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OfflineCyber
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creating new strains? * 1
    #2926043 - 07/25/04 03:35 PM (12 years, 4 months ago)

This question comes up from time to time and I have read all the responces to the possibility of creating new strains. Starting with the explanation of how they (The mushrooms) reproduce and going to the "It can not be done in a home setup." statement.
So with this information I am going to put forward a theoretical propition as to how it could be done and I am looking for some one to tell me why it can not be done.

We know that spoors within a strain are a crap shoot. We can use agar to isolate a substrain and then test the substrain for fruiting and growth, thus selecting a good substrain to use. This is all common knowledge.

We know (or should know) that mother nature has a way of making genetic abnormalities with in any species. It is survival of the fittest that allows that genetic information to proprigate to the next generation or in the case of a mushroom to be one of the genetic possibilities produced.

Now taking these bits of CK (Common Knowledge) and using them to our advantage to select the traits that we want. Then propagating them should with in 5 to 6 generations generate a noticeable change with in the growth parameters of the species. Thus creating a strain that has different growth parameters to the standard.

For example: I take 6 PDA plates and inoculate them. I then incubate them at 37C-39C and select the best growth (If any, this could take 100's of plates to find the best growth)

I then grow WBS jars of the best ones and fruit at 21C-23C. Collect spore prints and repeat the experiment 5 to 6 times. Each time Selecting the best growing and fruiting substrain with in our parameters. This should increase the number of spores produced with the genetics we are seeking to proprigate. Producing a new strain that has altered growth parameters allowing for higher temperature during myc growth and possibly expanding the thermal death point of the myc of the new strain.

This is similar to what is going on in hospitals with the new "Super Bugs" any disinfectant will only effect 99% of the germ population. It is that 1% that is now propagating and is immune to the disinfectants and antibiotics.

So what is wrong with this? This is selective prorogation to create a new strain!


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OfflineNNY
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Re: creating new strains? [Re: Cyber]
    #2926287 - 07/25/04 05:12 PM (12 years, 4 months ago)

gogo forced evolution!


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OfflineOpenminded
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Re: creating new strains? [Re: Cyber]
    #2926456 - 07/25/04 06:05 PM (12 years, 4 months ago)

Sure it can be done. Isn't that what people have been doing with weed for years - selective breeding of plants with desired characteristics?
And taking into account the much shorter life cycle of a fungus, strain enhancement could be very much more rapid.
What exactly is the definition of a strain, anyway? How different from the parent strain does it have to be before it can be called a new strain?


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InvisibleMykey
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Re: creating new strains? [Re: Cyber]
    #2927011 - 07/25/04 09:57 PM (12 years, 4 months ago)

I think you guys are assuming that the genetics of mushrooms are similar and as simple as the plant genetics present in weed. I have to tell you that ,with all due respect,that is an invalid assumption.

Making a totally new strain of shroom would actually be a much more daunting proposition than one might think. Their genetic code is very complex and somewhat difficult for the average person to manipulate.

The selective culturing that you speak of Cyber will certainly isolate a stong line of myc that will fruit consistantly and well.
What you describe is basically the isolation of one substrain that will not have to compete with others to colonize a substrate. You obtain the isolate through carefull observation of growth patterns on the plates and transfering the healthiest growth to fresh plates until you are left with a non sectoring,rhizomorphic myc that you can then use to innoculate your medium. You would then need to make either a master grain jar to do g2g or make more plates from the isolate.

Now here's the thing... Once you have grown out and fruited the isolate on your substrate there is only one way to preserve it and that is to clone from sterile tissue taken from the strongest fruit bodies. Simply taking a print and doing another multi-spore innoculation would put you right back at ground zero,as you would once again have many many little colonies competing for dominance again. The spores produced by the fruits wont all be genetic replicas of the parent shroom as one might believe. What you will find,however, is that once you have taken fresh,sterile tissue from teh strongest fruits you will have a strain that will perform very well in the environmental conditions that you are providing. They will be an "enhanced" strain,but alas not a new one at all.

Having said this I agree that you could ,in theory,develop an isolate that would fruit well at higher ,or lower,temps. This ,of course,assuming that you fruited them consitantly,over and over,in like temps and always cloned the strongest fruits for the next go at it.

Well, I did my best to explain this stuff as I understand it and I am sure that someone will gladly set me straight on any inaccuracies.
Hope this helps!

