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Re: Best chemical activators of spores for germination? [Re: bodhisatta]
    #25964169 -

Bod is correct, hydrogen peroxide neutralizes bleach resulting in pure o2 (which is still a deadly gas and highly flammable) and the remaining residue can be fully neutralized with white vinegar.


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Re: Best chemical activators of spores for germination? [Re: Professor X]
    #25964767 -

Oh damn, right... one of the few things you can mix with bleach i guess. :smile: my bad

Not the part i'm personally most excited about tho.

thanks for the help.


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    • Edited by Solipsis (04/30/19 05:21 PM)

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    Re: Best chemical activators of spores for germination? [Re: Solipsis]
        #25964948 -

    Simple sugars are probably the best activators, like monosaccharides. There apparently somehow linked to carrier proteins, that move ions around inside the Spore.

    If I'm losing my mind I apologize. Haven't slept much these past few days.

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    Re: Best chemical activators of spores for germination? [Re: vinnie boombotz]
        #25965546 -

    Is it only chemical signals? We know that temperature has an impact on germination. Below a certain temperature do spores even germinate? Tropical species don't seem to germinate at all if the temp is too low. I doubt there is one signal that will activate germination in all conditions. Except: https://www.sciencedirect.com/science/article/pii/S0007153678801284

    Paxillus involutus didn't germinate on agar (a really complicated recipe). Activated charcoal in the media promoted some germination. Chemicals produced by P involutus mycelia germinated even more spores, similarly for Leccinum scabrum.

    Here, the presence of living or growing (or whatever stage of growth produces those "volatile factors") mycelium promotes germination of some species. I wonder if inactivate mycelium mixed into an agar medium will promote germination of our species.

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    Re: Best chemical activators of spores for germination? [Re: teladi]
        #25967757 -

    You are right, it is not only chem signals, other types have been mentioned in the thread and/or the references linked and quoted.

    If the spores are in the tricky kind of dormancy where even normal basic nutrients dont activate them and trigger germination, you apparently need either some kind of stratification or at least certain particular changes in conditions and those do include high or low temp yes.

    Interesting and useful and i didnt necessarily mean to make the discussion narrow but i was looking for chemical activators which bypass dormancy issues and somehow just forcibly activate em and trigger germination. Simply because i am curious from looking at the example of plant seeds and hormones.

    But yea i do appreciate all this info on what helps to germinate. Some of these options are relatively easy to include if you have spores which are just unusually old and stored under unknown conditions. Even just as precaution, to just include that something extra and maximize your odds.

    Spores of e.g.:

    P. atlantis
    P. villarealia
    P. semilanceata
    P. friscosa

    And yes I realize these are old names.


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    Re: Best chemical activators of spores for germination? [Re: Solipsis]
        #25967818 -

    insect pheromones perhaps? maybe phorid fly pupae in the substrate, which eat mycelium

    also. from https://www.ncbi.nlm.nih.gov/pubmed/16907803/

    Germination in response to specific chemical nutrients requires specific receptor proteins, located at the inner membrane of the spore. After penetrating the outer layers of spore coat and cortex, germinant interacts with its receptor: one early consequence of this binding is the movement of monovalent cations from the spore core, followed by Ca2(+) and dipicolinic acid (DPA). In some species, an ion transport protein is also required for these early stages. Early events - including loss of heat resistance, ion movements and partial rehydration of the spore core - can occur without cortex hydrolysis, although the latter is required for complete core rehydration and colony formation from a spore.

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    Re: Best chemical activators of spores for germination? [Re: vinnie boombotz] * 1
        #25970091 -

    How did you figure insect pheromones? :smile:

    Bacillus however is a spore producing bacterium genus and I'd rather not try to glance from that even if there may be similarities.


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    Re: Best chemical activators of spores for germination? [Re: Solipsis]
        #25970137 -

    Okay forget the insect pheromones,can't figure how I came to that thought.... what about substances that would simulate the digestive system of mammals, since some saprotrophs are eaten then germinate. But they do need plant material after that.... what would keep a spore from germinating in the first place?

