Nouse luontoni lovesta
Haltiani haon alta havau
Veen emon lähteheestä!
Background
In 2024 I got fascinated by mushroom genetics. I was already familiar with the "cube is a cube" mantra and all the community drama about how discussion about strains is pointless and how the effects are a function of dosage, set & setting, and competency of the grower. A detail was bugging me however - there exists a widely accepted appendix to the aforementioned mantra: "...except for Penis Envy". To me this sounded like the full story is not as simple as I had previosly thought. Then I learned about the two famous PE hybrids: APE and Tidal Wave. Both matching or arguably exceeding PE in potency and both exhibiting some additional characteristics. Then the natalensis/ochraceocentrata boom started. Majority of ochra enjoyers reported consistently different trip qualities from cubensis. The cherry on top: being the closest known relative, ochraceocentrata is able to produce interpecific hybrids with cubensis.
At this point it was clear to me that there's so much more to uncover in the genetics department, but the full picture was still murky. Why so many cultivators were adamant about majority of the cubensis varieties(what many call "strains", I will refer to them as varieties or cultivars) being practically identical, yet these significant exceptions exist? (yes ochraceocentrata is not cubensis but sufficiently close still) What finally shed light to the mystery was this excellent paper. It outlines how majority of cubensis cultivars are indeed very close relatives and (the sample represented in the paper) can be divided to roughly 5 genetic clusters. Crucially, many of the established cultivars which are stated having a distinct geographical origin seem in fact to be Golden Teacher derivatives - possibly indicating a scheme by vendors to capitalize on a distinct phenotype isolate. Furthermore it offers details on genetic differences between cultivars and thus, their outbreeding potential.
So I interpreted the situation as follows: majority of the common cubensis varieties are very inbred, leading to low heterozygosity and known effects of inbreeding such as anomalous growth patterns and reduced resistance against contaminants and other environmental factors. Fortunately there seems to be genetic variance left between the aforementioned genetic clusters - groups of closely related cultivars which are at least somewhat distinct from each other. The paper above became basically my shopping list. I sought to get my hands on spores from each of the clusters. Wild spores would obviously offer even greater pool of genetic potential but according to the best of my knowledge the domestication process itself is a lengthy endeavour so I decided to omit that particular rabbit hole from the project - for now.
Motivation
I seek to contribute something back to this great community from which I've learned so much. My hope is that via a (somewhat) disciplined outcrossing program the genetic pool can be rejuvenated. I think there is sufficient evidence that a successful hybrid can be more than sum of its parts - by combining the best aspects of multiple domesticated varieties. At least initially, I will send these spores to those interested free of charge. I do only kindly ask that you 1) submit a post about your grow in this thread and 2) share the spores forward if possible, preferably from an early grow since the genetic variance diminishes after each generation. Beside those, feel free to do whatever. Isolate & stabilize your favourites, give them a name, sell the spores, whatever. Note: these spores are not to be considered strains or even cultivars or varieties. They are intentionally left unstabilized so I'd rather refer to them as "populations".
The Method because calling a multi-year process a "tek" feels wrong
The leading design principles for my breeding procedure were to maximize 1) the outbreeding potential and 2) its utilization for a given amount of labour. Hybridization of just two cultivars seemed too simple: in case both parents have low heterozygosity the best possible case for the offspring would be to inherit an allele from each parent in every locus. In practice this is not going to happen given the messy nature of meiosis. Using 4 origin varieties instead seemed to provide the best balance between genetic diversity and practicality. As established, there are 5 genetic clusters so 4 carefully selected cultivars would already cover a significant footprint of the commonly available genetic base. This approach would also make it practical to ensure equal chance of contribution for each of the origin varieties via 3 mating operations: (A x B) x (C x D) (two children, one grandchild).
Figure of the breeding procedure, explanation below:
Step 1: Four mutually genetically distinct varieties are selected and N instances of monokaryotic mycelium isolated from each one. This is done to A) extract any potential diversity and to B) hedge against sampling recessive and/or disadvantageous alleles. In my case I settled primarily on N=6 since that felt like a good balance between my current facility/time constraints and the goals above. Monokaryons are extracted via EG's drag & grab tek and confirmed (to the best of my ability) with a microscope.
Step 2: Monokaryotic mycelia are paired together to grow a series of 2-strain F1 hybrids. To ensure compatibility, one would ideally do the full N x N pairing matrix and check each pair for clamp connections. In practice however, I have found out that by virtue of having genetically distant parents the pairing succeeds with very high probability. (I'm yet to encounter a failure case) I usually make very small grows into takeaway containers since only a few prints are required.
Step 3: Practically same thing as in step 1, except for the fact that this time only one monokaryon per parent is isolated. Note: this is subject to further analysis, I suspect that lowering the number of monokaryotic samples in the step 1 and increasing it here would provide better results. I will perform some simulations on the matter later on.
Step 4: Identical to step 2. Since the dikaryons are again mated from mutually distinct varieties I consider them to be the "true" F1 for the hybrid population.
