Colchicine question???

[Agent-Smith]

I've been reading about breeding and I saw Colchicine mentioned a few times in cannabis breeding. They say that it is poisonous before administering it to the plant but they never say if it is poisonous after harvest when smoked or run through some form of extraction. So does anyone know?

[suzy cremecheese]

Odds are that the small amount used on the seed won't hurt you in the finished product but I have no research to prove it either way.

For colchicine to be effective, cause polyploids, you need to use enough to kill most of the treated seedlings. The only way to be sure you have a polyploid is to actually count the number of chromosomes. You need some lab equipment for that.

Let us know how it goes.

[plantbuilder]

agent smith,
that is kind of like asking if it's ok to eat seedless watermelon
you just treat the seeds
should be fine mon
peace.
pb

[Agent-Smith]

Well, I'm not going to be trying it anytime soon, just wanted to know if it was worth it down the road, and if it was non-poisonous. :teeth: Thanks guys/gal ;)

[smokinbasser]

I have read that you treat a mother plant and pollinate it and harvest the seeds which should all be polypoids and here is the cruel part dispose of the mother treated with colchicine since the buds are now not safe to smoke. I would not be willing to find out how similar colchine is to paraquat.

[clips]

found a lil information should be helpfull got to love cut and paste :wave: and as with any chemicals always research before use and be aware before ya venture . enjoy be safe and ill chat ya later clips

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Polyploidy (favorable traits in Cannabis) has not been shown to occur naturally in Cannabis; however, it may be induced artificially with colchicine treatments. Colchicine is a poisonous compound extracted from the roots of certain Colchicum species; it inhibits chromosome segregation to daughter cells and cell wall formation, resulting in larger than average daughter cells with multiple chromosome sets.

The studies of H. E. Warmke et al. (1942-1944) seem to indicate that colchicine raised drug levels in Cannabis. It is unfortunate that Warmke was unaware of the actual psychoactive ingredients of Cannabis and was therefore unable to extract THC. His crude acetone extract and archaic techniques of bioassay using killifish and small freshwater crustaceans are far from conclusive. He was, however, able to produce both triploid and tetraploid strains of Cannabis with up to twice the potency of dip- bid strains (in their ability to kill small aquatic organisms). The aim of his research was to "produce a strain of hemp with materially reduced marijuana content" and his results indicated that polyploidy raised the potency of Cannabis without any apparent increase in fiber quality or yield.

Warmke's work with polyploids shed light on the nature of sexual determination in Cannabis. He also illustrated that potency is genetically determined by creating a lower potency strain of hemp through selective breeding with low potency parents. More recent research by A. I. Zhatov (1979) with fiber Cannabis showed that some economically valuable traits such as fiber quantity may be improved through polyploidy. Polyploids require more water and are usually more sensitive to changes in environment. Vegetative growth cycles are extended by up to 30-40% in polyploids. An extended vegetative period could delay the flowering of polyploid drug strains and interfere with the formation of floral clusters.

It would be difficult to determine if cannabinoid levels had been raised by polyploidy if polyploid plants were not able to mature fully in the favorable part of the season when cannabinoid production is promoted by plentiful light and warm temperatures. Greenhouses and artificial lighting can be used to extend the season and test polyploid strains. The height of tetraploid (4n) Cannabis in these experiments often exceeded the height of the original diploid plants by 25-30%. Tetraploids were intensely colored, with dark green leaves and stems and a well developed gross phenotype. Increased height and vigorous growth, as a rule, vanish in subsequent generations. Tetraploid plants often revert back to the diploid condition, making it difficult to support tetraploid populations. Frequent tests are performed to determine if ploidy is changing.

Triploid (3n) strains were formed with great difficulty by crossing artificially created tetraploids (4n) with dipbids (2n). Triploids proved to be inferior to both diploids and tetraploids in many cases. De Pasquale et al. (1979) conducted experiments with Cannabis which was treated with 0.25% and 0.50% solutions of colchicine at the primary meristem seven days after generation. Treated plants were slightly taller and possessed slightly larger leaves than the controls, Anoma- lies in leaf growth occurred in 20% and 39%, respectively, of the surviving treated plants. In the first group (0.25%) cannabinoid levels were highest in the plants without anomalies, and in the second group (0.50%) cannabinoid levels were highest in plants with anomalies.

