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OfflineMuad.Dweeb
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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: Muad.Dweeb]
    #27993648 - 10/11/22 12:10 PM (1 year, 7 months ago)

The mold in the tub was, in fact, trichoderma!

I cultured it on water agar and eventually it formed conidia:



I'm not sure on the species, but my initial thoughts are harzianum or viride/aureoviride. My next move is to try and cultivate it without it producing conidia, maybe even conidiaphores.


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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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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: Muad.Dweeb]
    #27998743 - 10/14/22 02:04 PM (1 year, 7 months ago)

:popcorn:

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OfflineMuad.Dweeb
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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: Muad.Dweeb]
    #27998943 - 10/14/22 03:54 PM (1 year, 7 months ago)

Regarding the individual characters of monokaryotic mycelia contributing to the character of the resulting dikaryotic mycelia (P. ostreatus, but relevant):

https://scialert.net/fulltext/?doi=pjbs.2007.2334.2340

Scroll down for the pairings.


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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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OfflineMuad.Dweeb
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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: Stipe-n Cap] * 1
    #27998944 - 10/14/22 03:55 PM (1 year, 7 months ago)

Hey you!


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A beginning is the time for taking the most delicate care that the balances are correct. This every sister of the Bene Gesserit knows.



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The Weirding Way - Advanced Bene Gesserit Techniques

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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: Muad.Dweeb]
    #27998959 - 10/14/22 04:08 PM (1 year, 7 months ago)

Nice thread, not often I find something interesting to read here :rockon:

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OfflineMuad.Dweeb
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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: Stipe-n Cap]
    #28006121 - 10/19/22 09:31 AM (1 year, 6 months ago)

So, something I'm experimenting with right now has some interesting first results.

What you see here is mycelium (trich atm) who's cell walls have become porous on a supplemented MEA. I'm still very into the experimental/investigatory stage, so details are going to be absent for now. Suffice to say, I was trying a method of controlling conidia production, and while it doesn't appear to work, this other effect could be very useful.




[photos on agar @ 1000x and 250x]

While the pores are not large enough to liberate a protoplast, they are large enough to potentially pass nuclei and plasmids (and molecules...) through the cell wall. And the effects are reversible (this isn't venom or a lysing enzyme) by removing the mycelium and transferring to standard media (there are effects besides making the cell wall porous related to up and down regulation of gene expressions, according to some papers. One effect has been lower to no color expression in the conidia.)

I'll be transferring some cubes to the media later and we'll see if it translates.


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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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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: Muad.Dweeb]
    #28006187 - 10/19/22 10:14 AM (1 year, 6 months ago)

Are you saying that the structures within the hypha are pores? It appears as though trich hyphae are non septate, so perhaps the photo is showing free movement within the cytoplasm, Cytoplasmic inclusions?

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OfflineMuad.Dweeb
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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: Stipe-n Cap]
    #28007759 - 10/20/22 11:41 AM (1 year, 6 months ago)

1, the aerial mycelia and conidiophore don't have this pattern on the hyphae; it's only on the surface and submerged mycelia.

2, it definitely is septate, but the compartmentalization behavior is (I should think "obviously") different from basidiomycota.



Also, I have been able to observe nuclear or vacuole movement within other cultures' hyphae. These have been stationary.


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A beginning is the time for taking the most delicate care that the balances are correct. This every sister of the Bene Gesserit knows.



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The Weirding Way - Advanced Bene Gesserit Techniques

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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: Muad.Dweeb]
    #28007773 - 10/20/22 11:59 AM (1 year, 6 months ago)

Ah, very nice.  Nice snap of the septa.

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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: Stipe-n Cap]
    #28007877 - 10/20/22 01:47 PM (1 year, 6 months ago)

"Also, I have been able to observe nuclear or vacuole movement within other cultures' hyphae. These have been stationary."

What is the general meaning of this? What does it suggest?


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OfflineMuad.Dweeb
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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: smalltalk_canceled]
    #28012406 - 10/23/22 12:07 PM (1 year, 6 months ago)

Quote:

smalltalk_canceled said:
"Also, I have been able to observe nuclear or vacuole movement within other cultures' hyphae. These have been stationary."

