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Strain Creation


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#21 Guest_xxxsevxxx_*

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Posted 29 December 2005 - 08:30 PM

"No Im talking about taking a aussie pan print and mixing it with a goliath or a large florida print(have the original from years back put up) and createing a a large and abundant fruiter."

huh ? you have a lgflcy from years back ? my girl found 2 mushrooms in brooksvile fl (1 big and 1 small cyan) and i worked for a year from there to get them tame enough for mass consumption. i might be cumfuzzed here .

also mixing mono strains and seeing if you ended with a dikaryon myc. is the easy way of crossbreeding . a scope is a most have . the novelness wares off after the first success trust me . its a lot of work and its costly to eat threw as many perti dishes as you need for it .and the resulst are often not what you where looking for . with the phenotype leaning towards one side or the other . (in other words the changes or so subtle that a microscope is the only way to see if you actully did crosbreed anything.
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#22 mushaman

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Posted 30 December 2005 - 07:49 AM

Well this has been a pretty good discussion so far and have came up with an idea of my own. Say I took B+ spores and Golden Teacher spores and inculated one of each on different sides of the cake, then the myc grew together. Would this do anything other than just grow B+ on one side and Golden Teacher on the other? Just a thought.
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#23 the_chosen_one

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Posted 30 December 2005 - 09:57 AM

Well this has been a pretty good discussion so far and have came up with an idea of my own. Say I took B+ spores and Golden Teacher spores and inculated one of each on different sides of the cake, then the myc grew together. Would this do anything other than just grow B+ on one side and Golden Teacher on the other? Just a thought.
mush



Then you would have di meeting di almost guaranteed. It's highly doubtful any clamping would occur between the two. Ideally you would want one spore from each on each side of the cake. Or at least one spore from one and only a very few from the other to prevent the drowning effect. The dna exchange would be closer to 50/50 and continued on throughout the culture as it grows. Now that's all provided a 50% degree of transfer is allowed in the first place which is doubtful. This is what I mean by compatable strains and it's a big IF.

Keep thinking and asking mush! This is a really cool topic which I'm really not that knowledgeable about when you really start digging into it. For instance, I have no idea what transfer rate is allowed. Is it set for each strain? So many questions and so much to learn!

#24 Guest_dial8_*

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Posted 30 December 2005 - 10:02 AM

^^^^^^
Exactly. On both sides of the cake you would have germination of mono spores. These would mate with surrounding mono mycelium to create di mycelium which would grow through the cake only to run into the other side's di mycelium.

#25 mushaman

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Posted 30 December 2005 - 01:22 PM

Can mushrooms be grafted (spelling???) like a plant can be?
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#26 the_chosen_one

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Posted 30 December 2005 - 02:46 PM

Can mushrooms be grafted (spelling???) like a plant can be?
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Not that I'm aware of. The sample taken from a fruiting body would generate dikaryotic myc just as it would when cloned. Once again di meets di and it's too late. It all starts with mono meets mono which only occurs when the spores germinate. The closer the spores are together the quicker di is formed. Then the door is closed.

Diluting spreads the spores apart and improves the chance of mono A meeting mono B forming the desired di myc before A meets another A and B meets another B forming the undesired di myc. Sevens right. Usually what you end up with is mostly all A or mostly all B. Even if your mono mix is a perfect 50/50. Some genes are just dominant over others.

#27 Guest_Peter Cottontail_*

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Posted 30 December 2005 - 05:25 PM

Given the choice, I've found that spores almost always mate with like strain spores. Putting spores from two different strains near each other or mixed together rarely results in a cross.

However, dikaryotic/dikaryotic pairings are very possible and even common. Place two single sector dikaryotic isolates on the same petri dish, and allow them to grow towards each other until a line of isolation has formed between them. If a third sector opens up along the line of isolation, a new strain has been created. It can be tested by cutting it away, then placing both parent strains and the new strain on agar together. This time, if a three way line of isolation develops, you know you have a new strain distinct from both parents that was created by dikaryotic/dikaryotic pairing.
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#28 Bluejay

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Posted 30 December 2005 - 11:24 PM

seven yes I recieved a florida print from teonan on shroomery a few years back and fruited some very large pans for the time..Ralph wanted to buy them and produce them for him..but with young kids at the time i didnt want to focus so much energy into something so hush hush...

