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Mycena
mycoexplorer
Registered: 05/02/03
Posts: 270
Loc: indonesia
Last seen: 11 years, 9 months
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Re: Lets make them more potent! [Re: doc34]
#2676743 - 05/14/04 03:16 AM (19 years, 8 months ago) |
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I have plenty of seeds of P aquatica var Australia one of the higher potency strains
Where i live now is unsuitable for this grass (subtropics) so i want the seeds out there where they will grow
e-mail me at seedring@shaman-australis.com
we generally accept stamp donations with Oz or US/Euro/pounds 1 outside. Any extra appreciated as i run this as a voluntary non profit thing and there really is nothing left over, sometimes not enough. We usually chuck in extra species if you add enough to justify the extra postage and packing
Ideally we want you to grow the plant and then offer up seed for others and keep the planst in circulation
AFSR PO box 1417 Byron Bay NSW 2481 Australia
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suboriginal
enthusiast

Registered: 05/06/04
Posts: 278
Last seen: 19 years, 3 months
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Re: Lets make them more potent! [Re: suboriginal]
#2676757 - 05/14/04 03:28 AM (19 years, 8 months ago) |
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PS, has anyone used the 'series dilution' method before to derive pure strains from single mycelium ? Essentially it just involves making a series of dilutions from a master stock of unknown concentration, then plating a set volume from each dilution out on a different plate... at a certain dilution factor, you should get just a few cultures popping up (plaque forming units they are called when this method is used to derive pure strains of bacteria or viruses on, for example HB agar) on one of the plates, and these can be reasonably be assumed to be monocultures (pure strains derived from a sing mycelium)... should this work? Can a mycelia culture realistically be expected to develop from one single mycelium, or do they really only happy when there's a few of them around, like for example cultured human lymphocytes... Would it be too slow waiting for a culture to develop from a single mycelium ? Any thoughts appreciated... Lastly, though this may well be answered in the contam' forum where I'll visit next, can anyone advise on a broad spectrum antibiotic that I can add to my agar after autoclaving/ cooling, that will ensure no bacterial contamination, but have zero effect on the mycelial (p. subaeruginosa) growth ?
-------------------- Peace, love and organic brown rice...
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Mycena
mycoexplorer
Registered: 05/02/03
Posts: 270
Loc: indonesia
Last seen: 11 years, 9 months
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Re: Lets make them more potent! [Re: suboriginal]
#2677232 - 05/14/04 09:07 AM (19 years, 8 months ago) |
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re that last query no No antibiotic will inhibit all bacteria However i have found Chloramphenicol useful at a rate of 50mg/L It is heat sensitive but is added before sterilisation. The agar is sterilised for just the time needed, then cooled and pured As soon as possible to reduce the breakdown This is particularly well suited to slants as they cool rapidly why so much bacterial probs anyway? I find aside from those cryptic baterial infestations (cleaned up by reeated subs) that bacteria are not really an issue if the conditions in your workspace are favourable. Mould is more of a bugbear and yes they will reform from single cells in the manner you spoke of. just takes longer. This has been done both in successfully regenerating mushroom protoplast hybrids (eg shiitake x golden oyster) It was also the means by which high yielding Claviceps purpurea cultures were isolated for producing both the lysergic derivatives and ergopeptides and other ergotoxins. Single cells were plated up to form colonies. each colony was grown out and the level of biosysnthesis measured by culturing from half the colony. The other half of High yielding phenotypes were kept and subjected to the same treatment. Generation after generation it was repeated till they had achieved in some cases an increase in biosynthesis from less than 10 mg/l to greater than 2200mg/L. In some cases this happenned in as little as 5 generations with hard selection In the absence of selective pressure these clones revert back to lower yielding phenotypes The process is simple, so simple that when it was presented by the industrial mycologist to their academic colleagues it was thought to be incredible. The key was the large scale and many repetitions needed to carry out all the thousands of experiments If it works with Claviceps it might also work with Cordyceps, and if it worked for them too then maybe the phenomenon can be exploited in many more fungi, possibly even P cubensis, particularly in liquid bioreactor style. from looking at the acquired characteristics of Plant cell lines in Tissue culture it is also apparent that sometimes these traits selected for such as temp tolerance or increased biosynthesis do lead to higher yileding clones when regenerated, but many times they do not Sometimes the mutation is unstable or environment specific, other times there may be other limiting factors like the plants infrastructure acting as a limiting step to biosynthesis (nutrient and water supply for example) In light of this it would probably be best to keep things as whole as possible to avoid adverse selections, either ise dikaryotic mycelium or work with monokaryons Perhaps monokaryotic mycelium is capable of biosynthesis in culture? If so high yielding strains could be mated to inbreed such characteristics. This also has the benfit of heterosis once selection is done and may help the longe term stability and adaptability of the spore race
Edited by Mycena (05/14/04 09:23 AM)
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endokrin
Stranger

Registered: 02/13/04
Posts: 142
Loc: SouthEast
Last seen: 16 years, 1 month
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Re: Lets make them more potent! [Re: Mycena]
#2677566 - 05/14/04 11:10 AM (19 years, 8 months ago) |
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I think reed canary grass (phalaris) contains 5-OH-DMT, which from what I've heard is not desireable, as opposed to N,N,DMT & 5-Me-DMT.