What is the general meaning of this? What does it suggest?




Just what it says. The nuclei and other cellular components move around inside the cell, sometimes they even hop between cells through the septa. They get pulled around by things like actin and dynein, and if you have the proper refraction through the hypha, you can see them move about in live mycelium.

By contrast, I'm calling the apparent pitting on the trich mycelium I'm viewing "pores opening in the cell wall," both because they are stationary and because it is a possible side effect of my agar supplement.


--------------------
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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OfflineMuad.Dweeb
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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: Muad.Dweeb] * 1
    #28162841 - 01/29/23 12:41 PM (1 year, 3 months ago)

From a convo with Rhizy:

Quote:

it would seem like the mating type genes would be one of the things regulating fruitbody development




Mat1 and Mat2 are the mating loci names, and Mat1 is a simple allele, whereas Mat2 is a complex of various traits that generally seem to inherit as a unit. These traits relate to pheromone production, clamp coordination and often fruit body cellular orchestration, from what I understand.

Quote:

I’ve heard some really strange stuff recently as well like traits flip flopping between generations (apparently not due to heterozygotes), and lines that blob from spores and revert from cloning. I also think that environment seems to have a big impact on expression, and it seems that some varieties are a lot less stable than is assumed especially when grown in a multi-dik environment.




I have a few ideas regarding this. I have been assured by a couple vets that it isn't a real problem, but Same Species Cross Contamination is one possibility. Especially when it comes to any degree of open air fruiting and heavy target species spore load. One great example of this is when CronicR was fruiting John Allen Strain and CRS at the same time. Spores from the CRS produced fruit with purple spore (F1) that in turn produced mixed purple spore and red spore (F2), with similar happening in the descendants from JAS. Pasty assured me that CronicR has a massive sporeload in his grow room (visible on walls even), but I think it would still be quite possible with typical circumstances. It's one experiment design I return to repeatedly, I haven't quite figured how to simulate it effectively.

Another is partial expression, which could tie in to mitochondrial preference.

Quote:

how many prints you need to run from F1 to catch the offspring of different nuclei/diks?




Any time a cross happens, regardless of the sexual mode, the offspring present in typical Mendelian fashion. But, "how many prints" is a bit of a statistical variable. If you are only looking for one recessive trait, it likely will pop up within a grow or two. The more non-linked recessive traits you are looking for, the less likely a given dikaryon will match the criteria. there are also undefined variables in play, like how likely two adjacent alleles are to inherit as a unit, or separately. Don't quote me- math is not my strong suit- but the probability of stacking n recessive traits is an inverse exponential. Another variable is how diverse the grow is. I try to make my MS grows fairly limited, but I'm probably still getting around 1-3k spores in there. Others, like Fahtster, are using substantially more and getting far more varied results early on.

I'm sure as a Maths lecturer you've put together that more iterations in a limited set increases the odds of a hit.

Quote:

about di-mon. Do you look for new dik myc growing away from the interaction zone (maybe using morphology and heterosis), subculture the interaction zone, or would you wait until nuclear migration has happened and subculture from the area that was originally the mono?




Literally, I wait until the plate pins. There are details that indicate the Buller Phenomenon has taken place, like abrupt change in morphology (thickening, increased colonization rate, formation of clamps in new growth...), but those can be variable. Clamps normally form in new growth after 48 hours with cubes, for example, but some pairings don't develop clamps for a week or so. But there are only a few examples of basidiomycete monokaryons that can produce asexual fruit bodies (not oidia, but full on fruits), so I use that as a definitive marker most times. I have never seen a "third culture" grow out of the interaction zone using Di-Mon pairings. Mostly, that seems to be a response some dikaryons have in the presence of other dikaryons/competitors.

Quote:

and then there’s also mitochondria!



