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SoulButter
Joint Chief of Soul


Registered: 06/23/15
Posts: 312
Last seen: 3 years, 1 month
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Is there a limit to how many generations you should propogate via spores from the last generation?
#23550294 - 08/17/16 03:38 PM (7 years, 6 months ago) |
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What are the effects of eternal multispore reproduction? When should someone start over? Which propagation method leads to a decline in genetic stability over time?
Just some questions to better understand the long term genetics.
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cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 1 month
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Re: Is there a limit to how many generations you should propogate via spores from the last generation? [Re: SoulButter] 1
#23550433 - 08/17/16 04:25 PM (7 years, 6 months ago) |
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Inbreeding? 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
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 1 month
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Re: Is there a limit to how many generations you should propogate via spores from the last generation? [Re: cronicr] 1
#23550439 - 08/17/16 04:26 PM (7 years, 6 months ago) |
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I should point out that common sense dictates that inbreeding is bad since it can expose recessive genetic defects or undesirable traits. Ideally you want high heterozygosity to give a particular mushroom strain a wide range of available genes, which in turn makes it better adapted to unpredictable environmental conditions. This could be achieved by cloning a wild vigorous specimen or by crossing two very different strains. In my experience, the mushrooms resulting from such a cross are very vigorous and productive (hybrid vigor).
The problem is that to keep this strain going you have to keep the mycelium going. This isn't a problem with edible cultures but we need stable spores. The spores from a high heterozygosity strain will be genetically recombined and won't produce the original strain (although with aggressive isolation you could get something close). Multispore grows will be terrible, not because the newly generated strains are bad, but because they are so genetically different from each other. These different strains don't get along nicely in the same tray which reduces productivity. If we can reduce the differences between the strains generated by multispore to some minimal level, they tend to cooperate better with each other and act more like a single strain.
The point is that we are dealing with spores not clones. Many (most) Psilocybe growers use syringes to generate multispore grows with no isolation. In this context, stabilized strains are desirable. If everyone could trade/sell mycelium this wouldn't be an issue. workman
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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TheFinNewGuy
Stranger

Registered: 03/07/20
Posts: 42
Last seen: 9 months, 6 days
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Re: Is there a limit to how many generations you should propogate via spores from the last generation? [Re: cronicr]
#27336573 - 06/05/21 05:28 PM (2 years, 8 months ago) |
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I'm still trying to sort all of this out as well. I'm just a guy trying to keep a good thing going for as long as possible without having to re-source spores. I think it is safe to say, the next time I grow, I should print the hell out of it. I've also been told that grain to grain is a great way to keep things going. Unfortunately I've also been told to never exceed G4. I haven't tried slants yet, but there is varying advice on how long those are good for. It's all very confusing.
As an example, I just purchased liquid culture Lions Mane. Can you even print Lions Mane? Should I drop some on Agar and regrow it there?
The more advice the better.
Thanks in advance.
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