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7718


Registered: 12/21/16
Posts: 77
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Isolates, strains and genetics 1
#24126177 - 02/28/17 02:32 PM (7 years, 2 months ago) |
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Hey guys, I've got a project with ATL#7 that I'm working on right now where I intend to use agar plates to isolate the best sclerotia producer and the best mushroom producer. Once I have determined those two isloates, I want to 'breed' them and see if I can get both sets of genes into a single isolate. All of this thinking and planning has given rise to a few questions about genetics that I haven't been able to answer by searching.
Question #1: If I create an isolate, fruit and clone it...do I still have an isolate, or have I rerandomized everything? I know going back to spores rerandomizes, wondering if cloning does the same thing.
Question #2: In the image below, does AB1 = AB2?

Question 3: The images below show the first two steps in a hypothetical breeding program where I continually take the new growths from each side and breed them together. I'm wondering if doing this repeatedly would provide enough genetic homogeny to create a unique strain that produces spores that consistently behave and grow the same way...like with all the different cube strains.


Thanks for your help, guys. Forgive me if these are dumb questions...the more I learn the fewer dumb questions I'll have, but for now, they're all pretty suspect haha.
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7718


Registered: 12/21/16
Posts: 77
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Re: Isolates, strains and genetics [Re: 7718]
#24129744 - 03/01/17 08:10 PM (7 years, 2 months ago) |
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Bump
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Alan Rockefeller
Mycologist

Registered: 03/10/07
Posts: 48,392
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Re: Isolates, strains and genetics [Re: 7718] 1
#24129940 - 03/01/17 09:39 PM (7 years, 2 months ago) |
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If you clone a fruit, it will be the same individual. No genetic recombination happens until the spores germinate.
ATL7, P. tampanensis, P. mexicana, P. galindoi and various others are the same species - P. mexicana. You might want to try breeding with some of those as well. The way to breed them would be to make single spore isolates via serial dilutions, then mate those in a petri dish.
Edit: tampanensis and mexicana are two separate species. Atlantis is really tampanensis. Galindoi is really mexicana but what is going around the underground as galindoi is tampanensis.
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drake89
Mushroom Magnate



Registered: 06/26/11
Posts: 4,168
Loc: TN
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you can also try monokaryotic protoplast fusion. that's where you use chemicals to try and turn dikaryotic mycelium into monokaryons. it's more complicated and I could be wrong on the lingo but pop that into google scholar for some research.
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7718


Registered: 12/21/16
Posts: 77
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Re: Isolates, strains and genetics [Re: drake89]
#24132329 - 03/02/17 09:14 PM (7 years, 2 months ago) |
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A lot of good information here. Lots of reading to do...an entire different tangent on breeding I didn't even see coming haha. Monokaryotic protoplast fusion and single spore serial dilutions sound...extremely intense. I think I need to do a little bit more growing before I'll fit into my big boy pants.
Thank you guys, I appreciate it.
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BlueDisa
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Registered: 03/02/17
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Last seen: 14 days, 12 hours
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Quote:
Alan Rockefeller said:
ATL7, P. tampanensis, P. mexicana, P. galindoi and various others are the same species - P. mexicana.
What do you mean they are all the same species? Have we been misinformed for all these years to believe that these mentioned are all distinct species or does it just happen that they can crossbreed with each other? Is this because they are all in section mexicana?
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,891
Loc: Milky way
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Re: Isolates, strains and genetics [Re: BlueDisa]
#24133138 - 03/03/17 09:00 AM (7 years, 2 months ago) |
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They're all varieties of the same species. Like GT and B+ are varieties of cubensis.
It's common knowledge that those are all ps.mexicana. no one is misinformed. Some people are not informed
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drake89
Mushroom Magnate



