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Facts vs fiction - Conditions affecting mushroom growth and psilocybin production

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What are the proven facts that affect the growth of mushrooms and potency? A lot is said around different teks and posts but I was interested to find what was actually tested scientifically with controlled variables, and not just anecdotally.

Here’s what I found so far:


1- Ammonium succinate, glucose content and acidic pH of medium increase psilocybin production in mycelium of P. cubensis, but no mention whether it correlates with fruit psilocybin content later on (Catalfomo & Tyler 1964)

2- Darkness improved colonization of panaeolus spp. mycelium (Bustillos et al 2014)

3- No strong correlation between pH changes from 6-8 and mycelium growth of Panaeolus spp. (Bustillos et al 2014)


1- With Panaeolus fruits, horse dung was 10x worse in yield compared to cow or buffalo dung. (Bustillos et al 2014)

2- To induce pinning, light is essential, UV and blue wavelengths of 370, 440, and 460 nm were the most effective. Green and red wavelengths greater than 510 nm were ineffective. (Badham 1979)

3- Darkness post-pinning till harvesting increases psilocybin content by 100 times (!!!!!) compared to indirect or direct lighting (Riahi et al 2009)*

4- After pinning, darkness and light makes no difference to mushroom growth speed and size (Badham 1985)

(Scientific-minded experiments by home growers)

1- Straw vs coir vs straw+coir - Similar results but coir best results (Rahz@Shroomery)

What surprised me the most from this is the aspect about darkness.. Why aren't people talking about this 100 fold difference??? (edit: * Found this comment from Loveall saying the method used to extract shrooms to test might have not dissolved psilocybin. Still would be interesting to try and reproduce this. I might try and contact the authors and ask)

Any other variables you know have been tested before, either in published papers or at least in home experiments that followed the scientific method doing side-by-side comparisons and not comparing in different moments with other uncontrolled variables?
We need to get some experimenting and analysis on the go, this is most interesting.
How difficult is quantitative analysis for the everyman?
Well, this is very well timed, as I'm thinking about trying my hand at growing shrooms for the first time in over 10 years-- have you seen anything definitive regarding the gypsum admixture showing a consistent increase in yield? Some of the most prolific commercial (and recreational) growers buy it in bulk and swear by it...


endlessness, I tried to read the link for that amazing dark-after-pin thing, but this is what it said:

<Message>Request has expired</Message>
Xt, I think it can be done with TLC but I haven't tried it yet with mushrooms.x

coAsTal, I have not found a single experiment on gypsum vs no gypsum on cubensis.

I have found a few papers and patents describing the use of gypsum in growth of edible/medicinal mushrooms, and the reasons why it is used:

for example:

"addition of 1% gypsum in combination with 3% of horsegram flour increased the yield (of Plaetorus mushrooms)" ... "partially-filled paddy grains (PFPg) supplemented with calcium carbonate, horsegram flour and gypsum has been evaluated as the best substrate"

"Furthermore, chalk and gypsums act as buffer. Besides, the Ca2 obtained from gypsum (CaSO4) and chalk (CaCO3) neutralize oxalic acid produced during mycelial growth"

"Gypsum is added to minimize the greasiness compost normally tends to have. Gypsum increases the flocculation of certain chemicals in the compost, and they adhere to straw or hay rather than filling the pores (holes) between the straws. A side benefit of this phenomenon is that air can permeate the pile more readily, and air is essential to the composting process. The exclusion of air results in an airless (anaerobic) environment in which deleterious chemical compounds are formed which detract from the selectivity of mushroom compost for growing mushrooms. Gypsum is added at the outset of composting at 40 lb per ton of dry ingredients."

Counter-intuitively, this one paper talks about adding gypsum actually decreased speed of substrate colonization. Also it claims that gypsum presence changed the chemical profile found in the substrate, which could be beneficial for some mushrooms species but not to others. Unknown how this relates to cubensis.

This patent paper talks about how gypsum is used to improve physical characters of the substrate but the mycelium growth might slow down if you use too much gypsum

Regardng the broken link above, I changed it to link Loveall's post where the paper is attached, so you should be able to see it now.
Apparently if you mention the 'x100 psilocybin' on shroomery.org you will either get flamed or have your thread locked as that paper has been touted there many times.

It's one of those things that would be so revolutionary that I can't believe it hasn't been tried many many times regardless of the scepticism of would-be experimenters... and of course the information would spread through the internet like wildfire.

I'll give it a whorl just for the hell of it if I ever grow any.
Benzyme showed that adding tryptamine (10mM to 15mM) enhanced potency (6% to 9%). This jives with previous research by Gartz (paper is in the linked thread). Tryptophan doesn't help though, presumably because decarboxylation is too energy intensive (many home growers have looked into this as tryptophan is easily available).

Adding melatonin made the mushrooms in inactive and gave an unknown peak in MS. This needs to be repeated/confirmed as it is unpublished Nexus research (Dreamer042, Benz, and company).

