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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Re: differences betwen strain isolation by transfers and by cloning? [Re: creamcorn]
    #6293705 -

Phenotype
Quote:
1.
1. The observable physical or biochemical characteristics of an organism, as determined by both genetic makeup and environmental influences.
2. The expression of a specific trait, such as stature or blood type, based on genetic and environmental influences.
2. An individual or group of organisms exhibiting a particular phenotype.

phenotypic phe'no·typ'ic (-tĭp'ĭk) or phe'no·typ'i·cal (-ĭ-kəl) adj.
phenotypically phe'no


It goes on if you click the link, I would think isolates would be phenotypes.


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Re: differences betwen strain isolation by transfers and by cloning? [Re: Eraserhead]
    #6293743 -

Yes, isolates would be phenotypes.  But is the reverse true?  Are phenotypes *always* necessarily isolates?  I'm not just playing on semantics I promise, its a legitimate question if you follow me :wink:

If two mushrooms grow side by side and have some defining characteristic that makes them unmistakably the same phenotype (i.e., the dimple spots in the caps would be an example), is it safe to assume they have the same exact genetic make-up?  If I took a clone from each, then fruited them out under the same exact conditions, would I find undistinguisable results in separate grows from either sample?  Or can other characteristics come about that makes the two differ, because they weren't in fact the same to begin with?

To sum up all my babbling thus far, all I'm getting at is this: is there something about a proper agar isolate that makes it more "pure" in a genetic sense than the resulting culture from a cloned tissue sample?  (and if so, how  :confused:)


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Cloning the easy way with LC - newbie friendly pictorial tek

All your answers are here!

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Re: differences betwen strain isolation by transfers and by cloning? [Re: creamcorn]
    #6293759 -

I would think other charateristics would come out, if it was from a multi spore, but if you just had 2 mated spores, I would think they would both produce identically.

Like MJ, has strains, northern lights, kush, blah blah, but each seed is it's own phenotype, but if you take cutings from that plant, all the cuttings will produce _virtually_ the same, if the conditions they are grown under are the same, and they are the same size, and have the same nodes, ect.


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Re: differences betwen strain isolation by transfers and by cloning? [Re: Eraserhead] * 1
    #6293765 -

Why not call them what they are? Strains. The definition of a strain is not the geographical area the original print was taken from. That's marketing.

As for cloning vs strain isolation, they're not related. By the time a substrate fruits, hundreds or perhaps thousands of strains have exchanged DNA, either weakening or strengthening the mass. What you get is a 'heinz 57' that may or may not be that great because the weaker genes and the stronger genes(mycelium) have all combined. An example would be mixed breed dogs. We've all seen good examples and others that are dumber then hell.

Strain isolation on agar begins when the spores first start to germinate. I make the first transfers as soon as I can see mycelium growing from the point of inoculation, long before sectoring can be detected. By doing this, and by continuing to separate each individual growth, you can isolate mycelium prior to the process of anastomosis combining dikaryons into a single mass.

You don't isolate looking for one super rhizomorphic strain. You isolate down to single sectors and then fruit out each one to determine the best performer. When you transfer mycelium to a grain master, the original petri dish the mycelium was taken from is placed into a clean refrigerator. By doing this, when you find the best performing strain, you then go back to your well marked petri dishes, thus your original P1 culture. This petri dish can be used to inoculate a few test tube slants that can be incubated for a week, then placed in cold storage. Whenever you need mycelium, a tiny piece the size of a grain of rice can be taken from the test tube and put on agar to grow out, while the test tube is placed back into the refrigerator. These stored test tube cultures preserve the low P value of your isolated strain for years.

I have a complete video tek on strain isolation and master slant preparation and use already filmed. I'll release it when I get the rest of the teks filmed, and editing completed. Hopefully soon.
RR


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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 betwen strain isolation by transfers and by cloning? [Re: RogerRabbit]
    #6293777 -

"To sum up all my babbling thus far, all I'm getting at is this: is there something about a proper agar isolate that makes it more "pure" in a genetic sense than the resulting culture from a cloned tissue sample? (and if so, how )"

Yes. More than one strain can be present in an individual fruit. Thus, a clone is not an isolated strain, unless it was grown from an isolated line of mycelium.
RR


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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 betwen strain isolation by transfers and by cloning? [Re: RogerRabbit]
    #6293792 -

RogerRabbit said:
Strain isolation on agar begins when the spores first start to germinate.  I make the first transfers as soon as I can see mycelium growing from the point of inoculation, long before sectoring can be detected.  By doing this, and by continuing to separate each individual growth, you can isolate mycelium prior to the process of anastomosis combining dikaryons into a single mass.


