tryptkaloids said:

Look into muda bottles

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I did muda bottles but the flush was terrible, then I popped that block out of the jar and put it in a water tub, and now it's looking a lot better. When I was trying to fruit it from the jar, I had a very hard time dialing in the moisture. I'm wondering if this is actually a legit thing to try in the future: Let a sub+spawn mix colonize in a jar and then pop it out and use a water tub.

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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ThurmanMurman said:

What is the consensus on culture "sectors" being genetically different from one another? We had a discussion yesterday in another thread, and I guess I still dont know how to feel about it. Is every piece of an MS culture genetically identical or are we creating unique cultures through transfers?

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cronicr said:

ThurmanMurman said:

What is the consensus on culture "sectors" being genetically different from one another? We had a discussion yesterday in another thread, and I guess I still dont know how to feel about it. Is every piece of an MS culture genetically identical or are we creating unique cultures through transfers?

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As for sectors being genetically different it's case by case , you'll often see clones show " sectors " but if you put a wedge from two of them on a plate they won't form a line but rather connect with eachother without issue
Other times from ms you may try this again but this time they form a wall staying seperate to a degree , other times what seems like sectors could be based off water on the plate or contamination or damage to the wedge ( hot scalpel for example )

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This is all great, useful information for my overall understanding but I'm still not clear. Heres a pic of a plate I can use for an example.



Ok, so this plate was a T1 from an MS brf puck. As far as I can tell, there's no contamination present, and as far as what I'm asking, lets just assume that there isnt, regardless.

There's sectoring here, yes? Various triangular sections fanning out from the center that appear to have thin areas between, with each sector having slightly different characteristics. There's going to be a lot of genetic diversity on this plate, right? Lets just say there's 100 "strains" here, which to my understanding, would mean 100 "children", as we would say if we were breeding animals or something, yes? Now is every bit of that culture made up of the 100 strains, or are those sectors each just a percentage of that 100? I transferred from about 11 oclock, thinking that that was the thickest, healthiest looking myc on the plate. So will that transfer then have all 100 strains or did I just dramatically reduce the genetic diversity of the mycelium by doing that?

Ultimately, there's this idea of grabbing the healthiest, fastest growing sector and I dont understand if this is purely for the sake of cleaning the culture in that the best looking part should be contamination free, or if we're grabbing these sections bc they're performing well and we hope to increase the odds of favorable performance by reducing genetic variation.

ThurmanMurman said:

cronicr said:

ThurmanMurman said:

What is the consensus on culture "sectors" being genetically different from one another? We had a discussion yesterday in another thread, and I guess I still dont know how to feel about it. Is every piece of an MS culture genetically identical or are we creating unique cultures through transfers?

---

As for sectors being genetically different it's case by case , you'll often see clones show " sectors " but if you put a wedge from two of them on a plate they won't form a line but rather connect with eachother without issue
Other times from ms you may try this again but this time they form a wall staying seperate to a degree , other times what seems like sectors could be based off water on the plate or contamination or damage to the wedge ( hot scalpel for example )

---

This is all great, useful information for my overall understanding but I'm still not clear. Heres a pic of a plate I can use for an example.



Ok, so this plate was a T1 from an MS brf puck. As far as I can tell, there's no contamination present, and as far as what I'm asking, lets just assume that there isnt, regardless.

There's sectoring here, yes? Various triangular sections fanning out from the center that appear to have thin areas between, with each sector having slightly different characteristics. There's going to be a lot of genetic diversity on this plate, right? Lets just say there's 100 "strains" here, which to my understanding, would mean 100 "children", as we would say if we were breeding animals or something, yes? Now is every bit of that culture made up of the 100 strains, or are those sectors each just a percentage of that 100? I transferred from about 11 oclock, thinking that that was the thickest, healthiest looking myc on the plate. So will that transfer then have all 100 strains or did I just dramatically reduce the genetic diversity of the mycelium by doing that?

Ultimately, there's this idea of grabbing the healthiest, fastest growing sector and I dont understand if this is purely for the sake of cleaning the culture in that the best looking part should be contamination free, or if we're grabbing these sections bc they're performing well and we hope to increase the odds of favorable performance by reducing genetic variation.

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yes

edit: I mean no.

edit: not sure.turns out genetics is hard



edit: whoops my bad, didnt check the thread

ThurmanMurman said:

cronicr said:

ThurmanMurman said:

What is the consensus on culture "sectors" being genetically different from one another? We had a discussion yesterday in another thread, and I guess I still dont know how to feel about it. Is every piece of an MS culture genetically identical or are we creating unique cultures through transfers?

