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InvisiblePastywhyteMDiscord
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Re: Cubensis Lifecycle Questions [Re: c10h12n2o] * 2
    #23755358 - 10/20/16 03:58 PM (7 years, 3 months ago)

For the most part I'm not worried about sectors. I know some people believe that the different sectors of a clone will still have most/all of the different genetics bound up in them anyways. I have also seen sectoring with isolates to some degree as well (aloha cultures not mine). Because of that I don't really worry about isolating clones, in fact I suspect it might not even be possible though don't quote me on that.

Instead I focus on robust healthy growth. Rhyzo or tomentose doesn't matter, my best clone is tomentose as hell but grows as fast as oysters.  Instead I focus on sectoring as a flag for things like contamination or senscence. It can sometimes indicate something amiss in the culture, and if I think that it's getting worn out or possibly has something riding along I just go back to the slant.


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Re: Cubensis Lifecycle Questions [Re: Pastywhyte]
    #24203998 - 03/29/17 09:55 PM (6 years, 9 months ago)

i just updated the OP to include the latter questions

Since RR is floating around here now (god bless america), i figured id update and bump this thread in the hope that maybe he will see it and chime in on any of these questions/talking points. of course all other info/discussion is welcome as well

i am working on a ton of agar projects and it would be tremendously helpful for my future projects and goals to better understand these factors


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Re: Cubensis Lifecycle Questions [Re: c10h12n2o]
    #24204197 - 03/29/17 11:19 PM (6 years, 9 months ago)

Great thread. Hope more info keeps popping up.


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Re: Cubensis Lifecycle Questions [Re: bastard4life]
    #24204219 - 03/29/17 11:40 PM (6 years, 9 months ago)

:takingnotes:


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Re: Cubensis Lifecycle Questions [Re: van hatton] * 1
    #24204316 - 03/30/17 01:07 AM (6 years, 9 months ago)

I suspect in some cases spores land near a colony and germinate then if compatible will undergo anastomosis. Thus feeding a colony new fresh nuclei and genetic(even though it's selfing its still variation and freshness)

There's no combination of the nuclei in dikaryotic mycelium. This means many strains can be together in the same organism and eventually combine(karyogamy and meiosis). heterokaryosis is the name for this


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Re: Cubensis Lifecycle Questions [Re: bodhisatta]
    #24204591 - 03/30/17 06:49 AM (6 years, 9 months ago)

I will say this:

One full spore print in a single syringe, used to inoculate 4 identical sterilized grain jars.  Syringe was aggitated vigorously, between all innoculations, and in jar 4, 4cc's were injected.  In jar 3, 3cc's were injected, jar 2 got 2cc's, and Jar 1 got (you guessed it) 1cc.

Jar 4 showed visible germination after 3 days, with each jar following suit with a day or two between each...So by day 9 I was able to see one tiny isolated patch of wispy mycelium in jar 1.  It was so small that I had doubts that it would even be dikaryiotic as I suspected only one spore had germinated.  These are them at day 10 I believe:



The jars also colonized at rates dramatically magnified by the number of spores they received.  Each jar was then fruited to its own dish/tray in the fruiting chamber when it's time came.  Jar 1 took an entire two months to colonize.

Trays fruited in identical order of timeframewith every tray taking exactly 16 days from casing to show visible primordia. 

The kicker is this: tray 4 showed many vastly different sizes, shapes, and densities of fruits, some large and beautiful and solid, and some twisty and different sized and some with hollow stems...And diversity decreased accordingly down to tray 1 whose fruit were all astoundingly uniform and terrible.  Small, hollow stemmed retards, the whole lot.  I had achieved some level of isolation simply by virtue of using fewer and fewer spores, but sadly the most isolated jar had shit genetics.

The best fruits from trays 1, 2, and 3 were cloned to separate petris, and only one showed sectoring, which was selectively transferred for rhizomorphic growth.  These "isolates" were moved to sterile grain jars 1, 2, and 3, and again grown out.  This round, fruits were all similar, uniform, and much larger that the parents that were cloned, leading me to believe this was due to considerably less competition for nutrients for all three trays this time around.

Because of this, I suspect that in a given MS jar, cake, tub, tray there are actually numerous small separate mycelial colonies all with their own unique combination of genetics, never combining in a meaningful symbiotic way and all competing for their own chunk of the food, even though some may not even be fruiting strains or if they are, they never make it to the surface so they can see the needed environment to fruit.  I think it is all pretty much a repeat of the petri dish with three distinct strains refusing to combine.

My conclusion may be completely bogus.


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Edited by AtomHeart (03/30/17 08:16 AM)


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Invisibleamidogen
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Re: Cubensis Lifecycle Questions [Re: AtomHeart] * 1
    #24204607 - 03/30/17 07:00 AM (6 years, 9 months ago)

.


