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 Silven
Registered: 08/30/04
Posts: 2,072
Loc: El Mexicano
Last seen: 1 year, 6 months
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 A Mycelium Study?   1
#3323473 - 11/05/04 07:17 AM (20 years, 2 months ago) |
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So Discman's post about what exactly makes mycelium grow in liquid cultures got me wondering.. what exactly does make mycelium grow?
Are there any specifics as to why mycelium grows so well on grain, or what is present in grain/poop/straw/etc that makes it grow so well?
Is there a common link between each of these substrates? Is this link also present in karo or honey since it grows well in liquid cultures?
If we could pin-point a few of the main growing parameters wouldn't it be easier to eliminate contam risks, speed up colonization, perhaps increase potency of the mushrooms, as well as increase chances of a very healthy and strong looking fruit?
I'm unsure as to where to start testing, or if I should just get packages from each of the substrates/liquid culture ingredients and compare the make-up of each to one another?
Are there any tests that come to mind on how we could break these down to pinpoint any specific chemical or grouping of chemicals in the substrates that make mycelium really go crazy with colonization?
My wording might be a little confusing I realize, but if one of you more experienced or knowledgeable growers would like to rephrase some of what I wrote, feel free.
Maybe I'm shooting in the dark?
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hawksapprentice
Yearns to Snowboard
Registered: 06/06/03
Posts: 3,195
Loc: Oregon
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Re: A Mycelium Study? [Re: Silven]
#3323495 - 11/05/04 07:26 AM (20 years, 2 months ago) |
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Maybe this should be in advanced cultivation???
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"I celebrate the Earth, my home, my mother, my grave, and as long as men are Man they must, if they would preserve the integrated being, do the same---[and preserve]--this rank casual hungry smelly sweaty lusting transitory body, my oozy pulpy liquid-bag-swollen body, bones, blood, hair glands, my bejeweled sex; I love and celebrate it all. never to let men forget that they are animals as much as gods---that is one thing I shall say."
Edward Abbey
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Anno
Experimenter
Registered: 06/17/99
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Re: A Mycelium Study? [Re: Silven]
#3323526 - 11/05/04 07:39 AM (20 years, 2 months ago) |
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>what exactly does make mycelium grow?
A combination of various nutrients.
Carbohydrates, proteins, fats, minerals, vitamins.....
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Silven
Registered: 08/30/04
Posts: 2,072
Loc: El Mexicano
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Re: A Mycelium Study? [Re: Anno]
#3323542 - 11/05/04 07:48 AM (20 years, 2 months ago) |
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Yea this probably should be in advanced.
So, I guess what I need to do though in the experiments is find a way to artifically inject certain carbohydrates/protiens/fats/minerals/vitamins in the substrate one at a time to see if any certain one attributes to a more efficient grow? Is this possible?
I need to brush up on my chemistry I'm thinking if I go forward with any of this, as well as my history of what has already been tested/done with mushroom cultivation.
Anyway, if any of you mods want to move this to advanced cult, please do so.
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discman1
journeyman
Registered: 08/24/04
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Re: A Mycelium Study? [Re: Silven]
#3323553 - 11/05/04 07:51 AM (20 years, 2 months ago) |
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It should be possible, but I seriously doubt it hasn't already been done.  I am sure all the answers could be answered by a trip to the library + a week or ten of reading. 
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hawksapprentice
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Registered: 06/06/03
Posts: 3,195
Loc: Oregon
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Re: A Mycelium Study? [Re: discman1]
#3323573 - 11/05/04 07:58 AM (20 years, 2 months ago) |
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It would be more fun to just try it out. The more people who are experimenting the better a awnser we can get out of it.
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"I celebrate the Earth, my home, my mother, my grave, and as long as men are Man they must, if they would preserve the integrated being, do the same---[and preserve]--this rank casual hungry smelly sweaty lusting transitory body, my oozy pulpy liquid-bag-swollen body, bones, blood, hair glands, my bejeweled sex; I love and celebrate it all. never to let men forget that they are animals as much as gods---that is one thing I shall say."
Edward Abbey
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Silven
Registered: 08/30/04
Posts: 2,072
Loc: El Mexicano
Last seen: 1 year, 6 months
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I agree Hawk, but I'm still taking that suggested trip to the library to get some reading material while I'm doing these experiments.
