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Offlinejohnny_poke
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Why is a clone not an isolate?
    #9630676 - 01/18/09 08:12 AM (15 years, 2 months ago)

It has been suggested to me that mycelium grown from a tissue sample of a fruit body is not an isolate. However, if this statement from here is correct...

Quote:

When hyphae that emerged from two individual spores of the same species cross paths, if they’re compatible, they join together to exchange DNA between them. The resulting mycelium generally has two or more nuclei per cell, and is referred to as dikaryotic mycelium. This mycelium, formed from two separate hyphae, is the very definition of a strain.




...then was I misinformed?

I understand that when growing from multispore inoculation there will be several strains growing from one substrate. Do the strains somehow converge to produce multiple-strain-mushrooms? I was under the impression that each individual mushroom will only have 2 genetic parents (as per the quote above) and hence a culture borne from a tissue clone would in fact be a single and isolated strain.

Can anyone help with some facts please?


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[quote]But what our scientist does not realize is that every time he makes a measurement, the Flying Spaghetti Monster is there changing the results with His Noodly Appendage.[/quote]

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Offlinemushroomhunter10
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Re: Why is a clone not an isolate? [Re: johnny_poke]
    #9630694 - 01/18/09 08:16 AM (15 years, 2 months ago)

I think isolation of mycelium in a dish would contain several gene varieties.

When you clone a mushroom you clone that exact DNA and nothing else.

I'm not a pro so correct me if I'm wrong anybody!


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OfflineNibin
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Re: Why is a clone not an isolate? [Re: johnny_poke]
    #9630726 - 01/18/09 08:24 AM (15 years, 2 months ago)

Quote:

johnny_poke said:
It has been suggested to me that mycelium grown from a tissue sample of a fruit body is not an isolate. However, if this statement from here is correct...

Quote:

When hyphae that emerged from two individual spores of the same species cross paths, if they’re compatible, they join together to exchange DNA between them. The resulting mycelium generally has two or more nuclei per cell, and is referred to as dikaryotic mycelium. This mycelium, formed from two separate hyphae, is the very definition of a strain.




...then was I misinformed?

I understand that when growing from multispore inoculation there will be several strains growing from one substrate. Do the strains somehow converge to produce multiple-strain-mushrooms? I was under the impression that each individual mushroom will only have 2 genetic parents (as per the quote above) and hence a culture borne from a tissue clone would in fact be a single and isolated strain.

Can anyone help with some facts please?




Merging hyphal threads will form a patch of mycelia of a specific genetic strain. As they colonize the substrate the different strains which are compatible (some are not) will merge to produce a single organism made out of many strains.

You do not know if the portion of the organism that produces a specific pin contains 1, 2 or many strains in it.


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Invisibleseven
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Re: Why is a clone not an isolate? [Re: Nibin]
    #9630771 - 01/18/09 08:41 AM (15 years, 2 months ago)

when cloning tissue from a fruit to agar - sometimes it is only one substrain that grows off the sample. Most of the time it sectors into more than one substrain. One could conclude that fruits are, on average, made up of more than one substrain. A clone grow will ALMOST always produce fruits that look like the cloned fruit. not always. there is a chance that another substrain growing from the sample tissue can become dominate and the resulting fruit will be of a different substrain. Isolations on agar remove all other substrains and the chance of variation.


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OfflineRogerRabbitM
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Re: Why is a clone not an isolate? [Re: seven] * 1
    #9631378 - 01/18/09 11:21 AM (15 years, 2 months ago)

It's been shown from DNA testing that up to 200 organisms can be active in a single fruiting body.  If you're growing from multispore inoculation, several substrains can be active in a fruit.  If you're growing from a single sector isolate proved on agar in a petri dish, your clones will be isolated strains, but further down the cell division pool than the original petri dish, which is what you'd want to use for further growing.
RR


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Offlinejohnny_poke
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Re: Why is a clone not an isolate? [Re: RogerRabbit]
    #9631652 - 01/18/09 12:17 PM (15 years, 2 months ago)

OK, thanks for the clear and concise answers everyone. I think I understand. So there can be more than one sub/strain present in a single fruit body, meaning further isolation of sectors on agar is required, however there is a greater likelyhood that the clones may resemble the donor.

