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InvisibleATWAR
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Registered: 01/26/03
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Re: creating new strains? [Re: fastfred]
    #2931818 - 07/27/04 09:08 AM (19 years, 6 months ago)

Quote:

fastfred said:
What a silly thread.  Most of you guys don't even have the vocabulary to discuss this stuff, yet everyone has plenty of theories.

I see a lot of incorrect information.  PF is to blame for much of it, his knowledge of genetics was quite weak.




No information I have ever originated from PF. I am sorry my vocabulary isn't up to par with your level. I would very much like you to point out the incorrect information...

Quote:

In reply to the original post... Yes it is possible to create a new strain, it is not even that difficult. It's a shame that the vendors don't do a better job. You can have a new strain in as little as two generations.





I would very much like you to describe the method one would use for doing such a thing in only 2 generations without crossing different strains (races). It should also be noted that vendors do not create new strains, only spread the spores from those found naturally.

Quote:

I noticed that someone thought that hybridization would result in a new strain. That is not true. A F1 hybrid never breeds true. Hybridization is a way to introduce genetic variability, from there you've just begun.





This is a common conception used in breeding marijuana, and does not cross over into mushrooms. This is one way how new strains are developed in the commercial industry, it could easily be transferred over to Ps. Cubensis.

Quote:

If you're looking for some quick new strains, just throw your spores into a UV strata linker and give them 600-800 energy units. That should give you about 1-2% germination rates and plenty of mutants.




Oh, this is how you suggest we make new strains, by mutating them?

:rolleyes:

This has been discussed many times, and there is always somebody who just doesn't quite grasp the concept that mushroom genetics are not like crossing plants and breeding dogs. It is much more complex than that...


--------------------
To give is to live...



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Invisiblemycopsycho
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Re: creating new strains? [Re: ATWAR]
    #2931843 - 07/27/04 09:12 AM (19 years, 6 months ago)

of course it's complex atwar! gods aren't simple things ya know :mushroom2:


--------------------
I Am The Sickness.

Diploid: I think adults have a right to make stupid decisions and it's nobody else's fucking business.


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InvisibleMykey
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Registered: 04/07/04
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Re: creating new strains? [Re: fastfred]
    #2932816 - 07/27/04 01:54 PM (19 years, 6 months ago)

  WOW! I must say that I am taken aback by your intellectual omnipotance ,and am absolutely awestruck by your astute use of an obviously superior vocabulary. No, I really mean it, your post was so incredibly articulate,and full of deep and meaningfull insight into the subject that I just sat dumbfounded as my miniscule brain began to smoke from the strain it was under as I tried,in vain, to fathom what you were trying to say! Oh well,I guess I had better just give up and try to find my helmet before I accidently bump my head and do any more damage to my already inapt noggin!























NOT!!!
I actually understand fully what you are trying to say! You are so terribly distraught about your own inadequacies that you try and feel better by compensating with unwarranted attacks on others. It is simple psychology really,so you could get that fixed with some expensive therapy that might someday eleviate your inferiority complex!

  I could go on to pick apart your pathetic attempts at enlightening all of us with your infinite wisdom on the subject of mushroom genetics,but I must say that ATWAR did an outstanding job already in the eloquent manner that she is so very good at.

Didn't your mother ever tell you that if you dont have anything nice to say,or something intelligent to add,then you should just KEEP YOUR MOUTH SHUT! Well I'm sure she did,and IMO you should hede her advice and quit going out in public to make an ass of yourself!

Your Friend,
MYKEY :lol:


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OfflineOpenminded
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Re: creating new strains? [Re: Mykey]
    #2933021 - 07/27/04 02:43 PM (19 years, 6 months ago)

:lol:


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Anonymous

Post deleted by Papaver [Re: Cyber]
    #2933316 - 07/27/04 03:53 PM (19 years, 6 months ago)



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Invisiblemycopsycho
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Re: creating new strains? [Re: Anonymous]
    #2933323 - 07/27/04 03:55 PM (19 years, 6 months ago)

awwwwww FUCK! oh well....


--------------------
I Am The Sickness.

Diploid: I think adults have a right to make stupid decisions and it's nobody else's fucking business.