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4hodmt
AspiringMycologist
Registered: 04/06/04
Posts: 759
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Re: A Mycelium Study? [Re: Silven]
#3323617 - 11/05/04 08:11 AM (20 years, 2 months ago) |
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id say they all have simple carbohydrates.
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discman1
journeyman
Registered: 08/24/04
Posts: 962
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Quote:
hawksapprentice said:
It would be more fun to just try it out. The more people who are experimenting the better a awnser we can get out of it.
Oh, certainly.. but I mean, it gives you a head start so to speak. Not so much trial and error, etc.. can get more right to the point. It also might give you even more ideas.
Simple carbohydrates are certainly a very important aspect, but I think many of us are leaving a lot out.
Compost/manure generally gives the biggest shrooms, right? We should try and isolate exactly what causes that.
Nitrogen, phosphorus, potassium? Calcium, iron, copper, zinc, molybdenum, maganese? boron, cobalt, strontium? Humic acid? Fulvic acid?
What about growth hormones? Do mushrooms respond to auxins, cytokinins and betaines?
Amino acids? Glycine, alanine, valine, leucine, isoleucine, serine, threonine, aspartic acid, glutamic acid, aspartic acid, glutamic acid, asparagine, glutamine, lysine, arginine, histidine, cysteine, cystine, methlonine, proline, phenylalanine, tyrosine, and tryptophan?
How about vitamins?
Many of these things and much more are present in fertile compost...
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Silven
Registered: 08/30/04
Posts: 2,072
Loc: El Mexicano
Last seen: 1 year, 6 months
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Re: A Mycelium Study? [Re: discman1]
#3323766 - 11/05/04 09:03 AM (20 years, 2 months ago) |
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Thank you discman, that is the type of information I was looking for! 
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hawksapprentice
Yearns to Snowboard
Registered: 06/06/03
Posts: 3,195
Loc: Oregon
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Re: A Mycelium Study? [Re: discman1]
#3324081 - 11/05/04 10:46 AM (20 years, 2 months ago) |
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Acutally pound for pound grain give better yeilds.
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"I celebrate the Earth, my home, my mother, my grave, and as long as men are Man they must, if they would preserve the integrated being, do the same---[and preserve]--this rank casual hungry smelly sweaty lusting transitory body, my oozy pulpy liquid-bag-swollen body, bones, blood, hair glands, my bejeweled sex; I love and celebrate it all. never to let men forget that they are animals as much as gods---that is one thing I shall say."
Edward Abbey
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Tre_Selor
Stranger
Registered: 11/05/04
Posts: 35
Last seen: 19 years, 5 months
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Starch, and cellulose?
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 noxy
Dr
Registered: 09/20/04
Posts: 181
Loc: its more a time, not a pl...
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Re: A Mycelium Study? [Re: Tre_Selor]
#3324772 - 11/05/04 02:02 PM (20 years, 2 months ago) |
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the mushroom cultivator by stamets APENDIX 6
"analyses of basic materials used in substrate preparation"
a good place to start
but better yet is to isolate a strain from a culture
a multispore culture in a jar will have many many strains
consider that of all the strains of a tetrapolar system only one third of them will be capable of fruiting
your better off to isolate a strain that produces many big fruits given your testing conditions
Edited by noxy (11/05/04 02:10 PM)
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Search59
Yet anotherhobby for me.
Registered: 10/28/04
Posts: 116
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Re: A Mycelium Study? [Re: noxy]
#3324817 - 11/05/04 02:15 PM (20 years, 2 months ago) |
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Theres a few ways one can go about this study.
It will take ALOT of Petri dishes.
A single strain of mycelium in quesiton (as most mushrooms grow differently and require different conditions)
And the chemicals, starches, vitamins, protiens, and anything else you wanna test it with even coffee. Pure, uncontaminated liquid forms of these would be needed.
A sterile working environment. Fully sterile, clean room serile.
A optimized temp regulator for the petri dishes.
Equiptement, Microscope, quick injector sryinges.
Put medium in question in petri dish. Inject mycelium or spore.
Observe, make notes.
Then try using information gathered in the petri dish stage to make a fruiting stage experiment. I.E. the best way to get the nutrients to the fruit body. Via nano-carbon tubes??? Some cellular water/nutriend absorbing compound?
