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Darklight

Micropropagation of Lophophora spp- media

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Disclaimer-

Actions in this post should only be undertaken in places where the cultivation of Lophophora is legal

Clonal propagation of Lophophora spp is slow and not a viable way to produce mescaline. And for those unfamiliar with aseptic technique, it's not highly productive.

If it's mescaline bearing cactus you're specifically after, in a place where cactus is legal for the purposes of consumption, grow them from cuttings- the tek below will not help you or make it faster

However Loph cultivation in TC is an ideal way to learn about tissue culture. Seed is easy to obtain, easy to sterilise, the physical cactus, once sterile, is easy to work with and will greatly enhance your understanding of both aseptic culture techniques and the responses of cactus to hormones in-vitro

This media can be useful in conservation work where individual populations and plants are identified, kept separate and used for breeding purposes to re-establish diversity

I've used and keep Lophophora williamsii in culture for teaching purposes and it's response and ease of handling make it remarkably successful for these purposes

Proliferation media derived from the paper ( it's in Spanish, I can't remember where I got it from )

Propagacion in vitro de Peyote ( Lophophora williamsii ( Lemaire)Coulter ) JG Ortiz-Montiel y Alcantara Garcia R

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Lophophora proliferation by aseptic axillary culture:

Aseptically sterilise fresh Lophophora seed- 30 seconds in 1.5% bleach ( with no added perfumes etc )- rinse twice with sterile water and allow to dry on sterile filter paper- separate seeds so they can dry, and not cross contaminate.

Seed will germinate on MS media with vitamins under coolwhite at 16/8 light cycle at 24C. Better light is a single grolux tube at about 30cm distance- it seems to reduce red phenolic leaching. You can do this in a closed petri dish and dibble out cactus seedlings into individual tubes as they arise

Subculture to more freshly made sterile media about once every month or two. Larger plants have a better response than smaller ones to the following media:

To make 1L proliferation media

4.33g MS ( no vitamins )

100mg inositol

1mg Thiamine.HCl

1mg Kinetin

0.1mg NAA

30g sugar

8-10 g agar or 7g gelcarin ( pref, but does footle the pH around a lot more )

pH 5.8

Sterilise. Subculture as single or double axillary bud explants, using larger size cuttings ( ie don't excise too close to the ax bud, leave a bit around it )

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The above does not guarantee success, different populations and even individuals within the same population can have *very* different responses to the same media- a given for all plants in TC

However if you post your results or want to discuss problems or questions, the above process is an excellent place to start. There are a fair number of things I've left our, assuming some familiarity with the technology.

Happiest to answer questions here from ppl who have given it a go, or are seriously looking at undertaking learning the process, and who are prepared to share in this thread

Edited for spelling. The re-edited because it wasn't wrong in the first place

Edited by Darklight
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( My sudden post dearth is strongly linked to this godawful germination trial I'm running which is boring, labour intensive and has kept me chained to the lab bench all weekend where I have to be here to monitor conditions every hour. And it's not working. So much for returning to first principles. Someone get me out of here )

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Hello darklight I find this rather interesting! Was wondering if you can recommend a aussie supplier for tissue culture supplies MS media/vitamins hormones etc, this is kinda new to me..that is if your in oz! Thanks!

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awesome, would love to see pictures of such a process- where legal of course

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Hello darklight I find this rather interesting! Was wondering if you can recommend a aussie supplier for tissue culture supplies MS media/vitamins hormones etc, this is kinda new to me..that is if your in oz! Thanks!

Austratec in Melbourne

http://www.austratec.com.au/

Check their catalogue ( remembering to be aware that MS basal media and MS basal salts are two different products- one has vitamins, one doesn't )

You can buy the preprepared media in sachet lots to prepare 1L per sachet, and the hormones in 1mg/ml solutions which stay good for a minimum of 3 months if properly stored. Maybe buy a full suite of stuff and share the costs with a few people

It doesn't make any difference to them if you're just starting out and willing to learn, many businesses are happy to help a beginner as it's a good way to encourage long term clients- so let them know you're a beginner if you're comfortable telling them that. They're pretty easy to deal with, just remember to stay professional.

If you let me know what you're looking at starting with in terms of media and culture type I can help you put together a shopping list ( I don't need to know the species, that's your business ) ie. starting from seed for a species with a media protocol, looking to proliferate whole plants by axillary cutting

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Thanks darklight....I came across that catalogue afew weeks back the prices seemed abit daunting at first...but as ive begun to research this it is becoming more affordable, upfront starting costs are always a dilemma when first taking on any new hobbie, will prob send a PM in afew days with a few dumbass questions and seeking a little guidance on a shopping list when i wrap my head around this a little more..cheers!

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Happy to help, would prefer to answer questions here if possible- there are very few stupid questions at this point :) If you do it here you'll be helping other people learn, and teach me to explain stuff better

It can be a bit like learning to knit via youtube otherwise

Do you have basic myco technique? Do you play a musical instrument at all? I find people who do are amongst the easiest to teach- because they have a good idea as to where their hands are in space, and they're used to trying stuff for a while before they get it spot on

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Yes have myco experience, currently growing some oysters and ive been reading various literature on tissue culture for quite some time, purchased a copy plants from test tubes, managed to read half of it and been meaning to pick it up again for the last couple of months, always distractions about and my wandering mind doesn't help!.....

Just been trying to facilitate the space to attempt plants invitro and juggle the time to do so and tend to them(this is the hardest part so damn busy!) & yes i have been known to strum some guitar when i have some lax out beer time! & got a heap of loph seed that will be prime candidates for this kinda experimentation! :)

Edited by applesnail

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Excellent :)

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Bump

Did anyone give this a try?

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