Step 5: Here the objective is to express as much of the collected genetic diversity as possible. This is achieved by a wide multispore grow: germinating a large amount of spores and to each of them, providing an optimal environment to bear fruit. In practice I drop spores into a small jar of sterilized water and tween-20, shake it up and pipette it on agar. I then do drop & shake liquid inoculant of the plate and mix it well with the grain. I spawn to bulk by dividing the jar into 4 segments and mixing each of them with coir individually to decrease the likelihood of a scenario where a single particularly vigorous strain would dominate the entire tub. I might post a detailed overview with photos later on.
Step 6: From each MS grow select individuals with the best characteristics and collect spores from them. Currently I select for individual fruit size, cluster size, growth speed(days after spawning), bluing, stem solidity, albinism, and other visually distinct features. (Characteristics/traits indicated as alchemical symbols in the figure.) As the number of individual isolates grows rather large at this point(almost 50 for two of my hybrids), evaluation of "post-digestion" effects becomes impractical. According to my limited sampling however, it would seem that they are largely comparable between the grows. Splitting the fruit with a sharp knife and observing for bluing tends to provide a sufficiently reliable proxy at this point anyway.
Step 7: At this step the idea is to 1) finally mix together the N parallel populations and 2) specialize for each selected trait. From the spores collected in previous step, select the best performing individuals according to a characteristic metric (top 3 biggest fruits, top 3 fastest fruiters, etc...), mix their spores together and proceed with a MS grow in similar fashion to step 5.
Step 8: Further optimize for each desired characteristic by iterating the two previous steps. Doing this for too many generations will obviously lead to the very inbreeding issues we're trying to mitigate so it is important to stop if the population starts to show signs of regression. Of course there is no need for the process to be as linear as presented by the figure - backcrossing by introducing spores from previous generations is a good way to reintroduce lost diversity. The fact of having multiple selected traits also further shields against population collapse.
Step 9: The final step in Lovi™ method and the first one in new cultivar synthesis. Combine your favourite traits via MS grows or via mon-mon pairing. How about some abundant albino clusters? Beefy-legged big bois? Why not?
Step 9 alternative: Combine matching/best performing traits from two different Lovi hybrids. Now it's a hybrid of 8 origin varieties!
This is of course an idealized version of things. As you all know, growing shrooms is a messy process. Sometimes you come up with neat ways to cut corners, sometimes they fail miserably. Things contaminate, sometimes multiple phenotypes pop up from a supposed mon-mon grow. Sometimes such an exceptional individual blesses you with its presence that you're willing to make, well, exceptions. I make little variations to my approaches with each hybrid, while still staying true to the philosophy of the method. I follow the process strictly as outlined above where it truly matters, according to the principles defined in the first paragraph.
Current Status
There are 6 hybrid populations I'm currently working on:
- Lovi Koivu: (APE x Malabar) x (A+ x Ps.Ochraceocentrata) - Step 5/6, waiting for last tub to fruit
- Lovi Mänty: (APE x Ps.Ochraceocentrata) x (Thai Pink Buffalo x B+) - Step 7 starting shortly
- Lovi Kuusi: (Tidal Wave x Malabar) x (Mexican Dutch King x Costa Rica) - Step 4 done
- Lovi Leppä: (Mexican Dutch King x Thai Pink Buffalo) x Yellow Umbo - Step 4 (YU is Jack Frost x Ps.Ochraceocentrata)
- Lovi Paju: (Rusty Whyte x Treasure Coast) x (APE x TW) - Step 2 done
- Lovi Haapa: (Treasure Coast x Z-strain) x (Escondido Mexico x Malabar) - Step 2 done
As one can imagine, managing bookkeeping by hand soon got ...out of hand. So being the nerd I am, I developed a webapp for the task. The database and backend is hosted locally on my desktop but the frontend also features a neat mobile UI so I can add photos and notes directly with my phone. Super handy when cataloguing prints from 20 individuals in a single session. But most crucially now all the data, including the characteristics of interest and family relationships are stored in the same place. It enables me to track their properties through generations (I'll add more extensive analysis tools later on) as well as to produce neat visualizations.
Following is the the full family tree of ~everything I've worked on for the past year and a half. Unfortunately I wasn't very consistent about taking photos before I got the app running at the beginning of the year so many of the early entries are missing them.
Exported ancestral trees for single examples of Lovi Koivu, Lovi Mänty, and Lovi Kuusi populations:
  
The Lovi Koivu one shows a streamlined way of hybridizing cube and ochra: I essentially made a wide MS grow of both spores with the method described in step 5. My A+ spores consistently produced leucistic phenotypes with papillate caps and my ochra spores consistently produced pigmented phenotypes with convex caps. So any leucistic/albino phenos with a convex cap or pigmented phenos with a papillate cap would most likely be hybrids. Lo and behold, those are exactly what I got! It also features one extra generation in the APE x Malabar branch since it was a precursor project that got assimilated into this one.
The Lovi Mänty one shows a similar streamlined approach with the APE x Ochra hybrid with the exception of an extra F2 generation to express the diversity and further confirm successful hybridization.
The Lovi Kuusi tree is the most "canonical" of the three in terms of the method described above. Here the only deviation is addition of an extra generation before the step 4 2+2 mon-mon hybridization. It also highlights how, contrary to the popular belief, albinism isn't a recessive trait.
Any feedback or questions welcome! PM me if you'd like to contribute and receive some spores.
That's all I have at this point, I thank you for your interest.
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