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Invisiblec10h12n2o
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Cubensis Lifecycle Questions * 2
    #23675503 - 09/24/16 11:14 PM (7 years, 4 months ago)

hello friends, i have been ruminating on some things lately and wanted to see if any of yall could drop some knowledge on me :smile:

so, in our projects the standard lifespan of a fruiting colony is just 1-4 flushes, then we usually dispose/compost the substrate. Im curious about how things work in nature, and have been able to find next to no info on these questions so far. any info would be much appreciated!

a few quotes to set the stage:
Quote:

As with all long-lived plant and fungal species, no individual part of a clonal colony is alive (in the sense of active metabolism) for more than a very small fraction of the life of the entire colony. Some clonal colonies may be fully connected via their root systems, while most are not actually interconnected, but are genetically identical clones which populated an area through vegetative reproduction. Ages for clonal colonies, often based on current growth rates, are estimates.

"Humongous Fungus", an individual of the fungal species Armillaria solidipes in the Malheur National Forest, is thought to be between 2,000 and 8,500 years old.[33][34] It is thought to be the world's largest organism by area, at 2,384 acres (965 hectares).





1. what is the lifespan of a wild cubensis colony? is there a limit to the number of fruiting cycles a culture can sustain, beyond the restrictions placed on it by nutrition and environmental conditions? also, are there different phases in the lifecycle, beyond the basic reproductive cycle, where it has different potential characteristics and behavior? (possibly analogous to human infancy, adolescence, reproductive prime, old age, etc, and the way we can reproduce in some phases but not others, and are more susceptible to illness in some phases than others) 

2. in an "individual" such as "Humongous Fungus" or a wild cubensis colony, would the colony be likely to consist of numerous strains (like a MS grow)? or is it an individual strain, like a monoculture? i would suspect numerous strains to be present, but what i read is often confusing.

3. when a fruit is cloned, and multiple tissue samples are taken from different parts of the fruit, is the exact same variety of strains going to be present in each cultured sample? in my agar work, doing 6 plates of the same cloned fruit, i often see huge differences in the appearance and behavior of the myc. Is this the same collection of strains, expressing different characteristics? or are there a different cast of strains present in various clone cultures from the same fruit?

4. from year to year, if someone harvests cubes from the same place, it is likely from the same colony right? or overlapping, or subsequent colonies? if a wild colony is hundreds or thousands of years old, are there any effects of senescence or any other relevant phenomenon we could readily observe? does age affect wild cubensis colonies?

5. in MS grows, various colonies unite and consolidate somewhat, to network nutrients etc.. does this happen in the wild?

6. in an MS grow, are the same cast of strains present in all parts of the culture/substrate? or is the distribution less homogeneous?

7. in a clone culture, where the first plate shows several clear sectors, wouldnt a true clone culture continue to have all strains present in subsequent plates? how could one do this, since we nececarrily have to pick something to transfer, and cant pick all at the same time?


8. is there any indication that different strains within a colony might serve different functions/purposes within the colony? since some strains individually might not be able to fruit,  but could as part of a colony, are there other functions that might be distributed across different strains within the colony?

9. ultimately, in the beginning of an MS inoculation, monokaryons from spores meet up and become dikaryotic (forming a strain). but rather than having a million tiny colonies separated by the interface between them, we usually see a single, somewhat organized colony. So obviously at some point early in the process, the smaller MS colonies join together and build a network. but this doesnt really seem to happen if you put two established MS cultures together on a plate, since each colony remains distinct. is there some kind of phase, akin to adolescence or infancy, where the hyphae making up a colony are receptive to collaborating/consolidating/joining up with other colonies? 

10. also, since cleaning up clones involves picking a sector and transferring it, wouldnt that necessarily limit the number of strains present, and thus not be representative of all the strains initially present in the clone culture, much less the fruit?

