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InvisibleQM33
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Intergeneric Breeding through Protoplast Fusion
    #27606499 - 01/04/22 09:17 AM (2 years, 4 months ago)

Seems to easy to me. Seems like corners could be cut on a micro observational level to grow visually different macro fungi with relative ease.

Tell me why I'm wrong.


And maybe I'm just to new to the concept, but the implications of stuff like this, even for plants, just seems phenomenal. Sure with it's limitations but I mean we're talking Ligers all over. And if things couldn't be crossed maybe it could be crossed with something so it could..?


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Edited by QM33 (01/04/22 09:19 AM)

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OfflineNichrome
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Re: Intergeneric Breeding through Protoplast Fusion [Re: QM33]
    #27606609 - 01/04/22 10:54 AM (2 years, 4 months ago)

:laugh2:

Easy.... Cutting corners will work out great for you.


Quote:

QM33 said:

Tell me why I'm wrong.






Sounds like a fun way to start a long lasting argument...


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InvisibleQM33
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Re: Intergeneric Breeding through Protoplast Fusion [Re: Nichrome]
    #27606646 - 01/04/22 11:22 AM (2 years, 4 months ago)

Do you know how it works?


You don't think you could skip the viewing of clamp connections and just regenerate and fruit, look for visuals signs of mated fruit and continue to self the hybrid through spore grows?

In this case the cutting of corners would probably lead to a bit more work, but it's potentially something you could do without a miscrope even?

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OfflinePsiloscoby
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Re: Intergeneric Breeding through Protoplast Fusion [Re: QM33]
    #27610446 - 01/07/22 12:51 PM (2 years, 4 months ago)

Didn't RR say rattlesnake venom could accomplish this? I could be getting terms mixed up.

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InvisibleQM33
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Re: Intergeneric Breeding through Protoplast Fusion [Re: Psiloscoby]
    #27620098 - 01/15/22 05:07 PM (2 years, 4 months ago)

I think Protoplast Fusion technically makes something a GMO so I suppose I'm backing off of that route ..

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InvisibleCreonAntigone
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Re: Intergeneric Breeding through Protoplast Fusion [Re: QM33]
    #27620103 - 01/15/22 05:11 PM (2 years, 4 months ago)

Quote:

QM33 said:
I think Protoplast Fusion technically makes something a GMO so I suppose I'm backing off of that route ..




If you shop at a regular grocery store, you eat GMO foods every day.

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InvisibleQM33
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Re: Intergeneric Breeding through Protoplast Fusion [Re: CreonAntigone]
    #27620125 - 01/15/22 05:24 PM (2 years, 4 months ago)

Totally. Most of our energy consumption destroys the planet. Our clothes pretty much come from slaves. Farmed to shit or overfished. I mean there's alot of things that just come natural to us this day in age that are far from appropriate imo. But that's a whole nether thread, my opinions. The fact stands! And I'm not even going to get into my new breeding ideas now.

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InvisibleRaven44
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Re: Intergeneric Breeding through Protoplast Fusion [Re: QM33]
    #27620272 - 01/15/22 07:53 PM (2 years, 4 months ago)

If I’m not mistaken a 3% fusant percentage could be tested without viewing by simply making up some petris of the solution and see if you get any growth or not

I could def be wrong tho

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InvisibleCreonAntigone
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Re: Intergeneric Breeding through Protoplast Fusion [Re: QM33]
    #27620417 - 01/15/22 10:57 PM (2 years, 3 months ago)

Quote:

QM33 said:
Totally. Most of our energy consumption destroys the planet. Our clothes pretty much come from slaves. Farmed to shit or overfished. I mean there's alot of things that just come natural to us this day in age that are far from appropriate imo. But that's a whole nether thread, my opinions. The fact stands! And I'm not even going to get into my new breeding ideas now.




I think it is relevant because the process of making a GMO just quickly replicates a process that can be done naturally over many thousands of years through lots of selective breeding.

Look up 'horizontal gene transfer'. This article may be very much of interest to you: Horizontal gene cluster transfer increased hallucinogenic mushroom diversity.

One doesn't need to be monsanto to see genes being modified, it happens all the time in nature. It just takes a long process of evolution.

