Alkaloid-increasing methods

[Monk]

lol too many big replies!!!!

Haha. So true.

wtf? 1k = $1000? What am I missing here?

gamma, yeah the price of Trichs is getting to be ridiculous around here ! Just kidding, I meant to say "1kg." Refering to the fact that the mother of the clones would have to be quite large to supply nine cuts of this size.

That has long been thought for the Psilocybe genus of mushrooms. Personally, I think it is a bit ridiculous - what would have happened if humans had developed slightly differently and simply didnt experience the hallucinogenic effects? The mushrooms/cacti dont have brains or anything to suggest that they are capable of consciousness (other than a few people who might think mescaline visions are their way of communicating), so how could they decide to produce these chems for our use and their survival

teo responded nicely to this, Ace, but I wanted to mention that you might enjoy reading The Botany of Desire by Michael Pollan. It beautifully elaborates on how several plants have used the processes mentioned by teo to 'take advantage' of the capabilities of humans to coevolve to a remarkable level i.e.

They produce them because some enzyme doing another function malfunctions or changes and acts on a new substrate, natural selection kicks in and good adaptations are proliferated

It was recommended to me by a plant physiology prof and changed my whole perspective on plants. You'll love it If I remember correctly, the plants focused on were the apple, the potato, the tulip, and everybodies favorite - cannabis.

teo, let's talk other potential factors that could have led to SCBs experience since we're all taking this very seriously:

1. Injecting process stressed plants into greater alk. production (potentially ruled out by the second control group.)

2. Some tyrosine remained unchanged in the plant, but acted to boost neurotransmitters in the subject, enhancing the experience greatly.

3. Being grown in low light (window will) caused greater alk. production.

4. Cactus was dank as hell to begin with (stump should not have been gifted to a friendly hippy to grow on after experiment!)

As I see it, 1 and 3 will be resolved by said control groups. Number 2 would be eliminated as a possibility through A/B extraction/quantitative measurement or bioassays involving supplementation/no supplementation (I know, I know teo, the former option is better ) Number 4 is resolved by the whole experiment (except for the whole 'giving away the super dank cactus' part .)

Edited August 10, 2007 by FeloniousMonk

[nitrogen]

Ok, a few more thoughts...

1. First of all, the bugs go after and eat my potent cacti, and also the less potent pones... And a nooker posted awhile back saying a rodent was eating only his most potent lophs and trichos...

2. Stressing the plant - I still don't get why this would theoretically increase alkaloids? Stress is unrelated to predation no? - you can deprive the plant of water or poke it with something, and this will stress it, but it isn't being eaten on... And wouldn't a healthy robust plant be more likely to be eaten on than a dried-out and unhealthy plant?

So, if there is an alkaloid response to say, depriving cacti of water, that means the alkaloids are either serving an internal function for the plant, or created as a byproduct of a change in the way the plant is growing itself under those conditions...

3. Shade grown=more potent theory - I read above that placing cuttings in the shade causes the inner green layer to become darker green and more concetrated in that area - maybe the cactus isn't more potent, but simply the alkaloids have migrated to the dark green layer whereas they were previously more spread out through the green,white, and core layers... Since the core/white part is often discarded and those alkaloids lost, the cactus seems more potent because now more of the existing alkaloids are in the green layer... Just an idea...

[Monk]

Stress is unrelated to predation no?

In the most direct sense, this is true. However, when you or I are stressed about an exam or interview, our bodies produce the same chemicals as when something or someone is attacking us. This is an analogy for the theory of stress increasing alkaloid output. It is a totally different stimulus, but our systems are simple and produce the same reaction. This parallel leads us to your next question:

And wouldn't a healthy robust plant be more likely to be eaten on than a dried-out and unhealthy plant?

