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Muad.Dweeb
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Breed Like the Bene Gesserit - (AMA Mushroom Breeding) 20
#27865684 - 07/17/22 02:58 AM (1 year, 6 months ago) |
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Hi! I don't post here often, but I'm carving out the time to participate more. And yes, you profile crawlers, I have grown up a bit since my early posts, so humor me.
The past 4 years, I have been doing my own reading and testing of cube sexuality and patterns of inheritance, specializing in abnormal morphology. I have several unique (young) varieties as a result, most on the F3 or F4 at the moment, some in the F2 and F1 even. Some of these crosses were conducted just to confirm the method of crossing is effective. There are still several methods I haven't tested yet and some I haven't conducted enough to provide a compelling argument with substance.
My goal, and I believe I can achieve it, is to make novel and responsible mushroom breeding accessible to anyone with at least moderate competency in cubensis cultivation. This is important to me, and should be important to us all, I think, because as the domesticaters of the species, it's up to us now to care for their diversity needs in the genetics department. I can't be the only one to have noticed that some of these varieties have been inbred consistently for decades, right? Lots of unusual "mutations" lately, right?
And then we have, of course, Enigma. The ultimate sign to some people that the genes have degraded so much that it can't produce spore bearing structures (I dub these blobs "pseudocarps"). To others, Enigma is a mysterious, miracle mutation. I claim it is neither, and will shortly be demonstrating this. I already have one confirmed blob culture (I also had one I lost in a recent move), plus five in a test run right now to confirm they are actual pseudocarps, not environmental deformations. I will present the successful pseudocarps with my argument at a later date when I have all the data and can parse it for general consumption.
In full disclosure, I am writing a book on this concurrently. But since all of it started for me here, even if only bumbling blindly around the archives section for "teks" at first, I want the information itself as a consolidated resource for the community. Because we are all breeding if we realize it or not, with each spore print, every grow. Imagine what we could have if we were all doing it intentionally?
So, ask me anything. I'm no expert, but I'll try my best to offer real, accurate and meaningful information that you can try right now. I'll be checking back in every couple days to catch up with any questions.
Teks and Explainations:
* Multispore Crossing (The RustyWhype) * Three Streaking Methods * The Buller Phenomenon and Serial Dilution * Basic Mendelian Genetics (Inheritance) * Modified OG Print Tek (Small Scale) * In Vitro Pre-Germination * In Vitro BRF Cakes (The DubTube) * Recovering and Preserving Monokaryons/Basic Culture Hygiene 1 * Case Study 1: Observing Inheritance Between Nuclear Types * UV Mutagenesis (DIY UVO Box) (and a safer, cheaper version ) * Some Thoughts on Blobs (Meet Atodaso!) * Naming Conventions (How to Name Things) * Same Species Cross Contamination (Accidental Hybridization) * Ethics, an epistle * The Swinger Tek (Trauma Induced Promiscuity)
Edited by Muad.Dweeb (01/15/23 12:43 PM)
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Muad.Dweeb
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Re: Mushroom Breeder - AMA [Re: Muad.Dweeb] 3
#27865696 - 07/17/22 03:33 AM (1 year, 6 months ago) |
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 "Chimera" spores, "GOGH.Calico"
 Leucistic Gold Spore
 Gold spore x PF Red print from F1 fruit
 "Cypress" GOGH.Calico.F2.CL2
-------------------- A beginning is the time for taking the most delicate care that the balances are correct. This every sister of the Bene Gesserit knows.
How to Breed like the Bene Gesserit The Weirding Way - Advanced Bene Gesserit Techniques
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DERRAYLD
Constructus


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Re: Mushroom Breeder - AMA [Re: Muad.Dweeb] 2
#27865715 - 07/17/22 04:32 AM (1 year, 6 months ago) |
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I'm here for the mutations.
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fahtster
Now With 33%More Faht



Registered: 06/17/06
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Re: Mushroom Breeder - AMA [Re: DERRAYLD]
#27865719 - 07/17/22 04:37 AM (1 year, 6 months ago) |
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rumfor69
Bodhicitta Cultivator



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Re: Mushroom Breeder - AMA [Re: fahtster]
#27865761 - 07/17/22 05:48 AM (1 year, 6 months ago) |
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cozmyc
gentle modern ape



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Re: Mushroom Breeder - AMA [Re: rumfor69]
#27865825 - 07/17/22 07:27 AM (1 year, 6 months ago) |
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Oo welcome back
-------------------- You're conscious population 2 stardust ---------------------- and that's valuable
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anatomality
Nothern Counterpart



