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Offlinetr8orjohn
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Cross-breeding...is it possible, or impossible? * 1
    #9420446 - 12/12/08 06:51 PM (15 years, 1 month ago)

After searching, searching and searching and finding many posts about why something *isn't* a cross breed (since a cube is a cube is a cube, and substrains will cross, etc) and reading that snake venom has something to do with it...is there *actually* any way to cross breed mushrooms!?!

The impression I have so far is that by allowing two strains to grow towards each other on a snake venom agar plate, there is a possibility that they can cross genes. Is that effectively it? Isn't the reason we inject twice in a PF cake specifically so they *can* meet and *can* exchange genetic information?

If I missed the post in searching, I apologize ahead of time...but it seems like I have seen 20 different posts about why what the poster was doing *couldn't* have caused any cross in genetics, yet at the same time, with these being sexual(ish) organisms its seems impossible that *some* genetics won't get crossed...

Hopefully my questions will bring out the people that know about this subject to shed some more light on it...

jon


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Terry M said:
Looking for rules that other people use successfully. Sterile procedure is just a set of rules that work.


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Invisiblecreekfreek
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Re: Cross-breeding...is it possible, or impossible? [Re: tr8orjohn]
    #9420471 - 12/12/08 06:57 PM (15 years, 1 month ago)

I dont have an answer, I just want to see where this goes.
:peace:


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Offlinedenger
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Re: Cross-breeding...is it possible, or impossible? [Re: creekfreek]
    #9420628 - 12/12/08 07:31 PM (15 years, 1 month ago)

When you'r asking question in advanced forum, please organize your thoughts first and read up on the subject.
If you are asking whether it is possible to make a cross between species, then the answer is: sometimes, with a lot of effort. There are several methods, mostly by stripping the cell wall with enzymes and then forcing the cells to merge using certain chemicals. This is not a procedure one can repeat without access to a specialized lab.
Venom has been discussed at length here as an agent to cause cell wall perforations. It is not efficient, is very dangerous and is best left alone IMHO. Results of such crosses are usually unstable and rarely fruitable.
If you'r asking if it is possible to cross strains of P. cubensis or something similar, then yes, easily. You have to start with single-spore isolates, and put two of them from different strains on one dish. This has to be repeated several times since fungi have complex mating incompatibility system, and only one in several (4 to 16) of such crosses succeed.


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OfflineRogerRabbitM
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Re: Cross-breeding...is it possible, or impossible? [Re: denger]
    #9421175 - 12/12/08 09:16 PM (15 years, 1 month ago)

Strains of cubensis will cross without venom or any other agent. My biggest success with snake venom was to use it to degrade the cell walls of some 20+ year old spores that flat-out would not germinate. The spores were placed on venomated agar and monokaryotic mycelium from a different strain was allowed to crawl across the spores.  The result was a 'pairing' which then led to dikaryotic mycelium, and the result is still circulating as the Redboy strain. 

Crossing species, which would result in hybrids is another story.  It's been repeatedly done by professional mycologists, but so far all but one of my attempts has failed.
RR


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InvisibleHerbBaker
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Re: Cross-breeding...is it possible, or impossible? [Re: RogerRabbit]
    #9421256 - 12/12/08 09:31 PM (15 years, 1 month ago)

The term "hybrid" can be used to refer to both interspecies and intraspecies crosses.

Crossing unrelated strains can create hybrid vigor in the first generation(F1).


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OfflineRogerRabbitM
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Re: Cross-breeding...is it possible, or impossible? [Re: HerbBaker]
    #9421374 - 12/12/08 09:56 PM (15 years, 1 month ago)

So you're saying a child born to a man from Chicago and a woman from Peoria can be referred to as a hybrid?  Interesting.  The silly assed names given to 'strains' seems in most cases to be based on nothing but the geographical location the original print was taken from.  I know of no mycologists who would consider intraspecies crosses to be hybrids.
RR


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OfflineEmperor Reishi
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Re: Cross-breeding...is it possible, or impossible? [Re: RogerRabbit]
    #9421485 - 12/12/08 10:20 PM (15 years, 1 month ago)

Quote:

RogerRabbit said:
Crossing species, which would result in hybrids is another story.  It's been repeatedly done by professional mycologists, but so far all but one of my attempts has failed.
RR




what two species did you successfully create a hybrid of?


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Looking for:    Live Cultures: Maitake[sclerotia forming], Wood Ear[Auricularia Polytricha],            Prints: **Psilocybe mexicana,                                Reishi [other than red], Bioluminessent                                                                          Cubes & Pan Cyans

Available for trade:    Live cultures: Reishi[ganoderma lucidum], Shiitake[lentinula edodes], Chicken of the woods[Laetiporus                                            sulphureus], Enoki [Flammulina velutipes], Oyster [Pleurotus ostreatus], Lion's Mane [Hericium                                                            erinaceus]


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OfflineRogerRabbitM
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Re: Cross-breeding...is it possible, or impossible? [Re: Emperor Reishi]
    #9421501 - 12/12/08 10:24 PM (15 years, 1 month ago)

Cubes and Pan cyans. Three or four years ago.