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InvisibleQM33
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Re: Intergeneric Breeding through Protoplast Fusion [Re: Raven44] * 1
    #27620605 - 01/16/22 06:14 AM (2 years, 3 months ago)

Quote:

Raven44 said:
If I’m not mistaken a 3% fusant percentage could be tested without viewing by simply making up some petris of the solution and see if you get any growth or not

I could def be wrong tho




No your right, nichrome just thinks he knows something since he got a tag



And Creon, if I understand correctly, a GMO is one that cannot be replicated through nature
And although there are plenty of interspecific hybrids, I was going for a intergeneric hybrid, which would be behind the realm of possibility in nature. Cell fusion is not possible in nature. And when you start pushing walls like that, you leave potential for DNA and chemical synthesis to creat setting maybe so unnatural the earth just wasn't prepared for it, but again that's a philosophical debate for another thread. The GMO debate had plenty of spotlight other places I'm sure.



And I'll read the link, a quick skim seems to reveal it's right down my ally, thanks. And what your seeing primarily there, from a quick glance is interspecific. I don't want to get into my project yet, but maybe it's more along those lines;). Much more, organic feeling ha, more natural, more for lack of a better word, will of goddy.

But talking interspecific... I think things like spore capadability and chromosomes are going to play into it's potential ability to be fertile as well, which really just turns this whole thing into a super microscopic lottery, even moreso than it was ha. But it does happen in nature all the time, I've learned a bit about it. I mean all the Panthera species, lions tigers, jaguars, seem to have come from a common ancestor and even seem to have re shaded their DNA inbetween them selves at points, back crossing basically, but never stabilized.

I know I don't need to be Monsanto, that was my point. A heads up to everyone who considered Protoplast Fusion, it's not natural, at all.

That snake venom cell wall shit, idk about that. RR seemed to believe it could lyz a cell wall. It might be able to, I would think it would also kill the cells.. idk. I didn't do any research on snake venom ha. There is Cellulase available however.

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OfflineHotMess
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Re: Intergeneric Breeding through Protoplast Fusion [Re: QM33]
    #27623888 - 01/18/22 06:34 PM (2 years, 3 months ago)

Quote:

QM33 said:
Seems to easy to me. Seems like corners could be cut on a micro observational level to grow visually different macro fungi with relative ease.





i've read this like six times and can't figure out what you mean.  is it fusing cells from different genera of fungi to create a new genus?  what do you mean "corners could be cut on a micro observational level?"

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InvisibleCreonAntigone
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Re: Intergeneric Breeding through Protoplast Fusion [Re: HotMess]
    #27624092 - 01/19/22 12:38 AM (2 years, 3 months ago)

Quote:


And I'll read the link, a quick skim seems to reveal it's right down my ally, thanks. And what your seeing primarily there, from a quick glance is interspecific.





It deals primarily with intergeneric actually. It sequences three fungi of three genera - Psilocybe cyanescens, Gymnopilus dilepis, and Panaeolus cyanescens. It suggests there was horizontal gene transfer (HGT) of the same gene cluster between them. They have a bold theory, but if it is true it suggests information travels between mushroom genera without breeding.

My suggestion would be that HGT would be a potential mechanism to achieve some of these things you talk about without breeding.

Quote:

Phylogenetic analyses of PS homologs from a local database of 618 fungal proteomes yielded congruent gene tree topologies with respect to PS+ taxa, and clades of clustered PS genes from all gene trees excluded the PS− taxa in the database, suggesting the clustered genes are coordinately inherited (Figs. 3 and S2A–E). The gene trees also suggest HGT of the cluster from Psilocybe to Panaeolus and HGT of most PS genes between Atheliaceae and Agaricaceae when compared to a phylogenomic tree of related Agaricales (Fig. 3)




I may be misinterpreting the article, someone correct me if I'm wrong, but the suggestion seems to be that this particular cluster they exained for manufacturing psilocybin was transferred from Psilocybe to Panaeolus NOT by breeding but by HGT/

It also suggests HGT between Agaricaceae and Atheliaceae - two different families, even greater separation than two genera.

Tree from the article; on the right several lines marked in blue are putative HGT.