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    Re: Best chemical activators of spores for germination? [Re: vinnie boombotz]
        #25982934 -

    How about a basic wetting agent such as soap or Yucca. All you really need to do is re hydrate the spores to make them more viable.


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    You see what you want to see not what it makes you see. I see said the blind man to the deaf women .

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    Re: Best chemical activators of spores for germination? [Re: growmore]
        #25991772 -

    Simulating the digestive system may work for some spores which have a life cycle which specifically involves that sort of activation, interesting.

    Other than that you can read the thread, especially my later posts - there is some info and some references about how spores can go dormant and this would keep it from germinating even if basic conditions are right for germination. They then may need an extra trigger to stimulate and activate it, or it may be possible to force / bypass it through the trehalose story.

    Soap etc should work similar to but possibly a bit less effective than other surfactants such as polysorbate. It seems these are a few of many ways to alter the permeability of spores and a little of that can help to trigger germination normally (though possibly not with the hard type of dormancy).
    I think some of these surfactants may screw up the spore at higher concentrations than that, while other types of surfactants do seem to be tolerated and do more to separate and suspend spores than to force permeability too much.


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    Re: Best chemical activators of spores for germination? [Re: Solipsis]
        #26010192 -

    I love this thread. I have a lot of spores 10 years +  that I need a miracle for...

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    Re: Best chemical activators of spores for germination? [Re: murderlabz] * 1
        #26010374 -

    I highly recommend this paper for more information:

    https://link.springer.com/article/10.1007/BF03053077

    Fries, Spore germination in the higher Basidiomycetes,Plant Sciences 1984

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    Re: Best chemical activators of spores for germination? [Re: Buthoscorpio]
        #26011986 -

    Buthoscorpio said:
    I highly recommend this paper for more information:

    https://link.springer.com/article/10.1007/BF03053077

    Fries, Spore germination in the higher Basidiomycetes,Plant Sciences 1984



    Nice find! full text ->

    https://www.ias.ac.in/article/fulltext/plnt/093/03/0205-0222

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    Re: Best chemical activators of spores for germination? [Re: murderlabz]
        #26216410 -

    Success!

    For the life of me I couldn't get these 10yr old Ps. cubensis spores to germ on any Agar. So I went to LC(yeah yeah I know) Shot spores stupid heavy, 1 super dark spore syringe for 8 jars. Air syringed it and try to press as much air in the jar with the air syringe as possible. Maybe 1-2 psi if lucky? 2 of the 8 jars popped. The others either had the cloudy bacterial look or just a clean water look with sediment at the bottom. These two look gnarly, one jar has two colonies going and looked suspect so I worked with the other LC jar.

    I shot up cakes as I want spore prints ASAP also I can grab chucks off the cake to go to Agar and still fruit the cakes. So far the cakes look clean, which is crazy as I was ready for a battle.


    Here is the recipe I used

    1 cup = 240ml
    1/2 tsp = 2.5ml


    To make 4 pint jars at a time and have room for air:

    2 cups distilled water
    2 Tsp Honey
    2 Tsp Karo
    10 drops Trace Minerals ionic - Potassium
    10 drops Trace Minerals ionic - Trace Minerals
    BB size of Organic Activated Coconut Charcoal
    BB size of Blue-Green Algae

    I mistook the Potassium for the Trace Minerals, I just left it added in and kept rolling. This all could be luck, bullshit whatever just showing what I used.





    Edited by murderlabz (09/28/19 09:20 PM)

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    Re: Best chemical activators of spores for germination? [Re: murderlabz] * 1
        #26264684 -

    That's very interesting, thanks for sharing it. Looks good, be sure to check it on agar asap?

    So were you planning on diluting the spore suspension if this didn't work?

    And these were supposed to be cleanly printed non-wild spores, right?

    You pressurized the syringe to hydrate?

    Why the addition of the activated carbon here? And what about the algae?

    (I myself haven't tried a lot of special things yet for my old spores, i think possibly I had some drying out that didn't help my straightforward germination on half strength agar.)


