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Mushrooms, Mycology and Psychedelics > The Ethnobotanical Garden
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Re: Experiments with plant tissue culture [Re: smaerd]
    #10857792 -

Hey sushi and smaerd, thank you for such a good info!
This is my first attempt with cactus. The media i´ve used for germinating seeds was 1/2 MS, with no regulators and 30 g/L sucrose, and pH 6 before sterilization. If they germinate, i´ll try different recipes for elongate them. I would like to try your recipe, but it´s very difficult to find here (Spain) where to buy regulators. I bought kinetin and IAA in EEUU and the delivery took 3 months!:thumbdown: But i´m interested in BAP, it seems to be much stronger than kinetin and works well with lots of species. I also have a bit of 2,4-D. Very useful for callus induction.

About the sterilization of seeds,that´s what i did:

- 15min in water with 15% bleach
- three consecutive washes in sterile distilled water

I hope seeds resist and germinate, we´ll see..

But now i´m thinking about new recipes, what do you think about this one?

- 1/3 MS
- 5 mg/L kinetin
- 0,5 mg/L IAA
- 30 g/L sucrose/dextrose

Does PPM really avoid you to do sterile work or it´s just vendors advertising??


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Plant tissue culture
Crossbreeding experiments

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Re: Experiments with plant tissue culture [Re: troncotron]
    #10858370 -

Looks very interesting!:cool:

can someone explain to me how and why to do this culture?

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Re: Experiments with plant tissue culture [Re: troncotron]
    #10870317 -

Troncotron: Do you use a surfactant with your 15% bleach solution? That recipe looks good be sure to let me know if it works out! In regards to PPM it does really work wonders, I still follow all the normal sterile procedures but I get next to zero contamination with PPM, when working with valuable/rare plants this is important!

PsyAlien: I can't really explain to you how to do plant tissue culture, without writing you a book anyways! I suggest you do some googling + pick up a book called 'Plants from Test Tubes' it's a great resource. People do plant tissue culture for a ton of reasons, it's a great way to rapidly multiply plants, grow plants that wouldn't exist naturally [without chlorophyll perhaps], keep stock of important plants [For example if I have a plant I want to hold onto I can keep it in culture in a fridge for months and months without any sort of upkeep, cultures are completely self-contained so its a great way to hold onto valuable plants without much space], I'm particularly interested in secondary metabolite production in vitro - there is the potential to add biochemical precursors to your media, really the possibilities are endless!

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Re: Experiments with plant tissue culture [Re: sushi]
    #10874375 -

The book, or about half of it...

Plants from Test Tubes

Edited by Tom A Toes (08/17/09 06:31 AM)

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Re: Experiments with plant tissue culture [Re: Tom A Toes]
    #10880818 -

These are both very good, more detailed than "Plants from test tubes" (and more expensive too):

Introduction to plant tissue culture

Plant tissue culture concepts and laboratory exercises


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Plant tissue culture
Crossbreeding experiments

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Re: Experiments with plant tissue culture [Re: troncotron]
    #10883810 -

ooo man.. i'm too lazy to read books...

i want to try something simple for a start in my Dads kitchen, of course i don't have any laboratory equipments so i'll try to improvise.
the recipes you talking about are specific for some plant or it can be done with any plants?

p.s. the plant i plan to play with is mj.

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Re: Experiments with plant tissue culture [Re: PsyAlien]
    #10883927 -

Umm... no disrespect, but if you're not willing to read any books, and are going to be working in your dad's kitchen with improvised equipment... then expect to fail massively at tissue culture.  It's not exactly an easy thing to do. 

You'd be better off cloning from cuttings.  That's much much easier than tissue culture, and a lot more practical for someone who's beginning.  There's really no advantage to using TC unless you want to turn one plant into thousands within a single generation.

Beyond that, I wouldn't really recommend growing MJ to anyone who isn't willing to put a lot of time, effort, and research into it.  Unless you do it big, it's not worth the risk.


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The Plant Mage Guild

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Re: Experiments with plant tissue culture [Re: Horticulture]
    #10884512 -

^^I'd have been less polite but yea, what he said.


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Friendships have come and gone but I always regret the girls I didn't bang




I just dont really care what you think

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Re: Experiments with plant tissue culture [Re: Horticulture]
    #10891357 -

i think i will fail massively if i ain't gonna get some guidance to make this culture.

even if i will read and follow by the book i still will need someone to direct me to a proper materials or where to find them, oo yes materials... my primary problem:mad:!

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Re: Experiments with plant tissue culture [Re: PsyAlien]
    #10891421 -

If you don't like books and you want an introduction/overview to TC, my signature has a link to youtube videos about tissue culture... with sources for materials. Personally though, I feel like if you're not willing to buy at least a $20 book on the subject, you won't be dedicated enough to grasp it.

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Re: Experiments with plant tissue culture [Re: troncotron]
    #10895490 -

Very impressive work, Sushi! Quite possibly very valuable information too. Very nice thread!

