Cloning Psilocybin
- Thread starter SnakeByte
- Start date
Jogro
Well-Known Member
"Cloning" probably isn't the right word here.
If you have access to fresh mushrooms you should be able to get the still living tissue to grow into a culture.
That actually should be relatively easy; you just inoculate some tissue into agar under sterile conditions, making sure to isolate pure colonies for further growth.
If you have access to DRIED mushrooms, you probably won't be able to get this to work. . .but its not impossible.
Still, dried mushroom caps often contain viable spores, and you well may have luck getting those to grow. Mature dried mushroom stems well may have viable spores stuck on the outside, and if so, you may have a good chance getting them to grow out in culture.
In short, yes, if you have access to dried mushrooms, its worth a shot trying to culture them.
Note that mushroom spore prints contain no psychoactive substances and are also perfectly legal in most of the USA. There are any number of dealers who will ship them to you.
canndo
Well-Known Member
It is said that properly dried mushrooms will "come back to life" if soaked in sterile water and then placed upon a fertile substrate. I have tried this a number of times with no sucess. It is further said that dried sclerotia should do the same, considering tha sclerotia are a mushroom's method of banking it's genetics against a time of environmental adversity, and considering that sclerotia of other species does indeed re-spawn, the method should work. I have tried it, in an attempt to recover a very sclerotia forming species and failed a number of times. One of the problems however is that dried mushrooms are rarely prepared with regrowth in mind and the drying process will undoubtedly contaminate - often grossly (figuring that many folks dry their fruit over a fan, blowing lots of contaminated air across the sticky surface of a wet mushroom). If you should succeed, please post your process and results here, I'd love to find a way as I hahve species that I would love to duplicate.
As far as "cloning" live mycelium is concerned, this is the most common method - though you risk sencience depending upon the total length of hyphae.
As far as "cloning" live mycelium is concerned, this is the most common method - though you risk sencience depending upon the total length of hyphae.
SnakeByte
Active Member
The substrate would have to be one that is extremely easy for mycelium to grow in. Maybe changing the whole way MOST people do things.
www.shroomery.org/forums/showflat.php/Number/514231
http://www.shroomery.org/forums/showflat.php/Number/1470425
These threads gave me some good ideas. Talking about liquid cultures and using other nutrients as sort of "ferts" for Psilocybin.
"While the only real necessity for mycelium production is glucose or similar carbohydrate, alkaloid production requires the presence of essential nutrients not only as precursor components, but to effectively utilize carbohydrates, promote cellular development, and to regulate vital enzyme pathways."
Probably best to have a combination of the new and the old? Rye flour, Vermiculite, Cornstarch, Honey, and Water?
Furthermore, Crushing the dried mushrooms might allow for better results in this kind of substrate, to inoculate more with less? Though that won't take care of the contamination problem.
Do you guys think that adding very small amounts of isopropyl alcohol to the substrate help or hurt it? I know you can use it to make a tincture but would it kill the spores?
www.shroomery.org/forums/showflat.php/Number/514231
http://www.shroomery.org/forums/showflat.php/Number/1470425
These threads gave me some good ideas. Talking about liquid cultures and using other nutrients as sort of "ferts" for Psilocybin.
"While the only real necessity for mycelium production is glucose or similar carbohydrate, alkaloid production requires the presence of essential nutrients not only as precursor components, but to effectively utilize carbohydrates, promote cellular development, and to regulate vital enzyme pathways."
Probably best to have a combination of the new and the old? Rye flour, Vermiculite, Cornstarch, Honey, and Water?
Furthermore, Crushing the dried mushrooms might allow for better results in this kind of substrate, to inoculate more with less? Though that won't take care of the contamination problem.
Do you guys think that adding very small amounts of isopropyl alcohol to the substrate help or hurt it? I know you can use it to make a tincture but would it kill the spores?
Jogro
Well-Known Member
I can't see how that would help.
Do you mean you can't find that or that you live in a location that it can't be shipped to?
Have you ever done any sterile work?
