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Post by coldcoffee on Jul 12, 2011 13:21:21 GMT
Well, I started my little sphagnum experiment. I'll let you know how it goes. Used a very easy going disinfection so we will see who wins: the moss or the nasties
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Post by coldcoffee on Jul 13, 2011 5:49:34 GMT
If anyone is still following, The Sphagnum in the TC appears to have survived the disinfection protocol (barely...). I am still seeing Chlorophyll (I did mention that it was very a light disinfection protocol). Its too early to say whether or not a contamination exists (but I anticipate there may be, but I do not see evidence yet).
P.S. I prepared a photo, but it seems the upload link has disappeared....
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Post by bluemax on Jul 13, 2011 6:04:08 GMT
I'm certainly following, coldcoffee. Tissue culture is not something I have yet tried but I'm very interested. I'm not in a position to start right now but I can live vicariously. Good luck in your venture! - Mark
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Post by coldcoffee on Jul 13, 2011 7:04:56 GMT
Thanks, I will post pictures when I can. A bunch of my explants are really starting to show some good activity right now. I have an Dionaea explant which is covered in calluses (FYI, explant is just the term used for the portion of the plant that you use to start the culture, in the case of Dionaea I took a clipping of a young leaf). It kind of fun throwing a small leaf cutting into a jar of jello and watching tiny venus flytraps grow on top of it! The Drosera binata culture is particularly dramatic right now.
I'll post pix soon!
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Post by coldcoffee on Jul 15, 2011 5:55:30 GMT
Ok well attempt number 1 failed- contaminated like crazy now!!!
Looks like I need to titrate the disinfection protocol up a bit.... more news later....
On a positive note, I should know in the next week or two whether my stage 1 Cephalotus protocol works or not....
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Post by ahicks51 on Jul 30, 2011 17:25:56 GMT
Try starting spores on water agar, with or without antibiotics or antimicrobials.
We get moss as a contaminant in tissue culture now and again- the risk of growing in a lab associated with a greenhouse, I suppose.
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Post by coldcoffee on Jul 31, 2011 18:12:00 GMT
The question becomes how to get sterile spores. I had thought about disinfecting the spore heads (for lack of a better phrase) and then dispersing the contents onto agar.
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Post by peterhewitt on Aug 1, 2011 10:26:24 GMT
We often Initiate Nepenthes and Disa directly from unopened seed pods. When doing this, we just surface sterilize the pod, and then dispense the contents on to Agar. I'm thinking that the Sphagnum Spore capsule, might be a little too sensistive though. Would be an intereting Protocol, if you can get it to work.
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Post by coldcoffee on Aug 3, 2011 6:28:46 GMT
I had some ideas going, I just need to sit down as try again. I started a new job in an IT department and have been working long hours, so I have not really had the time to dedicate to TCing. I have a bunch of cultures that need to be subdivided. Maybe I will cogitate on this sometime this weekend and try again. Part of me thinks that TCing Sphagnum is sort of silly and more trouble than it is worth- part of me thinks the value lies in the challenge, and part of me suspects that it could prove to be an answer to the question of effectively propagating Sphagnum in a very sustainable way. Now, if only we could figure out how to TC Soylent Green, we would all be in business
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coline
Full Member
Life's essence: patience
Posts: 484
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Post by coline on Jan 3, 2013 1:14:22 GMT
Coldcoffee how did the cultures continue?
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Post by micropropagador on Jan 7, 2013 0:17:05 GMT
Hi there. I acidentally put Sphagnum in TC cuz I thought it was an aquatic plant that I received. To avoid contamination I removed all leaves and It remained just the stalk. After several days there were no contamination but no calus nor bud had been formed, but the stalk remains green. I used 30% bleach for 10´ and put the stalks in half strenght MS. Cheers Cleber
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