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OfflineBigJohnson
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Moved: Multi-Spore vs. Cloning
    #1452128 - 04/12/03 12:36 PM (18 years, 7 months ago)

I've been doing a lot of research lately on inoculating with mycelium water created by blending up a myc sample.

Now, we have all seen four distinct colonies radiating from the inoc points on a cake. These colonies are comprised of spores that have germinated in the substrate and formed a cooperative network comprised of numerous individuals.

My question is this: Can a pure (one set of genes) colony of myc be obtained from taking a sample of one of these colonies? Or, can one assume that this can only be achieved if the sample tissue is taken from a fruit stem?

If one took a small, rhizomorphic sample from one of these colonies, could he increase his chances of getting a colony with the same genes by doing or looking for something special?

From what I've read, yields can be almost doubled if your jars are colonized by a specific individual, as the individual gene set cooperates with "itself."

Your ideas and advice are appreciated


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OfflineBigJohnson
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Re: Moved: Multi-Spore vs. Cloning [Re: BigJohnson]
    #1464912 - 04/16/03 08:45 PM (18 years, 7 months ago)

Well, since no has commented, I'd thought I'd let you guys know what I've discovered so far.

There seems to be some debate as to whether or not colonies share genetic information between individuals, just as bacteria do through sharing phages through conjucation tubes. If so, then as the colony matures, the genes of each individual would gradually become more and more like those of the whole colony. Only the favorable genes would be shared most often as cell division would be in favor of better adapted individuals; more cell division = more cells = more genetic material shared. This makes sense, as one would not expect a colony of 1,000 individuals to cooperate well enough to produce anything but very tiny fruits.

So, if we assume that myc colonies are comprised of individual cells that share genes, to create a very cooperative colony, each individual cell should have the same genes as its colonial neighbors. However, as nature takes its course, the colony will become a genetically homogenous group of cells if given enough time.

Following this line of thought, if one cannot obtain a stem sample to clone, any substantial myc sample taken from a young colony may have several different individuals in the beginning but will eventually become genetically "one," for better or for worse.

What are your thoughts?


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Offlineamyloid
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Re: Moved: Multi-Spore vs. Cloning [Re: BigJohnson]
    #1465158 - 04/16/03 10:04 PM (18 years, 7 months ago)

well, there are some benefits in the long run for diverse myc. cultures, if a pure strain is used it is more susceptiple to disease/death by contam. because what will kill one part of your culture will kill it all, whereas with a diverse culture you have a better chance that one of the sets of genes will be more able to fend off or be immune to any said attack. in theory, a colony of myc. in a cake will NOT be genetically homogenous, where a fuzzy mess will sprout is where the spores settle/where the spawn lays, and unless it was a pure cultured cake to begin with it is highly unlikely, but not impossible, that the colony is a pure culture.

with spores, keep in mind that a single spore will produce a colony of cells called a monokaryon which will never produce fruits, it takes another compatible monokaryon to encounter, mate, and exchange cytoplasmic and genetic material to produce a dikaryon which IS able to fruit.

i suggest you get yourself an agar kit and play around, the best way for you to learn about this is to see it happening.

-doc


--------------------
"A human being is part of a whole, called by us the Universe, a part limited in time and space. He experiences himself, his thoughts and feelings, as something separated from the rest--a kind of optical delusion of his consciousness. This delusion is a kind of prison for us, restricting us to our personal desires and to affection for a few persons nearest us. Our task must be to free ourselves from this prison by widening our circles of compassion to embrace all living creatures and the whole of nature in its beauty."
-Al Einstein


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OfflineBigJohnson
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Re: Moved: Multi-Spore vs. Cloning [Re: amyloid]
    #1468225 - 04/17/03 07:52 PM (18 years, 7 months ago)

Quote:

well, there are some benefits in the long run for diverse myc. cultures, if a pure strain is used it is more susceptiple to disease/death by contam. because what will kill one part of your culture will kill it all, whereas with a diverse culture you have a better chance that one of the sets of genes will be more able to fend off or be immune to any said attack.




Hey, that's a damn good point. I hadn't thought of that. So, unless you're very sterile, you might just lose everything to that one particular contam that has your individual's number.

Quote:

in theory, a colony of myc. in a cake will NOT be genetically homogenous, where a fuzzy mess will sprout is where the spores settle/where the spawn lays, and unless it was a pure cultured cake to begin with it is highly unlikely, but not impossible, that the colony is a pure culture.




What do you think about the genetic exchange debate? Like, for example, inoc'ing with pans and cubes, same jar, to increase the overall potency?

Quote:

with spores, keep in mind that a single spore will produce a colony of cells called a monokaryon which will never produce fruits, it takes another compatible monokaryon to encounter, mate, and exchange cytoplasmic and genetic material to produce a dikaryon which IS able to fruit.




