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I'm looking for suggestions for testing the fruiting performance of isolates on agar.
    #27926864 -

I have some Mazatapec spores coming in, and I plan on trying to isolate a few different strains on agar.

My plan is to try to get (ideally) three different Mazatapec strains isolated, inoculate three-quart jars each with a wedge of agar, spawn each jar 1:1 to coir in a 27qt mono, observe colonization time and fruiting performance, compare dry weight, and compare potency/trip quality.

After I select the best performing strain, I would clone it with tissue from a fruit to agar and transfer those wedges to more agar plates to expand my stock of that particular Mazatapec isolate.  I'd probably also take spore prints from the second flush. 

Is this a good plan, or would you all go about selecting an isolate differently?  I could isolate more than three strains, but that starts to become more and more of a hassle.  Also, I don't like having to devotee lots of spawn to these projects, so mini monotubs or simple trays in an FC seem like the way to go for me.

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Re: I'm looking for suggestions for testing the fruiting performance of isolates on agar. [Re: uninc4life2010]
    #27926894 -

I believe the easiest and most straightforward way too narrow down too something that you want is cloning, whatever you do on agar or in whichever way you want too achieve that is totally up too. I'm not sure what you mean by isolates, but just transfer different sectors from agars, fruit out different sectors and goood luck noticing differences:D

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Re: I'm looking for suggestions for testing the fruiting performance of isolates on agar. [Re: Parafim]
    #27926900 -

The idea of isolating genetics is a little bigger than that. Even cloning a fruit can have hundreds of different genetics in it still and maybe more. Cloning is just more isolated than multi spore which has thousands/millions of genetics.

Testing 3 different cultures like this will probably get you some variety of results that will differ from each other. Size, clusters, potency, yield. But you may find the variations to not differ a lot from each other.

You could clone a fruit transfer it 40 times on agar and still watch it separate off into different sectors and become something different than that clone you munched on to test. And any time you take spores, they're always gonna be a genetic reset. Sure printing characteristics that you like is a good idea but still will be a billions of yr old million jillion sided dice roll when you use that print. Or whatever the numbers might be lol


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Links For Beginners And Beyond:mspoil:

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Re: I'm looking for suggestions for testing the fruiting performance of isolates on agar. [Re: rumfor69]
    #27926910 -

rumfor69 said:
The idea of isolating genetics is a little bigger than that. Even cloning a fruit can have hundreds of different genetics in it still and maybe more. Cloning is just more isolated than multi spore which has thousands/millions of genetics.




If I've already isolated a strain on agar and fruited that isolate, wouldn't cloning a fruit from the isolated strain yield mycelium that is still true to the isolated genetics?  I'm not talking about cloning a fruit that resulted from a multispore grow.  I'm cloning from the isolate that performed the best in fruiting. 

I understand that the prints taken won't be genetically identical to the original isolate, but shouldn't the gene pool should be narrrower than the original MS mycelium colony that came from the original spore print/syringe?  Isn't that the whole point of vendors selling spores of certain strains?  Penis envy spores grow penis envy-looking mushrooms despite the fact that there are a bunch of distinct genetics involved.  It's just that the penis envy gene pool has been narrowed so that all the resulting spores produce penis envy-type fruits.

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Re: I'm looking for suggestions for testing the fruiting performance of isolates on agar. [Re: uninc4life2010]
    #27926918 -

uninc4life2010 said:
rumfor69 said:
The idea of isolating genetics is a little bigger than that. Even cloning a fruit can have hundreds of different genetics in it still and maybe more. Cloning is just more isolated than multi spore which has thousands/millions of genetics.




If I've already isolated a strain on agar and fruited that isolate, wouldn't cloning a fruit from the isolated strain yield mycelium that is still true to the isolated genetics?  I'm not talking about cloning a fruit that resulted from a multispore grow.  I'm cloning from the isolate that performed the best in fruiting. 

I understand that the prints taken won't be genetically identical to the original isolate, but shouldn't the gene pool should be narrrower than the original MS mycelium colony that came from the original spore print/syringe?  Isn't that the whole point of vendors selling spores of certain strains?  Penis envy spores grow penis envy-looking mushrooms despite the fact that there are a bunch of distinct genetics involved.  It's just that the penis envy gene pool has been narrowed so that all the resulting spores produce penis envy-type fruits.



That's what I mean you can't really isolate genetics on agar. You can do hundreds of transfers on agar and there's still thousands of genetic strands all tangled around each other, connected to each other and helping each other. You wouldn't really want a TRUE isolate. It's better to have more genetics happening together that are getting their symbiotic groove on together. Multi spore just has so damn many that it's too random so we take clones.

Creating a cross hybrid variety has a lot more to do with how different variety spore germinate next to each other in the beginning germination stage when they first come out the spore they can genetically combine. So of course after that the prints from those selected fruits will share those combined genetics if they can be stabilized. Sometimes they fall apart like RRs PE6 variety did.