"Bufotenine (5-OH-DMT) is another DMT relative. This compound is vaguely referred to as "noxious" by Jonathan Ott. Apparently 10mg of pure 5-OH-DMT injected is enough to cause "dramatic circulatory crises." There appears to be debate over the psychedelic qualities of bufotenine. However, McLeod & Sitaram, Shulgin, and Fabins & Hawkings all report the presence of psychotomimetic effects. Bufotenine causes anxiety, circulatory distress, skin flushing, and percieved color distortions. Injected doses of 16 mg (over 3 min. IV drip) have been reported. At this dose, the symptoms of mild skin flushing to extreme purple cast appear. Subjects also report a great deal of psychological distress and fear at this dose. Doses of 8 mg produce mild skin flushing and increased anxiety. Doses of 2-4 mg of bufotenine do not produce hallucinogenic effects. The above discussed negative side effects at 16 mg last for approximately 10 minutes. Other side effects reported are sweating, nausea, yellowed vision, and perception of colored spots. So it appears that bufotenine is a nasty drug to be avoided, Not only does it tend to induce panic, it also appears to have the potential for a fatal overdose, although no case studies to this effect have been found for humans."--Erowid FAQ
-------------------- "If King Kong sells drugs, we'll put him in jail"
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Transplant
Janitor
Registered: 05/07/04
Posts: 77
Loc: Texas
Last seen: 19 years, 5 months
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Re: Lets make them more potent! [Re: endokrin]
#2679972 - 05/14/04 08:24 PM (19 years, 8 months ago) |
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The premises behind using a substrate such as phalaris, would be that the basic elements and elementary compounds such as tryptophan and tryptamine would be present, regardless of quantity.
An analogy to why any amount would be of interest and worth study would be: --would you rather build a house on concrete or on sand. Either one, in theory is feasible, but which one provides the basic foundation for future growth and longevity.
In case that analogy is more sound in my thinking I can relate it more to the topic: --We all have seen different degrees of potency in a mushroom and there are many debates on factors involving this, including substrate, casing, temperature, humidity, etc. But hopefully everyone agrees that a mushroom can form with very negligible amounts of resulting psilocin regardless of what caused it, it does happen. So this allows for further thought and exploration into WHY.
Prevailing belief of well known mycologists tend to believe that given an ideal growing environment, this does not include substrate, and a resulting low concentration is indicative that either the substrate lacked nutritional value to the mushroom, but not enough to hender growth either by design (using something low in the first place) or expended (possibly by competition or in ability to break down complex molecules present). So even though you could grown a fungus on a sand foundation (if this doesn't make sense or relate, then please read the entire post again) or it can grow on the concrete (more concentrated because there is plenty of foundation for cell growth as well as more reagents to cook up alkaloids inside).
The fear of a harmful reaction due to smoking the grass, as indicated on the vault, is arbitrary since fungus doesn't smoke to begin with and the natural filtration and usage of the fungus would be in place (meaning your not going to shit gold by having a gold ring on your finger as much as a mushroom is going to produce a chemical that it does not do in the first place just because it is growing in it, within reason)...does that make sense? There is a lot left to be done in the study of mycology, but some universal laws do exist that allow for some educated guesses towards an hypothesis and future studies.
-------------------- Will Screw For Shrooms
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Mycena
mycoexplorer
Registered: 05/02/03
Posts: 270
Loc: indonesia
Last seen: 11 years, 9 months
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Re: Lets make them more potent! [Re: endokrin]
#2681767 - 05/15/04 09:40 AM (19 years, 8 months ago) |
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Your notes on Bufotenine are in question now due to the experiences of so many smokers of Cebil/Vilca seeds (Anadenathera colubrina)
Bufotenine isnt necessarily the nasty thing ist made out to be The nastiest thinsg in toad poison are other cardiotoxic compounds
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