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Re: Mushroom Breeder - AMA [Re: cozmyc]
#27865831 - 07/17/22 07:33 AM (1 year, 6 months ago) |
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Very well written and interesting post.
You'd probably enjoy this book (I'm planning on reading it)
https://www.amazon.com/Botany-Desire-Plants-Eye-View-World/dp/0375760393
-------------------- “The strength of a person's spirit would then be measured by how much 'truth' he could tolerate, or more precisely, to what extent he needs to have it diluted, disguised, sweetened, muted, falsified.”
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Muad.Dweeb
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Okay, since there aren't any questions currently, I'll start with something simple:
Why RustyWhyte worked.
First, a brief overview of the sexual arrangement created by spores.
Success for a monokaryon is finding a compatible mate. Compatibility is determined by two gene locations (loci) inherited by the monokaryon as a spore nucleus when it was formed through meiosis. These loci absolutely cannot be the same in both monokaryons in order to be compatible. This system ensures that genetic diversity is maintained, even through extensive inbreeding. The sexuality can be considered exclusive, rather than inclusive, which is why mushrooms are sometimes said to have thousands of sexes. Harry Haplont really doesn't want to date his sister, if you catch my drift. Harry is looking for someone hot... Maybe someone like Nelly Nucleus.
When two monokaryotic strains meet, they attempt to fuse. Hyphae from either monokaryon touch and recognize each other through an exchange of pheromones. This weakens the cell walls between them and they attempt plasmogamy (“plasma marriage”). This creates a tentative fusion where the nuclear information can be shared to check for compatibility. Very much like going on a date, spending social time together to see if there is chemistry between Harry Haplont and Nelly Nucleus. If they are compatible, nuclei are exchanged and they move in together. During mitosis (cell division) they continue growing as a dikaryon, undergoing a process called nuclear migration where all the previously monokaryotic cells eventually become populated with the corresponding nuclear mate. If they are not compatible, the fusion fails, and they continue growing and dating.
Sharing living arrangements is a difficult task, and not everybody manages that in the same manner. In P. cubensis and many other Agaricales species, nuclear orchestration is maintained by the creation of “clamp connections” between cells, located at the septa between cells. Some other strategies exist in the mushroom kingdom. Certainly, though, any relationship needs a game-plan to remain cohesive. For the life of the culture, two distinct nuclei per cell are maintained, with the exception of when new spores are made by a successful fruit... or if they “break up” for some reason. For Harry and Nelly, who both feel very strongly about their own names, they decided to hyphenate their surnames. Since their plasma marriage, they go by the name Haploid-Nuclei.
But what if there was no Nelly Nucleus? Who would Harry marry? All the spores around him came from the same fruit as he did, now, and he can only hook up with one of his siblings. Well, it all goes the same EXCEPT for the probabilities involved for compatibility. When they were formed through meiosis, each spore's nucleus inherited two gene loci to test mating compatibility (AB and ab). Each spore has a chance of being AB, Ab, aB, or ab. Now, since the requirement for compatibility is NOT possessing either of the same allele, that means Harry Haplont will only be compatible with one out of four of his sister spores, and it happens that Harry would pick the sister that resembles him the least. Harry has AB mating loci, so he can't hook up with Sally, who is Ab, or with Susie, who is aB; but Maria has ab, and she's looking begrudgingly attractive to the socially isolated Harry. So they hook up and make inbred babies. Their babies look like them and their parents, and each inherit the same mating loci (AB/ab) and have the same odds of mating with each other, or any generation from the same inbred line, until a mating loci mutates for one of any of a number of reasons.
This limitation doesn't exist when the nuclei aren't siblings. Compatibility approaches 100% as you get less closely related. That's a 75% advantage over sibling spores for mating preference. Back to RustyWhyte, Pasty put spores from two varieties with distinct traits into the same agar plate, and mixed them. This is arguably the simplest way currently known to cross tetrad (tetrapolar, or 4 spore per basidia) mushrooms (anything "bisporous" or only producing 2 spores per basidia has pre-determined mating). If intimately mixed and distributed on the plate- say, using a cross streaking technique- you can expect a crossed genotype four times more frequently than a parent pair. So we do what is generally considered a Very Bad Thing® and drop a germination plate to grain and run it out. As a side note, anyone trying this should make several cross plates and only attempt to run the healthiest looking plate, since we're playing craps with contam here. You can opt for dropping T1's, but you are limiting your diversity if you do.
Now the really depressing part. You go through all that, and your mushrooms start to fruit, and they mostly look like Normal Fucking Mushrooms! This depressing sight is actually a good sign. See, Harry is from the MakTP line, and Nelly from Golden Halo line. So their babies should be giant dicks with gold spores, right? Well, no. While there is a small chance that the recessive traits can appear in the F1, both nuclei of the dikaryon have to have a copy of the same recessive allele per trait. So whatever is Dominant of either parent for any given gene is what we will see expressed predominantly in the first generation, i.e Normal Fucking Mushrooms.
The second generation (when inbreeding resumes) is when heterogygous traits have the chance to independently assort and stack as dominant or recessive homozygous traits. Once the trait is homozygous, it can only contribute that allele during meiosis, so careful selection is required to include all traits desired through each generation of inbreeding. Depending on the trait complexity (most of what we think of as "traits" are actually polygenic, using several genes that may not inherit as a unit), you will see variant phenotypes/genotypes emerge at different points in the stabilization process. For Harry and Nelly, I did see purple and gold spore co-mingled in the same print (picture above, "Calico"), in an otherwise Normal Fucking Mushroom package. When I cloned and ran it in isolation, the prints looked straight up dark purple, but under microscopy I could see intimately mixed purple and gold spores. I have repeated this by crossing PF Redspore and an F2 gold spore only isolate from the Calico (it grows better than my GH), producing an orange-yellow print (pictured above). So some traits have a potential to at least peek out at you in the first generation.
TL;DR:
Mixing spores on agar is a viable method for cross-breeding because fucking your siblings is gross.
-------------------- A beginning is the time for taking the most delicate care that the balances are correct. This every sister of the Bene Gesserit knows.
How to Breed like the Bene Gesserit The Weirding Way - Advanced Bene Gesserit Techniques
Edited by Muad.Dweeb (08/15/22 07:30 AM)
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Svetaketu
The Devil's Avocado 🥑


