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Intrepid
Stranger



Registered: 12/15/19
Posts: 83
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Back-crossing Tidal Wave 1
#27735907 - 04/15/22 12:32 PM (2 years, 1 month ago) |
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My last couple of grows have been disappointing in terms of potency. I use the shoebox technique exclusively and while the yields are quite good I find it very disappointing to go through all that effort only to end up with a bunch of beautiful but weak shrooms (I'm talking' to you, Amazon and Mazatapec). My new goal is to create and clone a strain which is easy to grow and predictably strong. Tidal wave seems to fit the bill so I recently obtained a spore syringe of that as well as PE.
My understanding is that this particular tidal wave is unstable I read that tidal wave can be back-crossed with PE and the stabilized over successive generations but I would like to get additional input before proceeding with the project.
1. Since this tidal wave is unstable should I assume that this is an F1 generation?
2. My preferred technique is to go spore syringe=>agar=>LC=>rye=>coir. My plan is to do 6 shoeboxes with 2 each of PE, tidal wave and a hybrid from a LC containing both strains. Would it be better to do 3 each of PE and tidal wave, clone tissue from the best looking fruits from each box and then use that to back cross the two strains? OR...
3. Should I just start with the tidal wave, evaluate the potency and then decide if it's worth it to back cross?
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Intrepid
Stranger



Registered: 12/15/19
Posts: 83
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Re: Back-crossing Tidal Wave [Re: Intrepid] 1
#27739055 - 04/17/22 05:05 PM (2 years, 1 month ago) |
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Anyone? Anyone? Bueller?
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Alan Rockefeller
Mycologist

Registered: 03/10/07
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Re: Back-crossing Tidal Wave [Re: Intrepid] 3
#27739291 - 04/17/22 07:44 PM (2 years, 1 month ago) |
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Use serial dilutions of spore solution to generate monokaryotic cultures of both Tidal Wave and PE and let them mate in petri dishes. Fruit each resulting cross and see if they have characteristics you like.
Here is a video I made on how to do serial dilutions:
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MycoWeek
Elementary Student



Registered: 07/05/21
Posts: 674
Loc: USA
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Re: Back-crossing Tidal Wave [Re: Intrepid] 1
#27739298 - 04/17/22 07:46 PM (2 years, 1 month ago) |
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aside from just doing it and isolate for the strongest one, no one can tell you which "strain" to back cross something with to get strong fruits.
you can try and see if you get strong fruits, or just grow a few different varieties and see if you can get something strong without doing the hard work.
Have you tried any True Albino Teacher varieties like Yeti or Ghost?
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Nillion
Nobody

Registered: 04/14/22
Posts: 1,002
Loc: Terra Firma
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Re: Back-crossing Tidal Wave [Re: Intrepid] 2
#27739700 - 04/18/22 05:00 AM (2 years, 1 month ago) |
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You can also use any dikaroyon culture (or fertile spore) of PE, Tidal Wave or B+ to fertilize any monokaryon of a compatible culture, which is one that has a different allele (gene) at the mating type locus (gene location).
Monokaryon + monokaryon mating is quite rare in nature, but if you practice serial dilution (as Mr. Rockafeller mentions) it is a simple matter.
Many cultures have distinct macroscopic forms for their monokaryon and dikaryon forms. Cubensis to the best of my knowledge cannot product rhizomorphic mycelia as a monokaryon.
Because spores can fertilize monokaryons as well, it is possible to mate strains by growing one as a monokaryon and then exposing the culture to fertile spores. This can give you more control than mon-mon mating attempts by allowing you to work with a single monokaryon culture and make several cross based upon it.
Most breeding attempts pursue a set goal, standard or seek to develop a previously identified a trait known in the population being developed.
Good luck.
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Intrepid
Stranger



