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Colchicine use in tissue culture.

Posted by Proteus none (My Page) on
Fri, Jan 6, 12 at 3:51

Hey guys.

Have any of you ever tried using Colchicine in a MS medium?The only source of Colchicine I can get here is in pill-form from the pharmacy,but I have no idea to what strength it should added.I know it can make plants polyploid,but I am aiming for variegation.

Does anyone have any experience with this?

thanks.


Follow-Up Postings:

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RE: Colchicine use in tissue culture.

i do know that you don't want the pill form. the rate of colchicine is way too low. http://www.sigmaaldrich.com/catalog/AdvancedSearchPage.do i haven't bought from them, but i do know people who have. much of the time dmso is also used in conjunction with colchicine. again, i'm not sure if it would cause variegation, but most likely if you used it on something with slight variegation, the doubling of the chromosomes should increase variegation. hope this helps


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RE: Colchicine use in tissue culture.

In addition to colchicine, caffeine has some value in plant breeding. The cells detect the caffeine but 'think' it is DNA. They may respond by forcing an additional cell division. This can be useful for getting diploids from tetraploids.

Even sugars can induce apparent mutations.

I've also included some suggestions for using solvents to reveal pigments that are only barely present.

Karl

Here is a link that might be useful: Chemicals in Plant Breeding


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RE: Colchicine use in tissue culture.

Hi,
I'm also curious about inducing ploidy in the plant breeding. What would be your opinion on use of raw Gloriosa tuber? I have some raw root tubers and am thinking about grating it or slicing very thin and use as a water extract for inducing ploidy. Could it work?


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RE: Colchicine use in tissue culture.

Chara2,
Gloriosa tubers do contain plenty of colchicine, but it can be tricky getting the dosage right. According to the following review, 20 grams of tubers contains 60 milligrams of colchicine.

If you do use colchicine, look for sports as well as polyploids. Sometimes chromosome doubled cells will revert to the original chromosome number by dividing again. The resulting cells probably won't be exactly like the originals. This is especially useful when working with sterile hybrids. For instance, a sterile diploid hybrid treated with colchicine can produce both tetraploid shoots, and diploid "sports" that may just as interesting.

I have added this info and links to some articles on colchicine use

Karl

Review: Croaking on a crocus
02 February 1991
From New Scientist Print Edition.
JULIE JOHNSON

Toxicity of Houseplants by David G Spoerke Jr and Susan C Smolinske, CRC Press, Boca Raton, pp 244, $124.95/100 Pounds.

Hours after cooking and eating a meal of sweet potatoes, a 28-year-old woman developed classic symptoms of poisoning. Four days later she was dead. She had mistakenly eaten tubers of the glory lily plant (Gloriosa superba), native to Africa and Asia but also grown as a houseplant in the US. Two years earlier, a 16-year-old girl died after eating 12 flowers from Colchicum autumnale, or autumn crocus. Just half a flower might have proved fatal. Both plants contain the poison colchicine (acetyltrimethylcolchicinic acid), which damages blood vessels and nerves, and stops cell division. A fatal dose of colchicine lies somewhere between 7 and 60 milligrams, not much considering that a dried seed of C autumnale can contain up to 3.5 milligrams, while as little as 20 grams of G superba tubers provide 60 milligrams.

Here is a link that might be useful: Chemicals in Plant Breeding


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RE:Inducing Polyploidy with 'raw' Colchicine

Hi Karl,
Thank you for responding to my post. So, basically the raw juice of the Gloriosa tuber has around 3% of colchicine in it, which is 0.03 g/L concentration? In order to get the needed 0.01 g/L, I have to divide it twice in distilled water? Is that is so simple or I am missing something? Of course, I am aware of the hazards and will do my experiments as safe as possible. One more question: how soluble the colchicine is in water? Would it be available for the plant cells absorption just in this simple water solution? Sorry if I sound naive or ignorant; it is been so long since my schooling that I forgot a huge part of it.
Thanks for the references also; something for me to read on the rainy Monday evening.


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RE: Colchicine use in tissue culture.

Chara2,
I don't know that all the colchicine is in the sap of the tuber, though it might be.

I took an organic chemistry class in college, but that was 35+ years ago, and I don't remember much. One gram of tuber = 3 milligrams of colchicine. But I don't know how much water would be needed to make a 3% solution.

There are times when we really need a chemist around.

Karl


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RE: Colchicine use in tissue culture.

Karl,
you already helped me so much! Last night I pre-soaked around 100 seeds of the Adenium Obesum. I plan to start treatment of the grow tips with the twice divided sap on the 6th day after germination. Hopefully something interesting will come out of this. Thanks again, Karl!


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RE: Colchicine use in tissue culture.

You have about as much of a chance as being struck by lightning and winning the lottery on the same day as getting Sigma Aldrich to send Colchicine to anyone but a recognized national research facility. (i.e., "Big Ed's Wholesale Nursery" isn't going to cut it) Especially now that it's a former orphan drug that's been hijacked and monopolized by a greedy company in Philadelphia and sold for 100X what it used to be sold for. Another chemical supplier moved it into their "Restricted" category normally reserved for controlled substances - obviously to stop the spouses or family members from helping reduce costs by ordering it from a chemical supplier.

Apparently, Oryzalin (aka Surflan) works almost as well, and is much safer to handle. I plan to start doing some experiments with it next spring, to induce polyploidy in rhododendrons.


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