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OfflineViper
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Amanita idea
    #6164958 - 10/13/06 08:23 AM (17 years, 5 months ago)

Ok, I don't know if anybody came up with this before, but I just got an idea: If you can create an edible medium for A. muscaria spores, and let them grow into a mycelium, would you still need to make it fruit, or could you just use the mycelium as an alkaloid source. Does anyone know anything about the alkaloid content in mycelium? Are the alkaloids present only in the fruitbody? Can we make edible substrates, and never need to grow fruitbodies. I know that myceliums od Psilocybes have alkaloids in them, and that they can be used for alkaloid extraction.
Anybody? :crazy:


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Invisibleelsig
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Re: Amanita idea [Re: Viper]
    #6165131 - 10/13/06 09:42 AM (17 years, 5 months ago)

I have been wondering about something similar, what are desired in fly alergic mushrooms? is it the alkaloid muscarine or the two isoxazols ibotenic acid muscamol. If you are going to make an extract you want a purified muscarine extract? I was under the impressiob that it was the isoxazols that were responsible for the hallucigenic effects to fly alergics.

Edited by elsig (10/13/06 10:02 AM)

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OfflineViper
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Re: Amanita idea [Re: elsig]
    #6165567 - 10/13/06 11:45 AM (17 years, 5 months ago)

Yes, the main hallucinogen is muscimole. But the question is wether or not muscimole is present in a growing mycelium. Would it be worth extracting it from an agar culture, or even eating the entire culture.


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Invisiblefastfred
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Re: Amanita idea [Re: Viper]
    #6167909 - 10/14/06 01:52 AM (17 years, 5 months ago)

Highly unlikely.

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OfflineViper
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Re: Amanita idea [Re: fastfred]
    #6168011 - 10/14/06 02:36 AM (17 years, 5 months ago)

Why not? Ok, I get the part about the highest concentration of alkaloids being just below the red skin of the fruitbody, but alkaloids are also present in the stem, and the stem is quite similar in metabolism to mycelium. And mycelium of A. phalloides is poisonous, is it not? It seems worth testing. Has anyone heard of a qualitative test for muscimol? I'd really like to check this out.


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Invisiblefastfred
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Re: Amanita idea [Re: Viper]
    #6174995 - 10/16/06 12:50 PM (17 years, 5 months ago)

Primary amines are fairly reactive so it shouldn't be too hard to figure out a test. Primary amines will usually react with ketones and aldehydes. So you might try to see if there is any reaction with acetone or MEK.

There is no indole present, so pDMAB probably wouldn't work.

Try looking here for more info...
http://en.wikipedia.org/wiki/Amine
"Primary amines react with ketones and aldehydes to form imines."

Imines are interesting. They have a triple bond to the nitrogen, so I would have to think there would be some sort of reaction you could use as an indicator.

The reason I think there probably isn't any actives in the myc is because that is mostly the case with psilocybe varieties. There is a tiny amount, but nothing worthwhile. I'm guessing the same would be true with amanitas. Since they use the ibotenic acid as a sort of poison it would make sense that they would produce this only in structures that need to be protected, i.e. the fruitbodies.


-FF

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Offlinenobhdy
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Re: Amanita idea [Re: fastfred]
    #6181432 - 10/18/06 06:52 AM (17 years, 5 months ago)

as novel as that idea seems, to grow the fruitbody would be much more economic. and all you would have to do is wait another few days.


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OfflineFraggin
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Re: Amanita idea [Re: nobhdy]
    #6181906 - 10/18/06 10:12 AM (17 years, 5 months ago)

Is amanita a mycorrhiza species? Or another form of a symbiotic mushroom?

I have read about products that contain mycorrhiza liquid inoculate with nutrient mixtures (including liquidized kelp) that are used to "inject" into the soil around the root ball of palm trees. Cant remember if they were Endodermal (is this the word) type fungus or not, but I do remember that it did not penetrate the cell wall of the cortex on the roots.
In any case, this may be a worthy experiment to see if one could inoculate a conifer tree with a similar liquid inoculate of amanita.