Again, do you think it would be possible to isolate a pan spore and a cube spore, introduce them to agar, and obtain a dikaryon from the colony? I doubt it, but I suppose it would depend on the number of chromosomes each have. Anyone know where this information can be found?

Quote:

i suggest you get yourself an agar kit and play around, the best way for you to learn about this is to see it happening.

- doc




Good idea!

Thanks for the response :smile:



--------------------
Should the US relegalize drugs?

http://www.lp.org/issues/relegalize.html


Edited by BigJohnson (04/17/03 10:50 PM)


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Anonymous

Re: Moved: Multi-Spore vs. Cloning [Re: BigJohnson]
    #1468245 - 04/17/03 08:00 PM (18 years, 7 months ago)

if i'm understanding mushroom genetics right, only one in 4 pure strains of cubensis is actually capable of fruiting. would this mean that obtaining a pure, fruiting strain of cubensis via cloning would result in substantially better flushes? would it also mean better pinsets?


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OfflineBigJohnson
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Re: Moved: Multi-Spore vs. Cloning [Re: ]
    #1468357 - 04/17/03 09:03 PM (18 years, 7 months ago)

Quote:

if i'm understanding mushroom genetics right, only one in 4 pure strains of cubensis is actually capable of fruiting. would this mean that obtaining a pure, fruiting strain of cubensis via cloning would result in substantially better flushes? would it also mean better pinsets?




That's the idea. A pure strain, one individual dikaryote, should be better able to cooperate with its own divided cells in the colony to produce some of the biggest fruits and largest flushes possible.

However, there seem to be a lot of factors that may or may not make the effort a success.


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Offlinemossie
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Re: Moved: Multi-Spore vs. Cloning [Re: BigJohnson]
    #1468592 - 04/17/03 10:47 PM (18 years, 7 months ago)

if you isolate a strand of rhizomorphic growth from one of your cloned isolates it will be better chances of bigger, even pinsets IMO.


--------------------
Better is a dinner of herbs where love is, than a stalled ox and hatred therewith. -proverbs 15:17


Edited by mossie (04/17/03 10:48 PM)


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OfflineBigJohnson
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Re: Moved: Multi-Spore vs. Cloning [Re: mossie]
    #1468603 - 04/17/03 10:50 PM (18 years, 7 months ago)

Quote:

if you isolate a strand of rhizomorphic growth from one of your cloned isolates it will be bigger, even pinsets IMO. 




Thanks for the tip :smile:


--------------------
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Anonymous

Re: Moved: Multi-Spore vs. Cloning [Re: mossie]
    #1468860 - 04/18/03 12:33 AM (18 years, 7 months ago)

but may not fruit. only 1 in 4 pure strains of cubensis is capable of fruiting. rhizomorphs are a good sign, but not a clear indication of fruiting potential. to obtain a pure fruiting strain from spores, one would have to isolate many strains on agar, then try fruiting each one to see if it could fruit. an easier and faster way to get a pure fruiting culture is to clone a fruitbody in my opinion.


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OfflineBigJohnson
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Re: Moved: Multi-Spore vs. Cloning [Re: ]
    #1468868 - 04/18/03 12:36 AM (18 years, 7 months ago)

I don't have one to clone from :frown:

I guess I'll have to wait! :smile:


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Offlinemossie
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Re: Moved: Multi-Spore vs. Cloning [Re: BigJohnson]
    #1470428 - 04/18/03 02:52 PM (18 years, 7 months ago)

if you isolate a rhizo from your cloned fruitbody is what i'm sayin, then it will be the same genetics only stronger.


--------------------
Better is a dinner of herbs where love is, than a stalled ox and hatred therewith. -proverbs 15:17


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OfflineBigJohnson
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Re: Moved: Multi-Spore vs. Cloning [Re: mossie]
    #1470597 - 04/18/03 03:57 PM (18 years, 7 months ago)

Quote:

if you isolate a rhizo from your cloned fruitbody is what i'm sayin, then it will be the same genetics only stronger. 




Gotcha :smile:


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Anonymous

Re: Moved: Multi-Spore vs. Cloning [Re: mossie]
    #1470678 - 04/18/03 04:39 PM (18 years, 7 months ago)

same genetics only stronger

how will two things which are identical not be identical?


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Offlinemossie
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Re: Moved: Multi-Spore vs. Cloning [Re: ]
    #1470751 - 04/18/03 05:01 PM (18 years, 7 months ago)

well its not scientifically proven to have variations in a pure strain's mycelium, but, IMO, you are promoting only the strongest rhizo growth. i see what you're sayin though.


