hu? (i don't get it) [Re: Doc_T]
    #10242157 - 04/27/09 01:24 PM (11 years, 10 months ago)

Odd


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Re: perfecting multispore strains... hu? (i don't get it) [Re: ChrisWho]
    #10242176 - 04/27/09 01:28 PM (11 years, 10 months ago)

What's odd?
That's how people have been breeding organisms for 10,000 years. Take the best of each generation, get them to breed. Make sure the culls don't. Repeat as needed.

Where do you think cows came from?
Or poodles? Ever seen a wild poodle?


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OfflineChrisWho
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Re: perfecting multispore strains... hu? (i don't get it) [Re: Doc_T]
    #10242187 - 04/27/09 01:30 PM (11 years, 10 months ago)

It's just strange to me for some reason that the spores would be like going back to multi spore... I get it and all, it's just weird.

I find alot of things involved with this hobby to be weird and interesting.

Either way tho, thanks for clearing that up.


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Re: perfecting multispore strains... hu? (i don't get it) [Re: ChrisWho]
    #10242207 - 04/27/09 01:35 PM (11 years, 10 months ago)

Spores are randomized in the same way that sperms and eggs are, if that helps. The parent organism splits its DNA and the chromosomes are assigned randomly. It's not quite the same as human reproduction, but the same concept.

If there wasn't any way for variants to occur, all mushrooms wold be the same. One dominant strain would have taken over long ago.


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OfflineChrisWho
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Re: perfecting multispore strains... hu? (i don't get it) [Re: Doc_T]
    #10242230 - 04/27/09 01:39 PM (11 years, 10 months ago)

That's what I was wondering to myself a few days ago, since I'm looking into starting isolations...

I wondered why spores from vendors were not isolated, and even thought about starting my own thread about it, but this one answered my question for me.


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Offlinekingjames488
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Re: perfecting multispore strains... hu? (i don't get it) [Re: Doc_T]
    #10242257 - 04/27/09 01:45 PM (11 years, 10 months ago)

Quote:

Doc_T said:
Not completely random- a multispore grow is going to have the genetics of the parent generations. In the same way that human children will tend to look like their parents. Two people with brown hair and brown eyes won't have blue-eyed children, right? Same principle.

Of course, it could very well be vendor BS also. Stick with sponsors.




actually im pretty sure ive herd of blue eyes kids from brown eye parents... and things like that.


I get the basics of cloning and isolating substrains in agar.
what i dont get is like... in some spore information it says that people did extensive work to isolate the strain or w.e...

but then the sites sell spores, so wouldn't that defeat the point of all the work?

or are spores a little less random then i thought...


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OfflineChrisWho
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Re: perfecting multispore strains... hu? (i don't get it) [Re: kingjames488]
    #10242283 - 04/27/09 01:51 PM (11 years, 10 months ago)

Quote:

Doc_T said:
Not completely random- a multispore grow is going to have the genetics of the parent generations. In the same way that human children will tend to look like their parents. Two people with brown hair and brown eyes won't have blue-eyed children, right? Same principle.

Of course, it could very well be vendor BS also. Stick with sponsors.




--------------------
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-Andrew Weil


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Re: perfecting multispore strains... hu? (i don't get it) [Re: kingjames488]
    #10242294 - 04/27/09 01:53 PM (11 years, 10 months ago)

Think of mushroom strains as similar to breeds of dogs.

Say I have a dog breeding site with poodle sperm and bulldog sperm and uh... Chihuahua sperm.
You buy a syringe of poodle sperm, use it to knock up your purebred poodle bitch. Poodle pups, right? Then there's some leftover in the syringe, so you use it on the neighbor's mutt... will you get poodles? They might have some poodle traits, but probably won't look like poodles.

Spores from a specific substrain will tend to have a narrower genetic profile than random spores, in the same way that a purebred dog will have less variance than a mutt.

(That's assuming you're dealing with a legit vendor, and not a bullshit artist.)

Quote:

kingjames488 said:
actually im pretty sure ive herd of blue eyes kids from brown eye parents...




Blue eyes are a recessive trait. Extremely unlikely that two brown-eyed parents could have a blue eyed child. Most probable explanation: introduction of outside genetic material.


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Offlinekingjames488
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Re: perfecting multispore strains... hu? (i don't get it) [Re: Doc_T]
    #10242414 - 04/27/09 02:16 PM (11 years, 10 months ago)

Quote:

Doc_T said:

Quote:

kingjames488 said:
actually im pretty sure ive herd of blue eyes kids from brown eye parents...




Blue eyes are a recessive trait. Extremely unlikely that two brown-eyed parents could have a blue eyed child. Most probable explanation: introduction of outside genetic material.




im actually pretty sure your right about the blue eyes thing... it is rarer (like albino shrooms hehe)
the other way around is a better example.

its not that important, the point im getting at is that a kids eyes arent always the same as the parents.

all this is just confusing me more lol

lemme find a good example.


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Offlinekingjames488
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Re: perfecting multispore strains... hu? (i don't get it) [Re: kingjames488]
    #10242447 - 04/27/09 02:22 PM (11 years, 10 months ago)

from strain descussion thread:

"This is a domesticated (3rd generation) version of the original Koh Samui strain brought back by researcher John Allen."

not exactly what i was looking for...

but how does this make sence either?

how can a strain change from just MS?

how can it stay like that?

am i missunderstanding what these people are doing?


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Re: perfecting multispore strains... hu? (i don't get it) [Re: kingjames488]
    #10242459 - 04/27/09 02:24 PM (11 years, 10 months ago)

That's actually a good example:
"aren't always the same as the parent"
but
tend to resemble the parents. Tall parents = tall kids. Dark skinned parents = dark kids. Deaf parents = deaf kids. Not always, but usually.

Light parent + Dark parent = range of potential colors.
One deaf parent, chance of deafness.
One tall, one short, etc, etc.

A mushroom strain is inbred and has reduced genetic variability. Think of people on a distant island all starting to look the same after several generations. Same thing with mushrooms.


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OfflineWorkmanV
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Re: perfecting multispore strains... hu? (i don't get it) [Re: kingjames488] * 3
    #10242476 - 04/27/09 02:26 PM (11 years, 10 months ago)

Mushrooom genetics are a little strange since a single mushroom produces spores that can then act as both parents for a new mycelium.  Essentially, you are selfing or inbreeding each time you do a multispore grow.

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?


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Re: perfecting multispore strains... hu? (i don't get it) [Re: kingjames488]
    #10242497 - 04/27/09 02:31 PM (11 years, 10 months ago)

Edit- disregard what I said, read what Workman wrote. ^^^


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Offlinekingjames488
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Re: perfecting multispore strains... hu? (i don't get it) [Re: kingjames488]
    #10242609 - 04/27/09 02:50 PM (11 years, 10 months ago)

the only way i can even come close to understanding this is by tossing what i knew about MS grows...

so MS grows are random, within a range of genetics?

so all strains wouldnt be the same, which i keep hear over and over they are.
seems SOMEONE has no idea what their talking about.

your explanation helped a little workman... would help if i knew what heterozygosity was... lol


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