Cube mycelium on the left, Pan cyan on the right.  The third sector at 12 o'clock is the cross.
RR


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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
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Offlinemisled
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Re: Cross-breeding...is it possible, or impossible? [Re: RogerRabbit]
    #9421579 - 12/12/08 10:37 PM (15 years, 1 month ago)

RR did you grow out the hybrid? what traits did it have? how did it fruit or did it not fruit at all?


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OfflineEmperor Reishi
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Re: Cross-breeding...is it possible, or impossible? [Re: RogerRabbit]
    #9421581 - 12/12/08 10:37 PM (15 years, 1 month ago)

That's very imnpressive. What protocol did you use to accomplish this and do you have any prints you'd like to give me for Christmas :smile: heheh


--------------------
EmperoR ReishI

Looking for:    Live Cultures: Maitake[sclerotia forming], Wood Ear[Auricularia Polytricha],            Prints: **Psilocybe mexicana,                                Reishi [other than red], Bioluminessent                                                                          Cubes & Pan Cyans

Available for trade:    Live cultures: Reishi[ganoderma lucidum], Shiitake[lentinula edodes], Chicken of the woods[Laetiporus                                            sulphureus], Enoki [Flammulina velutipes], Oyster [Pleurotus ostreatus], Lion's Mane [Hericium                                                            erinaceus]


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InvisibleHerbBaker
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Re: Cross-breeding...is it possible, or impossible? [Re: RogerRabbit]
    #9421625 - 12/12/08 10:47 PM (15 years, 1 month ago)

Quote:

RogerRabbit said:
I know of no mycologists who would consider intraspecies crosses to be hybrids.
RR




You should talk to Workman.
He has documented hybrid vigor in unrelated strain crosses.
If there was more crossing of mushroom strains taking place you would see term hybrid used more often.

Anything considered a cross-breed can be considered a hybrid, and that has nothing to do with Chicago..


You get any viable offspring from your "hybrid"?


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OfflineRogerRabbitM
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Re: Cross-breeding...is it possible, or impossible? [Re: HerbBaker]
    #9421686 - 12/12/08 10:59 PM (15 years, 1 month ago)

Yes.  The prints are viable, but the fruits far more resemble cubes than pans, with the exception that there's no veil.  One of these days I'll get the cultures out and work with them again.  My move last spring put everything on hold.
RR


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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison


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InvisibleHerbBaker
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Re: Cross-breeding...is it possible, or impossible? [Re: RogerRabbit]
    #9422036 - 12/13/08 12:17 AM (15 years, 1 month ago)

Cool! got any pics?

Here is one of many examples of the term hybrid being used for an intraspecies fungi cross.

http://www.freepatentsonline.com/PP05773.html

"The A. brunnescens isolates (D26 and L81WB) were from The Pennsylvania State University Mushroom Culture Collection (PSUMCC). The novel hybrid line was produced by crossing homokaryons derived from a golden white parent (D 26 , commonly used in cave culture) and from a smooth white parent (L81WB, commonly used for commercial cultivation)."



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Offlinetr8orjohn
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Re: Cross-breeding...is it possible, or impossible? [Re: HerbBaker]
    #9422206 - 12/13/08 12:56 AM (15 years, 1 month ago)

Ok, so *thats* why you used venomated agar to revive the red boy genetics...makes much more sense now

I am also curious how you crossed the cubes and pan cyans, if you care to share your protocol

jon


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Microbiologist turned mycologist, but still amateur enough to enjoy the occasional surprise...

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Terry M said:
Looking for rules that other people use successfully. Sterile procedure is just a set of rules that work.


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OfflineRogerRabbitM
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Re: Cross-breeding...is it possible, or impossible? [Re: tr8orjohn] * 1
    #9425402 - 12/13/08 04:48 PM (15 years, 1 month ago)

I'm going to hold off on the hybrid until I get time to work with it again.  I've moved into legal edibles only, and am working on a deal with local government to get all the recycle bin paper material from our county and the neighboring counties to set up a commercial mushroom farm.

The recycling plant that was accepting it has shut down, and they're now dumping several tons of clean paper and cardboard per week into the landfill.  If things work out, they'll be paying me $10/ton to accept the paper, and they'll deliver it right to my facility.  The county is working on getting me a grant and 100 acres of county land appropriated for this project.  That's about all I have time for right now.

