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bodhisatta 
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Quote:
Jakeoncid419 said: Even if they were both the same species (doubtful imo) both parents were dikaryotic Cultures so the venom reagent Would still have had to have done it’s job in order for breeding to take place PHV looks nothing like either parent culture and like I said I’ve ran both parent cultures at least 50 times each and have never gotten anything close to the PHV phenotype from them only the PHV hybrid produce is purple spores
People have been making crosses by putting two dikayotic cultures on the same plate with no venom. You don't need Venom to make varietal hybrids or even monokaryotic mycelium.
There's cubes with purple spores, brown spores, albino spores, and even cubes that look like dicks. All same species but big phenotype difference.
Alan knows more about speciation than I do. what I do know is venom and monokaryotic cultures are absolutely not necessary to cross varieties of the same species
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Jakeoncid419
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Re: Snake venom agar tek [Re: bodhisatta]
#26950965 - 09/23/20 12:07 PM (3 years, 4 months ago) |
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Well first of all they are not the same species, Second of all I have never had success trying to mate cultures by just throwing two Dikaryotic cultures on a dish next to each other, I did that along side my venom dishes as a control and you could clearly see the division between the growth of the two cultures I cut it out on the divide anyway transferred and grew out and the result was one of either parent strain, no cross or alteration of traits happened. A Dikaryotic culture Has already made clamp connections so it would not be able to make these connections with a different culture. I can’t say it’s 100% impossible because as of a year ago breeding different pan species was “impossible” But I have never seen it done and I don’t understand how it could happen either. However Most of my hybridization work is done with pans. cubes I’ve always just made monocultures and they have bred super easily for me so I haven’t really tried mating Dikaryotic cubes. However I have accidentally mixed two different cube strains of spawn in a tub together and I did not get hybrid fruits I got a mix of both strains, I’ve also seen other people do the same thing and get the same results I don’t see why it would be any different from trying to mate them on a dish. Again not saying it’s impossible however if clamp connections have already been made I don’t understand how the genetic material would exchange
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Mycostotle
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1 gram of western diamond back rattlesnake venom has been ordered now
Im doing this out of curiosity with very little expectations, the venom will used to attempt hybridize cubensis with semilanceata and the results will be posted here
I think I have a source for semilanceata spores
I have these cubensis trains: ape, pe6, red boy, mckennai, b+, GT, melmac and mexican
Choose the one I should use in my attempts 😆
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
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Quote:
Jakeoncid419 said: Well first of all they are not the same species, Second of all I have never had success trying to mate cultures by just throwing two Dikaryotic cultures on a dish next to each other, I did that along side my venom dishes as a control and you could clearly see the division between the growth of the two cultures I cut it out on the divide anyway transferred and grew out and the result was one of either parent strain, no cross or alteration of traits happened. A Dikaryotic culture Has already made clamp connections so it would not be able to make these connections with a different culture. I can’t say it’s 100% impossible because as of a year ago breeding different pan species was “impossible” But I have never seen it done and I don’t understand how it could happen either. However Most of my hybridization work is done with pans. cubes I’ve always just made monocultures and they have bred super easily for me so I haven’t really tried mating Dikaryotic cubes. However I have accidentally mixed two different cube strains of spawn in a tub together and I did not get hybrid fruits I got a mix of both strains, I’ve also seen other people do the same thing and get the same results I don’t see why it would be any different from trying to mate them on a dish. Again not saying it’s impossible however if clamp connections have already been made I don’t understand how the genetic material would exchange
You do know there's more than a couple shroomery made crosses that are circulating and some are even sold by vendors now that were made by people putting spores of two varieties on a petri dish than carefully breeding the resultant cross back with itself over several generations to stabilize.
Ask pasty where rustywhyte came from. Look up mudafuka's ESS. Etc..
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Jakeoncid419
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Re: Snake venom agar tek [Re: bodhisatta]
#26951448 - 09/23/20 04:47 PM (3 years, 4 months ago) |
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Well sure when you put spores on to a petri dish there is a chance that you will get two different spores to land side-by-side thus the monocultures will clamp connect with each other. it’s success rate is low because most of the time the spores cluster and they breed with themselves before they are able to breed with the other. But that’s not putting too Dikaryotic cultures That have already clamped side by side on a dish and breeding them. . you can ensure this will happen by making serial dilutions spreading the spores out then transfer the mono growth before it clamps to anything, The only difference here is you are lucking out and landing two different spores close enough that they clamp with the other strain before anything else
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bodhisatta 
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people put spores on opposite sides of dishes. There's nothing monokaryotic left after a milliliter of growth with a drop of spore solution let alone a few centimeters
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Alan Rockefeller
Mycologist

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Re: Snake venom agar tek [Re: Mycostotle]
#26951492 - 09/23/20 05:19 PM (3 years, 4 months ago) |
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Quote:
Mycostotle said: Im doing this out of curiosity with very little expectations, the venom will used to attempt hybridize cubensis with semilanceata and the results will be posted here
I think I have a source for semilanceata spores
Sounds like a good experiment!
PM me if your semilanceata spore source doesn't come through.