After many months of testing and trying, revew notes, make thesis, and post here.
If you don't have a few thousand bux, to convert an entire room into a clean room LAB, you might wanna borrow your schools lab. LOL
Have Phun.
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hyphae
born to grow
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Re: A Mycelium Study? [Re: Search59]
#3327265 - 11/06/04 10:22 AM (20 years, 2 months ago) |
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A synthetic nutrient solution that provides Psilocybe Cubensis with all essential nutrients is as follows:
Product: 60 g of concentrated synthetic nutrient solution
Concentration: 100 g will produce 8 L of aqueous nutrient solution.
Acidity: 7.5 g of concentrate should produce 1 L of solution with a pH of 5.5
Quality: Does not need to meet reagent purity, food grade is acceptable.
Make Up: Relevance of definition in the order of Formula, Name, then CAS RN:
1. Glucose (C6H12O6) [40 g] CAS: [50-99-7]
2. Ammonium Succinate (C4H12N2O4) [8 g] CAS: [2226-88-2]
3. Yeast Extract (Organic Extract) [4 g] CAS: [8013-01-2]
4. Magnesium Sulfate (MgSO4-7H2O) [4 g] CAS: [7487-88-9]
5. Glycine (C2H5NO2) [3 g] CAS: [CAS 56-40-6]
6. Potassium Phosphate (KH2PO4) [800 mg] CAS: [7778-77-0]
7. Thiamine Hydrochloride (C12H17ClN4OS HCl) [24 mg] CAS: [67-03-8]
8. Ferrous Sulfate,Heptahydrate (FeSO4-7H2O) [20 mg] CAS: [7720-78-7]
9. Cupric Sulfate,5-Hydrate (CuSO4-5H2O) [4 mg] CAS: [7758-98-7]
10. Manganese Chloride, 4-Hydrate (MnCl2-4H2O) [2.8 mg] CAS: [7773-01-5]
11. Zinc Sulfate, Heptahydrate (ZnSO4-7H2O) [2.4 mg] CAS: [7733-02-0]
12. Ammonium Molybdate, 4-Hydrate ((NH4)6Mo7O24-4H2O) [0.4 mg] CAS: [12027-67-7]
13. DiHydrogen Oxide (H2O) [146.4 mg - X] CAS: [7732-18-5]
14. Hydrochloric Acid (HCl) [X mg] adjusted to balance solution pH when properly diluted.
This solution is typically used for the production of psilocybin in submerged mycelium cultures of Ps.C. When well aerated this solution comes to complete submerged growth in about 11 days. It is typically ready for harvest and at highest alkaloid content in 7 days. Solution fermented for up to 50 days did not display any primordia or fruiting responses. Nutrient solution saturated agar surface cultures display rapid growth, primordia formation, and raised alkaloid content. Omission of essential nutrients suppresses growth and lowers alkaloid content, in some cases to undetectable amounts, in both submerged and surface cultures.
I believe the idea of nutrient depletion and deficiency acting as a triggering response to be false. Perhaps this is a misinterpretation of data, whereby a nutrient deficient mycelia culture was used to inoculate a nutrient rich substrate. The resulting mycelium is unable to begin fruiting until significant alkaloid creation has taken place. Nutrient rich mycelium cultures are capable of primordia formation with in as little as 3 days of exposure to an open-air interface.
The traditional triggers of the fruiting response are a drop in temperature, a drop in humidity, and an exposure to light, these stimuli correspond to mycelia surfacing in nature. I further purpose a set of internal triggers that directly relate to alkaloid levels within the mycelium, whereby primordia formation is seldom initiated in the absence of significant psilocybin levels. Further evidence to support psilocybin as a growth regulator comes from the evaluation of the organism's biomechanics.
Of most interest is this organism's management of phosphors, it is essential in the production of DNA, and this organisms spore reproduction method requires the production of allot of DNA. There is also an isolatable enzyme in the biomass that actively removes the phosphorus from psilocybin. We have a theory at "the lab" that assumes this enzyme is responsible for the acquisition of phosphorus to be used in the production DNA. The resulting levels of psilocin production and lowered psilocybin content are thereby directly related to the production of spores. The production of psilocybin seems to be a mechanism by which the organism can "sense" its environments chemical makeup for, as well as store, the nutrients necessary in the process of reproduction.