Adding Mexamine to the substrate would be a very interesting experiment. Mindlusion had this going but the experiment could not be completed.

This is not scientific and just an oppinion (sorry), but there is some consensus among growers that salts can help by being a humidity buffer. Also, using gentle alkaline material raises the pH a little to slow down growth of contaminants while not bothering cubensis too much. It could be that if the grow has optimal moisture and no contaminants these adjuncts don't help (or even could be detrimental if too many are added).
As already stated, adding tryptamine boosts potency. What about DMT itself? Anyone ever tried adding chacruna leaves to the substrate?

If the enzymatic steps are tryptamine >> dimethyltryptamine >> 4-hydroxy-dimethyltryptamine...?
Espurrr said:
can we have this stickied
Done. :thumb_up:

Several folks over at shroomery tried putting mimosa bark in the substrate and reported that it did not seem to noticeably increase potency of the resultant fruits. They deemed it a waste of bark.
I'm sorry to revive an old thread but I did find something to add this morning.

It's the last few sentences in bold.

Psilocybin biosynthesis requires four enzymes (Scheme 1): l‐tryptophan decarboxylation and hydroxylation at C‐4 are catalyzed by the decarboxylase PsiD and the monooxygenase PsiH, respectively, to form 4‐hydroxytryptamine. Subsequently, the 5′‐adenosine triphosphate (ATP)‐dependent 5‐methylthioribokinase‐like enzyme PsiK phosphorylates 4‐hydroxytryptamine.4a The biosynthesis is completed by the methyltransferase PsiM which installs the tertiary amine and requires two equivalents of S‐adenosyl‐l‐methionine (SAM). Psilocybin titers can reach up to 2 % of the fungal dry biomass.5 Consequently, SAM must be efficiently regenerated to keep up the high psilocybin production rates

It seems that more S‐adenosyl‐l‐methionine (SAM) means more psilocybin. At least up to the limits of the genetics.
This is far from being the oldest thread that's been dug up this week!

Thanks from the info - it makes me think that restricting the availability of SAMe would limit the production of aeruginascin - the substance hotly suspected of being responsible for 'woodlover paralysis'; not only that, it would also be interesting to steer the profile of mushrooms in the direction of baeocystin and norbaeocystin production in order to learn more about these little understood compounds.

I find the dephosphorylated version of norbaeocystin - which I like to dub 'norbaeocin' in analogy with the psilocybin/psilocin nomenclatural pairing - particularly interesting as it is a positional isomer of serotonin and amongst other things it raises important questions about the nature of legislation such as the controlled substances analog act. Is serotonin an analog of psilocin? Possession of a brain is likely to be illegal :?
downwardsfromzero said:
Is serotonin an analog of psilocin? Possession of a brain is likely to be illegal :?
Humans contain DMT and that doesn't make them illegal, which probably answers this particular question.
I was hoping that someone would reply to the effect that most of the serotonin in the body is present in the gut. And of course, one ought to remember Poe's law - that deadpan doesn't work on the internet. 😁

More on topic, does anyone know anything about the activity (if any) of norbaeoc(yst)in [4-HT]?
A small amount of data shows that 4-HT has some activity at serotonin receptors in the rat:
https://doi.org/10.1007/BF00633189 - see table on p.4, but 4-HT was not mentioned in the final discussion.

Some other animal studies were done in the 60's that vaguely demonstrated something going on and another one showed a small protective effect against radiation which, interestingly perhaps, was a greater (or more reliable) effect than that of either psilocin or baeocin.

Looking at Pubchem, the activity of 4-HT appears to be significantly higher at the 5-HT1A receptor (rat) than at the 5-HT2A. Some activity (Ki = 40nM) is present for human 5-HT2C receptors.

It would be great to get hold of a copy of the following:
Lloydia, 01 Nov 1977, 40(6):566-578
Sixty collections of ten species referred to three families of the Agaricales have been analyzed for the presence of baeocystin by thin-layer chromatography. Baeocystin was detected in collections of Psilocybe, Conocybe, and Panaeolus from the U.S.A., Canada, Mexico, and Peru. Laboratory cultivated fruitbodies of Psilocybe cubensis, P. semilanceata, and P. cyanescens were also studied. Intra-species variation in the presence of decay rate of baeocystin, psilocybin and psilocin are discussed in terms of age and storage factors. In addition, evidence is presented to support the presence of 4-hydroxytryptamine in collections of P. baeocystis and P. cyanescens. The possible significance of baeocystin and 4-hydroxytryptamine in the biosynthesis of psilocybin in these organisms is discussed.
endlessness said:

1- With Panaeolus fruits, horse dung was 10x worse in yield compared to cow or buffalo dung. (Bustillos et al 2014)

3- Darkness post-pinning till harvesting increases psilocybin content by 100 times (!!!!!) compared to indirect or direct lighting (Riahi et al 2009)*

These numbers seem totally exagerated and I'm sure will come to ligth these ratios are incorrect. as per horse dung with pan cyan and psilocybin content 100 times lol... :? :d
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