Ok, well this clears it up mostly for me. :smile:  There was in fact problems with that minor detail there in my procedure... like I mentioned you can see anastamosis take place right in front of you sometimes, when new sectors emerge from where two meet.  I had never considered that as something to avoid transferring (or transfer early to prevent happening), just thought of it as another choice to choose from. :wink:

So essentially you have more choices from an agar isolation process, but those choices are likely going to exhibit a wider range of characteristics in the end, because we (hopefully) captured a single dikaryotic strain from two and only two spores.  Seems reasonable then that the more mating that goes on, the more things tend to become "average" on you... and that's truly the pro then to isolation... you're starting at the top of the family tree and can travel any of those paths, rather than end up with some kid way down at the bottom. :wink:

To use your analogy, cloning leaves us with a "mutt" :smile:  But there still is that element of chance that says this mutt is actually a fantastic performer.  So again, maybe luck, but growing my mutts out has gotten the job done  :thumbup:  :smile:


--------------------
Cloning the easy way with LC - newbie friendly pictorial tek

All your answers are here!

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Re: differences betwen strain isolation by transfers and by cloning? [Re: RogerRabbit]
    #6293827 -

RogerRabbit said:
Yes.  More than one strain can be present in an individual fruit.  Thus, a clone is not an isolated strain, unless it was grown from an isolated line of mycelium.
RR


Ok, I guess this is where I need to do some research and make myself understand that... as in on a cellular level I guess.  I remember seeing pictures of yours a while back that was actually of tri-karyotic mycelium, and if i recall right the cells had 3 nucleii or something to that effect?  (and i realize that was to show trikaryotic mycelium happens, but isn't necessarily the norm.. besides the point really)... but i'm supposing there's some mixing and matching of those nuclei which is what allows for more than one strain to be present in the fruit body?

i know that's probably grossly over-simplified and i guess i don't really expect an answer, just mulling it over... have some more advanced reading ready to go when i find the time on the subject :smile:


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Cloning the easy way with LC - newbie friendly pictorial tek

All your answers are here!

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Re: differences betwen strain isolation by transfers and by cloning? [Re: Eraserhead]
    #6294293 -

Q - I understand that spores can germinate on agar with peroxide if enough spores are present on the one spot. i read that colonisation speeds of the agar are reduced but would it still work for isolation? or would the dominant substrain take over before the mycelium has time to stretch over the agar and for a workable sized piece to be transfered?

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Re: differences betwen strain isolation by transfers and by cloning? [Re: zeegos]
    #6294328 -

Peroxide should never be used when germinating spores. It's self defeating. If you use a massive amount of spores to find some that will survive the peroxide, it makes sense that you'll also be placing a massive amount of contaminants, some of which also will survive.

Save peroxide for control of cobweb mold. It has little other use in mycology besides possibly helping to control mold in humidifiers.
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 betwen strain isolation by transfers and by cloning? [Re: RogerRabbit]
    #6294374 -

hmm, ok. when doing the first transfer of mycelium, you say to transfer the first mycelium you see to a new dish. doesn't that basically mean transfer all of the visible mycelium? seeing as it'll be so minute. im just puzzled how this would isolate away from contams.
thats my last q, i promise  :grin:

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Re: differences betwen strain isolation by transfers and by cloning? [Re: zeegos]
    #6294389 -

No. The mycelium doesn't originate from a tiny dot. You 'swipe' the spores onto the agar so that the mycelium begins to grow from the zig zag pattern you make with your loop.(you can even sign your name in agar with spores) That means mycelium will begin growing in streaks. Transfer tiny pieces the size of grains of rice along these streaks to new dishes to begin the isolation process.
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]
    #9450232 -

I'm bumping this up 1 because its a great thread and 2 because i got a question !

would then isolating clone tissue transfers be almost a shortcut to achieve the best possible results rather than doing multiple agar transfers from ms prior to any fruiting at all ? if not a shortcut would it at least be the most surefire way to pin down preferred characteristics already tangible with less work before-hand ? not having to do control grows in essence

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Re: differences between strain isolation by transfers and by cloning? [Re: RoYaL_fLuSh]
    #9450295 -

No, isn't that clearly already explained in this two year old thread?