Only 1 last question... is there a difference between a strain and a substrain, or is it just terminology?


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[quote]But what our scientist does not realize is that every time he makes a measurement, the Flying Spaghetti Monster is there changing the results with His Noodly Appendage.[/quote]

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OfflineNibin
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Re: Why is a clone not an isolate? [Re: johnny_poke]
    #9632030 - 01/18/09 01:35 PM (15 years, 2 months ago)

Quote:

johnny_poke said:
OK, thanks for the clear and concise answers everyone. I think I understand. So there can be more than one sub/strain present in a single fruit body, meaning further isolation of sectors on agar is required, however there is a greater likelyhood that the clones may resemble the donor.

Only 1 last question... is there a difference between a strain and a substrain, or is it just terminology?




Substrain does not really exist. The correct term is strain.

But because the spores of slightly different subsets of the same genetic pool are named as different "strains" commercially (Golden Teacher, Amazon, etc) we sometimes use the word substrain to avoid confusion.


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Offlinejohnny_poke
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Re: Why is a clone not an isolate? [Re: Nibin]
    #9632363 - 01/18/09 02:46 PM (15 years, 2 months ago)

Thank you Nibin.


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[quote]But what our scientist does not realize is that every time he makes a measurement, the Flying Spaghetti Monster is there changing the results with His Noodly Appendage.[/quote]

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OfflineBasement Boy
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Re: Why is a clone not an isolate? [Re: Nibin]
    #9632375 - 01/18/09 02:50 PM (15 years, 2 months ago)

When it was said that a clone will often produce fruits of the same size and appearance let me ask this, If you were to take a clone of a fruit before the fruit was allowed to grow to its full stature will the clone only grow to the size of the original fruit when it was cloned? Or is the potential of the full fruit body to grow still locked in the DNA? I am thinking the second. For example lets say that when the clone tissue was taken the fruit was only 3" tall but could have grown to 6" tall will the clones only grow to 3" or if let to fully mature will they grow to the 6"?


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Offlinejohnny_poke
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Registered: 12/03/08
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Re: Why is a clone not an isolate? [Re: johnny_poke]
    #9632409 - 01/18/09 02:57 PM (15 years, 2 months ago)

I doubt it. I can't see that the fruit being immature when taking the sample would make any genetic change.

Edit: Didn't make alot of sense :gameover:

Edit1: Still doesn't make alot of sense.


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[quote]But what our scientist does not realize is that every time he makes a measurement, the Flying Spaghetti Monster is there changing the results with His Noodly Appendage.[/quote]

Edited by johnny_poke (01/18/09 03:00 PM)

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Invisibleseven
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Re: Why is a clone not an isolate? [Re: johnny_poke]
    #9633651 - 01/18/09 06:33 PM (15 years, 2 months ago)

depending on growing conditions- the clone batch from the imature fruit\ pin will grow to its full genetics past the maturity level of the tissue you took. so yes your clones can outgrow the pin you clone from.


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InvisibleRedBeerd
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Re: Why is a clone not an isolate? [Re: seven]
    #9633914 - 01/18/09 07:15 PM (15 years, 2 months ago)

Can you at least say that a cloned fruit only contains strains which WILL fruit, and especially under your specific conditions?

Or can the genetics of fruiting strains make up for and coexist with other non-fruiting or more fickle strains?