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Invisiblefastfred
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Re: creating new strains? [Re: mycopsycho]
    #2935210 - 07/28/04 12:16 AM (19 years, 6 months ago)

Sorry I don't know how to use the boards quoting system so bear with me.

> how many generations would it take for a lay person with limited technology to produce a new strain? how did pf create a new strain?"

As I mentioned above, as little as two in the case of a spontaneous mutation. You will need to do some agar work and be capable of obtaining single basidospore isolates (SBIs) from your spores. In the case of PF both of his mystery strains were the result of spontaneous mutation. I know that he supposedly was using a black light when it happened.

> No information I have ever originated from PF. I am sorry my vocabulary isn't up to par with your level. I would very much like you to point out the incorrect information...

Sorry, I wasn't referring to you. You seem to know what you are talking about, however I think you are overly pessimistic. PC is certainly not as hard to breed as A. bisporus.

> I would very much like you to describe the method one would use for doing such a thing in only 2 generations without crossing different strains (races).

1. Spontaneous mutation occurs.
phenotype: red caps (just an example)
genotype: Rr (dominant trait)

2. Mate with brother
phenotype: mixed
genotype: 50% Rr, 50% rr

3. Line breed, or breed two Rr brothers.
phenotype: Mixed
genotype: 25% RR, 50% Rr, 25% rr

4. Select an RR and you have a new strain.
phenotype: all red caps
genotype: 100% RR

This new strain will breed true, because you will have just bred out the old genotype.

> Oh, this is how you suggest we make new strains, by mutating them?

Well... It's certainly the quickest way. It beats waiting around for it to happen naturally. A better way is to create inter-strain hybrids to introduce genetic diversity and then follow through with selective breeding.

>This has been discussed many times, and there is always somebody who just doesn't quite grasp the concept that mushroom genetics are not like crossing plants and breeding dogs. It is much more complex than that...

Perhaps you should elaborate... Non-self-fertile species can be bred in exactly the same way that any other non-asexual species can be. It may be slightly more difficult since you have to isolate monokaryotic tissue, but it is more than made up for by the quick turnaround time between generations.

> I really mean it, your post was so incredibly articulate,and full of deep and meaningfull insight into the subject that I just sat dumbfounded[...]

Many books have been written on the subject. I obviously can't teach a whole hell of a lot in one post.

> You are so terribly distraught about your own inadequacies that you try and feel better by compensating with unwarranted attacks on others.

Ok, whatever. I saw people suggesting everything from mated siblings to F1 hybrids to be a new strains... What did you expect me to do? Praise you guys?

Oh, BTW thanks for adding absolutely nothing to the discussion dipstick.

> basically,yes,it can be done,but you'll need topnotch equipment and plenty of time

You don't need much in the way of equipment, but plenty of time never hurts.


Edited by fastfred (07/28/04 01:55 AM)


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Offlineragadinks
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Re: creating new strains? [Re: fastfred]
    #2935324 - 07/28/04 01:26 AM (19 years, 6 months ago)

Great flame war on such an interesting topic :wink:

But it would be really interesting to breed new stains that are adapted better to certain conditions like substrate, temperature, yield etc.

What would you suggest?
To mutate spores found in the wild and try to grow them in the new environment and then select the best?
How could you tell that it is a new strain and not another phenotype of the old strain?


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Invisiblefastfred
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Re: creating new strains? [Re: ragadinks]
    #2935433 - 07/28/04 02:48 AM (19 years, 6 months ago)

> But it would be really interesting to breed new stains that are adapted better to certain conditions like substrate, temperature, yield etc.

That's basically what's been happening ever since they have been domesticated. Their new habitat is BRF and rye. I think they've adapted quite well. Any genes that reduce their ability to grow on that substrate will breed themselves out fairly quickly.

Personally, I'd like to see low humidity and contam. resistant strains developed.

It's very hard to introduce traits that do not already exist in the genepool. The trick is finding them. For example if I were looking to breed in contamination resistance, I would not look to PF, B+, or any of those standard strains. Instead I would look for the trait in another strain. Hawk claims to have one...

"The Indian cubensis also grows around most green mold like its nothing."
http://www.thehawkseye.com/india/india.html

Of course, there seem to be other problems with this strain. What I would do is grow some of the India strain and subject it to different contaminants, and select for the resistant ones. Then take your desired strain (strain A) and breed it with the India cube. Take the offspring of this mating and select for contam resistance, then breed it back to strain A repeatedly, always selecting for contam resistant individuals. After a few generations you will have a contamination resistant version of strain A.

