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Offlinelobo121782
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anyone know about hybrid mushrooms??
    #561209 - 02/24/02 03:35 AM (15 years, 6 months ago)

I have been looking for information on hybrid mushrooms, and what it takes to make one. if any one has a link on this type of subject please let me know.


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OfflineAzure
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Re: anyone know about hybrid mushrooms?? [Re: lobo121782]
    #561526 - 02/24/02 02:04 PM (15 years, 6 months ago)

I think the general consensus is we don't really know much about mushroom hybridization. PF wrote an article somewhere(check out his site, use keyword Psilocybe Fanaticus), but his information is all based on conjecture(conclusions based on observation) and no scientific proof.


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InvisibleIncognito
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Registered: 02/06/02
Posts: 125
Re: anyone know about hybrid mushrooms?? [Re: lobo121782]
    #561644 - 02/24/02 04:58 PM (15 years, 6 months ago)

i used to graft my shrooms.  like orchard farmers graft their trees.  then i used fungus gnat larvae to seal the cuts.  the way that worked is that they would eat the part that died and secrete a fluid that acted as a cohesive between the the mushroom parts.  leaving everything to heal up and grow nicely :smile: 


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Anonymous

Re: anyone know about hybrid mushrooms?? [Re: lobo121782]
    #561706 - 02/24/02 06:37 PM (15 years, 6 months ago)

There are two ways to make a hybrid without the addition of BIOTECHNOLOGY.

1. Make a very dilute spore solution. Drop onto agar plates and isolate monokaryons. Do this for the two strains you want to hybridize. Place individual monokaryons onto plate with individual monokaryons. One from each strain, and see if they are compatible. Grow out all suspected crosses, and test for fruiting.

2. Take dikaryons that are pure, and of know fruitability. One from each strain you want to hybridize, and place them on a single plate. The two will grow towards eachother, and if you are VERY LUCKY, a third strain might appear sectoring out of the area of interaction between the two known strains. This is called anastomosis.

When viewing a single Spore race, from a multispore germination, on a single petri plate. You will actually see incompatibility between the Dikaryons originating from this single print. Anastomosis will occur at a higher rate between Dikaryons from the single strain, then between dikaryons from different strains.

The second one is easier to try, but will have less chance to succeed. The first type involving monokaryon breeding, is far more tedious, and time consuming, but will result in Higher frequency of hybridization. You have to isolate individual monokaryons!!! This means you have to really dilute the spores, if two compatible monokaryons are near eachother on a plate, they will mate. You have to transfer quickly, and label your plates VERY WELL. LOTS OF PLATES.

According to Stametes, most hybrids are less vigorous then their donor strains.


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OfflineAnnoA
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Re: anyone know about hybrid mushrooms?? [Re: Incognito]
    #562166 - 02/25/02 04:52 AM (15 years, 6 months ago)

"i used to graft my shrooms. like orchard farmers graft their trees. then i used fungus gnat larvae to seal the cuts. the way that worked is that they would eat the part that died and secrete a fluid that acted as a cohesive between the the mushroom parts. leaving everything to heal up and grow nicely "

And then you woke up all wet and with a slipper in your mouth...


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Invisibleaaron
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Registered: 10/18/99
Posts: 89
Re: anyone know about hybrid mushrooms?? [Re: lobo121782]
    #562364 - 02/25/02 11:24 AM (15 years, 6 months ago)

GEt this book

Genetics and Breeing
Of Edible Mushrooms
By
Shu-Ting Chang
John A. Buswell
Philip G. Miles
Copyright 1993


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OfflineAnnoA
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Re: anyone know about hybrid mushrooms?? [Re: aaron]
    #562540 - 02/25/02 02:21 PM (15 years, 6 months ago)

Untill the book arrives read this excerpt from The Biology and Cultivation of Edible Mushrooms by S. Chang, W. Hayes.

http://www.fungifun.org/sex.pdf

(Sorry, page 86 is missing)


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Offlinelobo121782
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Registered: 02/12/02
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Re: anyone know about hybrid mushrooms?? [Re: lobo121782]
    #562801 - 02/25/02 06:17 PM (15 years, 6 months ago)