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Re: Mushroom Breeder - AMA [Re: Muad.Dweeb]
#27866602 - 07/17/22 05:37 PM (1 year, 6 months ago) |
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Good stuff dude
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Muad.Dweeb
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Re: Mushroom Breeder - AMA [Re: Muad.Dweeb] 2
#27866611 - 07/17/22 05:40 PM (1 year, 6 months ago) |
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My F3 selection for the MakTP x Golden Halo cross, "Gold Member".

-------------------- A beginning is the time for taking the most delicate care that the balances are correct. This every sister of the Bene Gesserit knows.
How to Breed like the Bene Gesserit The Weirding Way - Advanced Bene Gesserit Techniques
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SGV
EssGeeVee



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Re: Mushroom Breeder - AMA [Re: Muad.Dweeb]
#27866630 - 07/17/22 05:59 PM (1 year, 6 months ago) |
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#PullingUpASeatForThisRide
I was completely immeshed in your explanation(s) and your cliffs brought the legit LOL's.
I look forward to reading more!
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Guerrilla
Bumbaclart


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Re: Mushroom Breeder - AMA [Re: SGV]
#27866649 - 07/17/22 06:18 PM (1 year, 6 months ago) |
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Bookmarking so I can read this properly when I got a min.
-------------------- Being pissed on does not make you a real man.
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smalltalk_canceled
Babnik


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Re: Mushroom Breeder - AMA [Re: Muad.Dweeb]
#27866675 - 07/17/22 06:44 PM (1 year, 6 months ago) |
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-------------------- Willpower is the one true virtue
  
Edited by smalltalk_canceled (07/17/22 06:46 PM)
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rumfor69
Bodhicitta Cultivator



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Re: Mushroom Breeder - AMA [Re: Guerrilla]
#27866676 - 07/17/22 06:44 PM (1 year, 6 months ago) |
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This is great! So question, if it's two varieties that say have purple spores, you do a cross plate, you fruit it and it looks like normal mushrooms, you then print those and cross spores from one print and then those will fruit out to be your hybrid? Am I understanding this correctly?
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smalltalk_canceled
Babnik


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Re: Mushroom Breeder - AMA [Re: Muad.Dweeb]
#27866680 - 07/17/22 06:49 PM (1 year, 6 months ago) |
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Okay one two relevant questions I guess
How do we determine if a trait/occurence is genetical in this process, or if a trait is dominant?
-------------------- Willpower is the one true virtue
  
Edited by smalltalk_canceled (07/17/22 06:50 PM)
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Muad.Dweeb
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Re: Mushroom Breeder - AMA [Re: rumfor69] 1
#27868946 - 07/19/22 01:39 PM (1 year, 6 months ago) |
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Quote:
rumfor69 said: This is great! So question, if it's two varieties that say have purple spores, you do a cross plate, you fruit it and it looks like normal mushrooms, you then print those and cross spores from one print and then those will fruit out to be your hybrid? Am I understanding this correctly?
If it's two varieties that have purple spores, you simply don't use spore color as your identifier. The spore-mixing method relies heavily upon having distinct traits that you can observe. It would be very close to impossible to use this method with, for example, vanilla Golden Teacher and PES Hawaii, since they are basically wild type to start.
But the basic structure is the same. In a multi spore setting, the preference will be for non-related spores to mate. However, it will be difficult to discern GTxGT or PESHxPESH versus the crossed GTxPESH and you will only be able to make uninformed selections.
And it doesn't stop at the F2, each generation of selection is moving more and more traits towards being homozygous, so variation tapers off and you get a "stable" variety anywhere from F5-F10, where similar fruit are consistently generated from spore.
-------------------- A beginning is the time for taking the most delicate care that the balances are correct. This every sister of the Bene Gesserit knows.
How to Breed like the Bene Gesserit The Weirding Way - Advanced Bene Gesserit Techniques
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fahtster
Now With 33%More Faht