Quote:

The stress of changing between vegetable and reproductive states, for example relating to cloning, must be greater than subculturing/asexual expansion, and is also really interesting. I was also thinking about what you said about enigma and cleaning up a culture, makes me wonder what was involved in cleaning it…




Yes, there certainly is. I asked my friend Sidnee about this a couple years ago when I first started trying to wrap my head around Enigma. I was pretty familiar with Boomer Smith and Moe Matteo, who brought Enigma into circulation. I can say pretty confidently that Enigma was isolated into a monokaryotic pure culture by Boomer before it was sent back to Moe, who fruited it, so repeat cloning wasn't a factor. Boomer also doesn't use any antibiotics or anything, so it was cleaned with serial transfers, so not a factor. Further, I know a lot of back story of that particular drama, and I'm pretty sure that Magic Myco (Doma, Tidalwave breeder) probably found another from spore, and they were arguing over 6 vs a half dozen. My own ability to track down blob-only cultures from spore certainly supports that.

Back to Sidnee, I asked if the mitochondria of that particular selection could play a part. His response was that while cytoplasm isn't exchanged in plasmogamy and you end up with two cultures with the same nuclear DNA but different mtDNA, it doesn't really matter. I didn't accept that answer, though, and have been designing an experiment where I can investigate this without access to DNA scanning. I have a good idea on how to go about it, but it requires competency in several lab techniques like dedikaryotization. I'm close to attempting it, I just need to carve the time out for it and select ideal candidates.

Quote:

It’s interesting that you mentioned about reversion as a type of mutation, as I was going to ask what you thought about it.




You can find more information on it with the term "synthetic recovery".

Quote:

about auxotrophs, or mutants that require some specific nutrient in the medium? I guess it’s hard to know without specifically trying different media, but these mutants seem to be useful in experiments as identifiers. I also wonder if any of the cube mutant iso/cultigens would fruit more normally with supplements or different environmental conditions.




Auxotrophs are interesting, but in typical cultivation practices we usually surreptitiously breed them out- at least, by my vague understanding of artificial and natural selection. One interesting example mentioned in (I think) A.C. Chang's "Genetics and Breeding of Edible Mushrooms" was an auxotrophic strain that required a nicotinic compound for normal metabolic health. Without that supplement, it doesn't fruit or even colonize effectively. I imagine that depending on the exact metabolic mutation, this could be survivable and present deformed fruits, but I don't know.

Quote:

it also interestingly mentioned that the mutated low spore trait in the oyster was stable for more than five gens. You could add the photoreactivation stuff to your UV tek, but from the papers it isn’t really clear when the mutant culture can be exposed to the ‘normal’ light (definitely by fruiting it seems).




Photoreactivation triggers DNA repair. From what I've read in a few AG science papers, it occurs mostly in the blue spectrum, so UVC treated fruits and veg should be kept in darkness or red spectrum light after sterilization. When I tried this with live cultures, it outright killed them, reddening the mycelium often and halting all growth. Samples would not culture on fresh agar. Exposure to light seems to be required to avoid a terminal outcome. And while this is a very accessible route to mutations, it will probably always require tinkering to get ideal results, and then a lot of fruiting to find beneficial mutations. This technique has been used with a good clutch of species already.

Quote:

It seems that for cubes and more complex fungi there are enough mating alleles that there are thousands of sexes, and incompatibility would be hard to find even if you wanted to.




True, until you start inbreeding. Then, Mendelian inheritance applies to the predictability of Mat1 and Mat2. In a wild setting, inbreeding happens on much lower scale, and homozygous recessive strains present less frequently. At least, that's my assessment. Ecological studies of anomalous wild specimens could prove me wrong. It's proving wrong with several species like Boletus edulis, which is showing that the environment has more to do with their phenotypes than the genotype does. Even transplanting European varieties in the USA, the European variety takes on the characteristics of local strains, even though the genome stays intact. [ https://attheu.utah.edu/facultystaff/a-tale-of-terroir-porcinis-evolved-to-the-local-environment/ ]

Quote:

You mentioned something about a sort of di-di mating through exposure to chemicals, could any mutagen/shock treatment be used, like heat, UV, electroporation? What is the mechanism for this, I’m guessing it would need to prevent or delay somatic incompatibility in the interaction zone.