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

might be stupid questions, but i appreciate any light yall could shed on them. thanks in advance!!

warm regards


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Edited by c10h12n2o (03/29/17 09:34 PM)


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Re: Cubensis Lifecycle Questions [Re: c10h12n2o] * 3
    #23675518 - 09/24/16 11:19 PM (7 years, 4 months ago)

1. The mushrooms sporulate onto nearby plants which the cows eat. Its all ms in the wild. The cycle continues. . .

The rest is stuff we humans came up with to produce the mushrooms in greater abundance than nature could. No way nature can grow mushrooms better than me

:drunkdriver2:


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Invisiblec10h12n2o
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Re: Cubensis Lifecycle Questions [Re: Pastywhyte]
    #23675541 - 09/24/16 11:28 PM (7 years, 4 months ago)

right right, just wondering about some of the specifics and big-picture stuff

does that entire cycle happen every single season? as in spores on plants are eaten by cows, then new cultures start in their manure; how long do you think it takes for that new culture to become established enough to fruit?

a tiny little culture wouldnt take as long as a large one of course. in our projects, the substrate being completely colonized is one of the most important factors for fruiting. but obviously this is a very different dynamic in the wild, since it has no set amount of pasteurized substrate to colonize. any insights on how this dynamic plays out in nature?

obviously it would depend on lots of variables, but in general, how many months/years do you think it takes for these spores to become a new fruiting colony? and how many years/hundred years would it take for a culture to die of old age, or show signs of senescence, or be out competed because of substantial lost vigor? can a wild cube colony remain productive for hundreds/thousands of years?

in our projects we have condensed this lifecycle down into a month or 2, but fungal colonies in the wild survive long periods of time. Have we artificially condensed a process that usually takes several years down into a few months?


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

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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."

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Edited by c10h12n2o (09/24/16 11:34 PM)


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Re: Cubensis Lifecycle Questions [Re: c10h12n2o]
    #23675546 - 09/24/16 11:32 PM (7 years, 4 months ago)

Shit man those r good questions. But I have little experience with wilds in the wilds . So I shall defer to those with b detter experience than mine.

:popcorn:


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InvisibleKenInVic
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Re: Cubensis Lifecycle Questions [Re: Pastywhyte]
    #23675571 - 09/24/16 11:47 PM (7 years, 4 months ago)

Quote:

Pastywhyte said:
No way nature can grow mushrooms better than me




Who's a good cultivator?  You are!  Yes you are, yes you are! :smirk:


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Re: Cubensis Lifecycle Questions [Re: KenInVic] * 1
    #23675728 - 09/25/16 01:55 AM (7 years, 4 months ago)

The world is almost covered in mycelium :smile: its how the trees get so big, it's why your veg rots in your fridge, champagne has its fizz and why we have antibiotics. Fungi outnumber plant species 10 to 1 and they are also some of the largest single living things on earth. I'm talking 2 miles big...

Imagine just how many spores are in the air?? We breathe that stuff all time, there are so many spores that all the scenarios you mentioned are happening at the same time.


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Re: Cubensis Lifecycle Questions [Re: berserkbadger] * 2
    #23675923 - 09/25/16 06:22 AM (7 years, 4 months ago)

Quote:

berserkbadger said:
The world is almost covered in mycelium :smile: its how the trees get so big, it's why your veg rots in your fridge, champagne has its fizz and why we have antibiotics. Fungi outnumber plant species 10 to 1 and they are also some of the largest single living things on earth. I'm talking 2 miles big...

Imagine just how many spores are in the air?? We breathe that stuff all time, there are so many spores that all the scenarios you mentioned are happening at the same time.





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Re: Cubensis Lifecycle Questions [Re: berserkbadger]
    #23675983 - 09/25/16 07:19 AM (7 years, 4 months ago)

Quote:

berserkbadger said:
The world is almost covered in mycelium :smile: its how the trees get so big, it's why your veg rots in your fridge, champagne has its fizz and why we have antibiotics. Fungi outnumber plant species 10 to 1 and they are also some of the largest single living things on earth. I'm talking 2 miles big...