If not, don't play with this until you have that down pat.
It requires isolation via multiple petri dish transfers, it will be contaminated, and you do not inoculate a substrate with something is guaranteed to be contaminated.
Have you ever grown a mushroom?
If not, do not think about varying ANYTHING in a proven formula.
Most of those substrate additives will favor contamination over your myc.
Keep it as simple as possible, and do not try to optimize anything until you have a few grows under your belt.
This hobby is ridiculously easy for those with a bit of patience and an ability to follow directions.
This hobby is incredibly difficult (and painful, losing 2 months of work overnight to contam is NO fun) for those who want something fast or who are smart enough to research the hell out of everything, and want to try it before getting the fundamentals down (another form of impatient).
If not, don't play with this until you have that down pat.
It requires isolation via multiple petri dish transfers, it will be contaminated, and you do not inoculate a substrate with something is guaranteed to be contaminated.
Have you ever grown a mushroom?
If not, do not think about varying ANYTHING in a proven formula.
Most of those substrate additives will favor contamination over your myc.
Keep it as simple as possible, and do not try to optimize anything until you have a few grows under your belt.
This hobby is ridiculously easy for those with a bit of patience and an ability to follow directions.
This hobby is incredibly difficult (and painful, losing 2 months of work overnight to contam is NO fun) for those who want something fast or who are smart enough to research the hell out of everything, and want to try it before getting the fundamentals down (another form of impatient).
SnakeByte
Active Member
First of all, nothing I'm discussing here has been proven.
Second, I do understand what the sterile work requires.
The Only contaminant in this idea I can see would be the dried mushrooms. Which is why I'm curious as to if ISO would destroy spores.
It's a well known fact that Honey is already naturally sterilized and can stay good for a decade if stored properly due to it very low moisture content. Though the idea here would be to buy new, fresh honey.
This would be added to an already sterilized basic substrate as we don't want to put the sugar in the pressure cooker and changes it's structure.
I was also thinking of substrates using gelatine. Any thoughts on that?
Second, I do understand what the sterile work requires.
The Only contaminant in this idea I can see would be the dried mushrooms. Which is why I'm curious as to if ISO would destroy spores.
It's a well known fact that Honey is already naturally sterilized and can stay good for a decade if stored properly due to it very low moisture content. Though the idea here would be to buy new, fresh honey.
This would be added to an already sterilized basic substrate as we don't want to put the sugar in the pressure cooker and changes it's structure.
I was also thinking of substrates using gelatine. Any thoughts on that?
First of all, nothing I'm discussing here has been proven.
What does this mean?
Second, I do understand what the sterile work requires.
Oh, yeah, we all understand what it requires. Until we do it. So I'm not going to address the "only contaminant" statement.
And any time anyone starts with "it's well known" is pulling a Cliffy. You are talking about using it as an additive in a mushroom environment. Nothing to do with it sitting alone on a shelf. And I just realized we haven't differentiated between building sterile jars vs laying out a substrate. Sterile VS pasteurized. Different steps in the project, different requirements. If you really want comments on your thoughts, detail step by step your intended process.
And these words: "Maybe changing the whole way MOST people do things."
Delusions of grandeur.
I was also thinking of substrates using gelatine. Any thoughts on that?
No, I will not engage. You are looking for an argument. Have it with someone else.
Think all you want.
What have you DONE?
canndo
Well-Known Member
Gelatine has inheren't problems staying in a semi-solid state thorough the range of temperatures from 60 to 90 degrees.
As far as your honey statement, no, honey is LOADED with contaminants - and is far from naturaly "sterile". It is so sweet that it does not host bacteria or fungus in it's undiluted state but I don't believe that is true otherwise. I do know that the organism that causes botulism is often present in honey. As far as offering a new or better way of doing mycological work, we who have been working in this field for 40 years tend to grow narrow sighted and have notions that ours is the only way a thing can be done, there may well be a superior method, on the other hand, some of the methods in use have been refined by decades of use and experimentation.
