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Workman's Psilocybe Cubensis Breeding Method


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#1 Irishlion

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Posted 26 July 2009 - 10:44 PM

I found this on another forums and I thought I would post it here,
all credit goes to workman



Workman's Psilocybe Cubensis Breeding Method



By multiple requests, the hybrization method. This is the highly simplified version with as few technical terms as I can muster. It still requires some agar work and only works within a single species. In this example we are crossing strains of Psilocybe cubensis

Classic mushroom breeding requires the isolation and germination of single spores of both parents and then letting the mycelium from both parents cross in a petri dish. This is a precise and controlled breeding method but it is tedious and time consuming. It requires the isolation of many single spores and many petri dishes in the hopes of a few viable crosses.


This simplified method still requires the isolation of at least one monokaryotic mycelium from a single spore. To get the single spore mycelium I just streak an agar petri dish with a small amount of spores (serial dilution as described in The Mushroom Cultivator (Stamets) also works). At the end of the streak there are usually just a few spores on the agar. As soon as the tiniest bit of germination is noticed, transfer the smallest bit of mycelium to a fresh plate. Select single isolated germination points far from any clusters. The mycelium from a single spore grows slow, isn't rhizomorphic and can't fruit. You can confirm that the mycelium is monokaryotic by looking at a bit of the mycelium under the microscope (monokaryotic mycelium lacks clamp connections), but its usually pretty obvious by the way the mycelium grows. If you don't have a microscope you can skip the confirmation step.


Now that you have your petri dish of monokaryotic mycelium, the hard part is over. The next step is to innoculate some spawn with this mycelium and let it grow to near completion. Once the spawn is fully run with mycelium but not completely white you can proceed to the next step. Don't expect the jar to colonize as fast as multispore or fruiting clones.


Once your monokaryotic jar is ready you can add parent #2 in the cross. The nearly colonized jar should be fairly contaminate resistant at this stage so you don't have to be exceptionally careful now. Open the spawn jar lid and scrape in some spores of parent #2 or make a fresh syringe and inject some spores. The syringe should be freshly made as spontaneous germination can deflower the spores within. Shake the jar and let incubate for at least a week or two.



What is Happening?

What we started with was a jar of monokaryotic mycelium but when we added the parent #2 spores, some of them germinated and fused with the monokaryotic mycelium which becomes a dikaryotic mycelium by nuclear migration. Essentially the entire monokaryotic culture becomes dikaryotic by replicating and moving nuclei in a sort of a chain reaction through the already existing mycelial network. Since there isn't any available uncolonized substrate left in the spawn, any spores of parent #2 that fuse with each other won't have enough resources to produce any mushrooms.


Fruit the spawn directly (don't add to bulk substrate) and all the mushrooms produced should be varietal hybrids. The beauty of this method is you (or a friend) only have to produce a single petri dish of monokaryotic mycelium to make as many crosses as you like. You will want to start with your best performer for parent #1 and then you can easily make crosses with any prints you have around for parent #2. Maintain the parent monokaryotic mycelium with periodic transfers to new plates. Its also a good idea to use a very distinct strain for parent #1 since many cubensis look similar and it may not be visually obvious if the cross worked. In my example I used the distinct Falbino strain as the monokaryotic parent #1. Its pretty obvious when the jar produces pigmented mushrooms that the cross was successful.


It is important to realize that the F1 mushrooms are 50% of each parent at this stage but the spores they produce are genetically recombined. This means the prints are not going to breed true and any prints you distribute at this stage won't often produce mushrooms that look like the F1 generation. Only clones of the F1 mushrooms will be the same. To stabilize a new strain you need to grow out the F1 prints to produce F2 prints with selection of the mushrooms that have the traits you want. You need to do this for 5 or 6 sequential generations with selection before the strain can be considered stable.


Edited by Sidestreet, 04 September 2016 - 02:20 PM.

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#2 Hippie3

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Posted 26 July 2009 - 10:47 PM

thx
:bow:

#3 weeeeeee

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Posted 26 July 2009 - 10:50 PM

very very nice sir thank you for posting that here

#4 Irishlion

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Posted 26 July 2009 - 11:15 PM

Here is a copy of workmans thread with pics of the cross and parent 1 and parent 2 and the final cross specimen known as F1




APE 2.0 progress report


Step 1. Hybridization of Falbino with Penis Envy strain. [Complete]
Falbino + Penis Envy = F1 generation

The top picture is the Falbino parent, the middle is the PE parent and the final picture is the F1 cross.


Psilocybe_cubensis_Falbino Parent # 1


Psilocybe_cubensis_Falbino.jpg



Psilocybe_cubensis_Penis_Envy Parent # 2

Psilocybe_cubensis_Penis_Envy.jpg


Falbino x PE = F1 Generation


FalbinoxPE  F1.jpg



As expected, the F1 generation appears normal which indicates that albinism and penis shaped caps are reccessive traits and won't reappear until the F2 or later generations.