The problem gets much harder if the trait is recessive... But it can be done, each step just requires an additional generation or two.

For example you have:
1. India cubensis
phenotype: contam resistance
genotype: cc

2. strain A-India cross
phenotype: no contam resistance
genotype: 100% Cc

3. A-India by strain A cross
phenotype: no contam resistance
genotype: 50% CC, 50% Cc

4. Now breed it with itself.
When two of the Cc from step 3 mate 25% of the offspring will be cc and actually display contam resistance.

Note that at several points the recessive trait is hidden, and you will have no way to select for it. You just have to realize that it's there and wait for your F2s so that you can select for the trait.

There are many ways that this simple strategy can go wrong... It could be two different genes responsible, the strains could fail to mate, you could end up breeding better contams, etc.

Despite what you may hear, standard genetics do apply. Each individual still gets half of their genetic material from each parent.

Has anyone noticed the "hybrid vigor" phenemon with inter-strain breeding? I suspect that they may be too closely related to exhibit it.


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InvisibleATWAR
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Re: creating new strains? [Re: fastfred]
    #2935743 - 07/28/04 09:00 AM (19 years, 6 months ago)

Quote:

fastfred said:
Sorry, I wasn't referring to you. You seem to know what you are talking about, however I think you are overly pessimistic. PC is certainly not as hard to breed as A. bisporus.




Please refer to where I said:
"This is one way how new strains are developed in the commercial industry, it could easily be transferred over to Ps. Cubensis."

While this is all fine in theory, the problem lies in that we cannot select the desired traits from each mushroom. We do not know what spores have the genetics we desire until mated and fruited. Isolating monokaryons and crossing them between different strains is a "shot in the dark". Since there is no way to tell the phenotype a monokaryon will exhibit (or pass on), strain testing must be done on each resulting cross. Then if the fruit body is capable of producing spores, there is no way to know the desired phenotype has been passed to the majority of offspring until you test those spores.

Indeed breeding between strains can be done quite easily (provided they are cross-fertile). But the complexity is very much greater than that encountered in the plant and animal kingdom. The vast amount of spores produced each with varying genetic information causes this complexity. The different sexes of the spores also adds to this complexity, as roughly two thirds of your matings would result in incompatibility (they mate but do not fruit). Then when you have a successful mating that produces fruits and spores, once you go back to them to grow the strain there is no guarantee that desired phenotypes will be expressed (passed on in the spores). No matter what, the genes in the huge mass of spores will possess many variations, each with its own preferred conditions of life, and phenotypic expressions. Isolating the monokaryons that have the genotypes you desire is the largest hurdle...

Add anastomosis into the mix (genetic exchange between dikaryons) and it gets ever deeper. This is why I say it is much more complex...
You say pessimistic, I say optimistic...

BTW, the PF Redspore has apparently been put on the back burner since his bust. The spores survived the raid but the person who has them is awaiting a name that fits its producer (PF) before they ever get released...


--------------------
To give is to live...



Edited by ATWAR (07/28/04 09:08 AM)


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InvisibleATWAR
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Re: creating new strains? [Re: fastfred]
    #2935748 - 07/28/04 09:05 AM (19 years, 6 months ago)

Quote:

fastfred said:
Has anyone noticed the "hybrid vigor" phenemon with inter-strain breeding? I suspect that they may be too closely related to exhibit it.




I have heard it is reversed, as in the crosses rarely produce as well as the original sources by themselves... I assume this takes further strain selection from these hybrids to choose a suitable candidate for cultivation.


--------------------
To give is to live...



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OfflineKonkeyDong
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Re: creating new strains? [Re: fastfred]
    #2942933 - 07/30/04 12:46 AM (19 years, 5 months ago)

What the hell is this UV strata linker? If it's not accessible, then it doesn't help much.


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Invisiblemycopsycho
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Re: creating new strains? [Re: Cyber]
    #2943478 - 07/30/04 03:57 AM (19 years, 5 months ago)

i heard that the only thing pf got by using a black light was an albino, not the pf red spore or anything else.


--------------------
I Am The Sickness.