Edited by CreonAntigone (01/19/22 12:45 AM)

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InvisibleQM33
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Re: Intergeneric Breeding through Protoplast Fusion [Re: HotMess]
    #27624210 - 01/19/22 06:32 AM (2 years, 3 months ago)

Quote:

HotMess said:
Quote:

QM33 said:
Seems to easy to me. Seems like corners could be cut on a micro observational level to grow visually different macro fungi with relative ease.





i've read this like six times and can't figure out what you mean.  is it fusing cells from different genera of fungi to create a new genus?  what do you mean "corners could be cut on a micro observational level?"





Okay so bassically how protoplastfusion works, iirc. You get your mycelium, add it to a cell lyzing solution you made, cellulase, driselase. The cell walls are removed from the mycelium cells. Strain the cell walls through a micron strainer, wash with water. Then you would add them to PEG, polyethylene glycol which is Iirc is what they use to promote fusion, electrodes can be used to polorize cells and encourage fusion as well. An osmotic stabilizer is added which pretty much just adjust the PH.
Finally regeneration occura when replated on a apparently sugar rich petri dish, sorbital or mannitol would be some good options there.

And now, nichrome, is where you would potentially be able to cut a corner. Here you would regularly take your microscope and look for clamp connections between the cells. Isolate the clamps, grow them out, wambam your done, genetic material was transfered between two genera AND YOU DIDNT EVEN USE YOUR MICROSCOPE!
Side note things seem to get tricky when you start talking about mols in measuring units of weigh. Thankfully I don't think I need to worry about this anymore, at least now, but I think that might have been my biggest hurdle, but it's probably just another equation.

Or you could just grow it out without doing that, hope you visually see the hybrid sporocarp, and continue to self/proof the genetics similarly from that point on.
Problem there is back breeds in the process if the two individual proto plast are not isolated, like spores.
So not only is it much more of a hoping game, but you could get something and then let it disappear to more sexuality before it fruits. But if you can get your spore/cell count to solution ratios you should be able to get a single cell/spore per .1microliter on average. You could potentially bypass the microscope and work solely on probability and vision.
I hope that makes since, I have a new procedure I'm working on and don't want to completely give it away.

There is a surprising amount of very nice information of Protoplast Fusion of fungi. Mainly bacteria for industrial purposes, which is cool. I was even telling my girl, well what's the difference between a GMO and a chemical? You know like why's it so bad? And she felt me there, but idk, we can't forget these organisms are alive, have been for thousands of years, and like us I believe have a purpose, and not one to be tampered with dramatically, especially when we put it in our bodies regularly. Fuck I really should not get into that stuff for sure ha.


--------------------
OmManiPadmeHum,OmManiPadmeHum, OmManiPadMeHum...
There are known knowns, there are known unknowns,
          there are also unknown unknowns.
With great privilege comes great responsibility.

Quantom Qups PROOF AND Soft Drops
Turn your Swab to a Syringe and Syringe to Multiple Syringes!
No Pours (QuantomStyal)
Magic Fruit Leather
DMT for IandI

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OfflineHotMess
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Re: Intergeneric Breeding through Protoplast Fusion [Re: QM33]
    #27624747 - 01/19/22 03:18 PM (2 years, 3 months ago)

i'm sorry but protoplast fusion is meddling with nature just like gmo.  personally i don't have a problem with genetically modifying things, but if you believe gmo is wrong, you should also believe fusing two cells from different genera is wrong. 

now, on to your thing, yes you could try using various techniques of cellular fusion on fungi of different genera, assuming you had the supplies and abilities.  what you would get from it might die instantly, be extremely fastidious and difficult to cultivate, or not fruit.  considering to do this you need to use individual cells, that's tons of work and supplies that, ultimately, probably won't pay off.

i'm not really understanding why you would want to do this or what your goal is, but the only reason it hasn't already been done is because molecular biologists pretty much ignore mushrooms.  they're trying to accomplish something in the simplest way possible which is almost never by using fungi, other than yeast.

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InvisibleQM33
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Re: Intergeneric Breeding through Protoplast Fusion [Re: HotMess]
    #27624813 - 01/19/22 04:22 PM (2 years, 3 months ago)

Proto plast fusion, cell fusion makes something GMO, I know, that my point

Might is half the battle. And probably won't pay off... Well I think that's subjective to input, the possibility is there....
?


Oh I know, most of the info I've been reading is in regards to bacterias.. but not all of it. Orchids seem to come up pretty often.

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