So, although it is entirely possible, it isn't very likely that a revolutionary method could be brought to the fore.
As far as your honey statement, no, honey is LOADED with contaminants - and is far from naturaly "sterile". It is so sweet that it does not host bacteria or fungus in it's undiluted state but I don't believe that is true otherwise. I do know that the organism that causes botulism is often present in honey. As far as offering a new or better way of doing mycological work, we who have been working in this field for 40 years tend to grow narrow sighted and have notions that ours is the only way a thing can be done, there may well be a superior method, on the other hand, some of the methods in use have been refined by decades of use and experimentation.
So, although it is entirely possible, it isn't very likely that a revolutionary method could be brought to the fore.
SnakeByte
Active Member
@ Testtime
Hey bag which one douches with, this is a hypothetical discussion. Derp! Maybe your fine with doing 7-15 years, I'm not. So what I have done or will do is no one business.
If you have nothing mature to say or useful to add to this topic, fuck off lol.
Delusions of grandeur are not defined by a thought for a discussion either. It's a conversation and nothing more, get over yourself.
Because yes I obviously want only to argue, that's why I asked if gelatine would be good... I'm sure that makes sense in the "I'm better than you" world you have in your lonely little mind but really, I was making the observation that it's been done in science labs for bacteria. Herp!
And if you want to discuss whether it would be a better plan to use jars or lay out a substrate vs doing one than the other, then go ahead! That's what a discussion is for, opinion, not put downs. You've contributed nothing useful since your first post and even that was not your original thought. So I think everyone would appreciate you keeping your asshole attitude to yourself.
To everyone else, keep up the good convo and thank you for your CONSTRUCTIVE criticism !
Hey bag which one douches with, this is a hypothetical discussion. Derp! Maybe your fine with doing 7-15 years, I'm not. So what I have done or will do is no one business.
If you have nothing mature to say or useful to add to this topic, fuck off lol.
Delusions of grandeur are not defined by a thought for a discussion either. It's a conversation and nothing more, get over yourself.
Because yes I obviously want only to argue, that's why I asked if gelatine would be good... I'm sure that makes sense in the "I'm better than you" world you have in your lonely little mind but really, I was making the observation that it's been done in science labs for bacteria. Herp!
And if you want to discuss whether it would be a better plan to use jars or lay out a substrate vs doing one than the other, then go ahead! That's what a discussion is for, opinion, not put downs. You've contributed nothing useful since your first post and even that was not your original thought. So I think everyone would appreciate you keeping your asshole attitude to yourself.
To everyone else, keep up the good convo and thank you for your CONSTRUCTIVE criticism !
canndo
Well-Known Member
The first article is a very old portion of a method taken from the growth of exudate forming mycelia such as penicillin. This approach was used or attempted to be used before Oss and Oric (sp) publicized their initial fruiting methods. I have done such work and it is, for the organism we are talking about here - pretty much a waste of time and resources. your yield is miniscule compared to simply fruiting a healthy mass of mycelium and then, if interested, extracting some portion of active ingredient from that mass.
This is a form of biogeneration where the exudate or the byproducts of growth are extraced from the nutrient liquid. The weight of the mycelium after a full growth cycle is quite small although the mycelium can be rather potent. I have had what I believe to be the proper results of this method which tend to look like small, light blue chalky spheres. Ny experiment, yielded spheres of the same color and size but they congieled into a wet sparse mass when I attempted to dry them - it was only marginaly more powerful than any other of a half a dozen other attempts at harvesting mycelial mass without fruiting. The fact remains in this case, with this organism that the best way to get yourself large amounts of pure, high potentcy mycelial mass is to bring the mycelial material either to fruit or to a sclerotial stage. this is very very old school stuff and meant for biogenerators of materials that do not have proper fruit I.E. fungi imperfecti.
so far as innoculation amounts - that is, innoculating more with less - it is a moot issue in biogen set ups and in active liquid cultures. When a liquid culture is continuously agitated either by bubbling air through the vessel or by continuous mechanical stirring - mag stirers and the like, your hypae quickly break, "bud" and break again, each "bud" and the site of each break bearing yet more growth so it doesn't take long at all to arrive at as large a mycelular mass as can be supported by the environment. In other words, the difference in time between full "colonization" of a given container of liquid from a single cell and that same container being fully "colonized" through the introduction of say - a thousand individual cells would be only a matter of hours.