---

As for sectors being genetically different it's case by case , you'll often see clones show " sectors " but if you put a wedge from two of them on a plate they won't form a line but rather connect with eachother without issue
Other times from ms you may try this again but this time they form a wall staying seperate to a degree , other times what seems like sectors could be based off water on the plate or contamination or damage to the wedge ( hot scalpel for example )

---

This is all great, useful information for my overall understanding but I'm still not clear. Heres a pic of a plate I can use for an example.



Ok, so this plate was a T1 from an MS brf puck. As far as I can tell, there's no contamination present, and as far as what I'm asking, lets just assume that there isnt, regardless.

There's sectoring here, yes? Various triangular sections fanning out from the center that appear to have thin areas between, with each sector having slightly different characteristics. There's going to be a lot of genetic diversity on this plate, right? Lets just say there's 100 "strains" here, which to my understanding, would mean 100 "children", as we would say if we were breeding animals or something, yes? Now is every bit of that culture made up of the 100 strains, or are those sectors each just a percentage of that 100? I transferred from about 11 oclock, thinking that that was the thickest, healthiest looking myc on the plate. So will that transfer then have all 100 strains or did I just dramatically reduce the genetic diversity of the mycelium by doing that?

Ultimately, there's this idea of grabbing the healthiest, fastest growing sector and I dont understand if this is purely for the sake of cleaning the culture in that the best looking part should be contamination free, or if we're grabbing these sections bc they're performing well and we hope to increase the odds of favorable performance by reducing genetic variation.

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cronicr said:

Honestly that's 100 percent correct , I was just talking with a fellow tc last night about his culture where what seemed to be the slower sector on agar actually ripped through grain and substrate and absolutely smashed it out of the park yield wise...we really don't know until we know we can't read mycelium plain and simple all we can focus on is clean spawn which is why that is in my signature

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cronicr said:

Essentially when we use ms we are inbreeding
Workman does a great job explaining it
nbreeding?  Well, it isn't generally good long term, but it is a useful tool for enhancing and isolating certain traits.

Each multispore culture is selfing (breeding with one's self).  This doesn't seem to be a problem initially, but sequential multispore cultures reduces heterozygosity.  What I mean by sequential multispore is using prints from your current cultivation to start the next cultivation and so on.  A better idea is to save prints from your earliest cultivation and use those to preserve your "strain" as long as possible.  Cubensis spores should last at least 10+ years if kept cool and dry, but viability drops yearly, so you may need to use large amounts of spores to revive a culture from very old spores.

The loss of heterozygosity means that the spores will produce fruits that show less and less variability from multispore, which sounds fine.  You get a nice uniform crop with little variability, so its similar in behavior to a clone.  But this also makes your culture vulnerable to random mutations.  Random mutations that can be recessive and invisible to the grower.  The PE mutation is a good example.

The PE mutation is recessive, so lets say you made a multispore culture from a print, maybe you selected out a single strain on agar or just injected tons of spores into a substrate.  You got a crop of nice looking mushrooms and printed some.  You gave away all the prints you made and just have one print remaining.  Unknown to you, the mushroom you saved the print from was generated by a pair of spores, where one had been hit by a cosmic ray that damaged a key gene necessary for proper cap development.  The mushroom looks fine because each of it's cells also contains the nuclei from the other spore with an undamaged gene.

You only have the one print, you want to grow it out again and expect the same results as before.  But something is wrong, several mushrooms look like PE, with weird malformed caps.  The selfing, or inbreeding has resulted in pairs of the damaged gene appearing in 25% of the generated strains.  Many of the mushrooms in the crop look fine, but most of them will also contain the damaged gene.  And this is just one trait.  More mutations can sneak in over time, almost all of them are bad, some can make the mycelium unable to even fruit.

So save your earliest prints, save slants of your best cultures.  Combining the spores of different varieties can increase heterozygosity which can then be selectively selfed to produce a new custom made variety.

I know it's confusing and I probably didn't explain it very well, but maybe I helped.

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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Smellyhobbit said:

Zorry said:

Using telephone brand and have not tried no color.

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Switch to living Jin. It’s the telephone brand.

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catnip40 said:

what if a rogue non-tc comes in here and starts answering questions? whats the repercussions

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