Imagine just how many spores are in the air?? We breathe that stuff all time, there are so many spores that all the scenarios you mentioned are happening at the same time.






Im really diggin this thread :fonda:


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Invisibleamidogen
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Re: Cubensis Lifecycle Questions [Re: spore-ty]
    #23676075 - 09/25/16 08:33 AM (7 years, 4 months ago)

:threadmonitor:


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Re: Cubensis Lifecycle Questions [Re: c10h12n2o] * 1
    #23676137 - 09/25/16 09:11 AM (7 years, 4 months ago)

Quote:

c10h12n2o said:
in our projects we have condensed this lifecycle down into a month or 2, but fungal colonies in the wild survive long periods of time. Have we artificially condensed a process that usually takes several years down into a few months?




I believe this to be the case. In the very least, several months-a year or so becomes a couple of months.

This is partly conjecture, but…
If you spawn outdoor species (I'm mostly talking woodlovers), they'll take their sweet time getting ready to go, often up to a year. A spent sub might pop a couple loads off, but it's often much later than it would have in one of our tubs. And that's with our help. In the wild, it often seems like the fruit will often grow towards the edges of the mycelium in an effort to expand their network further (like faery rings). But, by adding younger mycelium to the network, you can get your outdoor patches to be fuller through and through.

I know cubes are different, as I don't see them propagating the same way. They AFAIK seem to propagate mostly by germinating in digestive tracts. So I know their cycle to be shorter, but they also don't seem to go for a larger network like say, the giant Amarilla colony in oregon. A cow poop is only so big, after all.

Because of this cycle of the cubensis, they will never see senescence in the wild, as it's MS every time. In fact, I'm not so sure senescence happens in the wild, merely because of the way it all works.

Where the fuck is Adden? He'd have better info on this for sure. I'll post a link in his journal :lol:

Some good stuff here to think about. Thanks for posting :smile:


Edited by dankington (09/25/16 09:20 AM)


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Invisiblec10h12n2o
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Re: Cubensis Lifecycle Questions [Re: dankington]
    #23680014 - 09/26/16 02:19 PM (7 years, 4 months ago)

Quote:

Pastywhyte said:
The rest is stuff we humans came up with to produce the mushrooms in greater abundance than nature could. No way nature can grow mushrooms better than me




it made us laugh, but pasty makes what is actually a REALLY relevant point to these questions

i think we often forget or take for granted (i know i do) how radically different what we are doing is from what actually occurs in nature, and how the species evolved to survive and reproduce in conditions and methods COMPLETELY different from the way we do things

for example, in nature, i cant think of anything that would be analogous to using grain spawn to colonize bulk substrates. what we are doing is taking a sterilized substrate (grain) and inoculating it with a clean culture, allowing it to colonize in a controlled environment with virtually no competitors (which never happens in nature), and then breaking up colonized grains (i dont think cubensis colonizes grains in nature, certainly not as primary spawn) and mixing it into pasteurized bulk substrate (which would never happen in nature), and taken from colonization conditions to fruiting conditions in about a month or 2.

i doubt there is ever a point in nature where little bits of fully colonized spawn are distributed throughout a substrate. in nature colonies probably originate from a single point of original inoculation (though this brings up many of my questions) 

in nature, i know our best cow field for cubes growing up had not had cows in it for over 10 years, and a new field a couple miles up the road that had lots of cows never had cubes, for years. but now, i hear both fields grow cubes.

by having controlled artificial conditions focused on optimizing yields and compressing a multi year cycle down into 2 months, we are able to grow much "better" mushrooms than nature. but it makes me wonder about some of the other ways this artificial lifecycle differs from nature, and how these differences might affect our projects

its funny that with all our research and interest in cubes, we dont even know their lifespan in the wild! (or at least i dont, hopefully someone will share)

some of my other questions about strains and MS colonies are fascinating as well. i wish i had a better idea of how these things work

Also, ive been wondering about ways to track genetic markers and hopefully distinguish individual strains genetically. when DNA is extracted from fungi, they often extract the total nucleic acids present, and it can be done to mycellium or to spores. wouldnt this necessarily be the DNA of countless individuals, due to the spores' genetic diversity, and the MS nature of natural colonies?

if this is the case, does one need an isolate to be able to study the dna of a particular strain? or would there be any way to genetically differentiate individual strains within an MS colony?

also, add this to my list in OP:

8. is there any indication that different strains within a colony might serve different functions/purposes within the colony? since some strains individually might not be able to fruit,  but could as part of a colony, are there other functions that might be distributed across different strains within the colony?