The answer is actually 'no' for all plants that I can think of. In populations of plants, the individuals that show nutrient/water or other types of stress are usually the first to be attacked by insects/diseases. I am glad you asked this question, though, as it forces the analogy to the next stage. The initial stress reaction in humans allows them to better deal with the situation i.e. increases alertness/focus/reaction time. This may parallel the stage in a plant when the increased alkaloid content helps defend the organism by confusing/harming predators. After extended stress, though, the neurotransmitters/hormones of a person get depleted leading to a lack of focus/exhaustion/irrational thinking etc. that actually put the person in danger. This stage is what one sees when a...let's say water stressed solanum, gets spider mites and aphids sooner/worse than its well watered friends. I never considered how this later stage could apply to cacti, though...hmm. I wonder if a certain time period of stress is ideal prior to the 'defensive response' depleting the reserves. Now I'm just brain storming, but precursor supplementation really helps with this stage of the human stress reaction i.e. tryptophan/tyrosine/DLPA etc. I guess you can see where I'm going with this.....(scratching chin, hmmmm)

[teonanacatl]

1. First of all, the bugs go after and eat my potent cacti, and also the less potent pones... And a nooker posted awhile back saying a rodent was eating only his most potent lophs and trichos...

maybe so but have you measured the levels in the plants? your growing conditions might be such that there is no appreciable amount of alkaloids, or they are eating the tips where there is no appreciable amounts. Simple test to see feeding deterency of mescaline is to get some mice or rats and feed them pellets, then add some mescaline in incresing amounts to some pellets and see if they eat them.

2. Stressing the plant - I still don't get why this would theoretically increase alkaloids? Stress is unrelated to predation no? - you can deprive the plant of water or poke it with something, and this will stress it, but it isn't being eaten on... And wouldn't a healthy robust plant be more likely to be eaten on than a dried-out and unhealthy plant?

So, if there is an alkaloid response to say, depriving cacti of water, that means the alkaloids are either serving an internal function for the plant, or created as a byproduct of a change in the way the plant is growing itself under those conditions...

Stressing a plant means it is unable to grow properly, ie no water, no nutes, too much light etc. This means that what it has inside it could be all it has to function with. Therefore it wants to protect that store untill such time as it can grow again. Sort of liek if you were fat and ran out of food, if other people wanted to cut a bit off you to eat you would defend yourself with a stick. Alternatively it could have developed as an indirect response to stress, ie it gets dry and the cactus can survive but the rodents need water, oh look cousin larry that cactus has alot of water lets eat it, so its caused by stressors to protect against somthing which is stressing another animal.

3. Shade grown=more potent theory - I read above that placing cuttings in the shade causes the inner green layer to become darker green and more concetrated in that area - maybe the cactus isn't more potent, but simply the alkaloids have migrated to the dark green layer whereas they were previously more spread out through the green,white, and core layers... Since the core/white part is often discarded and those alkaloids lost, the cactus seems more potent because now more of the existing alkaloids are in the green layer... Just an idea...

The whole layer colour change thing I rationalised in another thread (doesnt mean im right lol) but to me it sounds like a good old shade plant vs sun plant leaf differences. In the sun there is normally a thick epidermis because the sun is strong and alot of radiation is present they want to block the nasty stuff. But also because it is strong photons penetrate the tissue of the cactus further meaning having chlorophyll 1cm beneath the surface will still functional and it will still be absorbing light, you would probably find different ratios of chl A to B from surface to centre. A plant in the shade receives little light, therefore it will have a thin epidermis and will have chlorophyll concentrated at the surface to make the most of any available light.

Whether the alkaloids migrate along with the chlorophyll that is a different story.

[trucha]

RE:"The mushrooms/cacti dont have brains or anything to suggest that they are capable of consciousness (other than a few people who might think mescaline visions are their way of communicating), so how could they decide to produce these chems for our use and their survival"

Why would anyone think they decided to produce any chemicals any more than thinking that we have somehow decided to produce our own neurotransmitters? Besides we EAT these plants, that hardly contributes to their survival it as it merely promotes the propagation of their kind when a person who likes the experience is the one who ate them.

As for mescaline and other hallucinogen visions I would suggest as others have before me that what we are watching is the firing of neurons in our visual cortex and other areas of our brain. hhLots of what we see can be directly correlated to brain structure morphology. Communication with cacti is not nearly so flat and three dimensional as that colorful display. It can certainly be beautiful but really is not much more informative than watching cartoons (imho)

You really might want to read Paul Stamet's Mycelium Running before further perpetuating an illfounded anthropocentric assumption about a brain being required for consciousness.

