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crossbreeding mushrooms(Psilocybin)


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#1 Ovoideocystidiata

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Posted 15 November 2015 - 08:01 PM

crossbreeding mushrooms, is it possible?  taking individual spores of two different strains and creating something of it?


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#2 wharfrat

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Posted 15 November 2015 - 08:14 PM

no, not that i am aware of. you can however isolate different characteristics of a certain strain or genus of fungi. but you cannot make a hybrid. Someone will correct me if i'm wrong.


Edited by wharfrat, 15 November 2015 - 08:15 PM.

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#3 Ovoideocystidiata

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Posted 15 November 2015 - 08:22 PM

if that is indeed true.  does anyone know why spores from two different strains of the same species will not mate?



#4 SteampunkScientist

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Posted 15 November 2015 - 08:28 PM

I found this by someone who knows on another site. You should Google these terms:

A hybrid would result from the mating of a monokaryon of one strain with a monokaryon of another strain. This paired Dikaryon would be a hybrid between the two strains. The true success of the hybrid would be if it produced fruits that had spores. Only at this stage would the true hybrid STRAIN be accomplished. No genetic information is exchanged between the two strains until their seperate haploid nuclei have fused and then undergone meiosis. Recombination would then occur between the two seperate strains forming a third strain. It's offspring(spores) would be new combinations of the two donor strains.

Hybrids can also be formed by Anastomosis between two dikaryons of different Strains, but it will happen at Far less frequency then mating monokaryons.

Anastomosis occurs at a higher frequency between substrains of an individual strain. I.E. between different spore matings resulting from a single syringe. When you multispore inoculate a jar of substrate, this is Happening with a high degreee of frequency. A dominant mating will fuse with other matings, incorporating them into it's mycelial network. It can even rewire a false mating into a good mating. It can overcome an incompatible mating, by replacing one of the nuclei within another strand with one of it's own.

A Fertile dikaryon A1B1 A2B2 can run into an infertile A1B2 A2B2 and replace the A1B2 nuclei with a A1B1 nuclei. Creating a fertile A1B1 A2B2 dikaryon that joins the colony. Subsequent fusion could replace the A2B2 nuclei with it's own A2B2 nuclei, which would completely rewire the hyphal strand to it's exact genetic makeup. Or it could leave it partially rewired.

This process can be seen on a Nutrient agar plate. Not all substrains within a strain will fuse, some are completely incompatible. There will be a zone of zero growth between them on the petriplate. They just will not fuse.

In essence hybrids can be formed between substrains of a single strain or between different strains of the same species.

This is the theory, this has been scientifically demonstrated on other species of Basidiomycetes that have been studied!!! I know of know studies that have been done on CUBENSIS, relating to this information. But it must exist. Because it has been Clearly stated in several texts, that Psilocybe cubensis is heterothallic and tetrapolar. And all the above information relates to that type of breeding system in the Basidiomycetes.

You ask why this is not being done, because not much MONEY goes into this type of research in the legitamite world.

Anastomosis has been studied extensively in the edible mushroom world. Agaricus bisporus is homothallic and two spored, not four. Each and every spore it produces already contains both haploid nuclei to make a dikaryon. But fusion(Anastamosis) between these dikaryons produces more productive Dikaryons!!! The majority of High yielding bisporous are a result of HYBRIDIZATION within a STRAIN or between Strains of this species. So if it occurs between DIKARYONS of this species,it has been viewed occuring between monokaryons of other species, WHY WOULD anyone Doubt that it can occur within the Species Psilocybe cubensis.

The major factor to overcome with mating monokaryons is the proximity with which they germinate. Spores tend to clump. So simply placing spores of two different strains in a single syringe, will not overcome the clumping of spores of like strains, and hence their close proximity to each other upon germination.

Probably still occurs, when injected into a substrate, and some of the resulting fruitbodies might actually be hybrids, between Strains, and the resulting offspring (spores) from that mushroom will be different looking then both the mushroom it came from, and all of the mushrooms from both the Donor strains.