Registered: 06/17/06
Posts: 9,266
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Re: Mushroom Breeder - AMA [Re: Muad.Dweeb]
#27868996 - 07/19/22 02:23 PM (1 year, 6 months ago) |
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Cool thread
Maybe you can answer this.. not sure if you’ve come across it in your research.. I feel like I’ve read that the buller phenomenon can happen between mono myc and even ungerminated spores but I can’t seem to find where I read it or if I even read it somewhere or maybe I was just curious if that can happen. Do you know if that’s something that can happen or is it just something that occurs between mycelium types?
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Muad.Dweeb
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Quote:
smalltalk_canceled said: Okay one two relevant questions I guess
How do we determine if a trait/occurence is genetical in this process, or if a trait is dominant?
That is a good question and not a simple one to answer completely. Morphology is a tricky bitch, and needs lots of validation. If I were a geneticist it would be work but doable to actually track genes and gene groups to observe variations of expression, but I'm not. What it boils down to in practice without those tools is observation and testing.
For example, the blobs I mentioned previously are going through trials right now because I found them in a MS grow. That context has a lot of pressures and stressors that can cause deformation or other morphologic anomalies, from competition between strains, to bacteria, exposure to petroleum derivatives, etc, usually blanket termed "rosecomb". These aren't typically persistent traits, so to make sure, all you can really do is take your clone, clean it up, run it in isolation and see if the trait persists. Two of the three blobs I've fruited as isolates have proven to be fairly normal mushrooms, one albino and the other apparently sterile, but certainly not blobs.
But, again, sometimes what we are thinking of as a trait is actually a compound of independent genetic traits. In instances where you can run spore from what you confirmed in isolation (can't really do that with blobs), you can look for the same trait or similar traits to pop up in the offspring.
Dominance is just whatever expresses in a mixed state. That's the definition. Most of what we would consider unique traits among varieties are homozygous recessive traits. Some will co-express when pitted against each other (Co-dominance), or have a dominant/recessive relationship where one expresses over the other. Spore color has the rare honor of being controlled (at least with cubes) by a protein encoded by just one gene, so it is a lot easier to observe it inherit in classic Mendelean fashion. Purple spore is dominant to red spore and gold spore, but so far, when I put red and gold against each other, F1 fruits are burnt orange, suggesting they are recessive generally but co-dominant to each other.
So why would I get a calico print from a purple/gold cross? My hypothesis is that with two copies of a given gene, and millions of cells, you get random errors and the recessive allele occasionally gets used over a dominant. And with a multi spore grow adding multiple pressures it may happen at a greater frequency. The clone culture never did produce a truly calico banded print, but I found gold spores mixed in very well @800x.
-------------------- A beginning is the time for taking the most delicate care that the balances are correct. This every sister of the Bene Gesserit knows.
How to Breed like the Bene Gesserit The Weirding Way - Advanced Bene Gesserit Techniques
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P.Nowhere



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Re: Mushroom Breeder - AMA [Re: Muad.Dweeb]
#27869007 - 07/19/22 02:29 PM (1 year, 6 months ago) |
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Pulling up a seat!
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Muad.Dweeb
Seer



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Re: Mushroom Breeder - AMA [Re: fahtster]
#27869038 - 07/19/22 02:48 PM (1 year, 6 months ago) |
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Quote:
fahtster said: Cool thread
Maybe you can answer this.. not sure if you’ve come across it in your research.. I feel like I’ve read that the buller phenomenon can happen between mono myc and even ungerminated spores but I can’t seem to find where I read it or if I even read it somewhere or maybe I was just curious if that can happen. Do you know if that’s something that can happen or is it just something that occurs between mycelium types?
I'm going purely off memory here, but I think that may have been one of Roger Rabit's experiments with snake venom, attempting to revitalize his Redboy spores.
In short, monokaryotic myc wouldn't be utilizing the Buller Phenomenon with spores, since the spore is an ungerminated mono. But spores can germinate on mycelium, mono- or dikaryotic, and form new strains through mon-mon and di-mon mating. The conditions need to be right for the germination, but TBH, I regularly germinate my spores in sterile distilled water, so it doesnt take much.
Adding spores to a grain jar mostly colonized with monokaryotic mycelium was Workman's approach with APE and another cross he never followed up on afaik.
-------------------- A beginning is the time for taking the most delicate care that the balances are correct. This every sister of the Bene Gesserit knows.
How to Breed like the Bene Gesserit The Weirding Way - Advanced Bene Gesserit Techniques
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