Quote:

berserkbadger said:
The world is almost covered in mycelium :smile: its how the trees get so big, it's why your veg rots in your fridge, champagne has its fizz and why we have antibiotics. Fungi outnumber plant species 10 to 1 and they are also some of the largest single living things on earth. I'm talking 2 miles big...

Imagine just how many spores are in the air?? We breathe that stuff all time, there are so many spores that all the scenarios you mentioned are happening at the same time.




Isnt it amazing? :laugh:  that largest one ("Humongous Fungus" is the one i referenced in the OP, and its 3.4 sq miles!!)

absolutely, fungi are a fascinating kingdom, closer to us than to plants. and there is so much to be learned from them. we already use them for the production of everything from citric acid to penicillin and beer, i suspect in the near future there will be many more amazing medical and industrial uses for fungi. we are only recently beginning to identify some of the various unique compounds produced by fungi, and only scratching the surface of what all can be done with those compounds. lots of interesting retroviral and cancer trials going on right now, all kinds of cool stuff

right, the scenarios i mentioned certainly happen all the time, thats why i am interested in learning more about them, especially as it relates and contrasts with our projects
Quote:

dankington said:
Quote:

c10h12n2o said:
in our projects we have condensed this lifecycle down into a month or 2, but fungal colonies in the wild survive long periods of time. Have we artificially condensed a process that usually takes several years down into a few months?




I believe this to be the case. In the very least, several months-a year or so becomes a couple of months.

This is partly conjecture, but…
If you spawn outdoor species (I'm mostly talking woodlovers), they'll take their sweet time getting ready to go, often up to a year. A spent sub might pop a couple loads off, but it's often much later than it would have in one of our tubs. And that's with our help. In the wild, it often seems like the fruit will often grow towards the edges of the mycelium in an effort to expand their network further (like faery rings). But, by adding younger mycelium to the network, you can get your outdoor patches to be fuller through and through.

I know cubes are different, as I don't see them propagating the same way. They AFAIK seem to propagate mostly by germinating in digestive tracts. So I know their cycle to be shorter, but they also don't seem to go for a larger network like say, the giant Amarilla colony in oregon. A cow poop is only so big, after all.

Because of this cycle of the cubensis, they will never see senescence in the wild, as it's MS every time. In fact, I'm not so sure senescence happens in the wild, merely because of the way it all works.

Where the fuck is Adden? He'd have better info on this for sure. I'll post a link in his journal :lol:

Some good stuff here to think about. Thanks for posting :smile:




excellent points you bring up, Im glad you joined the discussion :highfive1:

kinda makes you wonder what other effects such a radically unnatural lifecycle might have on the cultures/projects, other than producing tons of killer shrooms as quickly as possible

regarding fruits on the edge of a fairy ring, i think you are right. but wouldnt the new colonies be new MS colonies, and wouldnt they act as separate colonies? or would they meld?

which makes me want to add another question:

9. ultimately, in the beginning of an MS inoculation, monokaryons from spores meet up and become dikaryotic (forming a strain). but rather than having a million tiny colonies separated by the interface between them, we usually see a single, somewhat organized colony. So obviously at some point early in the process, the smaller MS colonies join together and build a network. but this doesnt really seem to happen if you put two established MS cultures together on a plate, since each colony remains distinct. is there some kind of phase, akin to adolescence or infancy, where the hyphae making up a colony are receptive to collaborating/consolidating/joining up with other colonies? 


