The old Colchicine idea

[homebrew420]

Hey all I am starting a job at a dispensary in Boulder CO on Monday. Part of my job will be conducting experiments on production lighting techniques new variety developement, building Led lighting, and a host of other misc other responsabilities. I am to say the least excited. One area I have been interested in since reasing RCCs "Marijuana Botany" has been the use of colchicine to induce mutation in a particular plant, and possibly its progeny. The hopes of course would be to develope something truly novel. And this novelty would hopefully be in way of cannabinoid ratio and profile. I would not however be disapointed with a verigated plant of malformed leaves.

Has anyone attempted this and/or have 1st hand knowlege of results from this type of treatment.

I plan on using autumn crocus juice and spraying the plants thoughout their lives and heavy doses when they begin to produce seed. No material treated would EVER be used as medicine. Triploidy and tetraploidy is the end goal if the results are favorable. For all I know is could do the exact opposite and ruin the plants if it doesn't kill them.

And yes I am fully aware of the genetic issues that can arrise if I am exposed and that this substance is mutigenic across the board. All precautionary measures will be taken. Nobody needs to get hurt.

Peace

 

[Bass Akwards]

Don't Waste Your Time

Don't Waste Your Time

Your time is much better spent on selective breeding.

Access to lab analysis will greatly simplify the chore.

In the first place, the best way to insure that genetic effect is to treat seeds, not plants. Inspection of the growing tip will show a bulbous growth spurt if the mutagen has done the job. However, even if it survives seedling stage, and most won't, the trait won't be passed on. More likely, it will be a low yielding runt with very dark foliage.

People have wailed away at that scheme for decades, without success.

If you really want to give it a shot, try soaking the seeds first in plain water until they just sink, then drain for an hour, and soak for 8 - 10 hours in a Colchicine solution. ( Trial & error on the dose. Practice with imported mersh We had access to USP, you probably won't. ) Fold in damp brown coffee filters, place in a baggie in a warm, dark, place. When 80%+ die, a large number of sprouts will have the bulbous tip. After planting, 90% of those won't grow past the first 2 leaves.

 

[homebrew420]

interesting. And I thank you for your info on this.

Peace

 

[mofeta]

In the first place, the best way to insure that genetic effect is to treat seeds, not plants.

The way I remember is that application to the meristem was more effective than seed treatment in producing tetraploids.

People get confused because Clarke said in Marijuana Botany that the "safest" way was seed treatment. He meant safest with regards to avoiding the ingestion of colchicine if you planned on actually smoking the plant that you appllied the colchicine to. I don't think this is true though. Colchicine is used to treat acute gout in humans at levels and order of magnitude larger than you would ever get by smoking a treated plant.

Clarke also said that seed treatment was also more effective than meristem treatment. His reason was something like "the whole plant would be tetraploid". Well sure, this is true, but so what? How hard is it to root a tetraploid shoot from a plant treated meristematically? As he points out, successful seed treatment kills most of the seeds. Treating a established plant is much less lethal. Seed treatment = <1% successful tetraploids, meristem = ~25% success if I remember correctly.

One way to combine the benefits of seed treatment with the benefits of treating post-germination is to treat the hypocotyl of the emerging seedling. It produces a "whole plant" tetraploid with less lethality than seedling treatment.

I think that inducing polyploidy is useless unto itself. It is a useful tool to meet the ends of other goals though, like doubling haploid plants, or other schemes that require ploidy manipulation as part of a greater goal.

 

[Midnight]

If anyone has old High Times mags there is an article about what you are asking. I don't have the edition but iirc it will be sometime between May 79 to January 80.

 

[rick shaw]

I remember this one,not sure why.

http:www.imdb.com/title/tt0681814

 

[OakyJoe]

Translated by Google:

Polyploid species - mutations with colchicine

Polyploid species differ by a double to multiple set of chromosomes in the plant. These plants are often larger. In general, the use of colchicine, the poison of the autumn crocus , a ässerst rare , protected species , is assumed. The seeds are placed in an extract or a solution of the venom and the seeds used then normal.

Little is known about the use of colchicine in hemp plants. Principle is to consider the use of colchicine, that the probability that you get a new , better - mutant type , not very promising . It certainly requires a large number of experimental plants , so the phenomenon of polyploidy is clarified with hemp. It is also not clear to what extent polyploid plants contain more THC , or whether they are of larger stature .

You can leave mutate your seeds with colchicine . Plants with mutations are negative and / or positive qualities of the plant . Colchicine is very toxic. 7 mg is a lethal dose. If you treat the seeds with colchicine, So you 're going a risk. Normally you survived the use of colchicine Weed , but how much you think of something you 're worth . The responsibility you bear ! .