Good Luck!
MYKEY


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OfflineSuntzu
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Re: creating new strains? [Re: Mykey]
    #2927055 - 07/25/04 10:23 PM (12 years, 4 months ago)

It can be argued that to a degree this has been done with the PF 'strain'.  I don't know the semantics, whether it is considered a different strain or not.  Hundreds/thousands of generations have given rise to something that most would agree is 'different' [try casing it :wink: ].  These characteristics are not undone by going back to multispore.  Perhaps the specific substrate/conditions it's been fruited with [PF tek] has caused it to lose a few genes here or there, or imparted some kind of differences in gene regulation. 

I'm no expert in this area, but I think it's a point for discussion.


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OfflineCyber
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Re: creating new strains? [Re: Mykey]
    #2927075 - 07/25/04 10:33 PM (12 years, 4 months ago)

Quote:

Mykey said:
I think you guys are assuming that the genetics of mushrooms are similar and as simple as the plant genetics present in weed. I have to tell you that ,with all due respect,that is an invalid assumption.




Ok, I did not make the comparison between weed and fungus, and that comparison is wrong. When you are dealing with a life form that gets it's genetics from two partners then it is easy to select the major traits that you are looking for and bread for those traits. Some traits can not be bread. For example on the dutch breed of rabbit. There are markings that are considered "GOOD". Those would be the butterfly on the nose and the saddle across the back. These are not traits that you can bread at a 100% accuracy. But by consistently selecting good parents you can statically increases the number of offsprings that have the favorable marks. This boils down to a genetic square of dominate and resistive genes. It is also off topic, sorry.

Quote:

Making a totally new strain of shroom would actually be a much more daunting proposition than one might think. Their genetic code is very complex and somewhat difficult for the average person to manipulate.




I am not average. :wink: Besides making a new strain is easy. You buy a common strain and rename it. :smile: Ok, kidding aside. What constitutes a new strain? 

Quote:


Now here's the thing... Once you have grown out and fruited the isolate on your substrate there is only one way to preserve it and that is to clone from sterile tissue taken from the strongest fruit bodies. Simply taking a print and doing another multi-spore inoculation would put you right back at ground zero,as you would once again have many many little colonies competing for dominance again.




That is the point. You are trying to give the substrains with the higher incubation temperature the opportunity to thrive. Each successive generation would be better equipped to handle the heat. I can not believe that the offspring get nothing from the parent. The reason is that nature is doing this right now. Otherwise there would be no difference between the PF strain and the Penis Envy strain. They are a living and growing entity they must be able to addapt or they would never survive!

Quote:


The spores produced by the fruits wont all be genetic replicas of the parent shroom as one might believe.





I am not looking for a genetic replica. I am looking to increase the number of spores that survive and thrive at higher temperatures. After a few generations of this genetic selection the number of viable spores at the higher temperatures should increase.

Quote:


  Well, I did my best to explain this stuff as I understand it and I am sure that someone will gladly set me straight on any inaccuracies.
Hope this helps!





You did a good job. I just think you misunderstood what I was saying. Your selection of a good substrain provides those with the right genetic information a chance to thrive. Continuing this through successive generations should increase the success rate of myc growth at higher temperatures.

I am half tempted to buy a DNA sequencer and find out just how different these strains are!


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OfflineCyber
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Re: creating new strains? [Re: Suntzu]
    #2927091 - 07/25/04 10:42 PM (12 years, 4 months ago)

Quote:

Suntzu said:
It can be argued that to a degree this has been done with the PF 'strain'.  I don't know the semantics, whether it is considered a different strain or not.  Hundreds/thousands of generations have given rise to something that most would agree is 'different' [try casing it :wink: ].  These characteristics are not undone by going back to multispore.  Perhaps the specific substrate/conditions it's been fruited with [PF tek] has caused it to lose a few genes here or there, or imparted some kind of differences in gene regulation. 

I'm no expert in this area, but I think it's a point for discussion.




Ahh there is a good point. The PF strain was the Amazon strain which is also the Matias Romero strain. I have read that the PF strain does better on BRF cakes than in casing, but the Amazon strain does well in casings and may do will in brf. The selective breading and harvesting of PF from BRF cakes has led to what would appear to be a statistical rise in the preference of growth medium.