Step 2. Sequential cultivations from spores with selection for albinism and reduced cap size as well as stability, carpophore density and size.



 


Edited by Sidestreet, 04 September 2016 - 02:19 PM.
re-inserted pics

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#5 Foster

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Posted 26 July 2009 - 11:18 PM

Great info Il, and thanks to Workman for all his hard work. Interesting stuff for sure.:bow:

#6 RestartLater

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Posted 26 July 2009 - 11:20 PM

yes thank you for the information Lion... More and more everyday to learn.

Liked this too "Only clones of the F1 mushrooms will be the same."




Made some agar so I will have to use learn to cut cloning pieces from the stem soon!!!


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#7 buteo

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Posted 26 July 2009 - 11:20 PM

diggin the info :thumbup:

#8 warriorsoul

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Posted 26 July 2009 - 11:24 PM

It should also be noted the F1 generation should exhibit hybrid vigor and will outperform both parent strains. :heart:

#9 Guest_cap_*

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Posted 26 July 2009 - 11:24 PM

thx for sharing it here, IL .

good job.

#10 cattlexing

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Posted 26 July 2009 - 11:41 PM

damn... looks like the species took a step back with that hy-breed
thats rad though.... so mycel from spores from that f1 cross could look like either parent or anywhere inbetween?

those parent strains are pretty

#11 Irishlion

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Posted 26 July 2009 - 11:44 PM

damn... looks like the species took a step back with that hy-breed
thats rad though.... so mycel from spores from that f1 cross could look like either parent or anywhere inbetween?

those parent strains are pretty



It is important to realize that the F1 mushrooms are 50% of each parent at this stage but the spores they produce are genetically recombined. This means the prints are not going to breed true and any prints you distribute at this stage won't often produce mushrooms that look like the F1 generation. Only clones of the F1 mushrooms will be the same. To stabilize a new strain you need to grow out the F1 prints to produce F2 prints with selection of the mushrooms that have the traits you want. You need to do this for 5 or 6 sequential generations with selection before the strain can be considered stable.



#12 cattlexing

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Posted 27 July 2009 - 12:18 AM

""stage but the spores they produce are genetically recombined.""

That doesen't make sence... genetically recombined?

The spores they produce only contain one half of a genetic code. It's not until the monokaryots hook up with each other that anything is recombined. And its When that happends that genes (traits) from the mother and herself are given the opporunity to re combine... the father and the father to recombine... and by chance- the mother's and the father's traits are recombined in a equal proportion- again.... You probably have a 1 in 4 chance of getting a strain that looks like the F1 from diferent dicaryotic mycelium produced from f1 spores. maybe 1 in 8 considering a curve... but they may be that much liekleier(more likley) to survive if that new combo of traits proves to be an improvement of the genetic drift of the paretn species. Or you get some good traits and a good mutation. Than you're really breeding wild style.
They do look wilder- those f1's


#13 mydarling

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Posted 27 July 2009 - 12:26 AM

genetic recombination is a process that occurs during the formation of sexual gametes (in this case, spores), which increases genetic variety in the spores. it does not refer to anything that occurs during mating.

clearly, workman is well versed in the study of genetics. :bow:

#14 cattlexing

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Posted 27 July 2009 - 12:34 AM

Very Nice
I didn't know things had a recombination prior to sex cell gammets. That makes sence though for variation! Is it only funguses that do this?

#15 mydarling

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Posted 27 July 2009 - 12:35 AM

damn... looks like the species took a step back with that hy-breed
thats rad though.... so mycel from spores from that f1 cross could look like either parent or anywhere inbetween?


the hybrid f1 generation is expressing characteristics that are genetically dominant. the pigmented cap phenotype is dominant to the albino phenotype, since the hybrids have pigmented caps.

#16 mydarling

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Posted 27 July 2009 - 12:36 AM

Very Nice
I didn't know things had a recombination prior to sex cell gammets. That makes sence though for variation! Is it only funguses that do this?


ummm nope. every organism that makes gametes (for example, humans producing sperm and eggs) undergoes genetic recombination to make such gametes.

#17 mydarling

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Posted 27 July 2009 - 12:39 AM

if you seek to understand the genetics behind this, i highly suggest you read the following:

meiosis

homologous recombination

genetic recombination

chromosomal crossover

mendelian genetics
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#18 cattlexing

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Posted 27 July 2009 - 01:00 AM

Thank you micdarling and irishlion

I knew there was a chapter in my biology book I skipped!
gamit!

Anyways... It exhibits the dominant trait of pigmented cap... but that doesen't mean that it doesen't have the non pigmented gene avaliable to a gammit...?

this thread rocks

#19 apokalypse

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Posted 27 July 2009 - 01:20 AM

Awesome find!!
I guess I've got a lot of reading to do :thumbup:

#20 catdaddy

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Posted 27 July 2009 - 06:46 AM

Bookmarked for later study- thanks Irish and Workman!




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