To be clear, Di-Di mating isn't a thing, in my observation and opinion. But certain events could appear to be Di-Di mating. Hypothetically, this would involve some sort of dedikaryotization (dedi) event to happen when two strains are in close proximity. There are natural events that can cause this, some of which I'm investigating (see me Swinger Tek post in Bene Gesserit). IIRC, chemical dedi can happen in the presence of things like cholates (biles), with a fair chance of inducing damage mutations to the recovered mono. Using high enough concentration to do this prevents one nucleus from replicating and the new growth becomes mono. On a plate with cholates, or even theoretically in an animal's GI, this can cause one or two dikaryons in close proximity to become available for a new di-mon or mon-mon pairing. And there are a host of events that can do this. It can even happen randomly for undefined reasons, particularly in new growth. My contention is that people who have successfully "crossed two dikaryons" have actually exploited a form of dedikaryotization.

Quote:

I saw a ‘gold member’ variety for sale online, is this your cross or just a coincidence? I guess the later as it says PE x GT.




Nah, not mine. That is a "variety" (unstable from what I've seen). Mine is a clone name, just because my labeling system isn't verbally friendly. But, I do know the person who made it (they are pretty active on facebook) and they named it Gold Member as an attempt to slight me. That is actually something I anticipated though, which is why I haven't named the variety beyond the project name (GOGH, just initials that mark the original parents)

:2girls1cup:


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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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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: Muad.Dweeb]
    #28163176 - 01/29/23 04:22 PM (1 year, 3 months ago)

.


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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: HFM] * 2
    #28164374 - 01/30/23 01:12 PM (1 year, 3 months ago)

Mitochondrial Investigations



To investigate the influence of mitochondrial DNA (mtDNA) in abnormal fungal behaviors, in particular the formation of fruit bodies, the following procedures have been designed to generate cultures with identical nuclear DNA (DNA) but disparate mtDNA, for conducting experiments in linear growth rate (LGR), biological effeciency (BE), carpophore formation (CF), and any other metric that could potentially be affected by mitochondrial activity.

Introduction

During plasmogamy, when two monokaryotic cultures exchange nuclei, only a small area where the anastomatic connection is formed contains the cytoplasm of  both monokaryotic cultures. In this small area, copies of respective nuclei are transferred to the other cell, and rapid nuclear division of the new DNA takes place, with the donated nuclei traveling through the culture via nuclear migration through the cell septa. The new DNA now operates in tandem with the host DNA and the native mitochondria. In vivo, this results in two cultures with the same binucleate DNA being utilized by separate mtDNA genomes.

Using the simple tools of serial dilution and dedikaryotization by mincing cultures, it is possible to replace the mtDNA of a given monokaryon or dikaryon to study the specific effects that mitochondria produce while utilizing DNA. Moreover, the procedure can be used starting with a wild or domestic clone, or starting with spores to produce monokaryotic cultures. Competency with agar, serial dilution and dedikaryotization is required to execute this procedure.


Quote:

Materials

An Oster/Waring/Eberbach
Graduated centrifuge tubes
Syringes/pipettes
plenty of agar/LC





Before anything, monokaryons need to be generated with serial dilution (or a suitable method that accomplishes the same). These monokaryons cannot have the same mtDNA as the target culture for the procedure to produce viable data. In domestic cultivation, this means selecting varieties with disparate pedigree- which will require some degree of knowledge regarding such. This is a simple matter when investigating specimens that have particularly unusual morphology or metabolism, as wild-type mtDNA is (theoretically) plentiful in the population. In such case, generating monokaryons from wild-type leaning varieties is important. These will become the “normalcy” benchmarks for later experiments.

Tandem Dikaryotic Cultures (TDC)

After isolating monokaryons from disparate lineages, pairing monokaryons from those respective parents results in two dikaryotic colonies, each colony retaining it's original mtDNA. After confirmation of plasmogamy, each colony should be subcultured and maintained separately. Agar is the preferred method for tracking LGR, and can easily be tracked by marking a graduated grid on the bottom of the plate. Expanding and fruiting each maintained culture to observe differences in CF and BE is possible now. Dedikaryotizing the cultures will produce both DNA types times both mtDNA types.