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Edited by amidogen (06/21/18 07:20 AM)


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OfflineAtomHeart
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Re: Cubensis Lifecycle Questions [Re: amidogen]
    #24204612 - 03/30/17 07:03 AM (6 years, 9 months ago)

I think one of the questions being asked, though, was if these strains ever combine into a single genetic amalgamation, or at least combine into one colony and serve other valuable roles such as gathering nutrition for the overall colony...And my conclusion was that they don't.  Sorry I wasnt very articulate about that.

I mean, of course there are numerous strains in each jar...You can see that just by looking at the differing types of mycelium in my photo above...But as far as I can tell, they don't combine even though they eventually always appear to become one solid white mass... and most of the bad strains serve only to consume your valuable substrate and do not contribute to the strains that are trying to fruit in a noticable way.  They only rob them of their food.

Sorry if this just ended up restating shit everyone already knows, but I was attempting to draw a conclusion from this experiment that could answer questions about what happens in nature.

Of particular value is the fact that the clones of the parents in all cases grew much larger with the only real variables being the number of genetic strains present to compete for the same amount of food.


Edited by AtomHeart (03/30/17 08:17 AM)


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Invisiblemynakedrat
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Re: Cubensis Lifecycle Questions [Re: AtomHeart]
    #24204652 - 03/30/17 07:33 AM (6 years, 9 months ago)

And I think there 8s no way a fruit on one side of a tub gets the exact same conditions as another. We are not robots. Grain distribution, micro air currents, watering, although specific and metered, are no perfect. I think perfect growing only occurs in labs or on Agar. And even Agar can have a somewhat different solution to it, unless done in a lab with computers.
I'm. Saying all this is why a clone turns out so many different fruits. Many many factors present. And even while growing, if a bit of myc is getting a little more food than a part a centimeter away, that myc will grow faster, and thereby consumers the food of some neighboring myc.
I'm saying all this as I start a clone with a huge fruit I got. It may have been huge simply because it got more food. Hell, the damn thing may just turn out to be a bunch of weak. And small fruits, and I grabbed one that had optimal conditions.


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Re: Cubensis Lifecycle Questions [Re: AtomHeart]
    #24204876 - 03/30/17 09:30 AM (6 years, 9 months ago)

Quote:

AtomHeart said:
I think one of the questions being asked, though, was if these strains ever combine into a single genetic amalgamation, or at least combine into one colony and serve other valuable roles such as gathering nutrition for the overall colony...And my conclusion was that they don't.  Sorry I wasnt very articulate about that.

I mean, of course there are numerous strains in each jar...You can see that just by looking at the differing types of mycelium in my photo above...But as far as I can tell, they don't combine even though they eventually always appear to become one solid white mass... and most of the bad strains serve only to consume your valuable substrate and do not contribute to the strains that are trying to fruit in a noticable way.  They only rob them of their food.

Sorry if this just ended up restating shit everyone already knows, but I was attempting to draw a conclusion from this experiment that could answer questions about what happens in nature.

Of particular value is the fact that the clones of the parents in all cases grew much larger with the only real variables being the number of genetic strains present to compete for the same amount of food.



Yes that's what happens I just said what it was in my post.
heterokaryosis
Many nuclei of many strains exist in the whole organism of mycelium


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OfflinePlantManBee
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Re: Cubensis Lifecycle Questions [Re: c10h12n2o]
    #24204896 - 03/30/17 09:42 AM (6 years, 9 months ago)

Quote:

c10h12n2o said:

11. i guess that brings up another thought: since we see monocultures express different characteristics as we manipulate variables, is it possible that what we see as multiple sectors could sometimes be the same strain group, just expressing different physical charactaristics? ive long suspected multiple sectors within the same culture to be the same group of strains, only growing in a different direction. but this carries that even further, if the same group of strains could express different phenotypes within the same culture. if so, i would love to identify the variable that would cause a sector made up of the same cast of strains as another to differentiate itself within the same dish




From a genetic standpoint, a single organism (stain in this case) can express its genes differently in different environments. I'll just use pH as an easy example: when the pH of a solution changes it can alter the way proteins fold, exposing different base pairs for tranduction and replication.


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Re: Cubensis Lifecycle Questions [Re: bodhisatta]
    #24205142 - 03/30/17 11:37 AM (6 years, 9 months ago)

Quote:

bodhisatta said:
I suspect in some cases spores land near a colony and germinate then if compatible will undergo anastomosis. Thus feeding a colony new fresh nuclei and genetic(even though it's selfing its still variation and freshness)

There's no combination of the nuclei in dikaryotic mycelium. This means many strains can be together in the same organism and eventually combine(karyogamy and meiosis). heterokaryosis is the name for this




dopesauce bodhi :highfive1:

that is killer info, im gonna have to do some digging into these topics some more, and i will be back shortly with more informed questions.

one obvious question is this (might be misunderstanding something):

if "no combination of the nuclei in dikaryotic mycelium" , wouldnt they be "eventually combine" through karyogamy (i gotta look this up) / heterokaryosis? i think im missing something fundamental

any idea what would trigger a group of distinct strains to undergo combination in this way? also, im familiar with meiosis as it pertains to spores and sperm, but im totally ignorant as to how it comes into play here

so would a well established wild colony most likely have already undergone these changes?

also, is there any way to 100% verify that a culture is a monoculture vs a highly organized group of strains? also is there any way to know if the multiple strains in a MS culture have exchanged genetic material or if they remain genetically distinct?

i know you can verify monokaryotic cultures by verifying the lack of clamp connections, but is there a simple way to verify any of these other things?

i will be back later with more informed questions :smile:

thanks a bunch to everyone keeping this thread alive, it could really bridge some of the major holes in my understanding


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

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"Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing."

