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QM33
(NOT A PUPPET!) ❤❤❤❤❤



Registered: 04/09/20
Posts: 4,739
Loc: Oregon
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Re: Dedikaryotization of Cubensis fruit body clones? [Re: Psilotoad] 1
#27750284 - 04/25/22 06:33 AM (1 year, 8 months ago) |
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It would be about just as difficult as obtaining. A single spore isolation as well.
You would need to blend it to expose the 2 nuclei that are present in dikaryotic myceliums. And really I doubt a regular blender could do this, I imagine you would need a laboratory grade, razor sharp blender to slice through cell walls.
I was talking with a buddy about this recently. You might even be able to use a cell wall degrading enzyme like in Protoplast Fusion.
Either way. Once you have your nuclei exposed from the cell walls, you may as well have a spore solution as you have a bunch of individual nuclei, monokayotes floating around your solution.
The solution would need to be diluted down to the extent you would dilute a solution for single spore isolation. You need a microscope and hemocytometer for this. And then you would need to go back and identify the growth that's worth transferring ebfor it interacts with another colony.
And really if your trying to cross 2 specimens from the same species you can literally just let the myceliums mate. If you want a higher likelyhood of traits from one than the other use more of that mycelium in the culture.
In theory if you have 2 same species you want to cross, and you want the traights from 1 more than the other, if you say did a 1 to 3 ratio, 3 bring the spawn of the traights you want primarily, say you g2g 2 different spawns of 2 same species specimens together, than in theory you would be more likely to obtain the traights of the one with the higher spawn rate as their ar more nuclei containing the genetic information, increasing the breeding probabilities for those traights.
-------------------- OmManiPadmeHum,OmManiPadmeHum, OmManiPadMeHum... There are known knowns, there are known unknowns, there are also unknown unknowns. With great privilege comes great responsibility.
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QM33
(NOT A PUPPET!) ❤❤❤❤❤



Registered: 04/09/20
Posts: 4,739
Loc: Oregon
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Re: Dedikaryotization of Cubensis fruit body clones? [Re: Psilotoad]
#27781520 - 05/17/22 12:12 PM (1 year, 8 months ago) |
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What kind of blender do you need? You say eberbach jar, that's just a standard lab blender right?
Have you attempted this? Do you know if it damaged the nuclei?
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QM33
(NOT A PUPPET!) ❤❤❤❤❤



Registered: 04/09/20
Posts: 4,739
Loc: Oregon
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Re: Dedikaryotization of Cubensis fruit body clones? [Re: QM33]
#27781529 - 05/17/22 12:21 PM (1 year, 8 months ago) |
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All I was able to read was thi, but it did answer my question
"" The dedikaryotization method proposed for Pleurotus spp and other basidiomycetes by Leal-Lara and Eger-Hummel in 1982 was modified in order to recover the monokaryotic components of seven Lentinula edodes strains. By decreasing the time that agar cultures were blended, mycelial death was reduced and more neohaplonts were recovered. If agar culture homogenates were plated on malt extract agar, L. edodes strains barely survived 30 seconds blending. Agar cultures of L. edodes blended for more than 5 seconds did not yield viable mycelium when agar culture homogenates were inoculated in dedikaryotization liquid media. Optimum blending time of agar cultures was determined for each strain. An attempt was made to recover neohaplont types at different stages of the dedikaryotization procedure either directly from agar culture homogenates or from dedikaryotization media inoculated with such homogenates and incubated for 14 days at 24oC or from homogenates incubated for fourteen days in dedikaryotization media. All seven L. edodes strains were successfully dedikaryotized. In all cases, both monokaryotic components were recovered and in six cases, they were isolated in a 1:1 ratio.""
So it doesn't say what type of blender they use.?
What does isolated in a 1 to 1 ratio mean?
And of course you have the perks of the parent fruits.
Your still going to need to make a serial dilutions with a hemocytometer and plate them out and make sure they don't have clamp connection.
-------------------- OmManiPadmeHum,OmManiPadmeHum, OmManiPadMeHum... There are known knowns, there are known unknowns, there are also unknown unknowns. With great privilege comes great responsibility.
  Quantom Qups PROOF AND Soft Drops Turn your Swab to a Syringe and Syringe to Multiple Syringes! No Pours (QuantomStyal)Magic Fruit Leather DMT for IandI
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