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InvisibleTheHateCamel
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Re: creating new strains? *DELETED* [Re: Mykey]
    #2927229 - 07/25/04 11:28 PM (12 years, 4 months ago)

Post deleted by TheHateCamel

Reason for deletion: jh



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Anonymous

Post deleted by Papaver [Re: TheHateCamel]
    #2927279 - 07/25/04 11:40 PM (12 years, 4 months ago)



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InvisibleATWAR
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Re: creating new strains? [Re: Mykey]
    #2927750 - 07/26/04 03:11 AM (12 years, 4 months ago)

Quote:

Mykey said:
Now here's the thing... Once you have grown out and fruited the isolate on your substrate there is only one way to preserve it and that is to clone from sterile tissue taken from the strongest fruit bodies. Simply taking a print and doing another multi-spore innoculation would put you right back at ground zero,as you would once again have many many little colonies competing for dominance again. The spores produced by the fruits wont all be genetic replicas of the parent shroom as one might believe. What you will find,however, is that once you have taken fresh,sterile tissue from teh strongest fruits you will have a strain that will perform very well in the environmental conditions that you are providing. They will be an "enhanced" strain,but alas not a new one at all.




I just want to point out that once you have an isolate, there is no need for successive clonings. The fruitbodies have the same genetics of the isolate they grow from. You will in effect end up with the same isolate...



As for creting new strains through selection:
Ahh, the age old quest to put evolution on "fast forward"....
Good Luck!

Isolating and breeding monokaryons would be a much more efficient means of creating a new strain...


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



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OfflineOpenminded
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Re: creating new strains? [Re: ATWAR]
    #2928516 - 07/26/04 02:12 PM (12 years, 4 months ago)

But ATWAR, monokaryons can not fruit. Therefore, they can not create spores, and so the genetic diversity that is essential to producing a strain with desired characteristics is not there... In order to get sporulating fungi (to provide subsequent generations with genetic diversity, thus allowing the best to be selected etc etc), monokaryons would have to be combined, and therefore isolating the monokaryons in the first place doesn't make sense, unless I've misunderstood something?
Mykey, I don't see why genetic complexity has anything to do with it. The principals that govern evolution are the same no matter what the organism. Only instead of breeding suitable parents together, we are relying on the chance combination of two monokaryons to provide a dikaryon with desirable characteristics, which is then allowed to fruit, and its spores are germinated, thus allowing the process to be repeated until you are happy with the outcome.
Have I missed something..?

Edit: but we still need to decide on what exactly constitutes a new strain...


Edited by Openminded (07/26/04 02:14 PM)


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InvisibleATWAR
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Re: creating new strains? [Re: Openminded]
    #2928769 - 07/26/04 03:37 PM (12 years, 4 months ago)

Quote:

But ATWAR, monokaryons can not fruit. Therefore, they can not create spores, and so the genetic diversity that is essential to producing a strain with desired characteristics is not there... In order to get sporulating fungi (to provide subsequent generations with genetic diversity, thus allowing the best to be selected etc etc), monokaryons would have to be combined, and therefore isolating the monokaryons in the first place doesn't make sense, unless I've misunderstood something?




Read the part about breeding monokaryons... When combined on a plate together they will form Dikaryons, which will have the ability to fruit... You take many, many of these breedings and test each one for desired characteristics.

Quote:

The principals that govern evolution are the same no matter what the organism. Only instead of breeding suitable parents together, we are relying on the chance combination of two monokaryons to provide a dikaryon with desirable characteristics, which is then allowed to fruit, and its spores are germinated, thus allowing the process to be repeated until you are happy with the outcome.
Have I missed something..?




Remember it takes nature decades, if not centuries or millenia to do this...
Each mushroom will have genetics that span a broad range no matter how many generations you selectively breed. In the end you will still have "broad range" genetics...

I retire from trying to explain this over and over... Just try it... After 20 or so generations of this you will see it is not as easy as you believe...


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



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InvisibleMykey
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Re: creating new strains? [Re: Openminded]
    #2928880 - 07/26/04 04:12 PM (12 years, 4 months ago)

Wow! I knew this would spark up some good conversation and it looks like I was right! Looks like you asked a good question for some debate Cyber! I know this has been discussed before,but it does make for much interesting debate and gives us all a good chance to learn from each other. --I have to admit it is a nice change from the usual questions about air exchange and potency etc.


I first want to make sure that everyone knows that I dont consider myself an authority on this subject at all,but I cant resist discussing it with all of you and learning by doing it. This aspect of cultivation is very intrigueing to me!!


Cyber-- First let me say that I wasn't trying to say that you made the comparison between weed and shrooms(I read it in the post following yours so I just thought I would mention it.) Second I also didnt mean to emply that you are an "average" person as I believe that expressing interest and making intelligent observations on this subject makes it obvious that you are not. Furthermore I believe that with this post other like minded above average minds have been attracted to post their remarks.

