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Re: differences between strain isolation by transfers and by cloning? [Re: arp180]
    #9453570 -

Yes and no to your original question. Yes it's a shortcut if you just want to isolate a single fruitable strain. Not necessarily if you want to produce the strongest/fastest growing mycelium or most potent fruit. This is because the combination of strains may make it possible for a set of DNA with the coding for 'potent' to combine with a fruitable strain that is not potent. While when you separate them out the potent strain may not be fruitable. Does that make sense?

The different characteristics that make up a fruit body may not be presented when separated out.

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Re: differences between strain isolation by transfers and by cloning? [Re: seven]
    #9453669 -

i think cloning to agar and transfering each sector that comes from a fruit would be a quicker method of getting some isolates. the isolates are somewhat chosen for you as the results of a MS grow, but you will make less transfers.Same could be done to a handfull of other different ms clones for genetics to choose from. this is sort of a question: I havent done this but it seems logical you can use clones to acheive isolates in less transfers? :shrug:


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grind

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Re: differences between strain isolation by transfers and by cloning? [Re: seven]
    #9453718 -

Yes seven, of course. Like I said if you just want something that will fruit and give you an even pinset with hopefully stronger mycelium, that will do it just fine.

When you isolate for a specific property, that is when you run into issues with a clone.

Also clones would need way less dishes/transfers.

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Re: differences between strain isolation by transfers and by cloning? [Re: Shroominit]
    #9453845 -

Shroominit said:
Yes seven, of course. Like I said if you just want something that will fruit and give you an even pinset with hopefully stronger mycelium, that will do it just fine.

When you isolate for a specific property, that is when you run into issues with a clone.

Also clones would need way less dishes/transfers.

A clone will almost always show phenotypes of the fruit its cloned from. Isolation of a substrain takes the variation of phenotypes from other substrains out of the picture. Only growth conditions will likely change results of an isolate's performance. :thumbup:


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grind

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Re: differences between strain isolation by transfers and by cloning? [Re: seven]
    #9454594 -

As I understand it is very rare for a clone to have several substrains.


--------------------
"The third eye. You spend years doing everything you can to open it and then the damn thing opens and your friends laugh at you when you tell them you can see their souls behind their eyes burning like rainbows."

Links:
:regularshroom:Nibin's Guide for Noobs
:regularshroom:some easy teks on bulk & grain prep. (my journal)

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Re: differences between strain isolation by transfers and by cloning? [Re: dead]
    #9454598 -

I've wondered how common it would be myself, but I don't think it's that rare.  Someone said they once had a clone sector into like 6 sub-strains.  I don't think it's that uncommon for multiple sub-strains to 'collaborate' on a fruit.

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Re: differences between strain isolation by transfers and by cloning? [Re: J3illy]
    #9454989 -

On the other hand I've heard several people say they have never seen a clone culture sector at all.


--------------------
"The third eye. You spend years doing everything you can to open it and then the damn thing opens and your friends laugh at you when you tell them you can see their souls behind their eyes burning like rainbows."

Links:
:regularshroom:Nibin's Guide for Noobs
:regularshroom:some easy teks on bulk & grain prep. (my journal)

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Re: differences between strain isolation by transfers and by cloning? [Re: dead]
    #9455086 -

i have had a clone sector, they defiantly do


--------------------
Back the tape up.  I need it again!
Let it roll!  Just as high as the
fucker can go!  And when it comes
to that fantastic note where the
rabbit bites its own head off, I
want you to  THROW THAT FUCKING
RADIO INTO THE TUB WITH ME!
    Not me.  It would blast you through
the wall stone dead in ten
seconds and they'd make me explain
VVVVV Free Myco Thread

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Re: differences between strain isolation by transfers and by cloning? [Re: grod31]
    #11017830 -

Has anyone ever measured specific chemical compositions while isolating a strain? Theoretically could you isolate any above average mycelium (that showed excellent fruits with higher biochemical composition) to create some type of super strain?

