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InvisibleAyePlus
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Re: Dedikaryotization of Cubensis fruit body clones? [Re: AyePlus]
    #28182194 - 02/11/23 11:16 AM (1 year, 3 months ago)

Iโ€™m getting even more confused.


@Alan

Are you saying
Quote:

LtLurker said:
Shrooms Dont have just 2 parents like a punnit square. They have many parent spores, some from the same fruit, so we're talking thousands of parents for each fruit.





Isnt true?


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Offlinewy35
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Re: Dedikaryotization of Cubensis fruit body clones? [Re: Alan Rockefeller]
    #28197415 - 02/21/23 12:27 PM (1 year, 2 months ago)

Quote:

Alan Rockefeller said:
Julian shared his writeup on Facebook ten days ago:






What makes the peptone and glucose solution more dedikaryotizing than just water?

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OfflineAlan RockefellerM
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Re: Dedikaryotization of Cubensis fruit body clones? [Re: wy35]
    #28201677 - 02/23/23 10:57 PM (1 year, 2 months ago)

Quote:

wy35 said:
What makes the peptone and glucose solution more dedikaryotizing than just water?





Maybe they are nutrients to help the mycelium heal after the physical destruction of the blender.

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InvisibleMead

Registered: 07/26/02
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Re: Dedikaryotization of Cubensis fruit body clones? [Re: Alan Rockefeller]
    #28202122 - 02/24/23 08:41 AM (1 year, 2 months ago)

Yea, I think it's the recuperation and regrowth, not helping separate nuclei.

Peptone are amino acids and nitrates(and lots of other good stuff,) which are some of the major building blocks of life, and they're mainly/mostly water soluble, so when your blending it's stripping away and diluting it into the water. Adding extra allows them to recuperate/regenerate. Glucose plus nitrates also creates amino acids, glucose is also converted into lipids, and lipids are used for all kinds of stuff, including cell walls. Stuff they normally have to obtain by breaking down stuff which takes energy(and they've already been weakened), this concoction allows it to be readily available.

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OfflinePsilotoad
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Re: Dedikaryotization of Cubensis fruit body clones? [Re: wy35]
    #28203660 - 02/25/23 08:25 AM (1 year, 2 months ago)

You could use just water it has no magnesium or phosphorus but the survivability would be lower and the recovery would be much longer. Like Alan said the peptone glucose helps the mycelium grow and recover. Just basically giving it nutes with low magnesium and phosphorus to prevent clamps from forming when submerged in the solution.

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OfflinePsilotoad
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Re: Dedikaryotization of Cubensis fruit body clones? [Re: AyePlus] * 2
    #28203674 - 02/25/23 08:38 AM (1 year, 2 months ago)

When you use multispore each spore that germinates produces monokaryotic hyphae and each successful pairing of monokaryotic strains produces a new dikaryotic strain so when you put down multispore you have many strains of your type. That is why this mentions you want to use an isolated dikaryon. An isolated dikaryon is comprised of 2 genetically distinct types of nuclei one from each parent monokaryon. In di-mon mating only one nuclei type is donated to the monokaryon and this is not a reciprocal exchange. Each dikaryon only has two parent nuclei from their monokaryon parents or mon and di parent but with multispore you are producing many dikaryons. Once you isolate a colony or clone a fruit you have a single dikaryon this dikaryon will be comprised of 2 nuclei one from each parent monokaryon.

Here this includes some more info about nuclei exchange in mon-mon and di-mon pairings

https://www.mdpi.com/2076-2607/9/6/1248/htm

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InvisiblefahtsterM
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Re: Dedikaryotization of Cubensis fruit body clones? [Re: Psilotoad]
    #28205446 - 02/26/23 01:00 PM (1 year, 2 months ago)

Not sure if you guys have seen This thread but I seemingly have crossed compatible clones by blending pulverized myc water of two clones and inoculating qts directly with the mixed GLC

It was pretty easy

I actually got the idea to try it based on Alanโ€™s post from FB

If tldr:

Yeti clone


Mak 118 clone


Mixed myc water of both


Itโ€™s almost done with the second flush where Iโ€™ll be getting spores and doing a heavy ms grow to show the variation in phenos between parents in the f2

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Offlinewy35
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Re: Dedikaryotization of Cubensis fruit body clones? [Re: fahtster]
    #28208092 - 02/28/23 12:11 PM (1 year, 2 months ago)

I found this patent on dedikaryotizing strains: https://patents.google.com/patent/US4242832A/en

Describes the same procedure, including the peptone and glucose solution. Might be helpful for someone looking for the specifics (e.g. what peptone to use).