Diploid: I think adults have a right to make stupid decisions and it's nobody else's fucking business.


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OfflinePsiloman
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If mutations is what you are after.... [Re: mycopsycho]
    #2943885 - 07/30/04 08:33 AM (19 years, 5 months ago)

...then there are many ways.

Chemically induced mutations is one way that can be used on agar work,interesting experiments would be cloning a fruit on an agar slate and then transfering mycelium to agar plates with different concentrations of known mutagens and ,if the specimen survives, proceed to try making it fruit and checking the phenotype as well as spore viability.Sounds very easy ,but as always practical applictation is not something easy.If someone is prone to experimentation and is very carefull about the chemicals he uses and has a tidbit of sanity in hir,can start on a project like that and see what comes out of it.

Known mutagens are substituted DNA bases ( bromouracil ,aminopurine),Intercalating agents (acridine orange, proflavin, ethidium bromide) and many others. Have a look at http://www.web-books.com/MoBio/Free/Ch7F6.htm and at http://www-personal.ksu.edu/~bethmont/mutdes.html#mutagens

Mutagens (read:Potential carcinogens) cause random mutations and in no way this implys that the mutation will be favorable for the organism or the traits you are looking for. In the event of succesfull viable mutations someone may come up with very interesting traits in the new organism,starting from morphological differences to "non-visible" phenotypic changes that affect metabolism but not phenotype in a..visible way!

Please be carefull.Tose research chemicals (yes,they are trully research chemicals ,not "fuck me up" chemicals) can cause some damage to people not careful. On the other hand the medical condition called "Life" is 100% fatal....


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OfflineCyber
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Re: If mutations is what you are after.... [Re: Psiloman]
    #2944100 - 07/30/04 09:55 AM (19 years, 5 months ago)

Wow, that was interesting!

I only have a few problems with this route of genetic alteration.

#1) Most of them are known carsinegens.
#2) All of them are controlled substances and would be difficult to get ahold of. One being a hazard class 9 and one being listed as a class 3 explosive.
#3) I would be weary of them transmitting to humans through consumption of the mushrooms.


As to the need to deal with the genetic alteration of the mushroom strain via a bonding of monokaryons. Man has been altering the mushroom through selection for 100's of years. It is only recently that the ability to select on a monokaryon level has been achieved.
Ill admit that this is a way to do it and that you can obtain the results quickly using this method. This method is just out of the reach of most of us.
ATWAR, IF every mushroom spore produced has a different set of genetic code and you can not select traits then proprigate the traits through selection, you are saying that a PF Classic can produce a Penis Envy because they have random genetics. So with enough time you will eventually get a Penis Envy through random growth and that Penis Envy can then produce PF Classic spores. I am sorry but some traits have to be passed from parent to child. Other wise every grow would be a compleat crap shoot. The genetic mutation of the species would be rampent and eventually degrade.

As to PF, From the reading I have done on him and interviews of him, I would venture to guess that he was experimenting with a variety of ways to make changes to the mushrooms. One of them being lighting. That is evedent from the experiment using black lights. There are a variety of ways to alter the mushroom. ALL give random alterations to the genetics and COULD eventually generate a genetically stable new strain, but again they are random. This could and would take a lot of experimentation and testing. Not to count 100's or even thousands of grows.
I do not have an unrealistic belief that someone can magically create a new strain overnight. If it was that easy then there would be 100,000 strains and new ones every day. Like Edison's work on the lightbulb it will take time and experimentation.

Now a NOTE to EVERYONE!

This is not here to flame. I started this to try to learn from those with experience. To open a dialog about the genetics of mushrooms and strain creation. Please do not flame. If you do not have something productive to add to the discussion then don't post!


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OfflinePsiloman
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Re: If mutations is what you are after.... [Re: Cyber]
    #2944188 - 07/30/04 10:30 AM (19 years, 5 months ago)

A quick reply to your commenting on the mutagens put forth :

1)Yes,they are carcinognes thats why one "works" with them and not "plays".This can be translated in a variety of ways.Dont use them while under the influence of substances,keep them in a safe place ,you only need to work with small quantities of them so one must study before acquiring this material so as he does not acquire unneccesary quantities.One should also work with gloves when handling even their bottles,and to be extra paranoid and safe use a small facemask as well just in case.Make sure you have no spills and if one has spills disinfect the area imediatelly.Always clean the area after the work and be sure to dispose correctly the contaminated items.For the love of Nature,do not empty solutions of these materials however weak down the sink!!!!!OCD patients could do very well at all these points,they can be the best followers of Lab Safety Rules!