--------------------
Better is a dinner of herbs where love is, than a stalled ox and hatred therewith. -proverbs 15:17


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Invisiblepoke smot!
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Re: Moved: Multi-Spore vs. Cloning *DELETED* [Re: mossie]
    #1471526 - 04/18/03 10:16 PM (18 years, 7 months ago)

Post deleted by poke smot!

Reason for deletion: x



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OfflineAnnoA
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Re: Moved: Multi-Spore vs. Cloning [Re: BigJohnson]
    #1472300 - 04/19/03 02:40 AM (18 years, 7 months ago)



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OfflineUnknown
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Re: Moved: Multi-Spore vs. Cloning [Re: Anno]
    #1472474 - 04/19/03 04:17 AM (18 years, 7 months ago)

ahahaha,once again,READ THE FAQ,I didn't even expect this one,I was reading the FAQ the other day and it has improved a lot since the last time I looked at it


--------------------
The above is just like,my opinion man


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OfflineAnnoA
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Re: Moved: Multi-Spore vs. Cloning [Re: Anonymous]
    #1472561 - 04/19/03 05:19 AM (18 years, 7 months ago)

As allways, work in progress.


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OfflineBigJohnson
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Re: Moved: Multi-Spore vs. Cloning [Re: Anno]
    #1474165 - 04/19/03 09:37 PM (18 years, 7 months ago)

Quote:

Substrains interact, and rewire each other




Here's another mention of genetic exchange among fungal colonies.

Can anyone point me to a good book on fungids? I'm looking for something advanced, as I've had all my bio and chem classes.


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Anonymous

Re: Moved: Multi-Spore vs. Cloning [Re: BigJohnson]
    #1474347 - 04/19/03 10:59 PM (18 years, 7 months ago)

The Mushroom Cultivator
by Paul Stamets

is a must.


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Offlineamyloid
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Re: Moved: Multi-Spore vs. Cloning [Re: ]
    #1476167 - 04/20/03 12:10 PM (18 years, 7 months ago)

yes, and yes, which is why many cultivators use 2 pure monokaryon strains, with good rhizomorphic traits, to innoculate spawn.


--------------------
"A human being is part of a whole, called by us the Universe, a part limited in time and space. He experiences himself, his thoughts and feelings, as something separated from the rest--a kind of optical delusion of his consciousness. This delusion is a kind of prison for us, restricting us to our personal desires and to affection for a few persons nearest us. Our task must be to free ourselves from this prison by widening our circles of compassion to embrace all living creatures and the whole of nature in its beauty."
-Al Einstein


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Offlineamyloid
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Re: Moved: Multi-Spore vs. Cloning [Re: BigJohnson]
    #1476170 - 04/20/03 12:11 PM (18 years, 7 months ago)

>>Again, do you think it would be possible to isolate a pan spore and a cube spore, introduce them to agar, and obtain a dikaryon from the colony? I doubt it, but I suppose it would depend on the number of chromosomes each have. Anyone know where this information can be found?

no, i fairly positive two differnt species wont "mate".


--------------------
"A human being is part of a whole, called by us the Universe, a part limited in time and space. He experiences himself, his thoughts and feelings, as something separated from the rest--a kind of optical delusion of his consciousness. This delusion is a kind of prison for us, restricting us to our personal desires and to affection for a few persons nearest us. Our task must be to free ourselves from this prison by widening our circles of compassion to embrace all living creatures and the whole of nature in its beauty."
-Al Einstein


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Offlineamyloid
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Re: Moved: Multi-Spore vs. Cloning [Re: ]
    #1476187 - 04/20/03 12:18 PM (18 years, 7 months ago)

>>but may not fruit. only 1 in 4 pure strains of cubensis is capable of fruiting.

what is a pure strain? not a pure culture, considering no pure culture(a single monokaryon colony) would fruit.

>>an easier and faster way to get a pure fruiting culture is to clone a fruitbody in my opinion.

what tek are you using that fruits faster then spore+agar - pinning stage???


--------------------
"A human being is part of a whole, called by us the Universe, a part limited in time and space. He experiences himself, his thoughts and feelings, as something separated from the rest--a kind of optical delusion of his consciousness. This delusion is a kind of prison for us, restricting us to our personal desires and to affection for a few persons nearest us. Our task must be to free ourselves from this prison by widening our circles of compassion to embrace all living creatures and the whole of nature in its beauty."
-Al Einstein


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Offlineamyloid
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Re: Moved: Multi-Spore vs. Cloning [Re: ]
    #1476195 - 04/20/03 12:23 PM (18 years, 7 months ago)

just as cells in your body can change genetically and become cancerous, growing myc can have variation in growth trends, making seperate cultures of rhizo myc. growth will definitly encourage more rhizo growth hence more pins.