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Offlineelemental762
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Re: Amanita idea [Re: Fraggin]
    #6229857 - 10/31/06 05:52 AM (17 years, 4 months ago)

Amanita is a mycorrhizal species on conifers.
A solution of amanita spores could work the same way. I've often wondered how well they would develop from some intentionally placed spores. The solution seems like a more effective method, in theory.


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OfflineFeelers
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Re: Amanita idea [Re: elemental762]
    #6231963 - 10/31/06 04:16 PM (17 years, 4 months ago)

Growing amanitas is not worth it - there are millions everywhere throughout the world, picking them would be far cheaper than growing them - (which is currently impossible).

If you want muscimol ir Ibotinic acid or whatever one it is, I'm sure a direct chemical symthesis would be very easy. Cut out the middleman and save! :grin:

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Offlineasci
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Re: Amanita idea [Re: Feelers]
    #6232191 - 10/31/06 05:26 PM (17 years, 4 months ago)

amanita muscaria is a ectomycorrhizal fungi. the mycelium creates a sheath of cells around the rootlets of certain species of vascular plants, creating a symbiosis between the fungi and the vascular plant. culturing and cultivating a ecto or endo mycorrhizal fungi is possible, but extremely difficult. many species need a complex mixture of micro flora to support spore germination, or the mycelium needs to be introduced to a host plant or tree, with in a certain amount of time. there has been limited success in cultivation, but current break through are being made at oregon state university, with tuber and cantharellus species. but no amanita.

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Offlinehjalmar
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Re: Amanita idea [Re: asci]
    #6257173 - 11/07/06 11:44 AM (17 years, 4 months ago)

make way for the chemist :wink:

Quote:


Primary amines are fairly reactive so it shouldn't be too hard to figure out a test. Primary amines will usually react with ketones and aldehydes. So you might try to see if there is any reaction with acetone or MEK.





No this will not work as a qualitative test. Your amine will likely form an imine, which is btw a reversible reaction (A + B <--> C + D), but don't think you'll actually see something happening. There will be no color change or exothermic whatsoever.

There are better quantitative tests on primary amines, for example NaNO2 which gives N2 gas evolution, but these test are of no value whatsoever. Because the mycelium will more than certainly contain all sorts of amino groups from amino acid and proteins present in there.

There is however a relatively simple way to check it, and that is using Thin Layer Chromatography.
http://en.wikipedia.org/wiki/Thin_layer_chromatography

Mix some mycelium with a dilute acid solution, filter, basify and extract the muscimol containing fraction using dichloromethane or chloroform. Do the same with a piece of dry Amanita mushroom, this will be your control standard.

Which solvent to use and Rf values I have to look up, detection can probably be done using iodine vapours or spraying with aq. KMnO4

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Offlinehjalmar
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Re: Amanita idea [Re: hjalmar]
    #6257191 - 11/07/06 11:50 AM (17 years, 4 months ago)

Quote:



If you want muscimol ir Ibotinic acid or whatever one it is, I'm sure a direct chemical symthesis would be very easy. Cut out the middleman and save!




Without a professional lab with all necessary equipment and chemicals I'd rather extract a couple of mushrooms.

The synthesis of muscimol/ibotenic acid isn't facile and certainly not something an untrained individual could pull off in the basement, even if one could buy the necessary equipment, solvents and reagents as an individual without an eyebrow being raised..

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OfflineViper
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Re: Amanita idea [Re: hjalmar]
    #6261336 - 11/08/06 11:17 AM (17 years, 4 months ago)

Ok, so we have a few ideas for the tests. Now all we need to do is get some spores, start a mycelium, and do the qualitative tests for ibotenic acid and muscimole. If these come out positive, growing a mycelium on rye would make some sense. We could even grow it outside on a root of a tree, it would be faster with a grown up mycelium.

Indeed, it is more interesting to go out hunting, but autumns in my country tend to be cold... and my personal lab is so warm...