I thank all of you guys and girls that are helping me on this topic, my major goal is to produce the best mushroom ever, sure that it is a long shot but its well worth it. And when I achieve my goal in producing a mushroom that gives spores with its own genes to carry on its traits, I will post it and to all that helped me acomplish this goal My treat to you is a print of your own. I know that this will take some time, and I am no pro at this but I have this as a goal to acomplish. any more info that you run accrost please let me know. Thanks Eric


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OfflineAzure
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Re: anyone know about hybrid mushrooms?? [Re: ]
    #563636 - 02/26/02 12:02 PM (15 years, 6 months ago)

Teonan, have you actually tried this procedure, or is this just theory? If so, do you have any pictures?
Aaron-have you read that book? Explain to me how they proved that it works, because it sounds very interesting. Thanks


Edited by Azure (02/26/02 12:08 PM)


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Invisibleaaron
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Re: anyone know about hybrid mushrooms?? [Re: Azure]
    #563728 - 02/26/02 01:30 PM (15 years, 6 months ago)

I have the book, and i have read the book. it is a good book.
They proved that it works by cell fusion of two monokaryotic lines make a dikaryotic that will fruit and a DNA test to.



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OfflineAzure
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Re: anyone know about hybrid mushrooms?? [Re: Azure]
    #563955 - 02/26/02 06:00 PM (15 years, 6 months ago)

Pardon my ignorance, but how do they run such a DNA test? What is the procedure? Thanks.


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Anonymous

Re: anyone know about hybrid mushrooms?? [Re: Azure]
    #564201 - 02/26/02 10:50 PM (15 years, 6 months ago)

Look, Once you understand what Heterothallic, and tetra polar are, it is all tedious lab work from there. Cubensis Prefer to outcross, as do all Heterothallic, tetra polar mushrooms.

The fusion between monokaryons occurs between disimilar Mating Types. The mating types of any cubensis can be labeled generically as A1B1, A1B2, A2B1,A2B2. As long as A1B1 comes in contact with A2B2 you have a fertile dikaryon formed. The problem with incompatability is A1B1 will mate with A1B2, and A2B1, will mate with A2B2. These result in non fertile dikaryons.

When crossing two Strains you still have to get A1B1 from one strain to mate with A2B2 from another, and vice versus. This mating will result in a fertile Dikaryon. Even though the spores originated from different strains. Compatability is controlled by differing A factors and differing B factors. This is just sexual compatability factors. Not all genes!!! Just the sexual mating type factors.

Nothing special happens different from monokaryons from a single strain mating with itself, and monokaryons from two different strains mating with eachother. As long as they are sexual compatable, they undergo plasmogamy. This is not a fusion of Nuclei, it is simply combination of two uninucleate hyphae into a single bi nucleate hyphae. The nuclei do not fuse. Fusion of the nuclei only occurs during Karyogamy inside the basidium. This is the only time a basidiomycete is Diploid. The rest of it's time it has haploid nuclei. While it is Dikaryotic, the two nuclei are maintained inside the same hyphal cell, but they are independent of eachother.

This isn't my theory, it is scientific Fact. Read any book on fungal genetics, morphology. The most thourough study of compatability factors in bifactoral organisms is with Schizophyllum commune. Raper, Baxter, and Ellingboe,1960 and Raper, Baxter, Simchen,1967, found approximately 450 A factors and 90 B factors. They found the B factors to be associated with nuclear migration, and the A factors to be associated with nuclear pairing and clamp connections.


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Anonymous

Re: anyone know about hybrid mushrooms?? [Re: Azure]
    #564230 - 02/26/02 11:09 PM (15 years, 6 months ago)

And no, I have never actually took the time to do it.

P.s don't forget that Hybridization can also occur between two dikaryons via anastomosis. Hybrids within a single sporerace, and hybrids between different spore races. The frequency will be much less, then with Monokaryon breeding.
If I could get paid to do such things, I would be doing it all day long. But I don't have a Ph.D. and I certainly don't know of any Research being funded to do so with our little friends.