SnakeByte
Active Member
Right, I was thinking that it would be better to just use it to get healthy samples and not really to harvest at this point. Using ground dried mushroom not for the purpose of fruiting but to grow healthy mycelium and then clone from there. AND THEN harvest.
canndo
Well-Known Member
Your best bet there will be (as I just posted elsewhere) a two dimensional medium - agar dishes if you are looking for healthy samples from questionable living material. Liquid culture has limited usefulness in isolation - it shines in mass innoculations. Your ground dried approach risks your destroying the organism at the cellular level - once this happens you cannot proceed.
Cloning is common - but as I said, most often in two dimensional mediums. Exotic mixtures of nutrients such as the one you posted are not needed with this organism because it is a primary and secondary decomposer and so it pretty much creates it's own nutrients from more complex ones - the exact opposite of plants. This is a tough one for pot growers to comprehend as they are perpetualy looking for some secret nutrient or some missing component in a situatio were what they are growing is not really a plant, in fact it is more akin to an animal - it breaths in oxygen and exhales CO2, and it takes in complex molecules and breaks them down to more elemental substances.
SnakeByte
Active Member
Ok So keep it simple.
Instead of jars, couldn't I use one foil covered bowl in a pressure cooker?:
-Move the cooker and bowl inside the sterilized environment.
-Use an airtight Rubbermaid (Or wtv you're final grow will sit in)
-Add sterilized substrate from bowl to final container
-Inoculate all over the container
-Add your water shield
-Put on the container cover, wrap in a dark bag, store in a dark warm place
And that way I wouldn't have to take the substrate out of jars...
And there was a mention of Agar, which I read is a red algae derivative. I also read the red Algae is beneficial for healthy growth of many mushrooms.
But the "Kanten" food grade agar available in Oriental stores would work just as well?
Instead of jars, couldn't I use one foil covered bowl in a pressure cooker?:
-Move the cooker and bowl inside the sterilized environment.
-Use an airtight Rubbermaid (Or wtv you're final grow will sit in)
-Add sterilized substrate from bowl to final container
-Inoculate all over the container
-Add your water shield
-Put on the container cover, wrap in a dark bag, store in a dark warm place
And that way I wouldn't have to take the substrate out of jars...
And there was a mention of Agar, which I read is a red algae derivative. I also read the red Algae is beneficial for healthy growth of many mushrooms.
But the "Kanten" food grade agar available in Oriental stores would work just as well?
polyarcturus
Well-Known Member
water shelid?
listen if you want to "clone" (i put that in quotes cause in the mushroom world thats a shaky term) you need to gather a fruit then split it in half quikly cutting away a peice of flesh from the exposed inerds into a jar of sterile substrate. so people have done clones to brf or grain, but most would clone to agar because it is much more likely to suceed, not to mention "cloning" is not the most sterile process, therefore agar will also give you the advantage of isolation from a contamination.
but all agar is, is a jelly like substance thats somewhat nutritious and the mycelium only grows across the top surface not really to be used for fruiting but rather for preservation, purification and inoculation.
listen if you want to "clone" (i put that in quotes cause in the mushroom world thats a shaky term) you need to gather a fruit then split it in half quikly cutting away a peice of flesh from the exposed inerds into a jar of sterile substrate. so people have done clones to brf or grain, but most would clone to agar because it is much more likely to suceed, not to mention "cloning" is not the most sterile process, therefore agar will also give you the advantage of isolation from a contamination.
but all agar is, is a jelly like substance thats somewhat nutritious and the mycelium only grows across the top surface not really to be used for fruiting but rather for preservation, purification and inoculation.