I have personally watched what was clearly interactive communication, responsiveness to that communication and deliberate cooperation by a 'wolf-pack' of predatory spirochetes in organizing and carrying out a successful attack on a paramecium at least hundred of times their size.

Its both clear and proven that mushrooms are not just conscious but capable of making decisions based on what occurs around them and responding. Successful maze navigation by mycelia is only one aspect of that. Read Stamet's then let's talk some more.

Its just too easy to think of ourselves as being the only form consciousness and awareness. I would suggest we are among the least conscious organisms on the planet or at least our actions towards the terrarium we live in would strongly suggest that.

[Ace]

You really might want to read Paul Stamet's Mycelium Running before further perpetuating an illfounded anthropocentric assumption about a brain being required for consciousness. Successful maze navigation by mycelia is only one aspect of that.

Do you mind elaborating Trout? I have not yet read Mycelium Running (though at the top of my of to-buy list ), but have read his accounts in Psilocybin Mushrooms of the World and one or two other ones (cant recall names now). I have not heard of the maze navigation thing - could you give some detatils? I cant see how mycelia running through a maze could prove they have a consciousness or awareness, any more so than say a venus fly trap (just going off your brief description above). What would be displayed is an organism performing at its peak, using what it needs in its environment to assist it to flourish and reproduce. There is no empathy, communication or other signs that might hint at consciousness, so you have got me quite interested in how that could be an example.

Its just too easy to think of ourselves as being the only form consciousness and awareness.

I'm not saying that Homo sapiens is the only species with consciousness - I believe that every animal (especially mammals) has a consciousness of some level, but plants/fungi cannot (IMO) have such as they simply dont have the structure to do so. Well, not that we can see at this point in time (there could be some other method of communicating/empathising/etc with others, e.g. via chemical reactions - like psilocybin on the human brain that may constitute a form of communication, which may very well be what you are getting at). But like you said, all that is produced is the equivalent of cartoons, so would you accept that as a level of consciousness?

Its both clear and proven that mushrooms are not just conscious but capable of making decisions based on what occurs around them and responding.

Ok, yes, I totally agree that every living creature (from plants to animals to fungi and whatnot) can make decisions based on their environment, but how does that mean that they have a consiousness? I guess what I should define is the word 'consciousness':

–noun

1. the state of being conscious; awareness of one's own existence, sensations, thoughts, surroundings, etc.

2. the thoughts and feelings, collectively, of an individual or of an aggregate of people: the moral consciousness of a nation.

3. full activity of the mind and senses, as in waking life: to regain consciousness after fainting.

4. awareness of something for what it is; internal knowledge: consciousness of wrongdoing.

5. concern, interest, or acute awareness: class consciousness.

6. the mental activity of which a person is aware as contrasted with unconscious mental processes.

7. Philosophy. the mind or the mental faculties as characterized by thought, feelings, and volition.

—Idiom8. raise one's consciousness, to increase one's awareness and understanding of one's own needs, behavior, attitudes, etc., esp. as a member of a particular social or political group.

Taken from HERE

So, can you explain how fungi (perticularly Psilocybe) are aware of their own existence (of course, the mycelia/fruiting bodies can collide with each other, thus forcing themselves to colonise another area)? How do they display signs of sensations, producing thoughts and awareness of their surroundings?

I will have to find Mycelium Running I think...

P.S. Sorry for the thread hijack!

[occidentalis]

1. First of all, the bugs go after and eat my potent cacti, and also the less potent pones... And a nooker posted awhile back saying a rodent was eating only his most potent lophs and trichos...

2. Stressing the plant - I still don't get why this would theoretically increase alkaloids? Stress is unrelated to predation no? - you can deprive the plant of water or poke it with something, and this will stress it, but it isn't being eaten on... And wouldn't a healthy robust plant be more likely to be eaten on than a dried-out and unhealthy plant?

