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HYBRIDIZATION


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#1 waylitjim

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Posted 03 November 2005 - 07:26 PM

The #14 TEO was mailed out several days ago. It is the Issue that has Rodger Rabbit's article on how he brought the original 20 year old spore samples back.......a new novel method using Rattlesnake Venom!


I don't know if RR followed this lead or not, but here is some interesting
research about using snake venom as a hybridization agent.

We used purified snake venom from the Western Diamondback Rattlesnake (Crotalus atrox) for our hybridization techniques. The snake venom is added to the agar medium in quantities that alters the growth but does not prove toxic to the strain in question. This range of snake venom is from 10 mg to 30 mg per 300 ml of agar medium. The venom is not heat stable and must be added aseptically after sterilization of the medium. The agar used for this hybridization consisted of malt extract, activated carbon, minerals and humus – the carbon-rich ash residue from a coal burning industrial process. Other agars could probably be used as well. This just happens to be our production agar that we use everyday, and once we found that it also worked with the snake venom for hybridization.

HYBRIDIZATION TECHNIQUE:

Petri dishes of this R7 agar medium are inoculated with mycelium from two different strains of the Cordyceps genus. These different strains when inoculated together onto one petri dish will normally grow towards each other until they almost meet, at which point they form a zone of inhibition, where neither strain can grow. Eventually, one strain may prove stronger than the other and overgrow the plate, but they will remain genetically distinct; two different cultures residing in the same petri dish.

With the addition of a sufficient quantity of snake venom to the agar, we found that what happens is the two cultures grow towards each other until they meet and form their mutual zone of inhibition. This period of inhibition is short lived however, for in only about 2 or 3 hours the colonies each start sending out mycelial strands into this no-mans land, the zone of inhibition. These strands grow together and exchange nuclear material through their venom-weakened cell walls. They form a hybrid strain at this point of mutual contact. A new strain, one that is distinctly different from either of the parent strains. Within about 4 hours after first forming the zone of inhibition, the hybridization is complete and the colonies resume rapid growth towards each other. They become three colonies rather than the original two. There then exist in the same plate the original two colonies and a genetically distinct third…The Hybrid.

A section of the newly formed hybrid is carefully removed from the original zone of inhibition at the precise time that the colonies begin to fuse. That is during hour 3-4 after the initial meeting of the colonies. The hybrid is transferred to a new petri dish containing normal (non-snake venom) agar.
Our quick method of determining hybridization is to inoculate a new dish containing normal agar with all three strains, the original two and the suspected hybrid. If the hybridization has in fact taken place, these are now three distinct colonies, and will form a mutual three-way zone of inhibition. If hybridization has failed to occur, then the suspected hybrid will readily fuse with either one or the other of the original colonies. This proves that our suspected hybrid is not genetically distinct from the original and we start anew.

Once a hybrid is confirmed, it is tested for growth parameters. If it appears to be a vigorous and hardy grower on our substrate of choice, we grow out a quantity of mycelium, harvest it and analyze it for active ingredients. Through repeated testing in this way we were able to create the hybrid strain shown in Plot 6; a hybrid strain that is easily grown in solid substrate culture, with a potency greater than any other cultivated strain and at least equal in potency to the highest quality wild Cordyceps.

http://mycotopia.net...=1&d=1131063965

John C. Holliday PhD
Phillip Cleaver BS
Megan Loomis-Powers BS

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#2 Hippie3

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Posted 03 November 2005 - 08:00 PM

there is a rather significant difference between this
and what rodger did.
rodger's claim is that the living tissue's DNA was able to fuse with
essentially dead DNA.
their example was two mycellial cultures
whereas rodger's was 1 mycellial culture + dead spores.

#3 waylitjim

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Posted 03 November 2005 - 08:14 PM

Perhaps a similar technique was used to cross Penis Envy and Tex to create PE6.

#4 suckerfree

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Posted 03 November 2005 - 09:12 PM

In the new TEO that Peele puts out, it claims to be rattlesnake.

#5 Omega 11

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Posted 03 November 2005 - 09:50 PM

That is so crazy. No way I'll be able to create my own hybrid for the simple reason that snakes make me practically shit my pants!!!

#6 Guest_Peter Cottontail_*

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Posted 03 November 2005 - 10:25 PM

Dead spores? Hardly.
They were old and no longer viable for germination, but not dead.
RR

#7 inexorable

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Posted 03 November 2005 - 11:08 PM

I've got that old brain going again--LOOK OUT!
Now, does this tech work only for fungi of the same species, or genus, I'm not sure which I'm getting at. (IE: Cordyceps genus vs. Cordyceps genus OR? cubensis vs. cubensis). If the answer is no, then we need to start getting an Azur. vs. Cubie experiment going and try to get a potent/ simple growing MADNESS EXPLOSION UNSTOPABLE strain going!!!:headbang: :greenboun :space: :space: Sorry, got excited. What thinks yee, Mycotopia??:eusa_thin Probably won't work--don't get the hopes up...
--inex.:reb:

#8 Hippie3

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Posted 04 November 2005 - 06:52 AM

Dead spores? Hardly.
They were old and no longer viable for germination,
but not dead.
RR


a seed that won't germinate is dead.
you put plenty of water in it,
it still never germinated.
why ?

if it was not dead,
why didn't it germinate ?

all life requires water.
your 20 year old bone-dry spores ran out of water long ago,
cellular activity stopped and what was left is
essentially a fossil.
if it had still lived
it should have responded to the water
and germinated but it did not.

i understand that you claim differently,
i just have not seen any proof.