Much easier to DILUTE spores from each strain seperately, plate them, isolate slower growing monokaryons, and try mating as many of these from each strain as possible, with as many as possible from the other strain. All matings that fruit, are hybrids!!! If they produce spores, you now have a new STRAIN. Simply cloning the original matings that fruited, will be a Hybrid as well, but not a true hybrid, becasue their has been NO Recombination between the two strains, NO MIXING OF GENES. There has simply been a successful coexistence of two haploid nuclei, one from each Strain, acting independently, but together to create fruits!!! The real genetic swap occurs during Karyogomay and the subsequent meiosis.

by Teonan


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#5 Ovoideocystidiata

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Posted 15 November 2015 - 08:42 PM

spores must be fairly technologically advanced, on a certain level?  who does the work associated with spores and thing likewise? microbiologists?


Edited by Ovoideocystidiata, 15 November 2015 - 08:42 PM.


#6 coorsmikey

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Posted 15 November 2015 - 08:47 PM

crossbreeding mushrooms, is it possible?  taking individual spores of two different strains and creating something of it?

Cross breeding of strains is possible. Crossbreeding of species, not so much but I cannot say definitively. Check out the history of F albino and APE cubensis strains, also look into the work of members TheChosenOne and Workman here. Lots of good info for those who seek it. Just do a lil digging in the new vaults.
https://mycotopia.ne...the-new-vaults/
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#7 Tenderfoot

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Posted 15 November 2015 - 08:56 PM

I believe the H3 strain is a hybrid!! 

 

Sadly,  H3 never produced fruits for me.

 

Either genetics or user error, I suppose.


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#8 Tenderfoot

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Posted 15 November 2015 - 09:05 PM

https://mycotopia.ne...+breed +workman


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#9 coorsmikey

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Posted 15 November 2015 - 09:05 PM

Here is some good reads from 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.


tenderfoot beat me to it!

Edited by coorsmikey, 15 November 2015 - 09:06 PM.

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#10 Ovoideocystidiata

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Posted 15 November 2015 - 09:17 PM

i imagine it would take some equipment to try and mate spores from two different strains.  given the right conditions it seems like it would work.

 

edit : it would probably be difficult to remove one spore from each strain onto slides, etc.  and to move forward.


Edited by Ovoideocystidiata, 15 November 2015 - 09:20 PM.


#11 MLBjammer

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Posted 16 November 2015 - 03:29 AM

I believe the H3 strain is a hybrid!! 

 

Sadly,  H3 never produced fruits for me.

 

Either genetics or user error, I suppose.

Yes, that one is a hybrid.  It always produced well for me.


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#12 Microbe

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Posted 18 November 2015 - 01:44 PM

if that is indeed true. does anyone know why spores from two different strains of the same species will not mate?

Spores do not mate, they germinate. I do not think it is possible to develop a hybrid of 2 seperate species similar to cats and dogs cannot breed with one another. You can complete hybridization from 2 diffrent strains through use of rattle snake venom. The concept is basic IMO buy execution of the process not so much. If your interested here is a abstract.

Attached Files


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#13 Heirloom

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Posted 18 November 2015 - 03:07 PM

But its possible for 2 of the same species to mate? 

  Some of the same species won't mate due to being separated for long times and distances , ah genetic at work. genetic recombination of alelles and chromosomal position changes, an example is ps cyans  from Europe & north America.
Same species but not able to mate , if you could mate they would be so weak they would not survive due to outbreeding depression.

 but hey I am just a newbie


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#14 Microbe

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Posted 18 November 2015 - 04:31 PM

But its possible for 2 of the same species to mate?