So, if there is an alkaloid response to say, depriving cacti of water, that means the alkaloids are either serving an internal function for the plant, or created as a byproduct of a change in the way the plant is growing itself under those conditions...

I haven't read the whole thread, but something that people seem to be forgetting is that plants have to protect themselves not only from macroscopic herbivory, but from disease caused by microscopic organisms. Many plant alkaloids have been shown to protect pants from pathogens.

An excellent article is this (pdf) which shows harmaline and harmine production by Oxalis tuberosa increases in response to pathogen elicitors (compounds produced by the pathogen which are recognised by the plant and elicit a defense response) and methyl jasmonate (a signalling compound involved in the defense response).

I don't know specifically whether or not mescaline is antimicrobial or whether it is involved in the resistance response, but I would not be at all surprised if it is.

Stress (and particularly wounding) can activate the same metabolic pathways as pathogen attack in some plants. So if mescaline or other alkaloids are involved in the resistance response, stressing the plant could lead to higher production of these alkaloids. I don't know if this is the case with cacti or not.

A good experiment for someone with TLC or HPLC () ability would be to treat ramets of a clonal cactus line under otherwise identical conditions with some or all of the following:

methyl jasmonate

salicylic acid

benzoic acid

BABA

drought stress

waterlogging

shade

wounding

pathogen elicitors

and of course a control

And measure mescaline levels after 1 hour, 12 hours, 3 days, 1 week, 2 weeks, 1 month... etc.

come on, who has nothing better to do?

3. Shade grown=more potent theory - I read above that placing cuttings in the shade causes the inner green layer to become darker green and more concetrated in that area - maybe the cactus isn't more potent, but simply the alkaloids have migrated to the dark green layer whereas they were previously more spread out through the green,white, and core layers... Since the core/white part is often discarded and those alkaloids lost, the cactus seems more potent because now more of the existing alkaloids are in the green layer... Just an idea...

This is a possibility and nice work on the lateral thinking

however a local ethnobot has recently been doing some work that hints towards confirming the 'shade grown=stronger" theory. However I will have to let him tell you more about it if he wants to as it's not my experiment.

[trucha]

There are all sorts of variables that can come into play.

Additional nitrogen can help but in at least other plants ammonium favors alkaloids and nitrates favor vegetative growth at the expense of alkaloids. Feeding with some blood or blood meal is a good approach. In the late 1950s Claudine Friedberg claimed that around Huancabamba traditionally only women did the harvesting after midnight under the full moon. This does not seem to be the case broadly as far as I can tell but time of day for harvest might be an important consideration due to the CAM cycle as might the addition of menstrual blood to the soil. Friends who give their menstrual blood to their cactus garden do have amazing plants.

Too much of any nitrogen source favors rotting just as much as it does anything else.

Injection of precursors is at best going to have paltry results due to a small conversion ratio and is mostly only going to add more of what you actually inject to what you ingest. Speculation to the contrary like in Gottlieb is illfounded as to the scope of its impact. Carefully read the papers he refers to for a better picture about just how much increase is going to occur under the best of conditions. Let's say you had a whopping 7% conversion on the local level and you managed to get the plant to take up 100 mg of tyramine, that would add an entire theoretical 7 mg. Localized enough delivery is also going to be an issue.

An important thing to remember about cacti is that alkaloids have been determined to be locally produced and stored in parenchymal tissues and are not circulating or migrating as is the case in some other plants.

Feeding experiments in poppies with tyrosine work due to the alkaloids being produced in the roots and brought up into the capsule rather than their originating in the capsule, only modification of alkaloids occurs once they reach the capsule. Many plants do have circulating or migrating alkaloids but not the mescaline producers.

Older plants can be stronger than younger ones but once they are woody and hard this seems to no longer be an issue although the calcium oxalate druses that can be recovered do increase dramatically.

Starving for water appears to help in some but not all species. A friend of mine has extensively explored this for quite some years and has become convinced that a happy plant is stronger than an unhappy plant. He now only works with bridgesii which is not a bad suggestion.

Some species thrive in more shade and some thrive in more sun.

That said taking cuttings several months before their use does seem to help but its also worth keeping in mind that so many factors can impact the results of an ingestion its hard to use bioassays for meaningfully assessing alkaloid levels.