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Offlinejohnny_poke
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Re: Why is a clone not an isolate? [Re: RedBeerd]
    #9636661 - 01/19/09 07:22 AM (15 years, 2 months ago)

Good question Redbeerd. I've alreay asked this in the advanced forum. The answer I got was this: It is more likely that the clone will be able to produce fruit, but not a certainty. So yes, it seems that there could be non-fruiting strains present in a mushroom tissue sample.

I have another question related to this but don't want to start another thread:

Is it a bad idea to isolate your master culture from a clone? I undertand that every organism has a finite lifespan. If you clone from a fruit body sample you are infact cloning mycellium which has already passed through the fruiting stage. Does this make any difference to the resulting strain once it has been successfully isolated?

Edit: Spelling mistake.


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[quote]But what our scientist does not realize is that every time he makes a measurement, the Flying Spaghetti Monster is there changing the results with His Noodly Appendage.[/quote]

Edited by johnny_poke (01/19/09 07:23 AM)

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Offlinemephisto256
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Re: Why is a clone not an isolate? [Re: johnny_poke]
    #9637028 - 01/19/09 09:23 AM (15 years, 2 months ago)

I dont think that would matter JP. For the simple fact is that all the myc growth from colonization to fruiting size, color, shape, all thats pre-determined in the DNA, like we were at the point of our first cell division. Consider the DNA code as a center, and the stages of shroom growth a hub around the center. Whatever point u want to look at the hub, its center is still the center no matter what, so no, i dont think isolating from fruit body, or taken from a patch of agar myc will make any difference to the strain.

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Offlinejohnny_poke
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Re: Why is a clone not an isolate? [Re: mephisto256]
    #9638005 - 01/19/09 12:46 PM (15 years, 2 months ago)

Quote:

mephisto256 said:
I dont think that would matter JP. For the simple fact is that all the myc growth from colonization to fruiting size, color, shape, all thats pre-determined in the DNA, like we were at the point of our first cell division. Consider the DNA code as a center, and the stages of shroom growth a hub around the center. Whatever point u want to look at the hub, its center is still the center no matter what, so no, i dont think isolating from fruit body, or taken from a patch of agar myc will make any difference to the strain.




Cool, I hope you're right :smile:


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[quote]But what our scientist does not realize is that every time he makes a measurement, the Flying Spaghetti Monster is there changing the results with His Noodly Appendage.[/quote]

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OfflineNibin
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Re: Why is a clone not an isolate? [Re: mephisto256]
    #9638384 - 01/19/09 01:52 PM (15 years, 2 months ago)

Quote:

mephisto256 said:
I dont think that would matter JP. For the simple fact is that all the myc growth from colonization to fruiting size, color, shape, all thats pre-determined in the DNA, like we were at the point of our first cell division. Consider the DNA code as a center, and the stages of shroom growth a hub around the center. Whatever point u want to look at the hub, its center is still the center no matter what, so no, i dont think isolating from fruit body, or taken from a patch of agar myc will make any difference to the strain.




Wrong.

Fungi is generally one mega-organism made out of many strains of the same species. It is not like animals where every single cell of the organism have the same genetic code.

Not only are individual cells polinucleate but they also have different areas with different nucleii with different genes.

This is why we isolate a strain on agar. That way you ensure the whole organism has exactly the same characteristics because every single cell is identical

(Remember we are talking about real genetic strains here, not the bullshit vendors try and sell you as strains and their properties)

So isolating a specific subset of the strains by cloning a fruit will give you different results from your original grow.

It will, obviously, have the risk of producing a non fruiting strain, but the chances of that are pretty low, not only are you selecting from a much lower strainpool that from starting from multispore, you also know that at least some of the strains in there are fruiters. So your chances of success are greater.

Think of it like this: Isolating from multispore is like buying a lottery ticket. Isolating from a fruit is like buying a lottery ticket when you already know half the numbers.

The biggest disadvantage of cloning from fruit is the age of the mycelium, as it will be a month older than the one isolated on agar and kept in the fridge (while a small piece of it is used to grow it out and test it).


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