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OfflineRapidRoy
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A couple cloning questions
    #27716834 - 04/01/22 05:02 PM (2 years, 1 month ago)

Hey all,

I just had a couple questions regarding cloning...

1). If I have tissue that I cloned on agar, and moved a section of that  mycelium to a new plate to keep it going, I'am guessing the cloned traits stay with it?

2) If I take agar from cloned fruit and make LC out of it, I'am guessing that the LC will have the cloned traits as well?

Just a couple little questions that I would like to bounce around

Thank you all

RapidR


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Invisiblejoze
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Re: A couple cloning questions [Re: RapidRoy]
    #27716945 - 04/01/22 06:18 PM (2 years, 1 month ago)

Quote:

RapidRoy said:
1). If I have tissue that I cloned on agar, and moved a section of that  mycelium to a new plate to keep it going, I'am guessing the cloned traits stay with it?

2) If I take agar from cloned fruit and make LC out of it, I'am guessing that the LC will have the cloned traits as well?




Hi. To answer your questions:

1. Yes, the traits will stay with the culture. Each clone on agar contains the same genetic information as the mushroom you cloned it from. The same is true for spores: if you germinate a spore on agar and then transfer the culture, each cell contains the same DNA as the spore.

2. Yes, you've got it right. LC can be a great way to "expand" a clone with desirable traits and then inoculate a bunch of substrate.


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OfflineRapidRoy
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Re: A couple cloning questions [Re: joze]
    #27718106 - 04/02/22 02:07 PM (2 years, 1 month ago)

Thank ya sir


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Invisiblehamloaf
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Re: A couple cloning questions [Re: RapidRoy]
    #27722642 - 04/05/22 09:36 PM (2 years, 1 month ago)

There is decay caused by continuous cell division, and living organisms can only divide cells so many times before senescence sets in, so instead of transferring your cloned culture from plate to plate to keep it going; properly maintain cloned culture in master culture slants, and you should be able to grow from a cloned culture for the rest of your life.

I would suggest using grain to grain transfers over liquid culture for the expansion of your spawn.


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OfflineMarkamello
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Re: A couple cloning questions [Re: hamloaf]
    #27722856 - 04/06/22 05:26 AM (2 years, 1 month ago)

At the expense of hijacking a thread. These responses raise a thought.
When a clone is put to agar, then a small section of the clone plate is transferred to a new plate to clean the culture up, do all the traits of the clone transfer to the new plate? Or do some of the traits remain on the original clone plate?

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Invisiblejoze
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Re: A couple cloning questions [Re: Markamello]
    #27723002 - 04/06/22 08:09 AM (2 years, 1 month ago)

That's actually an interesting question. It kind of depends if you started from MS or if you've been working with a clone for a while and it's truly isolated. Two monokaryotic fungi must meet and form a dikaryon before a mushroom can be formed (the reproductive body of the fungi). When you're growing from a multi spore solution, there's hundreds, thousands, of individual spores in the substrate that germinate, each with their own genetic code. When these mycelia have formed dikaryons and are ready to reproduce, many many different sets of mycelia can join in on the action so to speak, and so a mushroom can actually contain a lot of different genetics and traits inside of it.
That's why when you place a piece of mushroom tissue on agar, you get sectors that grow differently. So by selective transferring, you are able to isolate a specific set of genetics and leave behind some of the others.


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InvisibleTheConfluence
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Re: A couple cloning questions [Re: joze]
    #27735694 - 04/15/22 10:05 AM (2 years, 1 month ago)

This brings up a hypothetical for me.

Imagine a culture is brought to fruit, tissue cloned, and brought to fruit until some type of genetic degradation  becomes evident. Could those later generations be merged via "di/Mon pairing" provided the cultivator has the ability to isolate a mono carrion... And would the resulting offspring ditch their maladaptive recessive traits?


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InvisibleNillion
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Re: A couple cloning questions [Re: TheConfluence]
    #27736376 - 04/15/22 07:25 PM (2 years, 1 month ago)

Quote:

TheConfluence said:
Imagine a culture is brought to fruit, tissue cloned, and brought to fruit until some type of genetic degradation  becomes evident. Could those later generations be merged via "di/Mon pairing" provided the cultivator has the ability to isolate a mono carrion... And would the resulting offspring ditch their maladaptive recessive traits?




Yes you could use an existing dikaryonic culture (A) to fertilize a monokaryon (B) to produce a new dikaryon (C) and potentially transfer one of the nuclei from A to C, if they have compatible mating types. If the nucleus that fertilizes the monokaryon contains the genes considered undesirable, then it is quite possible for the traits to be passed on, including if they are recessive.

It is also possible that the issue with the line could be senescence, which is not the same as bad genes, in which case simply growing it from the spores it produces may restore it. Having spores and or culture stored can be a lot easier than breeding a bad gene set out of a line.

Mating factor genes are also involved with mushroom formation and do not exchange genetic information with others but rather act like sex chromosomes in a way and mutations or detrimental alleles (bad genes) in or near the mating factor regions cannot be bred out of a line. This can mean that with some bad lines the only realistic solution is to discard them.

Hypothetically speaking of course.

Edited by Nillion (04/15/22 07:28 PM)

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InvisibleTheConfluence
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Re: A couple cloning questions [Re: Nillion]
    #27737183 - 04/16/22 10:46 AM (2 years, 1 month ago)

my understanding of genetics and allele pairing is pretty basic. I guess if I better understood which traits are dominant and which were recessive (Which is now officially homework for me) I could make some more informed choices in my own lab.

My biggest take away was that it will be the mono culture that will be adopting traits from the elder dikaryon, which I guess kind of implies a bit less predictability in the outcome.

So a hypothetical...
Lets assume I have a very potent cubensis, but it has pretty low yields. Then I also have an isolated monokaryon from a parent that is both a fast colonizer as well as a high yield... the goal obviously being to achieve an offspring that remains highly potent while also achieving higher yields. So, am I then controlling for 3 factors, colonization rate, yield, objective potency. can I simply draw up a punnentt square and trust those predicted outcomes?

for example: 3x3 is obviously 9 possible phenotype outcomes, so then reliably I could achieve an ideal offspring within approximately 9 mated pairs?

Added*
I found a Dihybrid Cross Calculator... The answer is no... Quite a lot more that 9 outcomes

Edited by TheConfluence (04/16/22 10:53 AM)

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OfflineNaeblis
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Re: A couple cloning questions [Re: hamloaf]
    #27788628 - 05/22/22 07:02 PM (1 year, 11 months ago)

Quote:

hamloaf said:
There is decay caused by continuous cell division, and living organisms can only divide cells so many times before senescence sets in, so instead of transferring your cloned culture from plate to plate to keep it going; properly maintain cloned culture in master culture slants, and you should be able to grow from a cloned culture for the rest of your life.

I would suggest using grain to grain transfers over liquid culture for the expansion of your spawn.





Hi, sorry this comment was a while ago, but this made me think. How important is it really to worry about senescence? cultures like Enigma or omni get cloned and expanded from agar to agar lots of times, but it seems to grow just fine?

Edited by Naeblis (05/22/22 07:03 PM)

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OfflineEclipse3130
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Re: A couple cloning questions [Re: Naeblis]
    #27788644 - 05/22/22 07:20 PM (1 year, 11 months ago)

I've personally seen clones die off within a year or 2 depending how many times you transfer them on agar. Usually it's a slow process than a rapid one as it ages, the mycelium may even look different over time and the culture eventually slows down and could even entirely lose its ability to produce. I ran the wheels off a PE culture I had for 2 years and now it is dead, just aborts or produces 1/10th of what it use to


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