A lot more real and controlled work is needed on all of this as a huge body of mythology has been created due to a lack of rigor and quantitative isolations.

The best approach to maximize the content is to choose good plants to begin with. If people want to have only really potent plants they would do best to get rid of their nonpotent ones to make room for some new acquisitions.

Bugs seem to equally eat all of my plants and target the newest and most succulent growth most often rather than selecting for the most potent ones.

[trucha]

I cannot do Stamet's work justice and it would take too much time to really even scratch the surface.

This book is not just eye opening but will give people some true optimism for there being hope of a real future.

For sake of not sidestepping the question, among the things that he has shown though:

If given a maze with food down only some paths, mycelium will not colonize the wrong pathways. They apparently have remote sensory devices capable of signalling where growth should be directed.

When placing a dougfir under shade stress mycorrhiza was found to transfer food out of another tree species it did not prefer and feed the dougfir, a species it did prefer, to keep it healthy. Not enough to kill the other tree species but enough to keep the dougfir healthy even in inadequate light.

Stamet's take is that fungi are the elder consciousness and in charge of running life on the planet. He makes a really good case for it at least.

Quite off topic though the most exciting thing to me was his discovery that oyster mushrooms can completely digest petroleum sludge and find it a high energy food source they can render nontoxic due to their ability to break every carbon-carbon bond it contains.

The discovery that P. azurescens was found the most effective known degrader of VX nerve gas is a touch interesting in a poetic way also.

People should not just buy and read this book they should try to get copies of it into every public library they can.

[Monk]

This book is not just eye opening but will give people some true optimism for there being hope of a real future

I couldn't agree more. When I get home today, I think I'll pick up my dog-eared, worn out copy and refresh my memory! I love that the Department of Defense actually sponsored the work he did with the P. azures/VX analog. I also like the myco-insecticide section a great deal. I totally agree: Get this book!

[Monk]

The best approach to maximize the content is to choose good plants to begin with. If people want to have only really potent plants they would do best to get rid of their nonpotent ones to make room for some new acquisitions.

Couldn't agree more with this statement as well.

Speculation to the contrary like in Gottlieb is illfounded as to the scope of its impact. Carefully read the papers he refers to for a better picture about just how much increase is going to occur under the best of conditions. Let's say you had a whopping 7% conversion on the local level and you managed to get the plant to take up 100 mg of tyramine, that would add an entire theoretical 7 mg

Is this info from the said papers (I have not seen them-only remember Gottlieb being the inspiration for SCBs youthful experiment) or is this just 'speculation' from the other direction? Also, the process involved injecting this amount at several points along the cactus many times over several months. So, say 400 mg (or more) per treatment, twice per month over 5 months....4.0 grams tyrosine. Even if the conversion is around 7%, we're looking at a theoretical 280 mg increase in a small (28 g. dried) sample.

[Ace]

Thank you Trout/Felonious - you have twisted my arm! As soon as I have a few $ saved up I shall invest in a copy and have a bit of a gander

-End of thread-hijack-

[trucha]

McLaughlin's team was getting trivial incorporation returns. This was simply studying what precursors were favored and ranking their percentge of incorporation not deterimining how to increase alkaloid levels.

There is no evidence I know of that anyone has ever gotten such great absorption or results outside of with paper and pen.

I'm plucking that 7% number from ancient memory or perhaps out of my ass for sake of discussion only but will check and come back with something more solid. Its not a great conversion probably due to the alkaloids not being circulating or migrating.

They were also doing tiny injections not whopping great ones.

Local tissue necrosis at the injection site is a common report of people following Gottlieb's suggestions which I suspect were purely armchair extrapolations like much of the stuff he wrote. At least this is true of the people who wrote to ER or me about it. Interestingly most used dopamine and no one I can tell ever bioassayed their plants. One person expressed fear of doing so which leaves me wondering just what their point was.

It would be nice if someone actually followed through with some isolations or bioassays that had controls.

Weeding out any subjects who are not negative placebo responders is a critical first step in the latter of course.