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InvisibleAyePlus
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Re: Dedikaryotization of Cubensis fruit body clones? [Re: Psilotoad]
    #28213997 - 03/04/23 11:25 AM (1 year, 2 months ago)

Quote:

Psilotoad said:
When you use multispore each spore that germinates produces monokaryotic hyphae and each successful pairing of monokaryotic strains produces a new dikaryotic strain so when you put down multispore you have many strains of your type. That is why this mentions you want to use an isolated dikaryon. An isolated dikaryon is comprised of 2 genetically distinct types of nuclei one from each parent monokaryon. In di-mon mating only one nuclei type is donated to the monokaryon and this is not a reciprocal exchange. Each dikaryon only has two parent nuclei from their monokaryon parents or mon and di parent but with multispore you are producing many dikaryons. Once you isolate a colony or clone a fruit you have a single dikaryon this dikaryon will be comprised of 2 nuclei one from each parent monokaryon.

Here this includes some more info about nuclei exchange in mon-mon and di-mon pairings

https://www.mdpi.com/2076-2607/9/6/1248/htm




Thank you.


--------------------
Learn about breeding

C10โ€™s agar guide
Good surface conditions = Good pinsets
Read more, post less.
๐Ÿ…‚ ๐Ÿ„ฐ ๐Ÿ„ผ ๐Ÿ„ด  ๐Ÿ…ƒ ๐Ÿ„ด ๐Ÿ„ฐ ๐Ÿ„ผ
๐Ÿ…ƒ ๐Ÿ„ด ๐Ÿ„ฐ ๐Ÿ„ผ  ๐Ÿ„ฒ ๐Ÿ„ป ๐Ÿ„ธ ๐Ÿ„ฝ ๐Ÿ„ถ ๐Ÿ…† ๐Ÿ… ๐Ÿ„ฐ ๐Ÿ„ฟ

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Offlinetryptkaloids
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Re: Dedikaryotization of Cubensis fruit body clones? [Re: Psilotoad]
    #28437476 - 08/17/23 06:41 PM (8 months, 27 days ago)

Quote:

Psilotoad said:
When you use multispore each spore that germinates produces monokaryotic hyphae and each successful pairing of monokaryotic strains produces a new dikaryotic strain so when you put down multispore you have many strains of your type. That is why this mentions you want to use an isolated dikaryon. An isolated dikaryon is comprised of 2 genetically distinct types of nuclei one from each parent monokaryon. In di-mon mating only one nuclei type is donated to the monokaryon and this is not a reciprocal exchange. Each dikaryon only has two parent nuclei from their monokaryon parents or mon and di parent but with multispore you are producing many dikaryons. Once you isolate a colony or clone a fruit you have a single dikaryon this dikaryon will be comprised of 2 nuclei one from each parent monokaryon.

Here this includes some more info about nuclei exchange in mon-mon and di-mon pairings

https://www.mdpi.com/2076-2607/9/6/1248/htm




Bump.
Everyone should learn this stuff. :cheers:
:takingnotes:


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InvisibleMushroom love
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Re: Dedikaryotization of Cubensis fruit body clones? [Re: tryptkaloids]
    #28438194 - 08/18/23 01:01 PM (8 months, 26 days ago)

I have some single ascospore isolate of Cordyceps militaris:
If i use one isolate to colonize a substrat and after full colonisation i inocule the substrat with a ms liquid culture: only the FIRST  monokaryotic compatible culture from ms go to give his genes to my isolate? Is there the same diversity with a clone or a ms cross with a monokaryotic isolate culture ?
I hope my English is understandable, sorry for any errors (corrections welcome)


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Offlinetrippleblack
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Re: Dedikaryotization of Cubensis fruit body clones? [Re: Mushroom love]
    #28440266 - 08/20/23 07:59 AM (8 months, 24 days ago)

Its possible that clones of mushrooms are made up of multiple mycellium colonies; not just two parents. To my knowledge this has not been directly studied in cubensis. i think it's just an assumption that a clone is only comprised of two parents.