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    Re: Best chemical activators of spores for germination? [Re: Solipsis]
        #26264744 -

    Solipsis said:
    So were you planning on diluting the spore suspension if this didn't work?


    No, My hypothesis is that the spore death rate is about 90-95% or more on these. I believe this is why agar failed. With LC I shot the whole dark print equally-ish into 8 jars. Only 2 showed germinated so that gives an idea of the spore death rate. I don't see how you could pull this off with Agar unless your scrape the whole print and pepper/coat the entire top of your Agar(soft Agar) with spores. I think the goal for success here is power in numbers.

    Solipsis said:
    And these were supposed to be cleanly printed non-wild spores, right?



    clean - a print I make in 2009-2010 on foil

    Solipsis said:
    You pressurized the syringe to hydrate?



    I have a syringe that has a blood pressure cup ball on the end.
    I'll have to post a picture to explain. I used it on the LC jars. So maybe 1 psi? I don't think it did much haha



    Solipsis said:
    Why the addition of the activated carbon here? And what about the algae?



    Carbon - Said to aid in germination
    Potassium - Total accident
    Trace Minerals - I always add to my LC's
    Blue-Green Algae - I always add to LC's as well. Right now this is "hokus pokus" in short - Cyanobacteria has a super thin cell wall and is a great snack for Myc.

    Edited by murderlabz (10/20/19 08:41 PM)

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    Re: Best chemical activators of spores for germination? [Re: Solipsis]
        #26266206 -

    Solipsis said:
    That's very interesting, thanks for sharing it. Looks good, be sure to check it on agar asap?



    Yeah, I am trying to fruit them out ASAP for prints as some one is riding my arse for spores. This is F2-F3 away from the original he sent me back in the day.

    But I am a re-noob I used poly lids for my BRF cakes lol! A little dry but working through it.

    I do have a SHIT-TON of wild cube prints collected in Texas from the US circa 2009-2011. If interested PM me.

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    Re: Best chemical activators of spores for germination? [Re: murderlabz] * 1
        #26275345 -

    Fascinating..

    Any reason why the algae even with a thin cell wall, would be better than things like yeast extract and peptone for a pretty complete biochem diet without even any cell walls at all? (i mean you can lyse open cells if you wanna get the food out).
    Just trying to figure out whether this is more convenient, improvised or advanced so to speak :laugh:

    Yeah i've been reading a bit more about the uses of activated carbon, it's also relevant for plant tissue culture I'm doing, i should have received the carbon now on another address for experimenting.

    Idk what the physical limits are of the average syringe (I do have a good brand), will be fun to try out some improvised ways of putting plenty of pressure on it (but not too much haha).


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    Re: Best chemical activators of spores for germination? [Re: Solipsis]
        #26277388 -

    I can't prove it's benefit. I "believe" it helps but could just be bullshit placebo. Just a cool little super food. I like peptone but didn't have any :smile:


    Updated*

    I'm not 100% sure of the mechanism of the activated charcoal but I think that was the "ticket" I should of tested pH.

    They are ruff but got me to where I now have fresh spores.








    Edited by murderlabz (11/28/19 12:37 PM)

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    Re: Best chemical activators of spores for germination? [Re: murderlabz]
        #26393329 -

    murderlabz said:
    think that was the "ticket" I should of tested pH.



    Per Mary-Lou Florian from her "What it Takes to Activate a Fungal Spore" "Acid and alkaline treatments also cause activation" I guess you could swing the other way to be more acidic.

    Coconut shell based Activated Carbon(what I used) normally is specified for a pH of 9 – 11.

    On a side note, This will help with Morels and other ectomycorrhizas

    Later it was discovered that germination in various ectomycorrhizal fungi was prevented by an inhibitor in agar which is formed from agarose during the autoclaving of the agar medium. The inhibitor could be removed by activated charcoal (F¡ 1978)

    Other related papers

    Edited by murderlabz (12/21/19 12:12 AM)

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