Since submerged cultures are constantly at +100% humidity the fruiting response is usually suppressed, even with lowered temperatures and exposure to light. It is possible a mutant strain could "adapt" and attempt to fruit in an aquatic environment, but it is more likely the response was initiated at the open-air interface and then produced abnormal submerged growth from that point.
These solutions can be aerated using an aquarium air pump, an inline HEPA filter, and some sterilized tubing. Sandstones can be quite useful but difficult to sterilize ( submerge in alcohol then rinse in distilled water and H2O2 ). The substrate in a hydroponics setup is typically inert since all the nutrients are provided directly from the nutrient solution.
In a supportive nutrient solution, a slightly acidic environment, and raised glucose levels the submerged mycelium exhibits rapid and aggressive growth. By doubling glucose levels from 5 g/l to 10 g/l the resulting mycelium had slightly higher mass and the psilocybin content was doubled to 1.01% by day 7, unfortunately the glucose induced hyper activity resulted in a rapid degradation and psilocybin content diminished to .1% by day 11.
Aside from the addition of the essential nutrients needed for efficient growth and raised glucose levels the addition of glucuronic acid and homobrassinolide have shown to stimulate mycelia and fruit growth. The growth promoting effect of glucuronic acid at 2% may be that of an additional carbon source in regards to Ps.C. The strong growth promoting effect of pollen at 2% and 10% is possibly the effect of homobrassinolide, a brassinosteroid hormone, which was detected at concentrations of 100 ng/g in pollen. 10% pollen in a medium would correspond to a hormone concentration of 10 ?g/l, a level that has been shown to enhance the growth of Psilocybe cubensis. Glucuronic acid at 2% had a weak growth promoting effect and pollen at 2% and 10% had a strong growth promoting effect.
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YSr
Stranger
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Re: A Mycelium Study? [Re: hyphae]
#3327474 - 11/06/04 11:53 AM (20 years, 2 months ago) |
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So 5,5 is optimum ph for Cubensis?
YSr
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Tantalus
Beyonddescription.
Registered: 06/05/03
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Re: A Mycelium Study? [Re: hyphae]
#3327475 - 11/06/04 11:53 AM (20 years, 2 months ago) |
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Quote:
hyphae said:
Of most interest is this organism's management of phosphors, it is essential in the production of DNA, and this organisms spore reproduction method requires the production of allot of DNA. There is also an isolatable enzyme in the biomass that actively removes the phosphorus from psilocybin. We have a theory at "the lab" that assumes this enzyme is responsible for the acquisition of phosphorus to be used in the production DNA. The resulting levels of psilocin production and lowered psilocybin content are thereby directly related to the production of spores. The production of psilocybin seems to be a mechanism by which the organism can "sense" its environments chemical makeup for, as well as store, the nutrients necessary in the process of reproduction.
So after the enzyme strips the phosphorus from psilocybin, does that psilocybin molecule become psilocin? I'm thinking that's the case, but my chem's a little rusty.
If this is true, this could explain the closed cap/open cap debate about potency. Perhaps open caps have a higher ratio of psilocin to psilocybin because that phosphorus has been used in spore production. This would explain the impression that open caps are less potent -- they just have more psilocin, which is much more likely to break down on it's way to the consumer. Is there anything to suggest that this enzyme would be less active in a phosphorus rich environment? If so, the substrate could affect this ratio.
With the large difference in stability, this could be key in growing fully mature mushrooms just as potent and perhaps just as stable as closed caps.
If this enzyme route were the only way the phosphorus is acquired for DNA production (was your wording seems to suggest), this theory would go out the window. However, this enzyme/psilocybin route being the only one for phosphorus seems pretty counterintuitive, at least to me.
Has there been any research done on the psilocybin/psilocin ratio of mushrooms of different maturity levels?
Hmmm, I just found this from THIKAL, "Because of its greater molecular weight psilocybin is less potent than psilocin... It is believed that after taking, psilocybin is easily converted into psilocin and that the pharmacological properties of both compounds are the same." Anyone have an idea how great this potency difference would be? This is the first time I've read this.
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Anno
Experimenter
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Re: A Mycelium Study? [Re: hyphae]
#3523487 - 12/20/04 06:21 AM (20 years, 1 month ago) |
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