Registered: 06/26/11
Posts: 4,168
Loc: TN
Last seen: 5 years, 2 months
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Re: Isolates, strains and genetics [Re: BlueDisa]
#24133141 - 03/03/17 09:01 AM (7 years, 2 months ago) |
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Quote:
BlueDisa said:
Quote:
Alan Rockefeller said:
ATL7, P. tampanensis, P. mexicana, P. galindoi and various others are the same species - P. mexicana.
What do you mean they are all the same species? Have we been misinformed for all these years to believe that these mentioned are all distinct species or does it just happen that they can crossbreed with each other? Is this because they are all in section mexicana?
Alan is big into PCR and taxonomy so I'd say he's done his homework, and found not much genetic difference between the species. There's not a great deal of consensus among academics on where exactly to divide up species. A lot of things that were different looking, and different species, ended up being lumped together after DNA analysis revealed them to be very similar. The opposite is true, different things that looked very similar can have wildly different genetics and are split into different species. I think there was some lumping and splitting among the morels for example.
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Alan Rockefeller
Mycologist

Registered: 03/10/07
Posts: 48,392
Last seen: 2 days, 19 hours
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Re: Isolates, strains and genetics [Re: drake89]
#24133181 - 03/03/17 09:16 AM (7 years, 2 months ago) |
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Quote:
BlueDisa said: What do you mean they are all the same species? Have we been misinformed for all these years to believe that these mentioned are all distinct species or does it just happen that they can crossbreed with each other? Is this because they are all in section mexicana?
We have been misinformed all these years. Turns out they have identical ITS sequences, and when the microscopy is checked no consistent difference between the species can be found.
Quote:
drake89 said: There's not a great deal of consensus among academics on where exactly to divide up species.
It has been a couple years since anyone has paid attention to this group. I've sequenced several cultivated sclerotia collections recently and all matched P. mexicana 100%. Alonso and I have a paper in press that synonymizes many names into P. mexicana.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,891
Loc: Milky way
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Ever since I've been growing over in mush cultivation it's common knowledge all the truffel producers are Psilocybe mexicana. So at least 4 years it's been not a misinformed thing
Most of us point to your research if I'm not mistaken, hah
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catnip40
xฬ็



Registered: 03/09/12
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Re: Isolates, strains and genetics [Re: bodhisatta]
#24135314 - 03/04/17 06:46 AM (7 years, 2 months ago) |
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leschampignons
Biochemistry + Mycology


Registered: 08/30/13
Posts: 1,586
Loc: NY/NJ/ME
Last seen: 3 days, 17 hours
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Quote:
Alan Rockefeller said:
Quote:
BlueDisa said: What do you mean they are all the same species? Have we been misinformed for all these years to believe that these mentioned are all distinct species or does it just happen that they can crossbreed with each other? Is this because they are all in section mexicana?
We have been misinformed all these years. Turns out they have identical ITS sequences, and when the microscopy is checked no consistent difference between the species can be found.
Quote:
drake89 said: There's not a great deal of consensus among academics on where exactly to divide up species.
It has been a couple years since anyone has paid attention to this group. I've sequenced several cultivated sclerotia collections recently and all matched P. mexicana 100%. Alonso and I have a paper in press that synonymizes many names into P. mexicana.
Do you have a link to this paper?
Edit: Last time I posted about this subject I wasn't sure
Edited by leschampignons (03/04/17 05:16 PM)
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Alan Rockefeller
Mycologist

Registered: 03/10/07
Posts: 48,392
Last seen: 2 days, 19 hours
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Quote:
leschampignons said: Do you have a link to this paper?
It has not been published so I have not put it online - but if you email me at my full username all lower case with no spaces at gmail, I will send it to you.
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leschampignons
Biochemistry + Mycology


Registered: 08/30/13
Posts: 1,586
Loc: NY/NJ/ME
Last seen: 3 days, 17 hours
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Thanks I will do that
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BlueDisa
Friend


Registered: 03/02/17
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Funny, how always all the mentioned species looked so different from each other to me and all I could see was their differences.
Since reading this post and comments a few days back I have looked online for mentioned species and now can only see their similarities.
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