--------------------
"A human being is part of a whole, called by us the Universe, a part limited in time and space. He experiences himself, his thoughts and feelings, as something separated from the rest--a kind of optical delusion of his consciousness. This delusion is a kind of prison for us, restricting us to our personal desires and to affection for a few persons nearest us. Our task must be to free ourselves from this prison by widening our circles of compassion to embrace all living creatures and the whole of nature in its beauty."
-Al Einstein


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Offlineamyloid
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Re: Moved: Multi-Spore vs. Cloning [Re: poke smot!]
    #1476209 - 04/20/03 12:28 PM (18 years, 7 months ago)

maybe due to degreneration of the culture over time, also how many times have u seen that happen? i havent noticed a differnce in myc. growth with differnt spawn used to innoculate agar. also keep in mind that the starchy insides of rye grain encourages thick fluffy/cottonty (tomentose) type of myc.


--------------------
"A human being is part of a whole, called by us the Universe, a part limited in time and space. He experiences himself, his thoughts and feelings, as something separated from the rest--a kind of optical delusion of his consciousness. This delusion is a kind of prison for us, restricting us to our personal desires and to affection for a few persons nearest us. Our task must be to free ourselves from this prison by widening our circles of compassion to embrace all living creatures and the whole of nature in its beauty."
-Al Einstein


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Invisibletripndicular
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Re: Moved: Multi-Spore vs. Cloning [Re: BigJohnson]
    #1476314 - 04/20/03 01:08 PM (18 years, 7 months ago)

THE MUSHROOM CULTIVATOR will guide you and correct alot of misinformation you are hearing .


--------------------
Any information I give is not intended to aide you in the production of potentialy illegal substances !None of my exp comes from growing illegal varities , so take it as you will .
So with that said here is our mission statement .

Then the priest fell into a trance or swoon,& said unto the Queen of heaven ; Write unto us the ordeals; write unto us the rituals; write unto us the law !


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OfflineBigJohnson
Whoa! You guysneed to lightenup!

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Re: Moved: Multi-Spore vs. Cloning [Re: tripndicular]
    #1476551 - 04/20/03 03:41 PM (18 years, 7 months ago)

Okay, I have Stamet's book on the way :smile:

If anyone knows of a book that's a little more "cutting edge," please post it up. It doesn't even need to be about cultivating per say. I just want to learn more about fungidae; if dikaryotes exchange genes in a colony, the number of chromosomes typical for a haploid, etc.

If I had access to a microscope, I'd really enjoy germinating a pan spore and a cube spore on the same plate to see if the two form a dikaryote. I believe Anno said he had heard someone describing a multispore with the two producing very potent fruits, though that could be for reasons other than the two forming a dikaryote.


--------------------
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Anonymous

Re: Moved: Multi-Spore vs. Cloning [Re: BigJohnson]
    #1479721 - 04/21/03 07:22 PM (18 years, 7 months ago)

A pure culture is one free of contamination, both visible and latent. Pure culture is also used to describe a culture made up of a single Dikaryon.

A fruit as a clone source is going to result in a culture exactly like the fruit it was taken from, less any random mutations LEADING to sectoring.

A colonized grain from within a multispore jar may have more then one Dikaryon present. When this is placed on agar, it can result in more then one Dikaryon visibly sectoring away from each other. The result can be more then one Dikaryon being isolated from the single grain. If the multiple Dikaryons are isolated away from eachother and replated. You can wind up with Multiple pure Cultures coming from a single transfer.

To avoid any False Dikaryons(non fruiting, or Sterile Dikaryons), it would be more advisable to grow out the multispore to fruits, and clone the best fruit with the ability to sporulate, if you are after spores, or non sporulating if spores are not a priority.

Anastamosis describes the interaction between two distinct Dikaryons when they come in contact. They can and do exchange (intact) haploid nuceli with eachother. (The same Strain will do so at a higher rate, then the interaction between different strains).

Without overcoming many barriers(structural, chemical, genetic, etc...) breeding a Pan with a Cubensis will be impossible. Simply placing a spore of each on a plate will not result in mating.

There are many books that might interest you, but the more specific your interest the better the books. General books, that incompass all aspects of Mycology tend to be extremely limited in the depth they go into each section. Understanding which area of mycology you are interested in will help narrow your search for a good book. Taxonomy, morphology, physiology, genetics, or cultivation.




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OfflineBigJohnson
Whoa! You guysneed to lightenup!

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Re: Moved: Multi-Spore vs. Cloning [Re: ]
    #1479862 - 04/21/03 08:10 PM (18 years, 7 months ago)

Thanks teo! That's what I've been looking for! Shrooms for you :smile:


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