My gallery on the shroomery under the same handle still has the pix..

I quit mycology and went into the whole legal pedro/torch area..after learning how to extract crystals and getting way to large of a cactus garden it put a strain on my marriage. I kept a few rare cacti and got rid of the rest(mmm crystals)

So Im back into mycology for that one day a week release..I have always loved the pans hated the cubes..wont or didnt even keep any cube prints.. Only will mess with pans,mexi's and azure beds..

I really like this forum due to more exotics here...shroomery is mainly cubensis and thats all....Hell anno doesnt even post grows anymore of exotics..just a small handfull of others do there so I enjoy it over here..

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#29 mushaman

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Posted 31 December 2005 - 05:51 AM

Well Rodger i'm not familiar with the term dikaryotic but is this just myc from a certain strain? From what I got out of your post is that the myc from one strain meets with the myc from another strain and has a distinctive line where they met, but if a new sector (as you put it) forms on this line then it could be the two myc joined together thus creating a new strain?
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#30 Guest_lost_onabbey_rd_*

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Posted 31 December 2005 - 03:50 PM

when a single spore germinates what you get is monokaryotic growth.. mono meaning 1 set of DNA
when that monokaryotic growth meets other monokaryotic growth they form what is called a clap connection, sort of like a bridge.. and across this bridge they share their genetic information.. and thus become a Diakaryotic growth

monokaryots can grow in a vegatative state but they can't fruit
so to create a strain one would have to grow out 2 monokaryotic growths from 2 different strains and let them meet eachother and exchange dna
this is complicated because isolating a single spore is pretty hard
even if you can get them to grow.. and cross.. there is no way of knowing that the single spores DNA wasn't crap and you end up with a new strain that fruits poorly or not at all.. making all that work for nothing
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#31 Guest_Peter Cottontail_*

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Posted 31 December 2005 - 06:18 PM

Monokaryotic mycelium is mononucleate, meaning that there is one nucleus per cell. Dikaryotic mycelium is binucleate, meaning each cell has two nuclei. This can be easily observed with a regular light microscope. Mycelial cells are cylindrical and layed end to end to form a strand of mycelium. Think of cutting a pencil into ten pieces along its length and then sticking the pieces back in place. Clamp connections form around each of these cell walls where two 'pieces of the pencil' are attached together. This allows the genetic material to travel down the mycelium, causing the mononucleate mycelial cells to become binucleate, even though they may be far from the point of original contact where the mating first took place.
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#32 Cthulhu's Tentacle

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Posted 31 December 2005 - 06:40 PM

After reading PF's experiment that produced the albinos, I've been exposing some Amazonian cakes to a blacklight tube to see if/how they produce mutants

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#33 mushaman

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Posted 01 January 2006 - 05:49 PM

Well what do you guys think about trying to mate Golden Teacher with B+ ?? I think their pretty similiar.
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#34 Guest_lost_onabbey_rd_*

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Posted 01 January 2006 - 06:36 PM

honestly mush there really isn't any point in crossing strains..
unless you have a very specific goal in mind, like RR did with the redboys
if you want a good yeilder isolating a good substrain on agar is the way to get it..
almost any strain out there will produce just as well as any other
the best way to increas yeild is by cloning or isolating a good fruiter
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#35 mushaman

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Posted 01 January 2006 - 07:08 PM

yeah i'm just doing it for an experiment ... see what comes of it. I don't care for the yeild anyways afoaf supplies me with more than enough when i'm in need ... a trip or two a month is enough.
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#36 the_chosen_one

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Posted 03 January 2006 - 07:02 PM

Clamp connections form around each of these cell walls where two 'pieces of the pencil' are attached together. This allows the genetic material to travel down the mycelium, causing the mononucleate mycelial cells to become binucleate, even though they may be far from the point of original contact where the mating first took place.
RR


So it would be a 50/50 mix but the features of the fruiting body would be dictated by the chromozones?