Replacing mtDNA in a Dikaryon

With cloned, dikaryotic cultures, attempting to utilize the Buller Phenomenon (di-mon mating) will result in a novel dikaryon with the host monokaryon's native mtDNA intact. To achieve the desired replacement of mtDNA, the dikaryon must be separated into its component monokaryons (neohaplonts). Both neohaplonts will have the same mtDNA, but pairing them with other monokaryons (generated from spore or an unrelated dikaryon) as when starting from spore, the respective pairings produce a TDC. Subculturing the TDC from the non-neohaplont side will select the novel dikaryon with different cytoplasm and mtDNA than the starting dikaryon. Further dedikaryotization of the novel dikaryons will produce the monokaryotic components of the original clone with new mtDNA.




--------------------
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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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: Muad.Dweeb]
    #28212311 - 03/03/23 10:37 AM (1 year, 2 months ago)

First post, woohoo! Great to see some of our chat in the thread! (Looks like a typo about Enigma being isolated to a mono).  Have to say, as usual, loads of great original content! Really impressed to see the new mito-swapping experiment design, a lot of work though lol!

I’ve only been using a microscope for a few weeks, but I’ve seen movement of small black organelles, even moving through the septa as you said, and what seem to be various types of vacuoles in the hyphae at 1000x (see pictures below and clip of movement is on discord microscope channel). I had hoped that the small black dots were nuclei, but after speaking to someone more experienced, looking some more, and looking at micrographs in the literature, then I think seeing nuclei with a standard light microscope without staining is very rare, and these are more likely vesicles or some other type of vacuole moving around.

The following pictures are from a standard light microscope at 1000x, with a hand held phone camera using some zoom. Some possible vesicles circled in the first photo.




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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: Rhizy]
    #28216210 - 03/05/23 05:08 PM (1 year, 2 months ago)

P.S. I went back to Chang and Miles (Mushrooms: Cultivation, Nutrition Value, …) to see what they say about UV mutagenic treatment. It seems that UV mutation works by forming dimers that prevent correct DNA synthesis, and they seem to suggest that spores (or conidia), or mycelial fragments are treated with mutagens.

So, I’m guessing photoreactivation needs to be prevented until after DNA has been synthesised in the treated sample, like the spores germinate or the fragments start to extend/grow. They also say that yellow light can be used to illuminate the work area for UV without causing photoreactivation.

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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: Rhizy]
    #28225059 - 03/11/23 12:11 PM (1 year, 2 months ago)

yeah nuclei are pretty tiny, not nearly the scale they are usually shown in illustrated diagrams.

Right now, I'm doubling back to do a deeper investigation into "swinger tek" using my Gold Member (GM) culture and Enigma (EN). I figure the clear visual difference should highlight patterns of inheritance pretty well.

There are actually 2 possible outcomes if the procedure does produce novel dikarya, dependent upon if the mtDNA is significantly different. The first is 4 novel combinations, plus the two original dikaryons, with no significant variation of mtDNA. If variant mutation(s) exist in at least one of the mtDNA lines, phenotypic expression can be between 5-8, plus the 2 original. Concurrently, I'm dedikaryotizing Enigma to do controlled matings with GM and EN (GMEN), to produce 8 total subcultures (transferring from both sides of each TAD to subcuture) as reference/control.

Presently, I have completed a battery of grows using the swinger tek for pheno hunting. I have identified 4 general phenotypes from the fruit, with possible mitochondrial variations of each.

The base phentypes are Wild Type (WT), Hat Type (HT - wild type with a party hat), Variegated Type (VT - having varied pigment on one cap from none to tan), and Conical Type (CT - cap starts conical and retains a nipple upon maturity); the possible variant phenotypes are the base types, plus wrinkled caps and stipes, leaning towards thicker stipes, but generally still wild type with modifications. Both leucistic and carpophoroid/pseudocarp phenos (GM and EN) appeared in most of the grows at a low rate.


Given the dominant alleles of each- GM produces generally agaricoid fruits, while enigma has "cap pigment" alleles- the predicted permutations of their nuclei should produce WT leaning phenos, including pigmentation of flesh and spores, which is what is showing up in the swinger tek grows. The 4 base phenotypes are already on grains, and a MS of each is also on grain, while I finish dedikaryotizing Enigma and make my reference pairings. I cloned the wild type and variegated wrinkle-phenotypes for comparison, as HT and CT didn't present wrinkle-phenotypes. I figure if there is a noticeable difference it will play out with the controlled pairings anyway.