Overall, treated plants showed a 166-250% increase in THC with respect to controls and a decrease of CBD (30-33%) and CBN (39-65%). CBD (cannabidiol) and CBN (cannabinol) are cannabinoids involved in the biosynthesis and degradation of THC. THC levels in the control plants were very low (less than 1%). Possibly colchicine or the resulting polyploidy interferes with cannabinoid biogenesis to favor THC. In treated plants with deformed leaf lamina, 90% of the cells are tetraploid (4n 40) and 10% diploid (2n 20). In treated plants without deformed lamina a few cells are tetraploid and the remainder are triploid or diploid.

The transformation of diploid plants to the tetraploid level inevitably results in the formation of a few plants with an unbalanced set of chromosomes (2n + 1, 2n - 1, etc.). These plants are called aneuploids. Aneuploids are inferior to polyploids in every economic respect. Aneuploid Cannabis is characterized by extremely small seeds. The weight of 1,000 seeds ranges from 7 to 9 grams (1/4 to 1/3 ounce). Under natural conditions diploid plants do not have such small seeds and average 14-19 grams (1/2- 2/3 ounce) per 1,000 (Zhatov 1979). Once again, little emphasis has been placed on the relationship between flower or resin production and polyploidy. Further research to determine the effect of polyploidy on these and other economically valuable traits of Cannabis is needed.

Colchicine is sold by laboratory supply houses, and breeders have used it to induce polyploldy in Cannabis. However, colchicine is poisonous, so special care is exercised by the breeder in any use of it. Many clandestine cultivators have started polyploid strains with colchicine. Except for changes in leaf shape and phyllotaxy, no out- standing characteristics have developed in these strains and potency seems unaffected. However, none of the strains have been examined to determine if they are actually polyploid or if they were merely treated with colchicine to no effect.

Seed treatment is the most effective and safest way to apply colchicine. * In this way, the entire plant growing from a colchicine-treated seed could be polyploid and if any colchicine exists at the end of the growing season the amount would be infinitesimal. Colchicine is nearly always lethal to Cannabis seeds, and in the treatment there is a very fine line between polyploidy and death. In other words, if 100 viable seeds are treated with colchicine and 40 of them germinate it is unlikely that the treatment induced polyploidy in any of the survivors. On the other hand, if 1,000 viable treated seeds give rise to 3 seedlings, the chances are better that they are polyploid since the treatment killed all of the seeds but those three.

It is still necessary to determine if the offspring are actually polyploid by microscopic examination. The work of Menzel (1964) presents us with a crude map of the chromosomes of Cannabis, Chromosomes 2-6 and 9 are distinguished by the length of each arm. Chromosome 1 is distinguished by a large knob on one end and a dark chromomere 1 micron from the knob. Chromosome 7 is extremely short and dense, and chromosome 8 is assumed to be the sex chromosome. In the future, chromosome *The word "safest" is used here as a relative term.

Colchicine has received recent media attention as a dangerous poison and while these accounts are probably a bit too lurid, the real dangers of exposure to coichicine have not been fully researched. The possibility of bodily harm exists and this is multiplied when breeders inexperienced in handling toxins use colchicine. Seed treatment might be safer than spraying a grown plant but the safest method of all is to not use colchicine. mapping will enable us to picture the location of the genes influencing the phenotype of Cannabis. This will enable geneticists to determine and manipulate the important characteristics contained in the gene pool. For each trait the number of genes in control will be known, which chromosomes carry them, and where they are located along those chromosomes.

