Funnily enough Slippery Crossing was the one sample that I couldn't get a clean sample from. I remember checking and it might be this.

All of the other wild Australians are uniform but very tomentose (F2 still), has that been the case for you? Most rhizo out of Eungella so far, this next plate should be nice.

Right now, the best growth I'm getting is from Axman's Tico (Costa Rica) that I have on 2 grain jars that I will shoebox each, and Tanan Negara which is Malaysian and supposedly only collected in the past few years and from distinct genepool.

T. Negara



More recently I've been hyping about a cross I think I managed to make by running a swab + print rubbed together in a pastyplate container and wrapped in foil for 45 minutes in the instant pot. This is not from wild genes, but if the method works out, it would be a new way to germinate wild samples, and maybe even simplifying things by weeding out weaker genes.

I thought it was a bust when my spore syringes didn't work, but after finding a germinated clump and putting it on agar, I think I might've found a very interesting way of creating crosses from Stipe's thread on spore cleaning.

Huskies said:

So I've been doing some thinking and research, and I came across quite a few things that are relevant to this thread, and my own experience with the instant pot. I'll make my own thread on that once I do more experiments, but check these out (note, I did use chatgpt, asked it to source and quote the answers).

1. Spore Germination Triggered by Heat Treatment and Water

A common finding in mushroom cultivation is that a brief heat treatment (or “heat shock”) followed by re-hydration in water activates the spore’s internal metabolic pathways and breaks dormancy. For example, Paul Stamets in Growing Gourmet and Medicinal Mushrooms notes that:

    “Heat shock followed by water exposure activates the spore’s metabolism, leading to rapid germ tube formation.”
    This process is explained by the release and activation of enzymes and stored nutrients (e.g., trehalose) when the spore rehydrates. In essence, the heat treatment disrupts the spore’s static, dormant state, while water provides the necessary medium for biochemical activity.
    Source:

    Stamets, P. (2000). Growing Gourmet and Medicinal Mushrooms. (A practical reference that outlines how controlled heat treatments can trigger germination.)

2. Innate Preferences of Mycelia to Mate with Non‐Inbred Genes

Fungal mating systems are designed to promote outcrossing by using highly polymorphic mating‐type loci that act as “non‑self” recognition systems. As a result, mycelia formed from spore fusion tend to favor the union of genetically diverse nuclei. One review explains:

    “Fungal mating-type systems function as a non‑self recognition mechanism, effectively favoring outcrossing and reducing inbreeding.”
    This bias is not due to a conscious ‘choice’ by the fungus but emerges from the molecular machinery—pheromone signaling and receptor binding—that permits only compatible (and thus typically non‑inbred) nuclei to merge.
    Source:

    Billiard, S., Lopez‑Villavicencio, M., Hood, M.E., & Giraud, T. (2012). “Sex, outcrossing and mating types: unsolved questions in fungi and beyond.” Journal of Evolutionary Biology, 25, 2397–2411.

3. Survival of the Fittest Spores in Extreme Environmental Conditions

Fungal spores are remarkably resistant structures designed to endure adverse conditions; only the most robust spores survive prolonged stress. A review on spore resilience notes:

    “Fungal spores exhibit exceptional heat and desiccation resistance, ensuring that only the fittest survive until environmental conditions become favorable for germination.”
    This “survival of the fittest” aspect means that, under extreme environmental stress (such as high temperatures, UV exposure, or desiccation), the spores that persist generally possess enhanced protective structures and internal reserves. These hardy spores form the basis for future growth and colonization.
    Source:

    Guzmán, G. (2005). Psilocybe: A Global Overview. APS Press.