What would be better to accomplish the goal of "strain creation", a monokaryotic or dikaryotic pairing? Or is there really a difference.

The dikaryotic is still receptive to new dna....how much can it take on after recieving at a monokaryotic level?

Sorry, so many questions. Interesting stuff.

#37 BuckarooBanzai

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Posted 03 January 2006 - 08:13 PM

Man, I love this place...some of these threads are so stimulating...

How many clamp/DNA exchanges usually take place, from spore to fruit? I had the impression it was just clamps a plenty throughout the mycellial body. Perhaps they are actually rare(?).

Does just one clamp qualify the whole connected mass as dikaryotic?

Could anybody suggest a text book or primer they have particularly benefited from on this amazing "fifth kingdom?"

I feel like a flat-Earther trying to imagine DNA exchange amongst multiple cells along a shared protoplasmic conduit. I currently have the look on my face that most people give me when I talk about networks and servers.

I love to have that look...

#38 Guest_Peter Cottontail_*

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Posted 03 January 2006 - 08:51 PM

I would imagine there would be more possibilities with dikaryotic/dikaryotic pairings than mono/mono due to the fact that each cell in dikaryotic mycelium has two nuclei, thus twice the genetic material available.

Dikaryotic/dikaryotic pairings take place all the time in our multispore projects. What starts out sometimes as hundreds of substrains, usually ends up as three or four(or sometimes even one) substrain by the time it gets to the fruiting stage. Sometimes we refer to this as 'the most dominant strain taking over', but that is really inaccurate. The other substrains are assimilated into the network by dikaryotic/dikaryotic pairings as well as mono/mono and mono/dikaryotic pairings.

The advantage snake venom gives when creating strains is it breaks down the cell walls of the unlike mycelium, allowing the nuclei of the different strains to come into contact. Exactly what happens after that is still a mystery to me, but the pairings do indeed take place and the results, such as John Holliday's http://www.nwbotanic...idcordyceps.htm cordyceps experiments and my redboy work speak for themselves, and are easily verifiable by placing the new strain on a petri dish with the two parents, and observing the three way line of isolation that develops. Without a pairing, the new mycelium would blend seamlessly into one of the parents.

Clamp connections form all along the mycelium between cells. They look sort of like a blob of solder holding two wires placed end to end. At 1000X you can see the fluids and 'cellular matter' passing from one cell, through the clamp connection and into the adjoining cell, then right on down the line to the next cell, on and on. While mycelium looks 'still' to the naked eye, under the microscope with enough magnification to see inside the individual cells, there is a kaleidoscope of kenetic activity. I'm trying to make a video of this, but having a hard time seeing the movement on film. I saw a presentation by Tom Volk http://botit.botany....edu/toms_fungi/ a few months ago where he had inserted some sort of phosfluorescent chemical into the mycelium to light up the particles as they traveled and he had an awsome film of it.
RR

#39 mushaman

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Posted 03 January 2006 - 10:43 PM

amazing just amazing !!! Rodger I would love to see that video when you get it to work for you!
mush

#40 BuckarooBanzai

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Posted 04 January 2006 - 12:50 PM

I've been wondering about that "dominant strain takes over" thing since I first heard it. How does one mycelium take over another? How do you "occupy" another organism's cell? Some kind of rogue RNA?

This makes perfect sense, though. The occupation isn't dissimilar to the Borg of Star Trek TNG fame. The surrounding cells are absorbed into the collective, with their diversity intact. The stronger expressions of the "dominant" genetic code are simply carried more successfully in subsequent growth.

I suppose at the cellular level the same process of dominant/recessive genetic sequencing and expression is taking place. The tendancy of a cake innoculated by multispore to become one big mycelium is basically just particularly dominant traits expressing again and again throughout the reproductive process.

That also explains how something as comparatively wimpy as a black light could cause lasting genetic damage. If one chromosome is whacked even a little, it has the possiblity of affecting a huge number of cells very quickly.

Am I correct in hearing, though, that mature cells can have an ongoing process of genetic exchange?

This is an amazingly interesting thread. I may have to try and audit a course on the fungi biology. I am really intrigued by what little I know about how these organisms function.




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