Although a clone is a genetic match to it's parent, unless that parent was created from an isolated strain, you still don't have an isolate.

You are basically breeding a mut.


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"Given the choice between the experience of pain and nothing, I would choose pain."  William Faulkner

"That which exists without my knowledge exists without my consent."
-A quote from the Judge in the novel Blood Meridian; or the Evening Redness in the West by Cormac McCarthy

"Let there be light" My Quick Reference Guide to Lighting
My AutoMono (11oz First Flush)
My Monster Mushroom Mono (9.3oz First Flush)


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Re: differences between strain isolation by transfers and by cloning? [Re: arp180]
    #9450306 -

Well, cloning is still beneficial, because even if you get several strains you will know they will all be efficient fruiters. Or at least capable of fruiting.


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"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]
    #9450333 -

By no means am I saying cloning doesn't have it's place in mycology. I just answered his question.


--------------------
"Given the choice between the experience of pain and nothing, I would choose pain."  William Faulkner

"That which exists without my knowledge exists without my consent."
-A quote from the Judge in the novel Blood Meridian; or the Evening Redness in the West by Cormac McCarthy

"Let there be light" My Quick Reference Guide to Lighting
My AutoMono (11oz First Flush)
My Monster Mushroom Mono (9.3oz First Flush)


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Re: differences between strain isolation by transfers and by cloning? [Re: arp180]
    #9450347 -

arp180 said:
No, isn't that clearly already explained in this two year old thread?

Although a clone is a genetic match to it's parent, unless that parent was created from an isolated strain, you still don't have an isolate.

You are basically breeding a mut.


i understand that its not going to become a pure isolate .. what im looking for is the best possible results by getting a great preforming mutt and strengthening it ..

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Re: differences between strain isolation by transfers and by cloning? [Re: RoYaL_fLuSh]
    #9450811 -

You will definitely have benefits using a clone that are comparable to using an isolate.  You should get a very even, full pinset, w/ all the shrooms growing together w/ similar potency.  It could very well BE an isolate, or it could still be multiple strains - but all the strains will be productive fruiting strains.

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Re: differences between strain isolation by transfers and by cloning? [Re: J3illy]
    #9451776 -

Cloning is the easiest way to isolate from a multispore grow.  It works just fine.  Regardless if it sectors a little or not you at least know you are getting the genetics you're looking for.

There could be di-mon matings, other substrains, or whatever but it's a good first step.


-FF

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Re: differences between strain isolation by transfers and by cloning? [Re: fastfred]
    #9453180 -

agreed, but if you isolate many strains and then fruit, to find what fruits best in your conditions, it's an even better step


--------------------
"Given the choice between the experience of pain and nothing, I would choose pain."  William Faulkner

"That which exists without my knowledge exists without my consent."
-A quote from the Judge in the novel Blood Meridian; or the Evening Redness in the West by Cormac McCarthy

"Let there be light" My Quick Reference Guide to Lighting
My AutoMono (11oz First Flush)
My Monster Mushroom Mono (9.3oz First Flush)


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Re: differences between strain isolation by transfers and by cloning? [Re: arp180]
    #9453564 -

you can acheive simular results to isolation by taking clones from a couple diferent MS grows. Lets say you have four ms trays. Take two clones from each tray>grow each clone batch out. Now you have eight clone batches to pick from.choose the best clone culture to keep. This isnt isolation but it is growing out a selection of cultures with different genetics to pick from. Note - an isolate is more dependable and consistant. With cloning: there is a chance that the fruits will not turn out the same as the fruit you cloned from. its rare but it happens. Other substrains in the clone culture can become dominate and express different phenotypes the next clone grow. < its rare. point being an isolate has been choped down to a single mating- thus no room for variation except for growing conditions.


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grind

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