4. Do the “Fittest” Spores Give Rise to Strong, Resilient Mycelial Cultures?

Several sources indicate that the spores surviving extreme environmental conditions (thus being “the fittest”) tend to develop into robust mycelial networks. For example, Paul Stamets has noted in Growing Gourmet and Medicinal Mushrooms that:

    “Heat‐treated spores that survive usually yield mycelium with enhanced vigor, as they represent the hardiest individuals that have withstood environmental stress.”
    (Stamets, 2000)

Similarly, Guzmán has discussed that the spores from Psilocybe species which remain viable after exposure to adverse conditions often lead to strong and resilient cultures in later growth stages:

    “The surviving spores, having inherent stress resistance, give rise to colonies that are unusually robust, ultimately outcompeting less viable propagules.”
    (Guzmán, 2005)

5. Do the Nutrients from Dead Spores Serve as Food for the Survivors?

It is well recognized in microbial ecology that autolysis and recycling of dead biological material help support remaining living cells. In the context of fungal cultivation, several authors have highlighted that when an extreme environmental stress causes many spores to perish, the cellular components and nutrients released from the dead spores can be recycled by the surviving fungal elements. For example, reviews on nutrient recycling in fungal networks indicate:

    “Fungal mycelia are efficient at recycling nutrients from senescent or dead tissue. The decomposition of dead spores provides additional substrate—albeit in small quantities—that can be assimilated by actively growing mycelium.”
    (Fricker et al., 2012
    pmc.ncbi.nlm.nih.gov
    )

This recycling process is one reason why fungal colonies can persist in adverse conditions; rather than wasting potential nutrients, the mycelium reclaims them to support growth.

So, in my case with the IP, I heated enough to sterilize the water from the spore solution. No bacteria or mold showed on my test plates, only after it had been opened up and the germinated clump taken out in open air did the yeast form on the transfer plate. It is currently getting eaten by the incredibly strong looking mycelium. OJ are the transfer plates.



The IP only goes up to 13psi, so perhaps it wasn't enough for full sterilization (and perhaps cubensis spores can survive it). Things that might help, like an armor of sorts: plastic pp5 containers, wrapped in aluminum foil, and submerged in water.

Even if that run in the IP killed 99.99% of the spores, considering a spore print can contain billions (let's say one billion for easy math), that is still 100,000 surviving spores, the strongest ones, that will create stronger mycelium, and they will even be able to cannibalize the nutrients from the dead spores, which would be an extra trigger for germination.

Adding a flake of nutritional yeast in the container where the spores are could also help trigger germination even further, and will be something I test.

This has fantastic implications for crossing spores. If you mix two spore prints in the same amount in water, then run it through the dehydrator or even IP, a huge proportion will die, BUT, the strongest will survive, they will have triggers for germinating in the sterile fluid, AND on top of that, are per-disposed to create mating pairs with other genetics.

If germination regularly occurs in this container, and you have a macroscopic germinated colony, that will almost certainly contain crossed genetics.





I can say that this mycelium looks very robust, its overtaking what I think is yeast. I wonder if that means it will eat it, or if it will be hiding underneath? I'm going to put that germination plate to grains, even if it contams, I want to try to get some fruits from it.

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I really need to get that Tico Update underway... Jars spawned to bags late on the 14th, moving along ok. I'll post a bigger update when I get some pics and compile my now 3? pending log updates for LAGM Things slipped by me a touch there. So... I'll be back later today with some ok pictures of barely-ok work

Awesome to have you here, Rev! I've been reading on crossing because it seems fun, though I need to get a touch more comfortable with my agar and clean work before it makes sense for me to engage in it... Still, faht has a few methods that don't rely on finding a monokaryon first, namely this thread and RADS, the latter being summarized in his FP+ thread as:

fahtster said:

I found the best way was to let the swabs dry for at least a week (I just let them dry in the swab packaging) and then take both swabs from a pack and first dunk them into a jar of sterile water (I like to rub them together lightly a couple times in this first jar to remove any gill fragments and possible dust or mold spores on the surface) and then I move them to a second jar of sterile water that only has ~15ml of water (this is about the amount of water in a 4oz jar that’s enough to cover the tips of the swabs but not enough to over dilute the SS) and vigorously rub the two swabs together where the spores are (anyone that gets swabs from me will notice I swab gills on one side of the swab tip so that rubbing the swabs together is easy… it’s much harder to rub the very ends of swabs together lol) until the swabs look free of spores… then I suck that water up into a sterile syringe. 

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