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newrook
Sucks at bulk



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Best way to isolate/stabilize a trait so it is more consistent,
#22175482 - 09/01/15 07:51 PM (8 years, 4 months ago) |
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I wasn't sure exactly what to title this, my question is pretty much if I have a trait that I want to isolate from a mushroom that is growing, is it better to:
a) take a spore print from the mushroom, do a ms grow, and take a print from a mushroom sharing the trait in the subsequent grow and repeat this process until most of the mushrooms growing from ms share the trait
or
b) take a tissue sample on to agar and clean it up.
Thanks in advance.
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SynKyd
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Re: Best way to isolate/stabilize a trait so it is more consistent, [Re: newrook]
#22175508 - 09/01/15 07:57 PM (8 years, 4 months ago) |
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B.
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PoIIo
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Re: Best way to isolate/stabilize a trait so it is more consistent, [Re: SynKyd]
#22175589 - 09/01/15 08:15 PM (8 years, 4 months ago) |
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B
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iSmkGrnBud
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Re: Best way to isolate/stabilize a trait so it is more consistent, [Re: PoIIo]
#22175878 - 09/01/15 09:16 PM (8 years, 4 months ago) |
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You're answering your own question. You really wouldn't want to take a spore print and go MS, because that would be going back to MS(Multispore, multiple genetics) and giving you a crap shoot on which genetic traits would occur. Ideally would would want to clone from that fruit body to agar. Then monitor and watch growth and continue to isolate on agar from there.
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cronicr



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Re: Best way to isolate/stabilize a trait so it is more consistent, [Re: iSmkGrnBud] 1
#22176378 - 09/01/15 10:54 PM (8 years, 4 months ago) |
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you're all wrong...sort of. You want both A and B. Cloning is a good way to get the genetics but the op says stabilized which is done over genrerations not just simply cloning so you would indeed first clone the fruit and fruit that, take spores from that grow and fruit that, spot the same trait and clone that...repeat
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uncle_rico
my own worst enemy


Registered: 03/28/06
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Re: Best way to isolate/stabilize a trait so it is more consistent, [Re: cronicr]
#22176494 - 09/01/15 11:27 PM (8 years, 4 months ago) |
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we also need to watch the 'age' of our culture.
when a culture gets too 'old' (chronological age or cell division age), it tires out and yield goes down. It is also more susceptible to contamination.
culture slants are the answer.
when a grower hasn't properly stored his culture and it is old and compromised, he'll go back to spores to reset the age clock.
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Machiavelliavore
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Re: Best way to isolate/stabilize a trait so it is more consistent, [Re: cronicr]
#22176498 - 09/01/15 11:27 PM (8 years, 4 months ago) |
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I'm working on isolating some conical (lib cap like) caps.
The way I figure it is that there is no point in cloning. If you print from a fruit with the characteristic, wouldn't a spore print from that have the same genetic composition as a spore print from a clone?
In a single generation it went from like 1/100 to 50%+ of the mushrooms. Growing out the third print now.
Hopefuly I can get it up to 75% or so, then go ahead and start isolating to weed out staggling standard cap shape. It should be way easier to find a strain that has the lib caps, now that it's prevalent instead of rare. Then print from the isolate fruit, grow out, so on and so forth for a few generations.
P.S. I don't know shit about fungal genetics. Just regular genetics. I know this dichariotic shit makes breeding traits harder.
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I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister. I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave. I think it will be good cause B+ is so potent. Triggered yet? Only a square would say "a cube is a cube."
No, this does not look right...
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Psilosoulful

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Re: Best way to isolate/stabilize a trait so it is more consistent, [Re: cronicr]
#22176504 - 09/01/15 11:30 PM (8 years, 4 months ago) |
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Quote:
cronicr said: you're all wrong...sort of. You want both A and B. Cloning is a good way to get the genetics but the op says stabilized which is done over genrerations not just simply cloning so you would indeed first clone the fruit and fruit that, take spores from that grow and fruit that, spot the same trait and clone that...repeat
So it goes from cloning >spore printing> cloning> spore printing, etc…
Couldn't you just keep cloning over and over (with isolating on agar of course) until all the fruits have the same phenotypes. Why does spore printing have to be involved, couldn't that be done last to preserve the desired mushroom genetics?
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cronicr