Registered: 12/15/19
Posts: 83
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Re: Back-crossing Tidal Wave [Re: Nillion] 1
#27740740 - 04/18/22 06:36 PM (2 years, 1 month ago) |
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Thanks for the detailed responses. I have a pretty good science background so serial dilutions of spore solution makes sense. I don't have access to the lab equipment utilized in the video so I might have to put that particular technique on the back burner for now. Other people have talked about stabilizing a strain by growing from a spore print=>cloning tissue=>growing from a spore print, etc for seven generations or so.
I haven't tried Yeti or Ghost as they're not readily available from commercial sources. I decided to go with Tidal Wave thinking that I could further stabilize it with PE using the method I mentioned above.
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joze



Registered: 11/10/20
Posts: 1,056
Loc: PNW
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Re: Back-crossing Tidal Wave [Re: Intrepid] 2
#27742130 - 04/19/22 02:16 PM (2 years, 30 days ago) |
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You definitely don't need all of that equipment for a serial dilution. You can pull it off with a lot less stuff, you just might get a few contams in the mix. But that can be resolved with agar work down the road. The general principle of serial dilutions is rather easy.
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TheConfluence
Stranger



Registered: 12/13/21
Posts: 157
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Bless you!
-------------------- When two people dream the same dream, it ceases to be an illusion. Philip K. Dick Lies, Inc. (1984)
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trippleblack
Stranger

Registered: 12/01/19
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Last seen: 2 months, 5 days
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you get some really destroyed looking fruits. at least i did when i crossed tw2 with another dikaryon, strain was melmac. it threw out multiple recombinants -none where as good as the original, but some did produce spores from mutated fruits. i have not tried them yet, but they bruise heavy blue/green and are rock hard.
recombinant 1 - a few of these produced spores :
 
recombinant 2

I do have some B+ monospore and some ape monospores. the easiest route to take would be getting monospores of other strain and doing a di-mon cross. I have no idea how you are going to properly and confidently do this work without a compound microscope or a very strong stereo microsope. I had plenty of instances where the mycelium appeared to be a monokaryon, but scanned under magnification showed plenty of clamp connection.
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Mycoplex
Sporocarp



Registered: 10/09/21
Posts: 898
Last seen: 20 hours, 36 minutes
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Quote:
Alan Rockefeller said: Use serial dilutions of spore solution to generate monokaryotic cultures of both Tidal Wave and PE and let them mate in petri dishes. Fruit each resulting cross and see if they have characteristics you like.
Here is a video I made on how to do serial dilutions:
Hi Alan, thanks for sharing this video. I was wondering if you could recommend a lab pipettor (like the one you're using in the video) that might be accessible through Amazon or ebay.
A while back I used one of your recommendations for grabbing an entry level microscope so I wanted to follow suit with your suggestion for a tool that can be used for this.
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joze



Registered: 11/10/20
Posts: 1,056
Loc: PNW
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Re: Back-crossing Tidal Wave [Re: Mycoplex] 1
#27773124 - 05/11/22 08:02 PM (2 years, 8 days ago) |
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If you were to get a micropipette from a scientific supply company like Avantor, it'd cost $400-700 probably. I just did a search on Amazon and there are a bunch of similar products for much much cheaper. I can't attest to their precision relative to a micropipette from a more prominent brand, but I'm sure for the purposes of a hobby mycologist, these will do everything you need.
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Mycoplex
Sporocarp



Registered: 10/09/21
Posts: 898
Last seen: 20 hours, 36 minutes
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Re: Back-crossing Tidal Wave [Re: joze]
#27773290 - 05/11/22 10:12 PM (2 years, 7 days ago) |
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joze, thanks man. It looks like one of the variants of that product has a range of 10-100 microliters which would work with the workflow Alan described.
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joze



Registered: 11/10/20
Posts: 1,056
Loc: PNW
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Re: Back-crossing Tidal Wave [Re: Mycoplex] 1
#27773618 - 05/12/22 08:08 AM (2 years, 7 days ago) |
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You're welcome my friend. Yeah, a 10-100 uL micropipette is probably all you need to get by. It's nice having the different volumes of micropipettes, but since they're adjustable it's easy to make do with one.
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