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Post by DroseraBug on Jan 15, 2012 2:45:10 GMT
I've been sterilizing Sarracenia seed and germinating in-vitro for a while now. I'm looking for any tips on media prep for Sarracenia. Currenty I'm using 1/2 MS just because I prefer that with vfts, sundews, and pings. Sarracenia seedling growing on 1/2 MS with no cytokinins or auxins  Another seedling growing on both cytokinins and auxins. I like the way these are growing a little better. They are so much slower than the vfts. But I'm definitely going to use auxins from now on. Auxin does seem to initiate some roots, shoots, and size. I'm looking for any tips on Sarracenia micropropagation. I'm using 1/2 MS for everything. I'm thinking I should reduce to 1/3 MS for Sarracenia only. Both of the above have been replated twice. I placed the last of the two on both hormones because I was interested in multiplication and root/shoot growth. With vfts cytokinins slow growth an really aren't even needed unless you want large quantities. I would assume cytokinins would also slow growth with Sarracenia however help to initiate callus or multiplication. I've done a good bit of research and found very little on this topic. Notice the very bulbous growth on the root apical meristems on the last photo compared to the first photo. The hybrid in the top photo was replated a week prior to the hybrid in the second photo. I'm thinking if I remove the cytokinins and use auxins only and 1/2 or 1/3 MS it may help with faster growth. Any advice?  |
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Post by DroseraBug on Mar 29, 2012 21:45:02 GMT
Quick update. Was able to initiate callus on multiplication media using different ratios of cytokinins and auxins. Interested to hear from others regarding your experiences with Sarracenia tissue culture from seed. More specifically, any advice on using ppm to sterilize Sarracenia seed would be great! Also, it seems like some seeds from specific hybrids or species are easier to sterilize than others. Maybe its just more difficult to sterilize larger seeds or it seems that way from observations (larger surface area). I've read several threads of great success using ppm but no specifics regarding concentrations and preparation.      |
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Post by ajmiller on Mar 30, 2012 5:11:38 GMT
Amazing results, please keep posting as you progress.
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Post by DroseraBug on Jul 12, 2012 21:37:58 GMT
Amazing results, please keep posting as you progress. Thanks. A quick and successful update so far  |
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Post by paulbarden on Jul 12, 2012 22:15:26 GMT
Perhaps I'm missing an important detail, but I can't help wonder why you'd want to go to so much trouble to culture seeds of a genus that are among the easiest seeds on the planet to germinate? nitrogenseekers.wordpress.com |
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Post by peterhewitt on Jul 14, 2012 15:05:28 GMT
Because you can speed up the growth by a factor of ten when you get the right Ratio's of Hormones, and you can multiply a specific individual Clone as much as you like in a short space of time. Relatively. And it is fun...
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coline
Full Member
Life's essence: patience
Posts: 484 
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Post by coline on Jul 30, 2013 2:44:35 GMT
Just wondered, which were your dissinfection protocols for seeds? If anyone else also has had good results with a method, I'll really appreciate to be taught
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Post by DroseraBug on Aug 16, 2013 2:17:11 GMT
Just wondered, which were your dissinfection protocols for seeds? If anyone else also has had good results with a method, I'll really appreciate to be taught I've had success treating with giberrellins in food jar lids for 24+ hours, then swirling in 10% bleach for 8 min followed by 6 min with hydrogen peroxide. Rinse with sterile DI at least twice and then transfer to sterile media 1/3 to 1/2 MS without hormones. Replate to media with auxins and meta-topolin/or cytokinins. Sterilizing Sarracenia seed is more difficult than other carnivorous plant seed due to scabrous nature of seed. More glabrous seeds such as vft seed is easier. Some use ppm as the sterilization method and I would like to try myself if anyone could link me to a good thread regarding methods used. I'm interested in anyone with experience sterilizing Sarracenia from actual plant tissue. I've been researching this for a while with little to no results finding literature on this process. I've attempted the process from the apical meristems and flower stalks, both ended in fungal/bacterial contamination with many repetitions. Rhizomes are my next step and feel that either method may require some sort of stripping away epidermal tissue before sterilizing. Anyone with advice on the best methods of sterilizing Sarracenia tissue from adult plants? I feel this info could benefit Sarracenia conservation, kitchen culture hobbyists, and small cp nurseries. I've researched this a good bit with minimal results Someone suggested contacting Big Bella regarding the mature plant tissue sterilization. Any advice, anyone? Thanks. |
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coline
Full Member
Life's essence: patience
Posts: 484
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Post by coline on Aug 16, 2013 5:50:39 GMT
Well, some time has passed and I did have a success with the sterilation I made to seeds, even though, I stratified them in a normal way in a freezer first, as at that time I did not had AG3. They have 2 weeks in vitro now. 0% contamination What I had on hand at that moment was a 8%ppm 1/3MS solution, which I used for 3 hours on the seeds, as because that was the time I had available to make it in 1 night I was working.  So, you say, using AG3 in a soak for 1 day works to break dormancy? In what concentration would that be? Regarding to actual adult tissue. I have not done it, but I have succeeded in nepenthes tissue, use the protocol Medina used here in the forum, just adding PPM and had a 60-70% success with 6 explants. When I have some time, maybe in vacation or next semester I will try the sarracenia, with daina's delight that is my most common hybrid cultivar. |
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