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Jakeoncid419
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How are they clamping if they are already clamped? And they why couldn’t you just mix different types of spawn together to make crosses if this works? I’m not saying I don’t believe you I’m just curious as so how that is possible without clamps. It’s also interesting than pans will not do this at least not that I have seen. Making serial dilutions is pretty easy and you can then verify with a scope that it worked, that’s y I’ve done more traditional mating with pans because it’s easier to verify
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Alan Rockefeller
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Perhaps if they are already clamped they don't need to clamp more, and can exchange genetic information.
Maybe if they are already dikaryotic they are less likely to cross, but still do occasionally.
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Jakeoncid419
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How would that work tho? When I put two different Dikaryotic cultures On a dish I can see the clear division where the two cultures meet it’s not like they meld into each other... Perhaps certain cultures naturally suffer from weak cell membranes and “venom bleed effect” happens on its own
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Alan Rockefeller
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Quote:
Jakeoncid419 said: How would that work tho? When I put two different Dikaryotic cultures On a dish I can see the clear division where the two cultures meet it’s not like they meld into each other... Perhaps certain cultures naturally suffer from weak cell membranes and “venom bleed effect” happens on its own
Maybe sometimes a third mycelium emerges where the two meet.
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Jakeoncid419
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It would be interesting to watch it breed like this under a scope
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bodhisatta 
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How do all the strains you get when you shoot tens of thousands of spores into a single cake or grain jar get along? They don't know they're different if they're compatible mating types.
MS grows become homogeneous substrates not just little islands of individual strains.
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Jakeoncid419
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Re: Snake venom agar tek [Re: bodhisatta]
#26951605 - 09/23/20 06:15 PM (3 years, 4 months ago) |
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Well like I said they mate with each other when you inject the cake, this still does not explain how two Dikaryotic cultures would mate
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Jakeoncid419
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Also the cultures are somewhat divide with MS grows that’s why isolated Dikaryotic cultures do a better job of putting up full canopies
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jason9086
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Quote:
Alan Rockefeller said:
Quote:
jason9086 said: yes, snake venom breeding seems like a load of bullshit to me. What kind of scientist literally does no genetic analysis of their supposed hybrid to confirm it? I think RR drank the kool aid on that one, and a large number of people on the shroomery followed suit.
What sort of genetic analysis would be used to confirm it?
And what should be looked for in the sequence data to confirm that they have crossed?
The PHV cross has a spore size that is in between the size of the spores of the two parent species, spore color is in between as well. The fruits look macroscopically much different than either parent.
PHV spores measure (10.4) 11.5 – 12.9 (13.3) × (6.5) 9.2 – 11 (11.4) µm, which is midway in between the spore size of Panaeolus tropicalis and Panaeolus cyanescens.
PHV parent 1: https://images.mushroomobserver.org/1280/949765.jpg PHV parent 2: https://mushroomobserver.org/images/1280/1150852.jpg PHV fruits: https://mushroomobserver.org/images/1280/1188752.jpg
Well we would first have to establish a marker. For instance, say the its region is distinguishable between all the nuclei involved which it likely is at least in the spacer regions, then you could sequence this of both patents and resultant sector to see if there is a new pair of nuclei (one from each parent dikaryon) or if a marker cant be established perhaps more intensive next gen sequencing would be in order. Maybe mat genes would be good as well. but i think just seeing a new sector pop up and assuming breeding occured (when i am unsure is even possible with two dikaryons, anyone with knowledge of published research on this please inform me) is not very solid.
I dont know though.. im not an expert on basidiomycetes so info on this would be quite helpful to me. Im trying to figure out the process of these so called dikaryotic crosses.
And just to be clear, im not saying it didnt happen, im saying the evidence isnt clear. I want someone to dunk on me with info. The PHV cross was done with dikaryons? Whats the history on that one?
Edited by jason9086 (09/23/20 10:53 PM)
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Alan Rockefeller
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Re: Snake venom agar tek [Re: jason9086]
#26952086 - 09/24/20 12:51 AM (3 years, 4 months ago) |
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Quote:
jason9086 said: The PHV cross was done with dikaryons? Whats the history on that one?
https://mushroomobserver.org/404923
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Solipsis
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Fascinating! I'm looking into this but it seems to be possible when there is 1) vegetative compatibility in the first place [,similar enough cytoplasm and lots more] ;
then 2) also compatible interactions between mating factor expressions;
yet 3) having different mating types, and then bifactorial fungi should have 25% compatibility.
Apparently it is called hyphal anastomosis fusion, I would love to get a clearer visual image of the dance the nuclei perform in such an encounter.
But just scratching the surface here i may easily be wrong... idk what is required more exactly that makes it possible between those Pans but generally imposes limits on crossing of (e.g. cube variety) dikaryons being fine requiring no monokaryon at all although having but 25% odds. I cannot find the reason why you couldn't just cross isolates of different varieties of e.g. cubes with those odds - at first glance that seems to check all the boxes but I guess there is a lot going on and one of the compatibility tests prohibits it.
It has apparently been tested in oyster species and resulted in hybrids - somehow or another considerable difference could make it quite favorable. Perhaps also because in other words species relatively related to each other but growing geographically isolated from each other do not have the same incompatibility safeguards that are in place to carefully regulate incompatibility of relatively much more related strains of the same species?
Hoping to get insights from a more knowledgeable friend in the coming days.
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