Some of the same species won't mate due to being separated for long times and distances , ah genetic at work. genetic recombination of alelles and chromosomal position changes, an example is ps cyans from Europe & north America.
Same species but not able to mate , if you could mate they would be so weak they would not survive due to outbreeding depression.

but hey I am just a newbie

You can force the said-above strains of the same species to mate using the rattle snake venom method. Breeding is not an area im fluent on as far as mushrooms go and many here know i have 5 kids so im an expert on breeding the human species haha but you are correct. Even though it is possible to mate these, the hybrid would be of lesser quality and have less value then the 2 parent cultures making it pointless for cultivation. However creating a hybrid in itself is an accomplishment regardless of its productivity. HS i do not think your a newb! You have a excellent scientific approach which many newb do not have. I cant wait to get back at it. I lost my path there for awhile and started chasing yields and realized i wasnt having fun anymore. When you get knee deep in the science and experimenting, man alive that is what this hobby is about imo. Now if someone would kindly catch me a rattle snake and milk it, send me the venom in a vile and i will attempt to purify it and see if i can introduce to topia a new strain ;)

I havent read the abstract in awhile but i believe what happens is when 2 seperate strains grow towards eachother they stop and form a inhibition zone then remain there until 1 of the cultures proves to be more dominant and grows over the plate. There are still 2 strains in the plate at this point.

When adding snake venom to agar the 2 grow towards each other and still develop a inhibition zone but for a few short hours either culture could send out filaments (i think thats what they are called) and the snake venom does something to the cell wall and allows for them to mate. In order for this to be successful you need to montior your plate and as soon as it appears the 2 cultures mated, you need to isolate to a clean plate to verify that they did in fact mate and are viable. Even though they can mate, many times they are not viable and unable to complete cell division to grow.

I would like a mycologist to explain to me why is there only 2-3 time window for hybridization to be completed from the time the 2 cultures create the inhibition zone.

And why snake venom? Is it because of its ability to alter cells and tissue?

Imagine the intelligence and thought process of the Scientists who thought to use snake venom.
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#15 SteampunkScientist

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Posted 18 November 2015 - 10:29 PM

Interesting, I wonder if there are other substances that could be used. Probably anything that causes necrosis - which is the disintegration of the protein walls of the cell, but not so much that the cell cannot recover, but enough to allow genetic material to leak.

Sort of a chemical knife that makes the two strains "blood brothers".

#16 mattyfresh

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Posted 19 November 2015 - 04:33 PM

I read some where that they used rattlesnake venom on a Petri dish grown over with 2 seperate cultures on it and the venom causes the cellular walls to weaken and then the mycelium can enter the nuclia easier or something. I'll try and see if I can find it.
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#17 Heirloom

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Posted 21 November 2015 - 03:10 PM

the scientists were highly trained with more knowledge that can be stored in the conscious mind.

like kekule who dreamed of snakes biting each others tails  making a benzene ring, which turned out to be true.

the subconscious at work , some times psychedelics work the same way.

proteins and enzymes can do things that require knowledge from within.

 


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#18 Heirloom

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Posted 22 November 2015 - 04:04 PM

So it is written in stone that two or more cubes grown together from spore will not mate? Like EQ ,Brazi; and Maz? a 3 way.

 if they did of course they would be a combination in the first generation, call it f1 if you like, the second generation will show a variety of phenotypes to be selected from.

Matching phenotypes could lead forward, phenos might be big or potent  or unique in some way.
a person could make an open spored cultivar, for those seeking these things.

 the fasted way to see this might be using a microspore to see it they do indeed combine/mate

agar on a sterile microspope glass a bit of agar a sterile cover slip, a microp drop of 2-3 spore solutions. wait and watch over a time

edit : if possible then one might select their ideal mushroom and cultivate, based on you ideas of the best.
 I don't know if you clone or try to isolate


Edited by Heirloom Spores, 22 November 2015 - 04:08 PM.


#19 Heirloom

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Posted 23 November 2015 - 02:19 PM

could cubes related be interbred,  use a petri dish to view the x's then the remove the x's to another dish to grow out

would this damage the cube fungi gene pool? I want to only do good



#20 Lakegal7

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Posted 23 November 2015 - 03:28 PM

These two containers are from spore print
Golden teacher.The original print I received from fsre . Did one grow, took a few prints. This grow is from one of those prints. The light ones look like the Golden Teacher, but the dark ones..well I don't knowimage.jpg
Can't figure out why the pix is posting upside down
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