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Re: differences between strain isolation by transfers and by cloning? [Re: Bluemoondreamer]
    #11017945 -

Bluemoondreamer said:
And strains also share more than 50% of their genes too.

"We found the average nucleotide identity (ANI) of the shared genes between two strains to be a robust means to compare genetic relatedness among strains, and that ANI values of ≈94% corresponded to the traditional 70% DNA?DNA reassociation standard of the current species definition. At the 94% ANI cutoff, current species includes only moderately homogeneous strains, e.g., most of the >4-Mb genomes share only 65?90% of their genes, apparently as a result of the strains having evolved in different ecological settings."



i think this is quit clear as to the need to isolate then clone instead of just cloning.

some of the impure genetics from a clone of a multi spore will show up as were a isolate which was isolated for a specific purpose to a point of stable repeatable traits will not.

so the end game is there but you need real good luck to win on the first try. cuz the second or third might loose the game for you.


LMAO!!!


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some people just don't get it:spank:
http://www.shroomery.org/forums/showflat.php/Number/11241796
so here is a video or two or three for you guys:rolleyes:

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Re: differences between strain isolation by transfers and by cloning? [Re: ratdog]
    #11018049 -

Quote:
i think this is quit clear as to the need to isolate then clone instead of just cloning.


When you isolate, you don't need to clone because you save well-labeled mycelium from each culture under refrigeration.  Later, when you determine the best isolates, you go back to your culture collection and grab that specific one to grow out.  This way, you always have very young cell lines, not months old cell lines from a clone. 

These juvenile cell lines stored in master slants are how commercial mushroom farms stay in business.  If they kept cloning, all their cultures would fade out due to senescence.
RR


--------------------
Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Re: differences between strain isolation by transfers and by cloning? [Re: RogerRabbit]
    #11018107 -

RogerRabbit said:
Quote:
i think this is quit clear as to the need to isolate then clone instead of just cloning.


When you isolate, you don't need to clone because you save well-labeled mycelium from each culture under refrigeration.  Later, when you determine the best isolates, you go back to your culture collection and grab that specific one to grow out.  This way, you always have very young cell lines, not months old cell lines from a clone. 

These juvenile cell lines stored in master slants are how commercial mushroom farms stay in business.  If they kept cloning, all their cultures would fade out due to senescence.
RR




this is a good deff of senescence too thanks RR


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some people just don't get it:spank:
http://www.shroomery.org/forums/showflat.php/Number/11241796
so here is a video or two or three for you guys:rolleyes:

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Re: differences between strain isolation by transfers and by cloning? [Re: ratdog] * 1
    #11018130 -


SENESCENCE
The End is Near


--------------------
The above post is fictional, hypothetical, or downright nonsensical.


:moon:  :moon:  :moon:  :moon:    :moon:

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Re: differences between strain isolation by transfers and by cloning? [Re: anonjon]
    #11024051 -

I can also see where the cost of materials would come into play. It is cheaper to figure out you have a good strain to grow in bulk on a small amount of agar rather than a whole jar, bag, or tub. I am going to look into doing isolation once I get myself a mini fridge. I would prefer to keep my isolated strains separate from my leftover mexican food.

RR do you perhaps have an answer to my original question?

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Re: differences between strain isolation by transfers and by cloning? [Re: beandip]
    #11028561 -

took me about 30 seconds to find this:
'I'm going to throw something out here that contradicts the prevailing beliefs in cloning/isolation. I would like to see some of you in a position to do so, attempt to verify or disprove the following.

I have noticed when cloning fruits from multispore inoculation, that more than one strain is cloned. I have seen obvious sectors on the agar from a single piece of cloned tissue. At first, I thougt I must have gotten some spores that were stuck to the stipe into the mix, but later I was very careful to only select tissue from deep inside the stem from fruits that had not yet opened their caps.

My theory based on these observations is that single fruits on a cake or cased substrate from multispore inoculation, can and are sometimes made up of multiple strains(substrains in shroomy speak).