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Links For Beginners And Beyond:mspoil:

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Re: I'm looking for suggestions for testing the fruiting performance of isolates on agar. [Re: uninc4life2010]
    #27927131 -

rumfor69 said:
uninc4life2010 said:
rumfor69 said:
The idea of isolating genetics is a little bigger than that. Even cloning a fruit can have hundreds of different genetics in it still and maybe more. Cloning is just more isolated than multi spore which has thousands/millions of genetics.




If I've already isolated a strain on agar and fruited that isolate, wouldn't cloning a fruit from the isolated strain yield mycelium that is still true to the isolated genetics?  I'm not talking about cloning a fruit that resulted from a multispore grow.  I'm cloning from the isolate that performed the best in fruiting. 

I understand that the prints taken won't be genetically identical to the original isolate, but shouldn't the gene pool should be narrrower than the original MS mycelium colony that came from the original spore print/syringe?  Isn't that the whole point of vendors selling spores of certain strains?  Penis envy spores grow penis envy-looking mushrooms despite the fact that there are a bunch of distinct genetics involved.  It's just that the penis envy gene pool has been narrowed so that all the resulting spores produce penis envy-type fruits.



That's what I mean you can't really isolate genetics on agar. You can do hundreds of transfers on agar and there's still thousands of genetic strands all tangled around each other, connected to each other and helping each other. You wouldn't really want a TRUE isolate. It's better to have more genetics happening together that are getting their symbiotic groove on together. Multi spore just has so damn many that it's too random so we take clones.

Creating a cross hybrid variety has a lot more to do with how different variety spore germinate next to each other in the beginning germination stage when they first come out the spore they can genetically combine. So of course after that the prints from those selected fruits will share those combined genetics if they can be stabilized. Sometimes they fall apart like RRs PE6 variety did.



You got me confused now. What you said seems to go against this post -> Stro's Cleaning and Isolating On Agar

I am I simply not getting it :confused: ?


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Currently looking for nothing. You guys who sent me stuff are straight up awesome!. :mushroom2:

We don't own things - things own us.

Semi-solid liquid culture (SSLC)

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Re: I'm looking for suggestions for testing the fruiting performance of isolates on agar. [Re: Rusty2096]
    #27927151 -

OP seems like for your purposes you could just do an MS grow and pick three fruits to clone. that would save you some time


--------------------
Confused? Well now you can!
HHG - cheapest way to start - how i roll

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Re: I'm looking for suggestions for testing the fruiting performance of isolates on agar. [Re: bakedbeings]
    #27927358 -

:whathesaid:

You can't create an isolate on agar from doing transfers. You can transfer some 1,000 times and it'll still throw sectors sometimes. Even a beautiful perfect shaped disk on agar still has lots of genetic strands all intertwined together.


This is a dish pin clone of a different clone that I've been transferring for yrs now. Still isn't an isolate and never will be.


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Links For Beginners And Beyond:mspoil:

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Re: I'm looking for suggestions for testing the fruiting performance of isolates on agar. [Re: bakedbeings]
    #27927568 -

Bakedbeings said:
OP seems like for your purposes you could just do an MS grow and pick three fruits to clone. that would save you some time



Pick three clones, then compare their performance with three otherwise identical monotubs?  Realistically, I think you're right.  It wouldn't only take a 7-10 days to go from clone to agar to fully colonized jar.

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Re: I'm looking for suggestions for testing the fruiting performance of isolates on agar. [Re: rumfor69]
    #27927602 -

rumfor69 said:
:whathesaid:

You can't create an isolate on agar from doing transfers. You can transfer some 1,000 times and it'll still throw sectors sometimes. Even a beautiful perfect shaped disk on agar still has lots of genetic strands all intertwined together.


This is a dish pin clone of a different clone that I've been transferring for yrs now. Still isn't an isolate and never will be.



Isn't the benefit of multiple transfers that the gene pool is substantially narrower as compared to MS?  As you say, the genetics aren't pure, but there are far fewer genetic lines competing and interacting in a culture like the one you have pictured.

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Re: I'm looking for suggestions for testing the fruiting performance of isolates on agar. [Re: uninc4life2010]
    #27927630 -

If it's from a clone then yes significantly narrrower. Instead of thousands/millions it's more like hundreds/thousands or whatever the numbers are.

But say just an MS plate, that you keep transferring and transferring, you might as well just consider it an MS plate no matter how many times you transfer it.


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Links For Beginners And Beyond:mspoil:

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Re: I'm looking for suggestions for testing the fruiting performance of isolates on agar. [Re: rumfor69]
    #27927818 -

rumfor69 said:
If it's from a clone then yes significantly narrrower. Instead of thousands/millions it's more like hundreds/thousands or whatever the numbers are.

But say just an MS plate, that you keep transferring and transferring, you might as well just consider it an MS plate no matter how many times you transfer it.



Here is the thing, if your original MS culture has 1,000,000 different genetic lines and you do 5 transfers, assuming that you decrease the genetic diversity by a factor of 4 during each transfer (it may well be much more), by the fifth transfer, you'd have narrowed down the genetic diversity by three orders of magnitude.