i really appreciate the info and discussion guys, yall rock :rockon:


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

C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide


"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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Invisiblec10h12n2o
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Re: Cubensis Lifecycle Questions [Re: c10h12n2o]
    #23739780 - 10/15/16 11:28 AM (7 years, 3 months ago)

bump

been thinking a lot about some of this, and would love to clear up some of my questions, since it affects how i will move forward on some projects

warm regards


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

C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide


"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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Invisibleazur
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Re: Cubensis Lifecycle Questions [Re: c10h12n2o]
    #23739796 - 10/15/16 11:34 AM (7 years, 3 months ago)

Oh man. Lots of type.
I first hunted wild cubes 15 years ago. I would have sooner, but they didn't grow where I grew up. I have a pretty decent understanding of them. What do you want to know?


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Re: Cubensis Lifecycle Questions [Re: azur]
    #23739884 - 10/15/16 12:24 PM (7 years, 3 months ago)

Nothing to do with cubes, but I thought it is something cool to share. Psilocybe ovoids around here are spread by MS, but also flooding redistributing myc during the spring floods. Since they grow exclusively by streams and rivers, high water washes away colonized wood from one spot and it piles up in bends in another, rapidly colonizing the new spot. If we have a flood and I see where a bunch of debris collected, the next year often times I find massive amounts of mushrooms.


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Invisiblec10h12n2o
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Re: Cubensis Lifecycle Questions [Re: NDStepp84]
    #23739940 - 10/15/16 01:01 PM (7 years, 3 months ago)

thanks buddy, i would love to have your insight into any/all of this. here are just the raw questions, with the ones im most interested in in bold red:

1. what is the lifespan of a wild cubensis colony? is there a limit to the number of fruiting cycles a culture can sustain, beyond the restrictions placed on it by nutrition and environmental conditions? also, are there different phases in the lifecycle, beyond the basic reproductive cycle, where it has different potential characteristics and behavior? (possibly analogous to human infancy, adolescence, reproductive prime, old age, etc, and the way we can reproduce in some phases but not others, and are more susceptible to illness in some phases than others) 

2. in an "individual" such as "Humongous Fungus" or a wild cubensis colony, would the colony be likely to consist of numerous strains (like a MS grow)? or is it an individual strain, like a monoculture? i would suspect numerous strains to be present, but what i read about the "largest living organism" is often confusing

3. when a fruit is cloned, and multiple tissue samples are taken from different parts of the fruit, is the exact same variety of strains going to be present in each cultured sample? in my agar work, doing 6 plates of the same cloned fruit, i often see huge differences in the appearance and behavior of the myc. Is this the same collection of strains, expressing different characteristics? or are there a different cast of strains present in samples taken from various parts of the same fruit?

4. from year to year, if someone harvests cubes from the same place, it is likely from the same colony right? or overlapping, or subsequent colonies? if a wild colony is hundreds or thousands of years old, are there any effects of senescence, or any other relevant phenomenon we could readily observe as a result of such great age? does age affect wild cubensis colonies?

5. in MS grows, various colonies unite and consolidate somewhat, to network nutrients etc.. does this happen in the wild?

6. in an MS grow, are the same cast of strains present in all parts of the culture/substrate? or is the distribution less homogeneous?

7. in a clone culture, where the first plate shows several clear sectors, wouldnt a true clone culture either use this first plate (with the fruit tissue in the middle +contam risk) or continue to have the exact same variety of in subsequent plates? how could one do this, since we necessarily have to pick something to transfer, and cant pick all at the same time?

8. is there any indication that different strains (or groups of strains) within a colony might serve different functions/purposes within the colony? since some strains might not be able to fruit individually,  but could as part of a multi-strain colony, are there other functions, besides fruiting, that might be distributed across different strains within the colony?