(Taken from 'Marijuana Botany',R.C.Clarke,CH.3

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Colchicine--- Tetraploidy can be induced by the mutagenic alkaloid colchicine, which is found in the autumn crocus, (colchicum autumnale). Colchicine allows a cell to double its chromosomes, but prevents meiosis (the splitting of cells), thus forcing the cells to become polyploid. When applied to cannabis, colchicine produces tetraploid plants which tend to be taller, with greater stem diameter, seed and pollen size. The THC content can increase up to 250%.(27)

A. Zhatov, et al., reported these findings from their research:

"The greatest % of polyploid plants is obtained when hemp plants are treated with 0.5% colchicine solution for 2 hours in the phase of cotyledon leaves. The treatment with colchicine solution inhibits growth. This inhibition continues for 2.5-3 weeks, after which the surviving plants resume normal growth and development. The guard cells in the leaves and the pollen grains of tetraploid plants are larger and the number of pores on the pollen grains are greater. Tetraploid plants are taller and the diameter of their stems, seed size and weight of 1000 seeds are greater. The anatomical structure of the stems differs from the diploid plants in a greater amount of primary and secondary fiber. The pollen viability of the tetraploid plants is lower than that of diploid plants. Vegetation period in tetraploids continues 8-15 days longer than in control plants." (28)

Colchicine also can sprayed on the seeds while they are developing on the mother plant. The flowers of plants treated in this manner should not be smoked because the concentration of colchicine may be dangerously high. A third method is to soak seeds in the solution for 2 to 4 hours. Colchicine stimulates the development of the taproot at first, but this effect ceases within a week; then the seedlings go into shock. About 30% of the survivors will be polyploid.

Colchicine can be bought, or prepared by grinding 100 grams of colchicum seeds to powder and percolating with 2 volumes of ethanol and 1 volume of water. The solution contains approximately 4 milligrams of colchicine. Label the bottle and store it safely: colchicine is toxic. Always wear rubber gloves when handling colchicine.

taken fromHemp Husbandry
Robert A. Nelson
4.6 ~ Polyploidy http://www.rexresearch.com/hhusb/hh4bot.htm
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General
Synonyms:
Use: Alkaloid
Molecular formula: C22H25NO6
CAS No: 64-86-8
EC No:
Physical data
Appearance: pale yellow crystals or powder
Melting point: 142 C
Boiling point:
Vapour density: 14.0 (air = 1)
Vapour pressure:
Density (g cm-3):
Flash point:
Explosion limits:
Autoignition temperature:
Water solubility: moderate

Stability
Stable. Light sensitive. Incompatible with strong oxidizing agents.
Toxicology
Poison. May be fatal if swallowed. Note very low LD50s below. Causes severe irritation.
Toxicity data
(The meaning of any abbreviations which appear in this section is given here.)
IPR-RAT LD50 6.1 mg kg-1
IVN-MUS LD50 1.6 mg kg-1
SCU-MUS LD50 1.2 mg kg-1

Risk phrases
(The meaning of any risk phrases which appear in this section is given here.)
R26 R27 R28.

Transport information
Personal protection
Safety glasses, gloves, good ventilation.

http://physchem.ox.ac.uk/MSDS/CO/colchicine.html

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found a lil more for your reading enjoyment =)~
http://www.cannabase.com/cl/bcga/breeding/colchicine.htm

[Agent-Smith]

It just sounds like something that if you could make it work, you could keep that girl around in clone form and have an extra potent variety. :teeth:

[suzy cremecheese]

that is kind of like asking if it's ok to eat seedless watermelon

Not exactly PB. Seedless watermelon is created by crossing a diploid (2x) with a tetraploid (4x). The surviving results are triploids (3x) which for the most part don't make viable seed. That's why you have all those little white aborted ones in seedless watermelon. There is no chemical treatment involved in making seedless watermelon.

The amount that you need to use to treat a seed is very slight and I'd imagine that over the course of a plants life any traces of it they may be left would be quite insignificant. However somke it at your own risk and don't pass it to your friends without telling them it might be hazardous.

I have read that you treat a mother plant and pollinate it and harvest the seeds which should all be polypoids...

I've never heard that about treating the mother plant. From what I've read you treat the seeds. The majority will be killed by the treatment. If they are not then you didnt use enough to induce polyploidy in any of them. Of the surviving results many may not be polyploids but merely mutants that might or might not survive to maturity. You need to get a look at the cell during the proper stage of division, sorry I forget which one, in order to see the chromosomes and count them to see if you actually have a polyploid. I imagine you would kill the mother plant if you treated it.

I believe that this has been worked with a few times and that any potential benefits are overwhelmed by the hassel and extra energy that goes into maintenance. Any increases in potency have been slight and measured using lab instruments not smoke tests.