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Re: Best way to isolate/stabilize a trait so it is more consistent, [Re: Psilosoulful] 1
#22176532 - 09/01/15 11:41 PM (8 years, 4 months ago) |
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becvause by cloning over and over again your not doing anything other then expanding the original genetics, sort of like a chick getting fat...still the same chick just more of her but by using the spores you are also keeping he culture young and hepling to truly stabilize that trait even by spores
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Kizzle
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Re: Best way to isolate/stabilize a trait so it is more consistent, [Re: cronicr]
#22177262 - 09/02/15 07:41 AM (8 years, 4 months ago) |
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The best way IMO is to isolate a bunch of strains and test them out. When you find one with the traits you want you can preserve and use the culture it came from as a master almost indefinitely.
Trying to stabilize a trait over generations takes so much more time and resources it's not even a realistic option for most people.
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newrook
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Re: Best way to isolate/stabilize a trait so it is more consistent, [Re: Kizzle]
#22177267 - 09/02/15 07:46 AM (8 years, 4 months ago) |
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Thanks for the answers, I was almost positive that I had seen info about carrying a trait though generations using ms grows, but couldn't find it again in the search function. Thanks for clearing things up a bit guys.
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MetaRox


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Re: Best way to isolate/stabilize a trait so it is more consistent, [Re: newrook]
#28477222 - 09/20/23 09:58 PM (4 months, 6 days ago) |
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Both...but we have to understand the different terms before doing anything to actually learn and make a progress.
A strain is a set of genetic from a species.
A Clone is a set of strains that come from a piece of the fruit body.
An Isolate is just ONE strain and can only be obtained by multiples sector isolation in agar over and over until there is no more sectoring in the agar.
Stabilization is when you redoo all the cycle over and over again from the very start(spores) to finish(fruit body) selecting a fruitbody and picking spores from THAT fruit body.
Yes, spore are a crapshot but even a crapshot is poiting to somewhere.
As a example, if i cross a black human with a white human and they have 10 child from blackest to whitest and then i cross one of the blackest child with another black child there is a chance that some of the next 10 childs may be white because there is white genes in they but i repeat the process and again select the blackest child and cross it with another black and repeat and repeat and repeat in the long run the consequent childs the predominant phenotype will be black, there always will be chances from growing a white child but the the breed is stabilized as black i will sell that strains as "Black" (sorry for the terrible bad example) and when i growing it will be mostly all black.
Thats is how the mushrooms strain from the vendor are made and it take between 7-10 cycles to truly stabilize the strain.
Cloning and sectoring are tools the select the genes and phenotype that we want but i still need the mushshroom to drop spore to start a stabilization process because of senescence.
Almost all living being have something called telomeres that are things the are in both extremes of the DNA and with every subsequent cell division(mycelium in case of shrooms) telomere shortens and shortens and the DNA will start to "malfunction" and may bring mutations or simply the living being will not divide anymore so i NEED to start over with spores sometimes. This is mechanism that nature got because genes variability is the only way to ensure species survival.The strongest strain survive and reproduce.
So to truly stabilize a strain is a hard and looong process but hell of rewarding.
Thats why some folks clone a nice fruit and save them in master slant and store them then pick from that slant the cloned strains and expand it in grains to test it, the slant is stored in low temperatures enough to not kill the mycelium and enough to reduce at max the cell division and avoid senescence but eventually it will come and they will need to pick another nice fruit.
In the end the ultimate process to obtain the best strain is:
1-Cloning because you can SEE the phenotype(fruit body)
2-Isolate the most vigorous strains in agar from the cloned fruit body
3-Save in masterslants all the strains selected from isolation and cultivate them.
4-Discard the one that not generated fruit bodys and the one that are sterile(not spores production)
5-Select the one that actually grows quick, big and potent
6-Repeat
That is the ultimate best way but the other way the everyone use is just clone the best fruit and cultivate and repeat 7-10 times until the mayor part of the fruit bodys are similar and the crapshot from multispore are not that crappy.
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B Traven
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Re: Best way to isolate/stabilize a trait so it is more consistent, [Re: MetaRox]
#28477233 - 09/20/23 10:22 PM (4 months, 6 days ago) |
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It'd be fun to run side-by-side comparisons of this over multiple generations, starting with a wild print.
Personally, the way I look at it is that I' trying to maintain a population. Rather than seeking a specific trait in a specific line, I'm just growing everything out in the same environment, and selecting for whatever thrives over time. Clone lines can come and go, but the overall population moves in the direction I want it to.
I just take an average by grinding everything up, anyway.
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