As to the question of what constitutes a new strain. I am afraid that I cant answer that one either so hopefully someone can provide a definative answer for us. When I think of a new strain I think of two things---hybridization and the natural genetic process of adaptation to the environment creating a strain which harbors enough notable and consistent difference in characteristics to be considered a newly evolved entity. The first of which is pretty cut and dry and from what I understand would take an assload of trial and error experimentation to successfully complete. The second is more along the lines of where this discussion seems to center. Suntzu brought up a perfect example of this in the PF strain. * I cant help but picture a "Mad Shroomerite" standing over his newly created strain and proclaiming,"IT LIVES!!!!!" Ha! Ha!

ATWAR-- I was hoping you would come in on this one! I figured you couldn't resist.
I was referring to cloning the fruits as being a the way to preserve the substrain(if you dont have the origional culture or master grain jar),as apposed to simply collecting the spores. As I understand it this would put you back to square one unless you collected the spores from a fruit that was in fact the result of many succesive life cycles within a strictly controlled environment that resulted in unique characteristics constituting a "new strain". I do think that if you are trying to manipulate evolution that it would be necessary to collect and culture from the fruit bodies of each life cycle and move forward ,rather than fruiting over and over from the initial isolate.(EDIT:)-> I guess the DNA will be exactly the same each time so it probably wouldnt make a bit of difference when I think about it,but I wonder if the myc would produce a mutated fruit,in response to the consistantly controlled environmental stimulus, that would be more successfull in said environment.
By the way I have enjoyed reading all of your post on cloning and agar techniques,Good Work!


TheHateCamel-- I dont think it would be a quick process,but IMHO it is possible to develop a substrain that would posses the ability to grow in colder temps. I think it would be a very worthwhile undertaking indeed if one was to try it!

OpenMinded--I could be wrong but I think ATWAR was referring to isolating monokaryons from different strains and attempting to create a hybrid dykaryon. Success would mean a new strain overnight verses attempting to manipulate evolution which could take a long time.



MYKEY


Edited by Mykey (07/26/04 05:11 PM)


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OfflineCyber
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Re: creating new strains? [Re: Mykey]
    #2929010 - 07/26/04 04:50 PM (12 years, 4 months ago)

Quote:

Mykey said:
Cyber-- Second I also didnt mean to emply that you are an "average" person as I believe that expressing interest and making intelligent observations on this subject makes it obvious that you are not. Furthermore I believe that with this post other like minded above average minds have been attracted to post their remarks.





My responce was tung in cheek. Just a little levity to lighten the mood. :wink:

Quote:


As to the question of what constitutes a new strain. I am afraid that I cant answer that one either so hopefully someone can provide a definitive answer for us.




I am not sure that anyone has a good answer for this. That seems to be half the problem. To date it appears that new strains are (For the most part) just a new wild specimen that was picked in a field. Is there even a difference in area that would constitute a new strain or can I pick one down the street and then 1 mile later pick a new one and call it a new strain?

Quote:


I cant help but picture a "Mad Shroomerite" standing over his newly created strain and proclaiming,"IT LIVES!!!!!" Ha! Ha!





Ahhh there is a little mad scientist in all of us just waiting to get out. There is also the inherent want in all of us to make a mark on this world. Discoveries and exploration are one of the ways we can accomplish it. collaboration may be one of the ways to find answers to this and other questions.

Quote:


TheHateCamel-- I dont think it would be a quick process,but IMHO it is possible to develop a substrain that would posses the ability to grow in colder temps. I think it would be a very worthwhile undertaking indeed if one was to try it!





One of the reasons I chose to frame this around temperature variations is that it is one of the easyest (For a home experimenter) to identify and test. This could be done with potency but would require a GC/MS (Gas Chromatograph/Mass Spectrometer) to test for content. I seem to have misplaced mine in the garage and have not had time to dig it out.  :rolleyes:

Quote:


ATWAR said:
Remember it takes nature decades, if not centuries or millenia to do this...





This is true but mother nature has to wait between flutings, some times years. That is one of the advantages of cultivation, You can speed up mother nature's time table.


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InvisibleMykey
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Re: creating new strains? [Re: Cyber]
    #2929157 - 07/26/04 05:23 PM (12 years, 4 months ago)

I also agree with ATWAR's comments on the process of evolution in nature.

I also agree with you, Cyber,and tend to think that the presence of unstable variables in nature are a key factor in the time frame in which notable addaptations become evident. Therefore I think that by carefully controlling as many of these variables as one can,like temp and growth cycle time,a remarkable acceleration of this addaptation process could be achieved. Bearing in mind,of course,that this could still mean a great many cycles and a long time would pass before any results could be noted.