Colchicinpillen be prescribed gout and prescription. Colchicine is sold only on the pure poison book. It is easiest if you win the colchicine from the crocus winter ( Cholchicum major). The pollen of this plant contain about 3% colchicine .

Press several tubers in a garlic press and filter the liquid through a coffee filter . If you then verdünnst your Prdukt with the same amount of water , you have the right solution with 1.5 % colchicine

Put your seeds 8-24 hours into the brew , and I hope that after that the seeds are left to germinate .

Colchicine prevents the first cell division , so that the plant can survive , they must form a double set of chromosomes . The result is much stronger , bushier plants with more but deformed leaves.

This dramatic action has reached its peak and we can turn back to nature.

 

[foomar]

There are other chems available , but some are impossible to buy without a license.

Ethyl methane Sulphonate
Methyl Methane Sulphonate
Ethyl Ethane Sulphonate
Ethylene Imines

5 Bromo Uracil
2 Amino purine

Acriflavin, Proflavin

Nitrous Acid
Hydroxylamine
Sodium Azide



In the case of DES the mutagenic solution should be changed at every half an hour to get good results. Half life is the function of temperature and pH for a particular compound.

One should be extremely careful in handling alkylating agents since most of them are carcinogenic. Especially for ethylene imine, it should be handled under aerated conditions. EMS though not dangerous, it should not be pipetted out by mouth. Besides the alkylating agents, we are also having chemical mutagens like, Base analogues, Acridine dyes, Antibiotics and other miscellaneous chemicals.

Treatment of seeds with mutagenic chemicals:


Materials required:- conical flask, beaker, pipette, glass rods, measuring cylinder, stop watch, distilled water and phosphate buffer.

Method: - Mutagenic chemical is diluted to the required concentration by using distilled water. To prepare the molar concentration of DES, the method is

Molecular weight x a.i. (purity percentage)
Specific gravity (active ingredient)

154 100
Eg. DES = ---- x ----- = 131 CC.

1.18 .................99
131 CC dissolved in one litre will give 1 molar solution.

Seeds have to be soaked in the distilled water for different hours depending upon the seeds, to initiate biochemical reactions. The chemical action is found to be affected by the frequency and spectrum of mutagen depending upon the stage of cell division, during the process of germination. If the chemical treatment is synchronized with DNA synthesis stage (G1, S and G2) then we can get better results.

The presoaked seeds are taken in a flask and chemical is added. Usually the quantity of the chemical is ten times the volume of seeds. Intermittent shaking should be given to ensure uniform exposure of the chemicals. The chemical should be drained after the treatment time is over. The seeds should be washed thoroughly in running tap water, immediately for not less than 30 minutes. After washing, the seeds should be dried in between the filter paper folds. Seeds are to be arranged in germination tray with equal spacing. Trays are kept in a controlled environment of temperature and humity. Periodical observation on germination upto 10-15 days is needed. From the germination percentage, we can assess the LD50 dose.

If you search Mutation Breeding there are quite a few interesting documents around.

 

[Crusader Rabbit]

Can a polyploid plant be crossed with a normal diploid plant to produce viable offspring?


For a period of time I was receiving injections of colchicine as an "experimental" treatment to reduce the pain of ruptured spinal disks (disk herniations can develop a crystalline growth like gout). As long as the injection went into a vein everything was fine.

Then one time the doctor missed the vein. After the long drive back home I was planning to start typing up a big project but my arm started feeling odd. Soon it had developed into a nerve irritation that left me unable to type or hold a pencil. The lightest brushing of the hairs on my arm brought excruciating pain. Here I had this table covered with notes and outlines and I couldn't put a word onto paper or the computer screen. This disability lasted a week, and during this time all I could do was sit and watch the news of Jerry Garcia's death, trying to hold my arm so that nothing touched it. I can still feel it.

 

[OakyJoe]

Can a polyploid plant be crossed with a normal diploid plant to produce viable offspring?

A cross between a polyploid plant and a "diploid" plant will always be a "diploid" plant.

 

[Only Ornamental]

A cross between a polyploid plant and a "diploid" plant will always be a "diploid" plant.

Naw, a diploid and a tetraploid will give triploids (theoreticlally), though I recall someone (was it Charles Scott or Sam Skunkman?) saying they got mixtures of whatever is possible (and even fertile triploid plants, however that should work).
Uhhh.... got to hurry, dinner's ready

 

[Only Ornamental]

....the use of colchicine to induce mutation in a particular plant, and possibly its progeny, and possibly its progeny. The hopes of course would be to develope something truly novel. And this novelty would hopefully be in way of cannabinoid ratio and profile.

