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I must inform you with joy, that succeeded me to multiply an A. titanum from a leaf segment. He has still not roots, but after six weeks, he looks already quite promising.

Greeting
Andreas

Na clear let I you partake.

The idea came to me when I heard attempt from coal Creek. The report can be found in ACTA horticulturae, issue 226 of 1988, if the want to read someone. To my knowledge, this was the first successful reproduction of A. titanum in nutrient medium via leaf cuttings. I took his attempts but only as a guide and choose another medium.

This was my first attempt, so not everything went off well so perfectly.

A MS served as medium medium with 1.0 BAP and agar.

All Explanate come from my A. sown in October titanum. 15 I have taken from the first sheet, and 7 of the second sheet. For that I used the first two segments and used a segment of the second. The leaves were disinfected in alcohol and then in sodium hypochlorite. Then, I have used only the Middle rib with approx 1 cm sheet on the pages.

The sterilization has worked well. Of the 22 Explanaten, then only two were contaminated. In the Kohlenbach attempt only ca 50 uncontaminated remained of ca 200 Explanaten, so I am quite satisfied with my quota.

However, apparently my glasses were not clean enough, so then again seven Explanate were contaminated. I suspect it was because that I have filled the medium in clean but unsterilized jars and then sterilized by I have heated it in the microwave and cooked for 2 minutes. Apparently, mould spores survived it on the glass.

The difference between the first two was that the contamination on the border between glass and medium occurred here, and not from the Explanat. Next time I'll sterilize so even the individual jars before filling a medium and then again together with the medium.

The Setup, I can say that I a a closed room have worked. First, I have sprayed water with a sprayer in the entire room to bind as much dust. I have washed off the table and all instruments with which I have worked, with alcohol. As "clean box" I've just taken an IKEA SAMLA box that I put on the head and cut a hole to work in the page.

On the subject of rooting, I'm not sure yet. I think just darrüber, if I share my Explanat and put them on to the subcultures, and then these bewurzele. I read me books, just again through the issue of hormones in various Micropropagagation to find out what hormone combination I use sensibly for the subculture. You must grow in step one, step two make a shoot, and in step three to slips.

By the way, it looks that the sheet may be not too old. The Explanate of first leaf, which was about 5 months old at the beginning of the trial, wither all on one. The Explanate of younger leaf look like velversprechender.

Definitely an exciting topic. I'll make soon attempts with my other types. When I'm done, I'll describe the Setup again more detail so that others can do it.

What was because your problem that led to less successful? And where did you buy your hormones and other?

I have taken two scoops of the agar, I mean the American were "pinch" per glass. Theoretically, the 45 ml jars were filled. Since I had no suitable measuring cup but, I have filled that hold roughly the same after eye. For some, the agar was also very hard for the leaves. Before I start a new attempt, I'll buy first a suitable scoop me.

The sodium hypochlorite I have a canister with a I believe 13% solution bought, which I then 1 / 10 used diluted. How long they were in there I can not tell grad, since I'm in the Office, and my records are at home. But I think it was about 5 minutes.

In the Explanaten I chose ca 2-3 cm in length, where I have no longer used the blade tip because that was too soft for the agar.

I think you must have done already 3-4 times the whole thing to get a feel for. That is why I will try me now times across through my collection. As in recent weeks, many are driven out, I have enough young leaves to test. If my titanum then drives his third hand, I learned then hopefully enough to get more than a successful Explanat.



Has anyone here possibly else experiences with Micropropagation?


Andreas

First of all your questions, BAP, and NAA are plant hormones or growth regulators. You can reach through the addition of one or more hormones, that such callus and roots are formed by the Explanat. What happens depends on hormones.

BAP - 6-Benzylaminopurine

NAA - 1-Naphthylessigsäure

The MS medium containing salts. It is a fertilizer solution. See here: http://de.wikipedia.org/wiki/Murashige-Skoog-Medium

Experience shows that the microwave sterilization is usually (not always) sufficient. I would mention as Carol Stiff, who practiced in the American region for years tissue culture for domestic use and teaches.

I have a ca-three-month-old Explanat which shows so far no traces of contamination. It is useful here sure that is set to the medium PPM which contamination prevents. See here http://www.plantcelltechnology.com/about-ppm/

If you want to put you closer to deal with the subject, I recommend the following sites:


http://tech.groups.Yahoo.com/group/hometissueculture/
To my knowledge the biggest community, which is engaged in tissue culture.

http://www.kitchenculturekit.com/index.htm
The page of Carol. Here, you can buy the accessories.

http://www.Amazon.de/plants-test-tubes-I...from+test+tubes
The book of plants from test tubes. A comprehensive guide and introduction to the topic.

Greeting
Andreas