P.S. The idea with imines seems worth a shot, so I'll give it some more thought. In combination with my trusty spectrophotometer it could be great. I presume nobody has a spore print or two of Amanita? :crazy:


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Offlineasci
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Re: Amanita idea [Re: Viper]
    #6262577 - 11/08/06 05:24 PM (17 years, 4 months ago)

i don't think you understand the requirements for the growth of this species. you can not just culture a couple of spores from amanita muscaria, and spawn they on rye grain. this is not a saprophytic species. amanita species depends on soil micro flora to germinate the spores and a immediate symbiosis with a vascular plant to survive. it would be to your best interest to pick the fungi in the wild and perform the extraction that way. whole lot cheaper!! but there is a special type of agar to culture ecto/endo mycorrhizal, beyond that point of cultivation it becomes extremely difficult to keep the fungi alive or to maximize your mycelium output. good luck

Edited by asci (11/08/06 05:51 PM)

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Re: Amanita idea [Re: asci]
    #6263529 - 11/08/06 09:47 PM (17 years, 4 months ago)

Have you tried cloning any mycorrhizal species?

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OfflineViper
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Re: Amanita idea [Re: falcon]
    #6265322 - 11/09/06 12:35 PM (17 years, 4 months ago)

No, I know I can't get the fruitbody of the Amanita, that's why I'm asking about the alkaloid content of the mycelium. I read that the mycelium can be grown in petri dish, on agar, so the idea is to produce a large amount of mycelium and extract alkaloids from it. It works with psilocybes, maybe it could work with Amanita, if there is muscimole in mycelium... True, the best way IS to go out hunting...


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Invisiblefastfred
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Re: Amanita idea [Re: Viper]
    #6265462 - 11/09/06 01:09 PM (17 years, 4 months ago)

> It works with psilocybes, maybe it could work with Amanita,

It actually doesn't really work with psilocybes. Sure you can get some small but detectable amount, but it's completely unpractical. I think you'd have the same problems doing it with amanitas.

If you could get a couple dozen quart jars of amanita myc to grow up to a good concentration you might be able to extract a dose or two from them, but it's hardly worthwhile.


-FF

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Offlineasci
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Re: Amanita idea [Re: fastfred]
    #6265571 - 11/09/06 01:41 PM (17 years, 4 months ago)

amanita mycelium is able to be cultured on a specialized agar medium, but the mycelium growing on rye grain is a different story. to produce the amount of mycelium needed for extraction is impossible with this specie. due to the fact that this fungi need another nutrient base that is not supplied by the grain medium, instead supplied by the vascular plant. if you could figure out the exact nutrient needs for amanita muscaria you might be able to spawn the mycelium, and skip the symbiosis needed to form the healthy mycelium in the quantities needed to extract compounds, currently i do not think that would be an option.

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Re: Amanita idea [Re: Viper]
    #6266230 - 11/09/06 05:11 PM (17 years, 4 months ago)

I have no idea how much mucimol if any is in the mycelium.
Your best bet is to learn to identify  Amanita muscaria.

Amanita muscaria mycelium is pretty easy to clone on PDYA, if you have
a young button with no worm damage, at least that has been my
experience. It will also grow on grain.

Amanita muscaria mycelium
will probably grow on just about anything that any other mushroom
will grow on. <-(speculation) based on my observation of other cloned
mycorrhizal species growing on other stuff, chantrelles and Boletus
subcaerulescens, the mycelium of both will grow on various grains,
coffee grounds and supplemented sawdust.

Amanita muscaria will not however fruit without a symbiot, so far :hehehe:
and since the mucimol is probably
a defense mechanism to protect spores in the fruit body, it's
probably  a good bet that there isn't very much of it in the
mycelium.

I wasn't  able to get the spores to germinate on agar working
from gill tissue. I haven't tried working from a print. I ended up
with a lot of green mold on the agar. The mold spores were probably
ended up on the gills from the mushrooms being
air dried.

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