Share The Seeds
Gardening Area => Advanced Cultivation Techniques => Topic started by: chums of chance on March 26, 2014, 12:37:22 PM
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I was inspired to look into this topic after reading this thread (http://www.shroomery.org/forums/showflat.php/Number/1932583/fpart/all/vc/1).
(http://i.imgur.com/W0NrIyql.jpg)
(http://i.imgur.com/uJAUz92l.jpg)
Unfortunately, the original poster was glib about his methods. I'm going to attempt the micropropagation of L. Williamsii. At this point I am still in the research phase of figuring out what my exact recipe will be.
However, I have a few ideas. I am going to use Linsmaeier & Skoog medium. Cacti, being desert plants, have cells with a low water potential and should be cultured in media with negative osmotic potentials. So, thoughts, suggestions, critiques, links etc.?
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Wow, those look awesome. Big +1 for sharing this. Soilless propagation = epic. I love how you can see the entire root structure. :)
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I decided on:
LINSMAIER & SKOOG BASAL MEDIUM W/ 30g/L SUCROSE & 7g/L AGAR, pH ADJUSTED & BUFFERED (http://anonym.to/?http://www.phytotechlab.com/detail.aspx?ID=406)
It takes care of just about everything, except for
cells with a low water potential [which] should be cultured in media with negative osmotic potentials.
This will be remedied by increasing the sucrose concentration to 60g/L. 8)
I will use specimens from my in vitro thread (http://sharetheseeds.me/forum/index.php?topic=1229.0) for explants.
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Are you planning to use any antifungal/antibacterial add ons in your media?
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I have been thinking about the contamination issue, but, based on my research, additives to fight unwelcome organisms are not usually added to tissue culture media without cause. I don't know if that's because such additives are incompatible with healthy growth or because the researchers whose work I've read don't want anything unnecessary to compromise the media.
In the past, I have successfully avoided contamination while growing fungi, so my hope is that the aseptic procedures I used in that project are adequate for my purposes here.
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If you're used to working with agar you're probably good. If not I'd probably look into adding something to control contamination. When I've tried micropropagation in the past I added some antifungal/antibacterial stuff and used a glove box and still ended up with a good deal of contamination. In my experience its been much easier to innoculate grain jars or spawn mycelium to a bulk substrate without contamination, than it has been to work with agar with out it getting contaminated. If you don't have a flow hood (I really need to make myself one of those things) you'd probably be best off to add something to help prevent contamination.
I wish you good luck with this and look forward to seeing how it goes.
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A-mazing! Such beautiful root structures. What was the medium formula?
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Amazing, i very interested on grow on agar plants, never thing on do it with cactus!
Good luck, waiting for result