I plan on using autumn crocus juice....

Triploidy and tetraploidy is the end goal if the results are favorable. For all I know is could do the exact opposite and ruin the plants if it doesn't kill them.

Hey again,
I guess it has already been mentioned; colchicine, though regarded as inducer of mutations, usually causes tetraploide cells and to a lesser extent hexa-/octaploide cells but usually no genetic mutations (depending on how and what you treat). It has been shown by several breeders (also here aboard) that, in case of cannabis, polyploidy shows no improvement in whatsoever regard. Besides, most modern vegetables/crops have been tested as polyploids (and many cereals actually are polyploide) but there's not hemp variety with that trait.

True genetic mutations usually are inherited; many 'mutations' observed with meiosis/mitosis inhibitors are only perturbances of cellular regulations but not mutations. Though, polyploidy may result in altered epigenetic changes; they may look like mutations but are usually not inherited (the polyploidy will remain and hence the alterations as long as you don't cross with a different ploidy).

Concerning your wish for changes in the cannabinoid ratio and profile: AFAIK, that won't change no matter what you do (unless you find a new synthase). Due to specific enzymes, one always has a theoretically fixed THC+THCV v.s. CBD+CBDV ratio (2:0, 1:1 or 0:2), a fixed one between THC+CBD v.s. THCV+CBDV (same as before) and CBG v.s. THC+CBD+THCV+CBDV (idem). True ratios are not 2:0 but rathe 10-20:1 (always a few 'side products'). But you may find, already in the treated plant or its heterozygous offspring, plants with simply one copy of the corresponding synthases lacking (so half THC content). What you might get is a new cannabinoid (most likely inactive) or higher/lower overall cannabinoid concentration (e.g. more or bigger trichomes or flowers).

Besides, autumn crocus contains highest amounts of colchicine in the flowers and the seeds .

As foomar stated, you're better off with other substances which truly induce mutations (look up 'Mutation' in Wikipedia). There, you have principally the choice between two possibilities:
- Inducing point mutations with alkylating agents such as N-ethyl-N-nitrosourea or ethyl methanesulfonate. Both are often used in animal models.
- Cause frameshift mutations with DNA intercalating agents like ethidium bromide.
Point mutations are easier to make but are often without effect or result in loss of function or minor changes whereas frameshift mutations may really result (with a very small chance) in something entirely new. Which of the two suits your cause better is up to you. May I refer you again to Wiki concerning the two mutation mechanisms and their effects?
Commonly, these mutations aren't deadly when you treat a growing plant properly or are even invisible due to their heterozygous and also often recessive nature. So, you will either depend on extensive gene analyses (not feasible I guess) or you self the treated plant (male/female flowers growing as close one another as possible) to eventually obtain a now phenotype in the first generation. Else, you may even be forced to do F2's to make a new phenotype appear. In any case, finding something new and useful likely requires thousands and thousands of plants (seeds from treated plants) if you work blind (without PCR equipment). Good luck!

 

[foomar]

Increasing the natural mutation rate by radiation has been an interest , from 1950,s irradiated grain trials the family farm took part in to new colour mutations developed commercially for annual plants.


Looking at what has been found in comparable species , there should be potentially useful non lethal variations , at one in a thousand rather than one in a million.


A number of research labs will irradiate seed with high energy proton beams , I could definitely arrange to have 5000 pelargonium seeds done but don't know if canna would be a problem legally.

But when you look at the numbers required it would be a daunting task finding the new Blackberry let alone trying to stabilise it , even with a lab .

 

[Stash]

I can remember way back when and this was big many years ago. I never got any where with it. However I do have a large patch of hundreds of Colchicum autumnale. They sure are some very pretty flowers.Later, Stash

 

[RoadRash]

I was just wondering about this, looking at a picture of Querkle, Urkle x Space Queen.

If I recall, Vic High @ BCGA bread SQ as a cross between Chemo & C99 (Killer Queen = G13 x C99, I think).

I had the impression that Chemo may have been a genetically altered strain.


However today's variety of high-THC strains came about - I'm glad they did.

I have a coffee (Arabica) plant that has stems growing in 3's instead of 2's, it makes me wonder if it's polyploid or special or if that is more common with coffee.

 

[foomar]

Some of the highest yielding food crops and fibre plants have resulted from radiation induced mutation , there should be value in treatind canna seed or pollen if you could run enough to find them.

Don't see any objection to using radiation as its the driving force behind evolution anyway.

And has to be less contentious than stuffing fish dna into tomatoes and the like.

 

[habeeb]

interested..

start growing Dj. I will tell you CK has the highest mutant ratio I've seen in his seeds..