Multiple strains within the same fruitbody? That is how it appears to me. Before anyone answers "impossible", do a bit of research. If there's another explanation, I'm all ears. I'm not talking about different types of mycelium such as cottony, rhizo, airy, etc., but actually separate sectors that when transferred to different petri dishes will not mix with other sectors from the same frutbody. Sometimes fact is stranger than fiction. I'd love to see a few of you myco-geeks jump on this. I know I am. Have fun.
RR '

originally here: http://www.shroomery.org/forums/showflat.php/Number/4345100#4345100


--------------------
The above post is fictional, hypothetical, or downright nonsensical.


:moon:  :moon:  :moon:  :moon:    :moon:

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Re: differences between strain isolation by transfers and by cloning? [Re: anonjon]
    #11029016 -

Nobody has ever shown that sectoring is in fact proof that there are different substrains involved.  There are a lot of different possible explanations.

It could be differing life stages, environmental conditions, differentiation of the tissue, haploidization, etc., etc..

While it's not impossible that different individuals have combined to create a fruit body, it's only one possible explanation.  It's also quite common to have myc colonize the surface of fruit bodies.  So it's quite possible that the fruit body simply had a different substrain grow on the surface of it, contaminating the culture.

Anyhow, more research is needed.  Someone who's isolated and identified mating types could haploidize and type the resulting tissue from these sectors to see if they are different mating types or what exactly is going on.


-FF

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Re: differences between strain isolation by transfers and by cloning? [Re: fastfred]
    #11029838 -

Weird. Dikaryotic tissue can 'unmate' back? This can be forced? When the cell nucleus loses chromosomes, does it only lose the dna from one spore or the other? i.e. is the resulting tissue exactly like one of the original mating partners, or can it be lose some chromosomes from each side, resulting in a new unique haploid sequence?


--------------------
The above post is fictional, hypothetical, or downright nonsensical.


:moon:  :moon:  :moon:  :moon:    :moon:

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Re: differences between strain isolation by transfers and by cloning? [Re: anonjon]
    #11029935 -

DE-evolution???  i think not! try mutation.


--------------------
some people just don't get it:spank:
http://www.shroomery.org/forums/showflat.php/Number/11241796
so here is a video or two or three for you guys:rolleyes:

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Re: differences between strain isolation by transfers and by cloning? [Re: ratdog]
    #11035973 -

> Dikaryotic tissue can 'unmate' back?

Yep.

> This can be forced?

Yep.  Stress or the use of haploidizing agents can cause this.  It can also happen spontaneously.


Quote:
When the cell nucleus loses chromosomes, does it only lose the dna from one spore or the other? i.e. is the resulting tissue exactly like one of the original mating partners, or can it be lose some chromosomes from each side, resulting in a new unique haploid sequence?


Usually the parental types will be recovered.  However, if recombination occurs then the recovered types will not be a parental type.


-FF

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Re: differences between strain isolation by transfers and by cloning? [Re: fastfred]
    #11036895 -

So this thread is saying that cloning from a fruit is a waste of time?Using the heinz 57 dog analogy, when the cloned myc. starts to fruit you will not know how its going to react due to all of the different genetic material available to it from the original multispore inoculation and myc. growth resulting from it?

Even when you use the iso method it still may be a crap shoot due to great rhizo. growth but crappy fruiting/potency?

If you happen to get lucky enough to find the "golden ticket"(all the great qualities) of spores have it fruit, the spores that drop from that fruit body is going to be as random as when you started originally from the iso?

Then if you take a chunk of that fruit and grow it in an lc the subsequent quality of the fruits there after will degrade if you repeat the process over and over again.

I would think the biggest challenge would be to have every iso tested for potency.It makes you wonder how everyones crappy multispore shrooms even grow or make you trip:smirk:


--------------------

Any post I post here is pure speculation from other read posts
Best tip for cultivation!

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