Another possible cause of the variation is overall sub volume and spawn ratio. Notably, and perhaps obviously, the half pint cake/mono tube produced the smallest fruit, while the large filter patch bag was about 5 qt volume with a 1:4 ratio and the largest fruit. Wrinkled variants did occur in all but the half pint grow, though. To eliminate this potential, the clones will be standardized 1:3 in large grow bags sealed after spawn, as will the controls.

The prediction for the MS is that each of the novel phenotypes will produce a diverse fruiting population with more individualized recessive traits of either parent presenting in 3:1 or 1:2:1 ratios, perhaps some looking strikingly like either parent; this diversity will be drastically higher than the initial swinger tek grows and not lean so heavily towards wild types. Increased homozygosity respective to selection(s) is also predicted in the F3.

Phenos:




Wild Type - there appears to be a nipple, but it smoothed out in maturity and wasn't conical as a pin.




Hat Type




Conical Type - retains cone shaped cap until maturity, retains a nipple once the cap is (near) planar.




Variegated Type - while all caps with pigment will have a hygrophanus color gradient, I am only considering it variegation if it is present as a pin.


Swinger Grows in situ:




Most of the grows were 1:3 rye to CV in domed pint cups (dub tubes)




One cup (half pint) of mixed 1:3 spawn/sub was fruited in a single unmodified quart cup (mono tube).




In a bag, at 1:4 with CV. Note the distinction between conical and wild type here.


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The Weirding Way - Advanced Bene Gesserit Techniques

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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: Muad.Dweeb]
    #28225068 - 03/11/23 12:19 PM (1 year, 2 months ago)

...as such, I haven't circled back to UV experiments yet.


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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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OfflineMuad.Dweeb
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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: Muad.Dweeb]
    #28299066 - 04/28/23 03:53 PM (1 year, 21 days ago)

I have some time, so I thought I'd update a couple things.

Apr. 1

Stage 2 of my Swinger Tek tests is underway. Here are the neohaplonts from dedikaryotizing Enigma and Gold Member, clone cultures I used in the initial GMEN blender experiment. They are much more flat and translucent than either dikaryotic culture. I'm waiting for the controlled pairings of GM×EN from their respective monokaryotic cultures to compare to phenotypes from the blender (and other) experiments.

GMEN.F1 [Control Set]




The respective nuclear types were paired on agar and I waited for clamps to show. Clamps were confirmed with visual alterations of macromorphology and in situ microscopy @100x. Here is the most dramatic change in morphology from the GMEN.AA plate:



Apr. 14-28

Proceeding, the "TADs" were subcultured from respective colony sides and labeled according to cytoplasm type (mtGM or mtEN). Comparatively, we can see significant differences in mycelium character:







The result about to go to grain is 8 unique cultures that should show similar results to the GMEN swinger experiment. These were subcultured again and will go through several serial transfers to continue observing mycelium habit.

In the meantime, I only ran 2 of the GMEN.F1 clones (Wrinkle and Hat), which showed opposite/swapped phenotypes (not a mix up, they were cloned on different days). They were 1:3 in sealed grow bags. Otherwise, same general (wild type) phenos.



Finally, I found 2 gold spore squat specimens in the GCKS.F2 (plus a couple other phenos I'll be pursuing), named Hotei and Goku. Forgot to grab pics of Goku, but they are both being cleaned and headed to grain soon. GCKS, if you've been following along, was also started with Swinger Tek. These two finds (after approx. 10 F2 multispore grows) completes the first demonstration/trial of Swinger Tek, combining the gold spore descended from Golden Halo (Golden Child F2 clone from my GOGH breeding project) and Koh Samui Squat's rotund, short stature. Proof of concept. Now with GMEN, we will have a deeper look at what's happening, hopefully.



--------------------
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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OfflineYoshiTrainer
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Registered: 04/30/22
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Re: The Weirding Way - Advanced Bene Gesserit Techniques [Re: Muad.Dweeb]
    #28322295 - 05/16/23 06:32 PM (1 year, 3 days ago)

Some great reading, happy to be following along!

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