MudaFuka said:

There are millions of different genetics in every spore drop. Even from a mono culture. starting from spores is basically like hitting the reset button.

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Workman said:

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?
If I may jump in here on the breeding true from spores aspect.

You lose on average 50% of heterozygosity with each strain generated from multispore (analogous to selfing in plants). What this means is, the more sequential generations you do via multispore, the less variability you will see in the later generations until the variability is nearly undetectable. There will come a point that the only variability will be from new random mutations.

OK, so what is this point, or how many generations until you can be confident you have a true breeding strain? With a wild strain you would assume 100% heterozygosity. Previously domesticated strains will obviously have less heterozygosity but since that is unknown it doesn't hurt to be conservative.

Mathematically you can see the reduction in variability with each generation. It drops off quickly and then around generation 5 or 6 the gains in stability drop off dramatically. After generation 7, with heterozygosity less than 1%, the rate of random mutations will outpace any small amount of remaining variabilty loss in later generations.

Wild 100%
F1 50%
F2 25%
F3 12.5%
F4 6.25%
F5 3.125%
F6 1.5625%
F7 0.78125%

Generate several strains from the spore print and you should expect to see huge differences between these new strains if the heterozygosity is high. Choose the strain(s) with traits you want (in this case, cap color and size) and take prints. Repeat with these new prints. You can probably feel pretty confident that after doing this 5 or 6 times that the strain is stable, especially if
you don't see any new variants in the later generations.

Good luck and I hope this is understandable.
one of my favs
breeding

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Mr.PhilCybin said:

From the DNA.  Every spore has half the genetic info, and every two 2 compatible spores that find each other make a new culture.

when you germinate by MS, there's lots of different cultures growing together.

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MudaFuka said:

Thank you that's the one.

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Jkinder313 said:

Shit man, that goes deep. Do you know of any selective breeding process? Breeding Monocultures in petri dishes?

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PirateSwazey said:

This might be of interest to you OP....

Workman said:

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?
If I may jump in here on the breeding true from spores aspect.

You lose on average 50% of heterozygosity with each strain generated from multispore (analogous to selfing in plants). What this means is, the more sequential generations you do via multispore, the less variability you will see in the later generations until the variability is nearly undetectable. There will come a point that the only variability will be from new random mutations.

OK, so what is this point, or how many generations until you can be confident you have a true breeding strain? With a wild strain you would assume 100% heterozygosity. Previously domesticated strains will obviously have less heterozygosity but since that is unknown it doesn't hurt to be conservative.

Mathematically you can see the reduction in variability with each generation. It drops off quickly and then around generation 5 or 6 the gains in stability drop off dramatically. After generation 7, with heterozygosity less than 1%, the rate of random mutations will outpace any small amount of remaining variabilty loss in later generations.

Wild 100%
F1 50%
F2 25%
F3 12.5%
F4 6.25%
F5 3.125%
F6 1.5625%
F7 0.78125%

Generate several strains from the spore print and you should expect to see huge differences between these new strains if the heterozygosity is high. Choose the strain(s) with traits you want (in this case, cap color and size) and take prints. Repeat with these new prints. You can probably feel pretty confident that after doing this 5 or 6 times that the strain is stable, especially if
you don't see any new variants in the later generations.

Good luck and I hope this is understandable.
one of my favs
breeding

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MudaFuka said:

If you want to breed certain traits in to a variety you will need to start from a wild print or mix the spoes of two domesticated varieties. You have 6 or 7 generations to get the traits you want bread in with the first couple generations being the most important. Even then there will still be plenty of variation since it takes very little difference in genetic code to produce very different outcomes. consider the fact that humans and chimps share like 97% of there genetic code. In order to get predictable results from each grow you need to clone or isolate and keep your culture going. Once you go back to spores there are tons of possible outcomes.

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Jkinder313 said:

So where do the new genetics come from? I'm really interested, because I thought I had the reproductive cycle of the organism figured out.

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the genetics in the spores from a monoculture will contain only the genetic information from the parents involved in creating the culture...

but

they wont be replicas of the fruit-body the spores was taken from just lots of combinations of the original genes involved. ( a multi-spore of them set genetics) new genes will not be in the multispore prints and genes cannot appear from knowhere.

think of two people having kids- same genetics being mixed but you get lots of expressions/combinations in the next generation 

PirateSwazey said:

There's more than one way to skin a cat, that is just how I have been doing it for the last year or so because it keeps things interesting and it always puts out. Obviously you can put spores to a plate and transfer it until you have a monoculture but there is a high probability of ruling out the strains you want in doing it that way. Another good method is to take a clone and then transfer it out to a few different plates... then isolate from those plates. Just a few of the methods I have used and had fun with. If you're interested in playing with genetics I would highly recommend getting some test tubes!

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