Next Year's Colchicine Experiment

I’ve tried surflan, and as many you know, I may have success with Rosarie d’lhay, but it hasn’t bloomed since the two years I’ve applied it. I’m thinking this is something that needs to be grafted…

This year I have obtained 500 milligrams of Colchicine, and bought all necessary supplies to keep me and the family safe. I have a mini refrigerator, new and sterilized doppler bottle, eye glasses, etc… Let’s just say everything is ready for Spring. I’ve fixed the door knobs of the backyard, so that when I do apply colchicine, the gates will be locked so that the kids won’t go in the backyard and potentially rub up against stuff.

Now I am going to germinate seeds from my openly pollinated R. foliolosa. Looking the amount I’ve cleaned, it is easily over a thousand seeds. I am thinking that obviously there will be many selfs as there maybe many hybrids. And hopefully when a few do germinate, I will apply the 5 percent solution of colchicine to many of the selved seedlings.

I will be using Dr. Basye’s method as described in his 1990 article in the ARS, using only half a drop overnight (I’m going to say around 8 hours). However, I am very intrested in trying to make Nigel Hawthorn fertile, a hybrid between a rugosa and hulmutha. It’s very sterile, but the problem I am seeing is how to maintain colchicine the buds long enough before it evaporates. Last Spring I applied a DMSO/surflan mixture without any success. Although I think I may had made too strong of a mixture…

Since I am not using DMSO, my thinking is probably using a cotton, or even using flavorless jello or algar. Has anybody else heard of any other methods?

In another experiment, I will be using the Autumn Crocus bulb, from which colchicine is made. During my research, it seems that many Marijuana users also attempt to make Marijuana polyploids. The most common method I heard was to mash a corm of the Autumn Crocus and mix it with an equal amount of water. After a careful extraction with a coffee filter, seeds from Marijuana are then submerged into the solution for many hours and then planted. Most of the survivors are usually polyploids.

Now I am thinking of using this method, but only with seeds of R. foliolosa. However the seed coat may present some trouble, and I’m thinking of stratifying them first to weaken, and to nick a few with sand paper, just like a gardener does with Morning Glories. But the fact the seeds of R. foliolosa all float presents difficulties, but perhaps I can fasten it to a tea bag or something…

Hi Enrique,

Cotton is a great idea. That is the method RHA member (prehaps inactive now)and retired geneticist Dr. Leo Dionne developed in the late 1960’s. The “Dionne” method decapitates the main growing stem forcing the axils into growth. After a little bit (? a day) carefully take a razor and try to remove some of the leaf primordia over the growing point so the chemical seeps in better. Next place your cotton ball around the axil securing it on the other side of the stem and then saturate it with your solution. I used this method to double diploid potatoes, just as Dr. Dionne did.

Another application method is to put the doubling agent in lanolin and place that heavier mix onto the bud.

For seedlings and recovering the most polyploids, allow them to germinate and then place a drop of solution between the cotyledons. As the cotyledons are just opening there is a crack between them usurally the right size to hold a drop of solution. Perhaps you will need to transplant seedlings at the same growth stage and treat them together so they are uniform. I needed to do that for my study. After treatment I put a plastic baggie over the pot for a day to prevent the solution from drying prematurely. It is detrimental to soak the seed or apply it in another way where the root tips become stunted. The goal of most of us is to convert the shoot and then take cuttings or graft to preserve the polyploid. Stunting the diploid roots just causes more mortality and hinders doubled shoots from surviving.

The Basye’s article I have from 1990 which is in the American Rose Annual states 0.5%, not 5%n colchicine. From studies I have read no one reported going higher than 1%.

Good luck Enrique.

Sincerely,

David

P.S. I don’t know if you have a scale on hand, but if not perhaps a good way to do your mixing is to mix all of your powder with the appropriate amoung of solution to get your final percent. After it is in solution I usually feel better. Trying to weigh out a portion of the powder in the lab even with a dust mask makes me nervous.

opps… I meant .5%, if 500 milligrams of colchicine to 100 millimeters of water is correct… (If I recall correctly, Basye wrote “one half gram colchicine” and “100 cubic centimeters of water”)

Thanks David, you’re the ones whom I look up to as inspiration.