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Re: Experiments with plant tissue culture [Re: Knoa6]
    #10899850 -

I thought I'd share some more photos, three more peyote cultures and two San Pedro cultures. I'm also happy to report that Epithelantha micromeris responds well to *my* peyote media [although I don't have pictures of this right now]:



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Re: Experiments with plant tissue culture [Re: sushi]
    #10899947 -

Looking awesome!

TC's never cease to amaze me!


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Re: Experiments with plant tissue culture [Re: sushi]
    #11476633 -

I am bumping this thread, despite its age, because the projects described will have had a couple of months to develop and I'm probably not alone in being curious how they've turned out.

Sushi and Troncotron: amazing work! My question to both of you, regarding both Troncotron's cannabis work, and Sushi's work with peyote, have you managed to achieve root formation? In all the pictures posted above, I don't see any roots... have either of you moved your plants to soil? Also, Troncotron, did those seeds ever germinate?

Thanks for posting the recipes! I don't have the science background of you guys, so I would be lost without a recipe to follow. One thing that I wonder about is the reports I see on the net about switching the media at different stages of plant development to trigger the next phase (which is partly why I ask about roots; here you can see different chemicals used for the cannabis root generation, if you read the text for figure four). Also, in terms of the cactus, some of Sushi's amazing pictures (although an indication of scale would be awesome) show a lot of rapid growth in strange forms, with some very promising-looking bulges on them. Do you then cut those nice growths off the mass and graft them to root stock, or do you place them in another medium to produce roots? Or do roots just eventually form?

I hope it's not too obvious that I haven't read any of those books listed above; maybe these are really simplistic questions. But I am fascinated by the work you're doing, and had to ask.

Best of luck! I'm about to place an order with a TC supply house, and want to work with Salvia d., Lophophora w., and Peruvian torch (can't recall the latin), and possibly Cannabis as well.

In terms working with Salvia, I'd be curious to know if any of you have succeeded in this area, and what medium you used. On the internet, I found one scientific paper concerning Salvia divinorum which contained the following passage: "...Then placed in sterile glass culture tubes containing Murashige and Skoog medium supplemented with 2% glucose, Gamborg’s Vitamin solution, 0.2% Phytagel, and 5 μM 6-benzylaminopurine

Keep up the good work! Thanks in advance for any update you're able to provide.

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Re: Experiments with plant tissue culture [Re: Puma]
    #11477066 -

Im gonna start playing with salvia TC in the new year!!! where did you get the above info?


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Friendships have come and gone but I always regret the girls I didn't bang




I just dont really care what you think

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Re: Experiments with plant tissue culture [Re: Fleadh]
    #11486473 -

Fleadh said:
Im gonna start playing with salvia TC in the new year!!! where did you get the above info?


Hey Fleadh, I'm looking to start playing with that too, we should compare notes. Here's the link to that info and further discussion (there's mention of a filter method of extraction of the salvinorin compound. {{{link}}}

Good luck!


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Re: Experiments with plant tissue culture [Re: Puma]
    #11501521 -

Hey Puma sorry it took me so long to reply to this but I've been pretty busy! Firstly, root formation can be achieved by moving the plants to either media without hormones or media with auxins (depending on the plant). I haven't actually tried to do this with my peyote cultures yet because I'm just working with callous cultures at the moment but I have had success in grafting buds that have appeared spontaneously out of the callous tissue in vitro to pereskopsis and then degrafting after awhile and moving to soil. Everything has been going well so far, I've subdivided my peyote cultures twice since my last post and I have way more plant mass! I've also recently had success growing Epithelantha micromeris callous in vitro using the same media as I use for my peyote (although there is still some annoying vitrification). I'll try to get some pictures and post them here soon for you with an indication of scale. The next step will be trying to induce the callous tissue to form buds and then roots/grafting. As for salvia I have had some success but I wasn't able to produce callous tissue, or regenerate plants from anything other then meristem cuttings, in the end I gave up since Salvia is so easy to propagate normally!

Also, here's a link to the actual (and much longer) pdf of the article you referenced:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2065853/pdf/nihms-27595.pdf

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Re: Experiments with plant tissue culture [Re: sushi]
    #11515314 -

These are the best photos I could get for now!

Here are some peyote callous cultures with a Canadian quarter for scale:



Epithelantha micromeris callous:


Rooting a small Pedro:


Some better photos of Epithelantha micromeris, peyote being moved from culture to soil, and a very well established plant in soil from culture:



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Re: Experiments with plant tissue culture [Re: sushi]
    #11515639 -

:congrats:


Great work sushi!


It's been years since I done any of this but your post does tempt me to try again.


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My Ethnobotanical Growing TeksWhere to Start Flow Chart, QnA, and helpful links

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Re: Experiments with plant tissue culture [Re: karode13]
    #11516741 -

WOW!

I am in awe.

Thank you sushi, from one loph-loving Canadian to another. Fascinating, insightful, and incredibly well-documented. I'll let you know down the road what sort of results I can attain with your recipe. Thank you so much for this great contribution!


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Mushrooms, Mycology and Psychedelics > The Ethnobotanical Garden
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