"Convictions are more dangerous enemies of truth than lies"
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Edited by c10h12n2o (03/30/17 11:38 AM)


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InvisiblebodhisattaMDiscordReddit
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Re: Cubensis Lifecycle Questions [Re: c10h12n2o]
    #24205160 - 03/30/17 11:43 AM (6 years, 9 months ago)

Combination happens in the gills to make spores.

Monokaryotic mycelium is hard to obtain. You would have an easier time with a micro manipulation scope rather than trying to dilute spores down enough


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Invisiblec10h12n2o
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Re: Cubensis Lifecycle Questions [Re: bodhisatta]
    #24205402 - 03/30/17 01:26 PM (6 years, 9 months ago)

aaaaaahhhh ineresting....

i undersand that monokaryotic cultures are exceedingly difficult to obtain  (even prohibitively so), I was just pointing out that there is a way to verify 100% whether a culture is monokaryotic  (by checking for the distinctive clamp connections prevalent in dikaryotic cultures), and wanted to know if there are a any comparable methods for verifying that a culture is indeed a monoculture vs many strains, especially when the strains are well organized and not forming distinct sectors

on the same page, is there any way to distinguish highly compatible strains from each other when they are part of an organized culture (not distinguishing themselves by forming sectors)?

also, I've never worked with a micro manipulation scope. what exactly are some of the ways in which this could be used to obtain monokaryotic cultures, etc (in terms of theory and technique)? identifying and germinating single spores I suppose?


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"Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing."

"Convictions are more dangerous enemies of truth than lies"
― Friedrich Nietzsche


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Re: Cubensis Lifecycle Questions [Re: c10h12n2o]
    #24205596 - 03/30/17 02:35 PM (6 years, 9 months ago)

Micro manipulation scope you could grab a single spore and put it on its own dish.


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Re: Cubensis Lifecycle Questions [Re: c10h12n2o]
    #24414099 - 06/18/17 04:03 AM (6 years, 7 months ago)

I know flys an other bugs have to be spreading spores for the mushrooms.
Specially cubes growing from a cowpie.

Idk if some1 said that already, just woke up so sorta skimmed.


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Re: Cubensis Lifecycle Questions [Re: LemurLemur]
    #24414186 - 06/18/17 05:44 AM (6 years, 7 months ago)

What is monokaryotic tissue used for?


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Re: Cubensis Lifecycle Questions [Re: LizardWizard]
    #24414498 - 06/18/17 08:55 AM (6 years, 7 months ago)

Creating isolates. Isolates made that way are also free from anastomosis having had any influence along the way and are a true genetic pair.


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Re: Cubensis Lifecycle Questions [Re: Pastywhyte] * 1
    #24414597 - 06/18/17 09:28 AM (6 years, 7 months ago)

I highly recommend this book.. The Fifth Kingdom.. Alan recommended it to me when I started picking and getting super into fungi.

First 5 chapters are free online.

http://www.mycolog.com/fifthtoc.html

Looks like the free parts have been expanded. It's a very good read.


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Re: Cubensis Lifecycle Questions [Re: Adden]
    #24415158 - 06/18/17 01:40 PM (6 years, 7 months ago)

I will read all the comments later but i just have to put my two cents in. I lived in Iquitos, Peru where cubes grow naturally in cow fields, when i first got there every cow field along the highway was FULL of Cubes , however LOTS of people went picking and nearly picked them out of existence. I also worked at a retreat center with a cow field next door, one that used to produce a lot but was a rather popular picking spot and was picked out of production. I then started regularly going there finding the old mushrooms that were too old to eat and I wold shake them up in a ottle of water and find cow pies at the right stage of maturity and pour the spore water into little holes poked into the cow patty, i also fed the cows spores and after 6 months production was back to normal.

From my observations I have concluded that the cubes are mostly living as individual colonies based in and around individual cow patties they are established in and not an integrated mycelial network connecting the entire field. Like in captivity each colony has a limited amount of flushes before the nutrients are exhausted. That is how they got over-picked because the cycle of spores to more cow pies was interrupted by people constantly picking each and every flush. now sometimes they do go underground and even fruit from the ground but I believe this is an occasional occurence when a food source happens to be readily available in the soil adjacent to a cow pie the once lived in and not the fruiting of a pervasive mycelial network.


Edited by Lennybernadino (06/18/17 03:17 PM)


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