The only Psilocybin mushroom with it's mating system already figured out is P. cubensis. At least in all the Text's that I have read. Stametes mentioned Gartz found reproductive barriers between P. bohemica,cyanescens, and azurescens, but I have not seen that journal article. Did he actually work out each mating type for each monokaryon of each species first? How detailed of a study was it?

Stametes does not mention compatability information for any species other then cubensis!!!

If any information is out there, and someone has access to it, please let me know!!! I would be very interested in seeing anything that relates specifically to Mating systems in Psilocybe and Panaeolus species.


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OfflineAzure
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Re: anyone know about hybrid mushrooms?? [Re: ]
    #565241 - 02/27/02 11:47 PM (15 years, 6 months ago)

Is Schizophyllum commune a mushroom or lower fungus? Do the mating tendencies in lower fungi work the same way as mushrooms? I'm stil more interested in in how to do a DNA test to prove that you end up with a dikaryon that originated from two different strains(ie. a cambodian crossed with a palenque strain of cubensis). I don't doubt that this has been done based on what people are saying, but I just want to know an overview of how they ran the DNA test. Your explanation does not reveal how they proved that it works; it just explains how it works in theory. You don't need to trip out because of my curiosity:)
In science, our understanding of logic changes constantly, and laws that we once thought were true can sometimes be modified...Math that we call fact is what has been proven to be true, and has not been disproven. It is most likely correct, but just because we cannot disprove it doesn't mean that it can be disproved!


Edited by Azure (02/27/02 11:50 PM)


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Invisibleaaron
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Registered: 10/18/99
Posts: 89
Re: anyone know about hybrid mushrooms?? [Re: Azure]
    #565314 - 02/28/02 01:09 AM (15 years, 6 months ago)

DNA Fingerprinting, method of identification that compares fragments of
deoxyribonucleic acid (DNA) It is sometimes called DNA typing. DNA is the
genetic material found within the cell nuclei of all living things. In
mammals the strands of DNA are grouped into structures called chromosomes.
With the exception of identical twins, the complete DNA of each individual
is unique.

A DNA fingerprint is constructed by first extracting a DNA sample from
body tissue or fluid such as hair, blood, or saliva. The sample is then
segmented using enzymes, and the segments are arranged by size using a
process called electrophoresis. The segments are marked with probes and
exposed on X-ray film, where they form a characteristic pattern of black
bars?the DNA fingerprint. If the DNA fingerprints produced from two
different samples match, the two samples probably came from the same
mushroom.

DNA fingerprinting is expensive!!


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Anonymous

Re: anyone know about hybrid mushrooms?? [Re: Azure]
    #565396 - 02/28/02 02:43 AM (15 years, 6 months ago)

Sorry, wasn't trying to bug out!!

Yes it is a basidiomycete. It is tetrapolar and heterothallic, like cubensis.

If you have one monokaryon from one strain and put it next to another Strain monokaryon on a petri plate, and they grow together, form clampconnections, and can fruit, It has to be a hybrid. Monokaryons of cubensis are self sterile.


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Invisibleaaron
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Re: anyone know about hybrid mushrooms?? [Re: ]
    #565426 - 02/28/02 03:18 AM (15 years, 6 months ago)

Teonan, is right on the money!

A hint: use a surfactant when making a spore dilution. (the spores will not stick to each other.)


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Anonymous

Re: anyone know about hybrid mushrooms?? [Re: aaron]
    #565453 - 02/28/02 04:05 AM (15 years, 6 months ago)

Very helpful tip!!! I have read that surfactants also can be used to by themselves or in combinations with heat to stimulate germination of some spores. Increasing membrane permeability.


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OfflineAnnoA
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Re: anyone know about hybrid mushrooms?? [Re: aaron]
    #565467 - 02/28/02 04:48 AM (15 years, 6 months ago)

I allways use one drop of the dishwashing liquid (the one against limestains) per pint of water when making spore suspensions.


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OfflineAzure
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Re: anyone know about hybrid mushrooms?? [Re: ]
    #566198 - 02/28/02 09:08 PM (15 years, 6 months ago)

Thanks everyone! I know with Tissue culture of plants, we use tween 20 (polyoxyethylene (20) sorbitan monolaurate) to break the surface tension of water so that a piece of tissue can be fully sterilzed in a bleach solution.


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