There is much more interesting work with plants that do have migrating alkaloids where feeding raw opium latex with tyrosine or glucose showed conversion into morphine.

To me a much more amusing thing about the DOD funding the VX degradation work is Stamet's submitted assorted mycelium with only code numbers and when they learned the ID they were not only quite annoyed with him but did not pursue azurescens use.

The work Chemical Warfare by Ketchum suggests VX is an effective antidote for belladonna not just vice versa. Its dose that makes the poison of course.

His take on it is quite fascinating considering all the scare hype we get dumped on us. His take is that if you get VX on you just scrape it off and find some soap and water. If you are exposed to it in the air get to fresh air and you are unlikely to die if you did not ingest enough to die right away. Its dispersed as an aerosol not a gas so is apparently much harder to use than the powers that be would like us to believe.

Edited August 15, 2007 by trout

[Phytotechnic]

Thanks to Creach for informing me of this thread. I can provide some very preliminary data on alkaloid concentration of some different Trichs, including 2 clones, gown at the same locality but under different conditions. This data is from trials of a rapid method of quantitative analysis. This method is still under development, but should be published by late 2008.

6 different samples were carefully analysed:

1. T. bridgesii, mature , healthy, full sun grown, although with little recent fertiliser application, dark greenish blue 1.2 %

2. T. bridgesii monstrose, healthy, half shade grown, no recent fertiliser application (over 5 months), more green than blue 0.7 %

3. T. pachanoi, healthy, full shade grown, dark bluish green, with very broad ribs, little recent fertiliser application 0.7 %

4. T. scoplulicolus, mature, healthy, rich green, little recent fertiliser application ,3/4 sun grown 0.2 %

5. T.pachanoi, not the best looking sample, was grown in +/- full sun then moved to shade then back into full sun, it became very elongated, with the top 8 cm or so of growth around 2 cm in diameter, bright green, with narrow ribs, no recent fertiliser application. 0.4 %

6. T.peruvianus, fairly healthy, green, little recent fertiliser application, half sun, 0.2 %

Notes.

sample number 2, bidgesii monstrose, appeared to have been over saturated, causing dampening of the signal. its content appears to have been somewhat higher than sample 1. much work remains to be done on this method, but based on some later examinations, a figure twice as high as sample 1 seems possible.

percentages are as dry weight and may include dimethoxy-pea if present.

In an earlier experiment, under less controlled conditions than above, a sample from a mature plant, from which sample number 3 was cloned, was estimated to contain around 0.2 % dry weight tmpea, it was pale green and grown in full sun, and the sample did not contain any woody core. Also analysed in this batch was a very robust T. pachanoi-like cactus with slightly longer spines and a faster growth rate, it had an estimated concentration of 0.1 % dry weight.

After the next growing season more detailed information will be available to more objectively measure the influence of several environmental factors on cactus alkaloid concentration. The information will be posted here as soon as it becomes available.

will add some photos soon.

[teonanacatl]

what was your method?

was only the 2 pachanoi samples clones?

Edited August 15, 2007 by teonanacatl

[Mr Griffin...]

For anyone interested:

BIOSYNTHESIS OF MESCALINE AND ANBALAMINE IN PEYOTE. II'

Jan Lundstrom and Stig Agurell

Department of Pharmacognosy, Royal Pharmaceutical Institute

Tetrahedron Letters No.42, p p. 4437-4440, 1968.

mescaline_biosynth.pdf

A COMPLETE BIOSYNTHETIC SEQUENCE FROM TYROSINE TO MESCALINE

IN TWO CACTUS SPECIES.

Jan Lundstrom and Stig Agurell

Faculty of Pharmacy, Department of Pharmcognosy

Tetrahedron Letters No.39, pp. 3371-3374, 1969.

Tyrosine_to_mescaline_biosynthesis.pdf

Tyrosine_to_mescaline_biosynthesis.pdf

mescaline_biosynth.pdf

Tyrosine_to_mescaline_biosynthesis.pdf

mescaline_biosynth.pdf

[Teotzlcoatl]

Nice thread.

Rather than doping up our weak clones, we should just breed new potent ones.