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InvisibleKauaiOrca
Waterman

Registered: 08/12/08
Posts: 3,131
Cloning Questions ... on Genetics
    #23975099 - 01/02/17 06:00 AM (7 years, 28 days ago)

Suppose I was to do a MS grow and take tissue (generation #1) from a good fruit from that ... clean the tissue and then grow it out.  If I was to take tissue from a fruit from that grow (generation #2) would it have the exact same genetics as the fruit I took the first tissue from and could I expect roughly the same growing results from the tissue from fruit generation #2 as generation #1.

Thanks for your help!


--------------------
"The universe is endless, limitless and infinite.  Any effort to define it's boundaries is an attempt to overcome ignorance.  We are physical, mental and spiritual beings ... there is no beginning and there is no end.  There is only memory.  Our repeated loss of memory experiences create the illusion of beginnings and ends.  Immortality is the ability to retain full memory through all consciousness transformations.  Loss of memory is man's greatest curse and, in very real terms, death."

-- Ancient Taoist Master


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Offlineenlightenment
alchemist
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Registered: 08/09/09
Posts: 1,647
Loc: Europe
Last seen: 8 months, 21 days
Re: Cloning Questions ... on Genetics [Re: KauaiOrca]
    #23975160 - 01/02/17 06:35 AM (7 years, 28 days ago)

Quote:

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?




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InvisibleKauaiOrca
Waterman

Registered: 08/12/08
Posts: 3,131
Re: Cloning Questions ... on Genetics [Re: enlightenment]
    #23975173 - 01/02/17 06:44 AM (7 years, 28 days ago)

Quote:

enlightenment said:
Quote:

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?







Thanks ... that's a fantastic post.  Answered my question and also gave insight into something I've been thinking about and noticing from the MS grows I've done with from a print I had couple of years ago ... each MS generation gets better and better and better.  I knew there had to be a reason for it.  The MS grows now, although slower to get started obviously, are as good and sometimes even better than good clone cultures I've got.


--------------------
"The universe is endless, limitless and infinite.  Any effort to define it's boundaries is an attempt to overcome ignorance.  We are physical, mental and spiritual beings ... there is no beginning and there is no end.  There is only memory.  Our repeated loss of memory experiences create the illusion of beginnings and ends.  Immortality is the ability to retain full memory through all consciousness transformations.  Loss of memory is man's greatest curse and, in very real terms, death."

-- Ancient Taoist Master


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OfflineEarhman
Stranger
Male
Registered: 01/08/00
Posts: 26
Loc: NorthEast US
Last seen: 1 year, 3 months
Re: Cloning Questions ... on Genetics [Re: enlightenment]
    #23975251 - 01/02/17 07:26 AM (7 years, 28 days ago)

What a stellar post. While I'd had a generalized intuition of the process you've outlined you've done a superb job clearly explaining it.

Well done.

Earhman


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InvisibleKauaiOrca
Waterman

Registered: 08/12/08
Posts: 3,131
Re: Cloning Questions ... on Genetics [Re: Earhman]
    #23975645 - 01/02/17 10:38 AM (7 years, 28 days ago)

Based on this information, would it be a fair deduction that the spores from a clone that has come from several generations off the same multispore grow are going to show similar growing results as the clone itself?


--------------------
"The universe is endless, limitless and infinite.  Any effort to define it's boundaries is an attempt to overcome ignorance.  We are physical, mental and spiritual beings ... there is no beginning and there is no end.  There is only memory.  Our repeated loss of memory experiences create the illusion of beginnings and ends.  Immortality is the ability to retain full memory through all consciousness transformations.  Loss of memory is man's greatest curse and, in very real terms, death."

-- Ancient Taoist Master


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InvisiblebodhisattaMDiscordReddit
Smurf real estate agent
 User Gallery
Folding@home Statistics
Registered: 04/30/13
Posts: 61,889
Loc: Milky way
Trusted Cultivator
Re: Cloning Questions ... on Genetics [Re: KauaiOrca]
    #23975745 - 01/02/17 11:07 AM (7 years, 28 days ago)

Not much more at all, because the variety you started with is very homozygous so those spores were predisposed to being similar anyway


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