2)Unfortunatelly i have no legal knowledge on the matter conserning the states (presumably where you live from your "loc" under the avatar).If all of them are controlled then the only way of acquiring some is to try to find access to a lab.For me it is quite easy because i work for a lab and here in Greece,chem supplies are pretty lenient if someone looks trustworthy enough.Hmmm..There must be someway of acquiring small quantities of them....

3)Thats why one should take extra steps to insure safety! Just to be on the safe side i will state common sense "knowledge".Using it straight on a substrate that is going to fruit and its fruits used is a definite NO NO (assume that one will instantly die ,a horrible death and litle Satan is going to torture him forever by letting Hitler exhibit freely infront of him his pantyhoes for an eternity).Always when making an agar to substrate transfer use "clean" agar.That is never transfer mycelium and agar straight from the plate you got the mutagen in.Instead transfer mycelium on a clean plate first and when it grows select a "clean" area and use this to innoculate.To be extra paranoid and extra carefull ,do not use the results of your experiment straight away.Instead view their traits ,take spores,clone your favorite "mutated" fruit and from then on pursue indulging in the hobby in further "port grow" actions.

All one needs is to be very carefull,mature and sober.Please,those chemicals are dangerous,and i cannot stress it enough. Word of caution: People who have used agents like that usually develop signs of OCD in its "cleaning and tyiding up" version. Better OCD,than cancer sorry!


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Invisiblefastfred
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Re: If mutations is what you are after.... [Re: Psiloman]
    #2944363 - 07/30/04 11:30 AM (19 years, 5 months ago)

> We do not know what spores have the genetics we desire until mated and fruited.

This is easily overcome by keeping a culture of each of the SBIs that you use. You may have to go a few generations down the line, but you should be able to figure out the phenotypes you are looking for one or two generations down the line.

> Add anastomosis into the mix...

Don't add it to the mix, use monoculture.

> BTW, the PF Redspore has apparently been put on the back burner since his bust. The spores survived the raid but the person who has them is awaiting a name that fits its producer (PF) before they ever get released...

How about if you old hands put some pressure on this fella to release the spores. It's really an amazing mutation. And what's wrong with simply calling it "PF redspore" I think that would make it pretty clear who takes the credit.

> What the hell is this UV strata linker?

It's a device that produces a measured ammount of UV radiation. It's used for linking (not ligating) two pieces of DNA. It's nice because you can get reliable 1-2% germination rates so that you're ensured plenty of mutants. I would use a tanning lamp if I didn't have the strata linker available.

> i heard that the only thing pf got by using a black light was an albino, not the pf red spore or anything else.

I don't know about the redspore, but the albino was found under the blacklight. PF foolishly put his breeding line under a blacklight to the point where his entire breeding stock was ruined.

That is NOT the way to do it. Irradiate the spores or blenderize the mycelium and then irradiate, and if you do get a good mutant, start breeding it back to healthy stock as soon as possible.

> ATWAR, IF every mushroom spore produced has a different set of genetic code and you can not select traits then proprigate the traits through selection, you are saying that a PF Classic can produce a Penis Envy because they have random genetics.

ATWAR is correct. Each spore has a completely different genetic makeup. Look at human genetics. You could have hundreds of children and each of them would be different. The chances that you would have identical twins born seperately is astronomical. They may all look a lot like you (the parent), but they each look different from you and each other.

With nearly infinite combinations there are nearly infinite different ways to look. However if we are talking about blue eyes or red hair, that's a different story. Look at it that way. We aren't trying to find one that looks exactly like the parent or some sort of ideal mushroom, we're just trying to pluck out the blue eyes or the red hairs.

A note on mutations... Stay away from the mutagens. They may produce usefull mutations or even different types of mutations than you can get with UV, but I don't think that anyone here is advanced enough to need them. Use UV, it's much safer and highly effective. When you guys are talking about how best to store your 100+ collection of SBIs, then we'll revisit mutagens.


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