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Offlinemegtheabyss
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Monoculture Isolation - Early Identification of Genetic Traits
    #27106648 - 12/24/20 10:43 PM (3 years, 1 month ago)

I've done a few cube grows over the last year with good yields, and now looking towards isolating a monoculture for better predictabiliy of fruit size, overall yield & potency. I've cloned three of the biggest fruits from the first two flushes and got them onto agar.

However, I'm at a loss when trying to figure out how to go about the isolation. I understand there may be upto 10 transfers before I reach a monoculture, but I'm wondering how I would be able to identify the (genetically) best sector on the attributes of the mycelium alone (faster / ropey / ?). By the time I reach a monoculture, it will more than likely not be the best monoculture, and I may have eliminated better fruiting genetics due to improper sector selection during the earlier transfers. As far as I know, it is not possible to identify fruiting capability from the visual traits of mycelium itself.

The only way I see around this is to start from one clone, distinguish the most distinct sectors during each transfer, and grow them out on micro tubs to identify the best genetics. This does not seem too feasible however, because it could take years to identify the best monoculture from even one clone, with the senescence clock ticking along the way. So two questions basically:

1. Is this the only way to isolate the best genetics?
2. How big of a concern is senescence when the monoculture isolation takes years. The reference cultures from individual sectors will stay refrigerated during the tub grows so I assume the culture won't technically age because the mycelium will not expand in the fridge.

Thanks in advance and Merry Christmas to y'all!


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abltsandwich said:
Gypsum adds sterilization to the pressure cooker.  If you PF Tek some bulk substrate and proper sterilization pasteurization, then mushrooms and gas exchange (GE) plus FRESH AIR EXCHANGE (brown rice flower [FAE]).


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OfflineShroomoisseur
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Re: Monoculture Isolation - Early Identification of Genetic Traits [Re: megtheabyss]
    #27106714 - 12/24/20 11:49 PM (3 years, 1 month ago)

I’m not that experienced but I’ll offer my opinion:
I believe since you’re starting on agar from clones, rather than print/syringe, plates should grow consistently/symmetrically from a clean cut of clone tissue. That’s the purpose of taking clones, to try to replicate whatever traits that fruit had.
Yes, most would say to take transfers from the furthest reaching section or whichever has very distinct rhizo/“ropey” strands as you described. This trait is what some would consider signs of a fast or “aggressive” colonizer.
Yes you will have plates that look great and perform shit and ugly plates that end up performing pretty well. Your assessment is correct. I think everyone should get some plates going, then take them to grain and from those fruit take clones and repeat until you get desired canopies/mutants, etc.
If you are getting serious about preserving genetics that you isolate, read up about slants
I hoped some of this could help


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OfflinecronicrFacebook
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Re: Monoculture Isolation - Early Identification of Genetic Traits [Re: megtheabyss] * 1
    #27106716 - 12/24/20 11:51 PM (3 years, 1 month ago)

if you're starting from a clone just make sure it's clean, that's your isolate.
if your starting from spores or clones the amount of transfers doesn't matter senescence happens due to numbers of cell division which is very very minimal on a 2 d surface in Petri dishes , it's more of a grain expansion to bulk type of issue


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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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Offlinemegtheabyss
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Re: Monoculture Isolation - Early Identification of Genetic Traits [Re: Shroomoisseur]
    #27106745 - 12/25/20 12:14 AM (3 years, 1 month ago)

Thanks for your input man.

I do have the clone tissues (from the best 3 fruits) preserved in slants already from the grow last winter. I'm planning to do three individual tubs from that. Depending on which of the 3 yields best, I'll be making the slant as the base for the monoculture selection. I'm thinking about doing upto 10 mini tubs (one per sector) in each phase and log observations for each tub. Then select the best sector for the next phase, and repeat the process again, until I reach multiple monocultures to select the best one from. Hopefully should not take more than 2 years!


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abltsandwich said:
Gypsum adds sterilization to the pressure cooker.  If you PF Tek some bulk substrate and proper sterilization pasteurization, then mushrooms and gas exchange (GE) plus FRESH AIR EXCHANGE (brown rice flower [FAE]).


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Offlinemegtheabyss
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Re: Monoculture Isolation - Early Identification of Genetic Traits [Re: cronicr]
    #27106756 - 12/25/20 12:21 AM (3 years, 1 month ago)

Quote:

cronicr said:
if you're starting from a clone just make sure it's clean, that's your isolate.





Sorry if I'm off base here, but isn't a first generation clone (from a MSS grow) an aggregation of multiple genetic lines, rather than being an isolate? My understanding was that because of that, even with a clean tissue from a large / potent clone, it is optimal to sub-select / isolate the dominant genes from the entire pool that contributed to the size or potency of the fruit.


--------------------
abltsandwich said:
Gypsum adds sterilization to the pressure cooker.  If you PF Tek some bulk substrate and proper sterilization pasteurization, then mushrooms and gas exchange (GE) plus FRESH AIR EXCHANGE (brown rice flower [FAE]).


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OfflinecronicrFacebook
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Re: Monoculture Isolation - Early Identification of Genetic Traits [Re: megtheabyss]
    #27106766 - 12/25/20 12:37 AM (3 years, 1 month ago)

not really because the genetics that make a fruit are one so let's say you put some albino a in with some pe( twonisolates) and your tub showed about half of each except one fruit off in the corner which looked like a white cock, in the setting where most believe a fruit contains multiple sets of genetics you would be able to clone that fruit and only see 2 sectors, transfer each of these separate and grow them out and ones normal or and ones albino a ...right?
however this is not the case because in order for multiple strains to make one fruit they would have to decide who makes the cap who makes the stem who makes the spores lol.
if somebody can pull off the scenario above I got a prize for u


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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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