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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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why isnt it a clone?
    #10798081 -

ok pardon my knowledge here on cloning..
i just made an LC clone using RRs dvd section on cloning. only instead of putting tissue on agar i put the tissue into an LC. i read somewhere that using the LC means it isnt a true clone because the fruitbody itself can contain multiple genetics..or something.
so why is it that when you clone to agar you are getting a true clone but not LC?
is it because you keep making transfers?


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Re: why isnt it a clone? [Re: prismism]
    #10798117 -

when you clone to agar, it still represents multiple strains. you have to further isolate in order to get a single strain isolate. when you take a piece of tissue from a fruitbody, it still represents hundreds of different substrains until it has been completely isolated. usually numerous transfers are required to obtain a single strain.

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Re: why isnt it a clone? [Re: prismism]
    #10798124 -

Right- as your mycelium sectors out on the agar, you can select a single sector to propagate.
With liquid, it's likely you have more than one genetically distinct organism, and no way to separate them.
Way better than multispore, though.


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Re: why isnt it a clone? [Re: Doc_T]
    #10798158 -

It is a clone just not an isolate.


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Re: why isnt it a clone? [Re: CH HELL]
    #10798184 -

CH HELL said:
It is a clone just not an isolate.


now i understand the difference between those two terms. thank you.


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Re: why isnt it a clone? [Re: CH HELL]
    #10798230 -

CH HELL said:
It is a clone just not an isolate.


True. And in that sense, Grain LC or even making new LC from existing LC is cloning too. Just that you are cloning multiple substrains.


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Re: why isnt it a clone? [Re: CH HELL]
    #10798261 -

So .... If I put tissue on agar and it grows without sectoring (monoculture) ... this tissue MUST have come from an isolate?

?Tissue from a MS or previously cloned fruit can never grow on agar without sectoring?

I've wondered about this forever.


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Re: why isnt it a clone? [Re: uncle_rico]
    #10798272 -

If it comes from an isolate fruit it shouldn't sector.


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Re: why isnt it a clone? [Re: uncle_rico]
    #10798280 -

I think it is possible-but-unlikely to get a monoculture from a fruit that wasn't previously isolated using lab techniques.


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Re: why isnt it a clone? [Re: Doc_T]
    #10798331 -

I wonder about this because we put tissue samples on agar from fruits that we can't remember (or never knew) if they were clones or isolates ....

So .... no sectoring: the culture is an isolate. sectoring: the culture is MS or clone.

For sure?


Another question:  I put tissue on agar to rejuvenate my culture (reset the clock) .... am I resetting the clock with tissue?  I wouldn't have to go all the way back to spores to reset the clock would I?


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Re: why isnt it a clone? [Re: uncle_rico]
    #10798360 -

uncle_rico said:
So .... no sectoring: the culture is an isolate.
For sure?


That's my understanding. It's a monoculture by definition.

uncle_rico said:
Another question:  I put tissue on agar to rejuvenate my culture (reset the clock) .... am I resetting the clock with tissue?  I wouldn't have to go all the way back to spores to reset the clock would I?


From spores = 'generation' 0. Depending on how you make grain spawn, your fruits may be 'generation' 2 or 3.

(Expansion generation, not spore-to-spore generation. Sorry I don't know a better way to state it.)


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Re: why isnt it a clone? [Re: CH HELL]
    #10798369 -

CH HELL said:
It is a clone just not an isolate.


Ha ha, I started reading the thread and was wondering if anyone was actually going to point that out. :thumbup:


Doc_T said:
CH HELL said:
It is a clone just not an isolate.


True. And in that sense, Grain LC or even making new LC from existing LC is cloning too. Just that you are cloning multiple substrains.



uh, okay.  But no one uses the word "clone" to describe those procedures.


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Re: why isnt it a clone? [Re: Psuper]
    #10798380 -

Psuper said:
uh, okay.  But no one uses the word "clone" to describe those procedures.


Not in mycology. But that sort of 'cloning' is common with plants, though. Take seedlings and chop them up for starters.
(I've always thought those would be monocultures, but I would have thought the same about mushrooms. :justdontknow:)


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Re: why isnt it a clone? [Re: Doc_T]
    #10800501 -

There are isolates and there are pure isolates. A pure isolate fruited and cloned will be your best change at maintaining common factors. To my understanding even a pure isolate can sector when placed on/in a new substrate such as MEA to PDA agars. In a sense this would not be considered a true pure isolate but it as good as it gets. Remember all life is diverse it's that way for a reason (survival).
A multispore fruiting can definitely contain multiple sectors/substrains.


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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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