#9 Guest_Peter Cottontail_*

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Posted 04 November 2005 - 08:08 AM

Snake venom has been used to cross different species as well as strains. We also now know that the mycelium itself is capable of penetrating into a spore to obtain the genetic material necessary to become dikaryotic. This tek might work where direct mycelium to mycelium hybridization has not. The possibilities are endless.
RR

#10 waylitjim

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Posted 04 November 2005 - 12:16 PM

Snake venom has been used to cross different species as well as strains. RR

It would be such a breakthru to cross species. Panaeolus cyanescens
and Psilocybe cubensis would be such an excellent hybrid. If you could get
the size of cubensis and the potency of Pans, that would be a huge of
advancement in psilo-mycology. Pan Goliath x South American!

One question, if a woodlover was crossed with cubensis, what would be the
ideal fruiting substrate? I guess you'd need to mix manure and woodchips.
I'd love to see what an interspecies hybrid would looks like, and how stable
it would be, whether it would drop spores etc...

#11 shedthemonkey

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Posted 04 November 2005 - 05:28 PM

Silly Jim! Haven't you heard that song by Journey? "Pig and Elephant DNA, Just Won't Splice!"

Since you guys are being all Mad-Scientist I thought I would just throw that from one of my favorite classic SouthPark episodes! :)

#12 vitamin_d_added

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Posted 05 November 2005 - 09:32 AM

Silly Jim! Haven't you heard that song by Journey? "Pig and Elephant DNA, Just Won't Splice!"
Since you guys are being all Mad-Scientist I thought I would just throw that from one of my favorite classic SouthPark episodes! :)


I thought that song was by "Loverboy"! Ha!

#13 fucgubarn

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Posted 05 November 2005 - 01:31 PM

Most mushroom species posses two mating-type loci that control their breeding. The genes encoded in the A loci lead to the formation of transcription factors that belong to the class of homeodomain proteins.
Active transcription factors are formed by heterodimerization of two proteins of different allelic specificities. In nature, this is only the case if two cells of different mating type have fused to combine the different proteins in one cytoplasm. While fusion in homobasidiomycetes is found irrespectively of mating type, exchange of nuclei between mating mycelia is dependent on the products of the B mating-type loci. The B genes form a pheromone and receptor system that enables the fungi to initiate nuclear migration. The molecular details of the two genetic systems controlling breeding in basidiomycetes are presented in this review.

.... dont have the full article

#14 slp

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Posted 05 November 2005 - 04:39 PM

waylitjim.....your wood eating Ps. cubensis already exists. It is pictured with an
article. Reference #14 "TEONANACATL", November 2005. The Electronic Issue
of #14 "TEO" will be going up shortly at www.mushroomsfmrc.com
then, just click "TEO Journals" off of the main menu.....until then #13 "TEO" will
come up.... slp/fmrc

#15 inexorable

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Posted 07 November 2005 - 06:37 PM

Hmm, spore vendors shoud get on this Hybridization idea. Sounds like a great opportunity to start speciallzing in hybrid strains. Might be good for the community, too, if it actually started having some note worthy results or competition between hybrid vendors. Actually, this could get a bit rediculous as many hybrids wouldn't produce anything better than what nature already has. I just want an Azure/cubensis cross. I'd die happy.

#16 slp

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Posted 08 November 2005 - 12:20 AM

Species cannot be crossed. Cross-over can only occur if the number of
chromosomes match. A hybrid is a cross between two species, not of the
same species. True hybrids do naturally occur in the Amanita species, but
this is because many of the species have the same number of chromosomes.
There will be no Azure/cubensis cross. To cross any vendor's name of a
certain strain (Psilocybe cubensis) with another vendor's strain (Psilocybe
cubensis) does not produce a true hybrid. You are only crossing Psilocybe
cubenis with Psilocybe cubensis. What you get is Psilocybe cubensis...grin.
slp/fmrc

#17 Guest_freakachino_*

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Posted 08 November 2005 - 12:31 AM

What would be a really neat hybrid is a psilocye oyster mushroom. Or a Magic Morel. See if the magic can get hybrid into an edible that tastes good :D


*note, I don't mind the taste of psilocybes, I mention the above because its the taste most don't like.*

#18 waylitjim

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Posted 08 November 2005 - 12:38 AM

Species cannot be crossed. Cross-over can only occur if the
number of chromosomes match. A hybrid is a cross between two species,
not of the same species.


Snake venom has been used to cross different species as well as strains.
RR


So is it possible or not?

#19 Guest_Peter Cottontail_*

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Posted 08 November 2005 - 05:46 AM

Different Cordyceps species have been crossed. Whether it works with other species or not is doubtful, but under investigation. :reb:
RR

#20 Hippie3

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Posted 08 November 2005 - 08:23 AM

it bear mentioning
that some 'species' are very closely related,
and surely it's much easier to 'cross'
two closely related species that have only a few micro-differences
than to try to cross a tree with a fish.




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