Id still be curious to see what you find.

[plantbuilder]

the most common way of getting tetraploid donors is to 'freak' em
http://cuke.hort.ncsu.edu/cucurbit/wmelon/seedless.html
yes that is one step away from seedless watermelon
i can't see why you would spray the bush when you could remove the seed and treat it
peace.
pb

[suzy cremecheese]

Seedless cultivars are produced by crossing a tetraploid (4X=44) inbred line as the female parent with a diploid (2X=22) inbred line as the male parent of the hybrid. The reciprocal cross (diploid female parent) does not produce seeds. The hybrid is a triploid (3X=33), and is female and male sterile. Triploid plants have three sets of chromosomes, and three sets cannot be divided evenly when they go into two daughter cells during meiosis (the cell division process that produces the gametes). Since the triploid hybrid is female sterile, the fruit are seedless. Because the triploid is also male sterile, it is necessary to plant a diploid cultivar in the production field to provide the pollen that stimulates fruit to form. Usually, one third of the plants in the field are diploid and two thirds are triploid. Cultivars should be chosen that can be distinguished easily so the seeded diploid fruit can be separated from the seedless triploid fruit for marketing.

Stage 2 is the production of tetraploid plants. Many methods have been used effectively in other crops to produce polyploids, including tissue culture regeneration, temperature shock, and X-rays. However, colchicine (C22H25O6N), a product from the seeds and bulbs of Colchicum autumnale L., is probably the most widely-used chemical for induction of watermelon tetraploids.

Stage 3 involves tetraploid line development. Tetraploid plants are selected (using methods such as leaf guard cell chloroplast number) in the T1 generation (plants from colchicine treated diploids) from the greenhouse flats where they were treated with colchicine. It is then necessary to plant the T2 generation in flats to verify that the plants are tetraploids in that next generation, and transplant the selections to greenhouse pots for self-pollination. Seeds from those selections can then be increased in larger plantings such as field isolation blocks to get sufficient numbers of seeds per tetraploid line to use in triploid hybrid production.

While the tetraploid parent is usually developed from a colchicine treatment that isnt the melon that is being consumed it is the offspring of this melon and a diploid. So in this case we are at least one generation away from the treatment. Tetraploid lines are developed so you could be many generations away from the treatment.(this is what I edited

Stage 3 involves tetraploid line development. Tetraploid plants are selected (using methods such as leaf guard cell chloroplast number) in the T1 generation (plants from colchicine treated diploids) from the greenhouse flats where they were treated with colchicine. It is then necessary to plant the T2 generation in flats to verify that the plants are tetraploids in that next generation, and transplant the selections to greenhouse pots for self-pollination. Seeds from those selections can then be increased in larger plantings such as field isolation blocks to get sufficient numbers of seeds per tetraploid line to use in triploid hybrid production.

Good link. I think there is something in here for those to consider when attempting to make their own polyploids. This also relates to smokin bassers earler commnet about treating mother plants.

Of all the methods of colchicine application, shoot apex treatment at the seedling stage was found most effective.

The bottom line is that the most dangerous part opf making polyploids is the treatment process. Take the proper precautions when working with the colchicine. The remnants left over in the plant are secondary to this.

[madscientist]

For those of you worried about ingesting colchicine from treated plants and seeds, don't. It was once used medically to treat gout. It inhibits the formation of the metabolic by-product of purine metabolism, uric acid, so that crystals of it aren't deposited in your body's joints. You'd have to smoke many pounds of bud from the plants to get what a gout suffer used to take each day. That's not to say it isn't detrimental in large amounts (but so is water!), especially the bulb extracts, since there are other alkaloids in the solution. I first used colchicine that a buddy of mine got from his dad who was a pharmacist. Just use caution as you should with any chemical procedure, such as wearing rubber gloves.

[Roach]

if you want to make tetraploid plants use Surflan instead, alot safer and cheaper (and easier to get)

http://members.tripod.com/~h_syriacus/oryzalin.html

[j6p]

... It was once used medically to treat gout. .

still is - anyone want some I got a bottle sitting in front of me...