1,000,000/(4^5) = 976.6

This means that you'd need about 10 transfers before you got down to a single genetic line. 

1E6 / (4^10) = 0.954

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Re: I'm looking for suggestions for testing the fruiting performance of isolates on agar. [Re: uninc4life2010]
    #27927836 -

:popcorn: Will listen/read the debate carefully :takingnotes:


--------------------
Currently looking for nothing. You guys who sent me stuff are straight up awesome!. :mushroom2:

We don't own things - things own us.

Semi-solid liquid culture (SSLC)

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Re: I'm looking for suggestions for testing the fruiting performance of isolates on agar. [Re: Rusty2096]
    #27928101 -

And that math is great I love thinking that way myself with things too. Ok so we know spores are crazy small. If you can see the spores on your agar there a crazy amount of them there. They all germinate touching each other through membranes touching and exchanges I just don't have the terminology for.

All their mycelium strands combine some, wrap around each other, twist, in braids, just microscopic insanity mesh. What we are able to see as white mycelium is billions of single cell microscopic ropes so twisted and piled and numerous you can see it with the naked eye. And all these different genetics are expanded out in every direction.

You make a cut from that..sure you're narrowing it down. But it's still a massive amount of different genetics, they'll be tons of them in every transfer you can see with the naked eye. They're laying on top of each other woven together so tight they look like one rope, that rope though is tons of genetics still.

If you google image search mycelium structures and germinating spores it might help this intricacy I'm trying to describe.


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Links For Beginners And Beyond:mspoil:

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Re: I'm looking for suggestions for testing the fruiting performance of isolates on agar. [Re: uninc4life2010]
    #27928115 -

:popcorn:

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Re: I'm looking for suggestions for testing the fruiting performance of isolates on agar. [Re: rumfor69]
    #27928118 -



Like this for example. This is a tiny segment under high magnification. Each of these strands could and probably is slightly different genetic variants. You couldn't scalpel cut out a piece this small to transfer even.


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Links For Beginners And Beyond:mspoil:

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Re: I'm looking for suggestions for testing the fruiting performance of isolates on agar. [Re: rumfor69]
    #27928142 -

the moral of the story is if the plate looks organized to the human eye, the flush will be uniform enough


--------------------
Confused? Well now you can!
HHG - cheapest way to start - how i roll

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Re: I'm looking for suggestions for testing the fruiting performance of isolates on agar. [Re: rumfor69]
    #27928156 -

rumfor69 said:
And that math is great I love thinking that way myself with things too. Ok so we know spores are crazy small. If you can see the spores on your agar there a crazy amount of them there. They all germinate touching each other through membranes touching and exchanges I just don't have the terminology for.

All their mycelium strands combine some, wrap around each other, twist, in braids, just microscopic insanity mesh. What we are able to see as white mycelium is billions of single cell microscopic ropes so twisted and piled and numerous you can see it with the naked eye. And all these different genetics are expanded out in every direction.

You make a cut from that..sure you're narrowing it down. But it's still a massive amount of different genetics, they'll be tons of them in every transfer you can see with the naked eye. They're laying on top of each other woven together so tight they look like one rope, that rope though is tons of genetics still.

If you google image search mycelium structures and germinating spores it might help this intricacy I'm trying to describe.



No, you're making yourself quite clear.  I can see how the different genetic lines may be intertwined three-dimensionally.  It's really a question of how many genetic lines started out and the percentage reduction in genetic diversity with each transfer.  There may be millions of genetic lines to start out with, there may be thousands.  Each transfer may reduce genetic diversity by 10%, or it may reduce it by 90%.  I'm not informed enough to know.

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Re: I'm looking for suggestions for testing the fruiting performance of isolates on agar. [Re: bakedbeings]
    #27928163 -

Bakedbeings said:
the moral of the story is if the plate looks organized to the human eye, the flush will be uniform enough



:whathesaid:

So taking clones is great cause tremendously cuts down on the genetic variants and gets you to uniformity faster or at least more efficiently and preserves physical traits you wanna replicate the best you can.


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

Links For Beginners And Beyond:mspoil:

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Re: I'm looking for suggestions for testing the fruiting performance of isolates on agar. [Re: rumfor69]
    #27928175 -

rumfor69 said:
Bakedbeings said:
the moral of the story is if the plate looks organized to the human eye, the flush will be uniform enough



:whathesaid:

So taking clones is great cause tremendously cuts down on the genetic variants and gets you to uniformity faster or at least more efficiently and preserves physical traits you wanna replicate the best you can.



this is so key. why narrow down genetics without an idea of the phenotype and potency youre selecting. tbh i dont like getting my cultures too organized if im starting from spores i just get it clean and go. maybe thats whacky but i want a lil diversity if im gonna be picking clones


--------------------
Confused? Well now you can!
HHG - cheapest way to start - how i roll

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