9. ultimately, in the beginning of an MS inoculation, monokaryons from spores meet up and become dikaryotic (forming a strain). but rather than having a million tiny colonies separated by the interface between them, we usually see a single, somewhat organized colony (eventually). So obviously at some point early in the process, the smaller MS colonies join together and build a network. but this doesnt really seem to happen if you put two established MS cultures together on a plate, since each colony remains distinct. is there some kind of phase, akin to adolescence or infancy, where the hyphae making up a colony are receptive to collaborating/consolidating/joining up with other colonies? at what point do they become established enough not to combine with other colonies?


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

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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


Edited by c10h12n2o (10/15/16 01:05 PM)


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Re: Cubensis Lifecycle Questions [Re: c10h12n2o]
    #23754587 - 10/20/16 11:37 AM (7 years, 3 months ago)

Quote:

c10h12n2o said:
3. when a fruit is cloned, and multiple tissue samples are taken from different parts of the fruit, is the exact same variety of strains going to be present in each cultured sample? in my agar work, doing 6 plates of the same cloned fruit, i often see huge differences in the appearance and behavior of the myc. Is this the same collection of strains, expressing different characteristics? or are there a different cast of strains present in samples taken from various parts of the same fruit?

6. in an MS grow, are the same cast of strains present in all parts of the culture/substrate? or is the distribution less homogeneous?

7. in a clone culture, where the first plate shows several clear sectors, wouldnt a true clone culture either use this first plate (with the fruit tissue in the middle +contam risk) or continue to have the exact same variety of in subsequent plates? how could one do this, since we necessarily have to pick something to transfer, and cant pick all at the same time?





here are some pics to elaborate on these 3 questions a bit:



these plates are all clone cultures taken from a recent fruit i harvested that weighed nearly 300 grams. the tub i harvested it from was an AA+ clone, first flush weighed 3,498 grams. i know its bad form to clone a clone, but there were clearly lots of strains still present



so, like most times i have taken many clone samples from the same fruit, the cultures look and behave quite differently on agar, and often seem to be made up of a different cast of strains, or at least a different distribution of prominence within that cast of strains

it really makes me wonder, are any of these cultures really genetically identical to the fruit, if different strains are present in different cultures (and maybe different parts of the fruit; probably not, but it makes me wonder about the distribution of strains and functions of various strain groups within the colony)

also, since cleaning these up involves picking a sector and transferring it, wouldnt that necessarily limit the number of strains present, and thus not be representative of all the strains initially present in the clone culture, much less the fruit?

anyways, would love some feedback from you guys, it would be great if i could connect some of these dots, or at least broaden the context

warm regards


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Re: Cubensis Lifecycle Questions [Re: c10h12n2o]
    #23754597 - 10/20/16 11:40 AM (7 years, 3 months ago)

Gene regulation and expression can make limited genetics or even isolates have some variance in performance. That's my hunch considering brewing yeast can behave within a range even though brewers use isolates for the most part with all the yeast bought in a tube of some strain being identical twins.

Just like with edibles. If you buy shiitake 75 you get an isolate. An actual strain(not a variety like cubensis spores)

But shiitake 75 won't perform exactly the same from grower to grower because of the variables outside of genetics as well as what the environment does to genetic expression and gene regulation as well


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Re: Cubensis Lifecycle Questions [Re: bodhisatta]
    #23754861 - 10/20/16 01:20 PM (7 years, 3 months ago)

that makes a lot of sense, especially when we think in terms of groups of strains, and all the potential ways the genetic variation and conditions can affect the phenotypes expressed

but in a case like this "monster" clone, i took great care to make the plates exactly the same and keep them in the same conditions, the only difference being which part of the fruit i took the cloned tissue from, and how much tissue was used.

i know a lot of this could be the same principle we use in isolation: hoping the various strains and strain groups will differentiate themselves from the rest of the colony by growing in a particular direction on the agar while others grow in a different direction, allowing us to differentiate phenotypes

makes me wonder about a lot of things, particularly how stain groups act within the bigger picture of a colony, and the smaller picture within a fruit, and if their roles differ in any particular context

i dont have anything that i would bet on being an isolate; some of the edibles ive received in trades probably are, but i wouldnt assume so. id love to work with a known monoculture like Shitake 75, for these exact reasons (among others). my main drive towards obtaining isolates is so that i can use them to collect meaningful data points to use for some research projects (big data, yummy), and in developing some computer controlled systems and sensors