Just my 2 cents!
MYKEY


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InvisibleATWAR
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Re: creating new strains? [Re: Mykey]
    #2929242 - 07/26/04 05:41 PM (12 years, 4 months ago)

I noticed somebody asked for what the true definition of strain was, and just to clarify:

Strain: A race of individuals within a species sharing a common genetic heritage but differing in some observable set of features, which may or may not be taxonomically significant. - Stamets, GGMM.


Quote:

Mykey said:
ATWAR-- I was hoping you would come in on this one! I figured you couldn't resist.
I was referring to cloning the fruits as being a the way to preserve the substrain(if you dont have the origional culture or master grain jar),as apposed to simply collecting the spores. As I understand it this would put you back to square one unless you collected the spores from a fruit that was in fact the result of many succesive life cycles within a strictly controlled environment that resulted in unique characteristics constituting a "new strain". I do think that if you are trying to manipulate evolution that it would be necessary to collect and culture from the fruit bodies of each life cycle and move forward ,rather than fruiting over and over from the initial isolate.(EDIT:)-> I guess the DNA will be exactly the same each time so it probably wouldnt make a bit of difference when I think about it,but I wonder if the myc would produce a mutated fruit,in response to the consistantly controlled environmental stimulus, that would be more successfull in said environment.
By the way I have enjoyed reading all of your post on cloning and agar techniques,Good Work!




Fist of all, Thank you Mykey, but I am not done yet. More to come on Agar... :wink:
I believe that over time of repetitive clonings, the genetics will change. Whether it will change in your favor or not is the question at hand. The only type of genetic change I can say for certain will happen is the loss of viability due to far too many cell divisions. The genetics simply mutate or degrade... Basically you lose the isolate...

And to address the issue of speeding up evolution, I will say this one last thing:
Take this situation for example:
Say sometime in the future (or past), high temperatures have dropped 10-20 degrees below the normal for say, 20 years. Cubensis continues to be found growing, although sparse and slow, spreading its spores each year (forget about spores that were there previously). Then you collect spores from a specimen on year 20 of a huge mushroom growing in the cooler temperatures. Do you have spores that will be adapted to the cool temperatures? No. You have millions of spores, with huge genetic diversity between them. The spores will have genetics that favor growth under all different kinds of conditions; each separate substrain is not exactly the same. This is nature, this is how life lives on, and this is the genetic complexity of mushroom reproduction...

This is why we must isolate our desired characteristics. If we wish to try and breed these new characteristics then we must cross monokaryons from different strains (I.E. - GT and B+), possibly combining desirable traits of both strains. But when you go back to spores again, you may or may not end up with what you want consistently. The genetics will still vary...

This is the nature of the life of mushrooms. This is the nature of their survival...

Edit: Clarified on a point, and checked spelling...  :blush:


Edited by ATWAR (07/26/04 05:54 PM)


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OfflineOpenminded
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Re: creating new strains? [Re: ATWAR]
    #2929510 - 07/26/04 07:36 PM (12 years, 4 months ago)

I'm just not sure why you have to isolate at the monokaryon stage rather than letting them form dikaryons on their own, and then isolating from, say, the fruitbodies that grow best? Is it just to have more control over the combinations formed, or is there some reason why it can only be done that way?
Maybe I'll try it when my spores arrive :laugh: (j/k!) Mmmm... a super-strain forming perfectly tetrahedral sclerotia..? That'd be fun :smile:


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InvisibleATWAR
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Re: creating new strains? [Re: Openminded]
    #2929799 - 07/26/04 09:02 PM (12 years, 4 months ago)

I speak of isolating monokaryons for breeding different strains together such as GT and B+. You isolate the monokaryon from GT and mate it with a monokaryon from B+. If you were to use the same strain then isolating dikaryons is how it would be done. Or, you could grow out multispore and clone the best specimen. This is the best and fastest way IMO...


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To give is to live...



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Invisiblefastfred
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Re: creating new strains? [Re: ATWAR]
    #2931555 - 07/27/04 07:35 AM (12 years, 4 months ago)

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.

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.

In addition to Stamets definition of a strain, I would add that to be a strain it must breed true. That is to say that it maintains the phenotype for which it was declared a new strain.

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.

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.

BTW what ever happened to PF (and others) albino strains? How about the PF redspore? It's a shame that PF knew little about breeding and genetics. I don't mean to dis him, because in many other aspects he was a great man.


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Invisiblemycopsycho
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Re: creating new strains? [Re: fastfred]
    #2931729 - 07/27/04 10:25 AM (12 years, 4 months ago)

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?


--------------------
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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Mushrooms, Mycology and Psychedelics >> Advanced Mycology

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