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Old 02-20-2013, 07:42 AM   #1 (permalink)
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Default TC experimentation

I'll post pictures today but I thought I'd make a thread about my TC experience as I go along with it. took some time to gather the materials but I finally got them all!
First things first, basic info regarding how to do TC
primarily I used the following sites: (http://kryptokoryne.aquaticscape.com...gation-part-2/......http://www.flytrapcare.com/phpBB3/ti...faq-t1426.html)

and Carol M. Stiff (owner of kck http://www.kitchenculturekit.com/index.htm)
was nice enough to help me with gathering my materials and also emailed me tons of info on the process and scientific articles of TC done on aquatic plants. If anyone needs those feel free to pm me, or i'll just upload all of them onto mediafire/dropbox later to make it more convenient.

List of materials gathered:
pressure cooker
magenta B caps (similar to phyto caps for baby food jars)
6oz glass baby food jars washed and with the sticker removed
IBA hormone
BAP-6 hormone
PPM (i have like 40 ml of this lol... bought 30 from ppm and carol gave me 10 ml)
MS basal salt mixture with vitamins
Agar (I have two types of agar, agar that carol gave me and agar from ebay (http://www.ebay.com/itm/18-grams-of-...item20ce3e9549))
new stainless steel pincettes also from carol
bunch of plastic pipettes

i think that's about it....
I plan on experimenting with some rotalas first and if those are successful I want to try my hand at propagating my anubias variegated and stardust.
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Old 02-20-2013, 09:00 AM   #2 (permalink)
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Default Re: TC experimentation

Good luck! I played around with P helferi in tissue culture in the lab at UF. Sterilizing the plant prior to culturing was the hardest part. I finally got 1 small piece to grow after several weeks of trying. After that, it was a piece of cake to subculture over and over.
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Old 02-20-2013, 10:14 AM   #3 (permalink)
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Default Re: TC experimentation

forgot to mention that instead of using a ph meter I plan to use beer making ph strips (http://www.ebay.com/sch/i.html?_odkw...strip&_sacat=0)
the usuable range is perfect for the range that TC requires.
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Old 02-20-2013, 10:52 AM   #4 (permalink)
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Default Re: TC experimentation

You should have said something, I have a bunch of stuff sitting here collecting dust. Used it once and then never again!
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Old 03-04-2013, 06:39 PM   #5 (permalink)
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Default Re: TC experimentation

shoot i got too busy but anyhow, here are the pictures of the materials
here are the baby food jars (6oz) with the regular lids and the magenta b caps (same thing as the phyto caps)

as you can see... the caps are actually like push on. At first i thought they screwed on but i guess not, it slides on very snugly and feels legit.

beer ph testing strips. these were cheap, 8 dollars shipped from ebay and since i need a ph of 5.7 this is perfect because the color differences are VERY obvious and it's a narrow range

vinegar, 70% rubbing alcohol, new SS forceps from carol, and graduated plastic pipettes for the hormones

here's laboratory grade agar i bought from ebay but apparently it's much more "professional" than what's needed for TC :P it was pretty cheap so it doesn't really matter. the purpose of agar is to thicken the MS solution and keep the explant from sinking. People are reported using coffee filter paper to be successful as well since it still delivers the medium to the explant and surports it.

plant hormones and the MS+vitamins, PPM, and agar from Carol
I bought 30 ml of agar from the company that makes ppm (plant preservative mixture) and it was 1 dollar/ml. very expensive x_x so as a note to those who want to try TC, just buy it from Carol on kitchen culture kits. she rebottles them into smaller 10 ml leak proof bottles as pictured. The hormone concentrations are 1 g/L and are likewise bottled in leak proof bottles. The hormones and MS and ppm and Agar MUST BE refrigerated.







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Old 03-05-2013, 09:18 AM   #6 (permalink)
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Default Re: TC experimentation

Wow nice work, I will like to see all the process. Keep up the good work.
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Old 03-05-2013, 09:37 AM   #7 (permalink)
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Default Re: TC experimentation

What do you plant to use for a hood? There's a special clear tape you can wrap tight around the lid to make an even better seal. I forget what it's called though.

I know Ghazanfar was successful with Lagenandra meeboldii 'Pink' and the Crypt nurii that everyone has now. He mentioned it took about 10 plants to get one clean explant. As mentioned above it seems the hardest part is getting the plants sterile to begin with, which is why I think it's wise to start with stems because you don't have to use a part of the plant that is near the soil, such as with Crypts or other rosette plants.

Heck, if you can get ahold of some of the ones already in production, i.e. Tropica 1-2-Grow you could start by simply splitting one of those out as it's already sterile.
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Old 03-05-2013, 07:28 PM   #8 (permalink)
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Default Re: TC experimentation

yeah too bad most micro propagation dealing with aquatic plants are not in the US. e.g. anubias, aquaflora, and tropica. I've been talking to some people who've done it and they all said it was very very hard. I'll continue to read the documents Carol Stiff sent me and hopefully they will help. anyone interested in those?
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Old 03-06-2013, 12:13 PM   #9 (permalink)
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Default Re: TC experimentation

The sterilization part is all that's holding me back from trying to TC my more unusual plants. I'm not much one for having sterile lab conditions... keeping a few germs around builds immunity!

Still, I'd really like to try it out with some of my Buces or something similar one day... I don't much see the point of using tissue culture to propagate stem plants, since they can be grown quickly and effectively using normal propagation techniques. IMO, it's only worth it for slower-growing, more expensive plants.

Anyway, let us know how things go, Neil! Literature only tells you so much, it's much cooler to see these things in action with photos and a firsthand account.
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Old 03-06-2013, 01:50 PM   #10 (permalink)
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Default Re: TC experimentation

Quote:
Originally Posted by asukawashere View Post
I don't much see the point of using tissue culture to propagate stem plants, since they can be grown quickly and effectively using normal propagation techniques. IMO, it's only worth it for slower-growing, more expensive plants.
I think the idea is to practice with something easier to sterilize. If done properly you can still propagate thousands of stems over the course of a year. That's a lot more than I can grow conventionally.
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