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Re: Cubensis Lifecycle Questions [Re: c10h12n2o] * 1
    #23754977 - 10/20/16 01:57 PM (7 years, 3 months ago)

From my limited work with known isolates like Hericium ABM or shiitake 3782 they do indeed have a great deal of stability but small variables can still affect performance. My last run of 3782 had 5 substrates perform identical but one did not. There was no obvious contamination that ever showed up and the sub mix was the same but it didn't popcorn properly and yield suffered.

I should also say that most of my clones have been extremely stable, my RW F2 has only showed variable results when I changed obvious variables. It was consistent in its response to those as well. Cased vs not was a big one as was cased grain vs bulk. Each time the performance for the controls was the same and the response to each specific varable was the same. To that end I'm confident cloning can result in stable results compared with isolates for the most part. I certainly wouldn't consider a clone to be uncontrolled unless it proved itself already to be very unstable. However those seem to be pretty rare, I only had an instance of that once.


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Re: Cubensis Lifecycle Questions [Re: Pastywhyte]
    #23755042 - 10/20/16 02:21 PM (7 years, 3 months ago)

excellent info pasty, thanks for sharing  :chugbeer:

again, that fits in quite well with understanding them in terms of strain groups

pasty, what is your take on #7? for a culture to be truly representative of all strains (and strain groups) found in a sample taken from a fruit, do we have to use the first plate (the one with the tissue sample) for it to be a true clone culture?

often, when i take a clone, there will be several sectors, including tomentose and highly rhizo. i typically transfer from the rhizo sectors, trying to get a more organized culture. but wouldnt this contain a more restricted cast of strains than the initial clone culture, by virtue of the transfers?

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


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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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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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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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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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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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Edited by c10h12n2o (03/30/17 11:38 AM)


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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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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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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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Re: Cubensis Lifecycle Questions [Re: Lennybernadino]
    #24415243 - 06/18/17 02:14 PM (6 years, 7 months ago)

:kaneclap:
great addition! I like the Peru variety I have... Much smoother of an onset, very giggly, and very clear headed the first 3 hours in. Also a very visual strain, IME.

I wonder if the mushies in your area have those characteristics too?


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Re: Cubensis Lifecycle Questions [Re: AtomHeart]
    #24513675 - 07/28/17 09:43 AM (6 years, 6 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.




But what about multiple strains working together to produce one mushroom? That demonstrates at least some cooperation between the strains. I always thought some work together better than others like people. The more that are present the more complicated the social situation becomes. Maybe they compete except when it come times to shed spores they are in it together? I can definitely see the evolutionary advantage to more than one strain combining to release a greater variety of genetic material in the spores. So maybe in terms of consuming nutrients and moisture it is a free for all, but the ones who are around when it is shroom-time drop the competition game and just focus on working together to get as many spores of as many genetic types as possible. Think about it one round of competition to weed out the weak but there is no advantage to withholding colonized mycelium energy when it's only desire is to fruit.
Maybe they form alliances and compete against other alliances?


Edited by Lennybernadino (07/28/17 09:45 AM)


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Re: Cubensis Lifecycle Questions [Re: Lennybernadino]
    #24513916 - 07/28/17 12:04 PM (6 years, 6 months ago)

https://www.shroomery.org/forums/showflat.php/Number/10839971
Different strains work together and make fruiting bodies they don't compete.  Even when non compatible they're not fighting each other off


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Re: Cubensis Lifecycle Questions [Re: bodhisatta]
    #24513975 - 07/28/17 12:54 PM (6 years, 6 months ago)

Fighting each other off and competing can be a different thing. The one may be better at speedy colonization that the other, and that could technically be viewed as competition, though friendly.


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