As for calling an intraspecies cross a hybrid, I've seen it done many times, but I must respectfully disagree.  However, it's not worth arguing over.  Workman and I have had the discussion in this forum before a year or two ago.  I simply disagree that's all.  It's just a term anyway.  In my opinion, 'strains' are more like races or breeds, thus I also wouldn't consider a cross between a German Shepard and a Collie to be a hybrid.  It's just my opinion, that's all.
RR


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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
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OfflineAlan RockefellerM
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Re: Cross-breeding...is it possible, or impossible? [Re: RogerRabbit]
    #9427071 - 12/13/08 09:36 PM (15 years, 1 month ago)

Quote:

The prints are viable, but the fruits far more resemble cubes than pans, with the exception that there's no veil.





Cool, got any pictures?


Quote:

My biggest success with snake venom was to use it to degrade the cell walls of some 20+ year old spores that flat-out would not germinate. The spores were placed on venomated agar and monokaryotic mycelium from a different strain was allowed to crawl across the spores.  The result was a 'pairing' which then led to dikaryotic mycelium, and the result is still circulating as the Redboy strain. 




Did you have some control dishes that had monokaryotic mycelium crawling over the 20+ year old spores which failed to pair?


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OfflineRogerRabbitM
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Re: Cross-breeding...is it possible, or impossible? [Re: Alan Rockefeller]
    #9427608 - 12/13/08 11:32 PM (15 years, 1 month ago)

Quote:

Cool, got any pictures?




Yes, but I'm going to sit on them for the time being.  They look too much like cubes, and I frankly don't want to see the inevitable arguments over whether they really are a hybrid or not play out here.  When I find the time and inclination to start working with this again, I'll send them to Accugenics for DNA testing for the final word.

Quote:

Did you have some control dishes that had monokaryotic mycelium crawling over the 20+ year old spores which failed to pair?




Yes.  None of the dishes without snake venom resulted in pairing.  After the initial crossing, I repeated the experiments with several strands of monokaryotic mycelium from the cross, resulting in what I assume to be a 75% Redboy/25% PR, although I'm no geneticist so I can't be sure it's truly 75/25.  A few hundred of those prints were sent out to shroomery members at the time.
RR


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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison


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Offlinerungi
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Re: Cross-breeding...is it possible, or impossible? [Re: RogerRabbit]
    #9427720 - 12/13/08 11:57 PM (15 years, 1 month ago)

Crossing species, which would result in hybrids is another story.  It's been repeatedly done by professional mycologists, but so far all but one of my attempts has failed.
RR

But then you said you crossed a cube and cyan was that fist post a mistake.


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OfflineRogerRabbitM
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Re: Cross-breeding...is it possible, or impossible? [Re: rungi]
    #9427961 - 12/14/08 12:56 AM (15 years, 1 month ago)

No.  Read my post that you quoted again.
RR


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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison


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InvisibleHerbBaker
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Re: Cross-breeding...is it possible, or impossible? [Re: RogerRabbit]
    #9429631 - 12/14/08 10:58 AM (15 years, 1 month ago)

lol.its all about limiting confusion.:grin:


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Offlinerungi
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Re: Cross-breeding...is it possible, or impossible? [Re: RogerRabbit]
    #9429730 - 12/14/08 11:20 AM (15 years, 1 month ago)

Excuse my ignorance, but I do not understand
Are these definitions accurate?

1. A hybrid is a cross between dikaryotic mycelia of two different species creating  a third dikayon the hybrid.

2. A hybrid is a cross between dikayotic mycelia of two different strains of the same species creating a third dykaryon the hybrid

3. RR says some clones sector. and can be divided into several different non-sectoring dikaryotic mycelia.  These fruits could be called hybrids using the definiton number 2 above.


Rabbit,  The fist definition is how you crossed the cube and cyan.  Your fist post or second post says you have failed at doing this.  I did re read.


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OfflineRogerRabbitM
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Re: Cross-breeding...is it possible, or impossible? [Re: RogerRabbit]
    #9429829 - 12/14/08 11:40 AM (15 years, 1 month ago)

Quote:

RogerRabbit said:
It's been repeatedly done by professional mycologists, but so far all but one of my attempts has failed.
RR




Bold for emphasis on the 'but one'
RR


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semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison


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Offlinerungi
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Re: Cross-breeding...is it possible, or impossible? [Re: RogerRabbit]
    #9436244 - 12/15/08 11:51 AM (15 years, 1 month ago)

I see sorry for my language weakness


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OfflineLittle Burdette
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Re: Cross-breeding...is it possible, or impossible? [Re: RogerRabbit]
    #28568404 - 12/04/23 02:28 PM (1 month, 23 days ago)

Good day. I was researching the Pan/Cube cross possibility, and ran into this old post. Do you have viable prints I could acquire ? I am interested in seeing how this one fruits. I would also like to know if you have had the fruit analyzed? This is very intriguing to me.

Thank you.


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OfflinePnin
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Re: Cross-breeding...is it possible, or impossible? [Re: Little Burdette] * 2
    #28568505 - 12/04/23 04:05 PM (1 month, 23 days ago)

14 year necro to ask for spores :kimjongil:


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