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[chums of chance]

I was inspired to look into this topic after reading this thread.



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.?

4 Likes Mandrake, New Wisdom, Sunshine and Greentoe like this.

[Sunshine]

Wow, those look awesome. Big +1 for sharing this. Soilless propagation = epic. I love how you can see the entire root structure.

[chums of chance]

I decided on:
LINSMAIER & SKOOG BASAL MEDIUM W/ 30g/L SUCROSE & 7g/L AGAR, pH ADJUSTED & BUFFERED
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. 

I will use specimens from my in vitro thread for explants.

1 Like Sunshine likes this.

[nobody]

Are you planning to use any antifungal/antibacterial add ons in your media?

[chums of chance]

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.

[Greentoe]

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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[chance1122]

A-mazing! Such beautiful root structures. What was the medium formula?

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[oriky]

Amazing, i very interested on grow on agar plants, never thing on do it with cactus!

Good luck, waiting for result

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