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InvisibleMushroom love
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Re: Dedikaryotization of Cubensis fruit body clones? [Re: trippleblack]
    #28440559 - 08/20/23 12:06 PM (8 months, 24 days ago)

Quote:

Psilotoad said:
When you use multispore each spore that germinates produces monokaryotic hyphae and each successful pairing of monokaryotic strains produces a new dikaryotic strain so when you put down multispore you have many strains of your type. That is why this mentions you want to use an isolated dikaryon. An isolated dikaryon is comprised of 2 genetically distinct types of nuclei one from each parent monokaryon. In di-mon mating only one nuclei type is donated to the monokaryon and this is not a reciprocal exchange. Each dikaryon only has two parent nuclei from their monokaryon parents or mon and di parent but with multispore you are producing many dikaryons. Once you isolate a colony or clone a fruit you have a single dikaryon this dikaryon will be comprised of 2 nuclei one from each parent monokaryon.






Quote:

trippleblack said:
Its possible that clones of mushrooms are made up of multiple mycellium colonies; not just two parents. To my knowledge this has not been directly studied in cubensis. i think it's just an assumption that a clone is only comprised of two parents.




Who is right?

So if i do a di-mon cross with a monokaryotic strain and a multispores culture, just the first fastest compatible dikaryons from the ms go to give a nuclei to my monokaryotic culture ?

will it be a means of natural selection of high-performance culture or is it only chance? (the first drop of inoculum to hit the monokaryotic strain)
I think this is the case if the substrate remains whole after colonization and there is a lot of combinations if the grain is broken before (for a multispores inoculum)


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OfflineZwinst
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Re: Dedikaryotization of Cubensis fruit body clones? [Re: Mushroom love] * 1
    #28482325 - 09/25/23 12:08 PM (7 months, 19 days ago)

Quote:

Mushroom love said:

Who is right?




First, my least convincing observation: In a tray there can be multiple different genotypes present. For example when you see both leucistic and coloured fruiting bodys in a tray that came from multispore.
But i guess there could be multiple, distinct dikaryons present. I have observed this multiple times, in different cubensis.

More convincing to me: A single fruiting body can display different traits that donยดt necessarily get expressed, when cloned. Those i got described as "chimeras" to me. For example a fruit that has "patched" gills with zones of coloured and coulerless spores. Or If you have a fruit with leucistic traits, clone it and you get different fruits both with and without leucism.

Of course i am talking about mutations that arenยดt necessarily stable or even well understood and there could be different mechanisms or factors involved. But judging from observations made by people here and having seen this myself multiple times, i would tend to agree, that multiple parents must be involved.


If you think about it, beeing able to accept more than just 2 nuclei into your organism can have big advantages.

Lets just look at the life of a monokaryon, freshly germinated, stretching itยดs hyphae for companionship...
Now it finds a compatible mate east and another one west at the same time. Why should it have to choose? I bet this one mono will just mate with both.
If someone wants to try this on a plate - the way i described it, would be a good starting point, i guess.
My hepa isnยดt finished yet and my own 2 leucistic strains arenยดt stabilized to the point, where i want to release them or start creating monos for crossing. But it is going to happen! And if noone has made this experiment till then, iยดll have to do it.

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InvisibleNillion
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Re: Dedikaryotization of Cubensis fruit body clones? [Re: wy35]
    #28599977 - 12/27/23 03:56 PM (4 months, 19 days ago)

Quote:

wy35 said:
What makes the peptone and glucose solution more dedikaryotizing than just water?



It is actually slightly deficient in nutrients, which stunts the growth of the mycelia and makes it weaker, leading to increased production of neohaplonts, which can appear and recombine rapidly and thus be difficult to recover in some situations. A key part of the method is also culture density, by slowing growth and avoiding excess density it becomes easier to isolate the neohaplonts.

Also, the blender is nothing more than an agitation method, hand agitation or another method of stirring can work just as well to create the sheer forces needed for this, the blender is just faster. Any blender you can keep sterile works just fine for this.

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InvisibleNillion
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Re: Dedikaryotization of Cubensis fruit body clones? [Re: fahtster] * 1
    #28599983 - 12/27/23 04:01 PM (4 months, 19 days ago)

Quote:

fahtster said:
Not sure if you guys have seen This thread but I seemingly have crossed compatible clones by blending pulverized myc water of two clones and inoculating qts directly with the mixed




Yeah, one doesn't need to isolate neohaplonts to use dedikaryotization for breeding, one can just agitate mixed cultures prior to inoculation and screen the results for the desired phenotypes. It's quite handy actually.

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