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 x7x_x7x
x7x, my problem child.
 
Registered: 05/05/07
Posts: 3,817
Loc: buenos aires 
Last seen: 7 days, 19 hours
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 Cannabis micropropagation project 2023  2
#28326663 - 05/19/23 06:43 PM (10 months, 22 days ago) |
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Namaste sisters and brothers, just started a series of experiments leading to get an entire cannabis plant from a little piece of tissue.
Here we go, let's start with roses.
day 1. agar media is tomatoe + amaranthus pigment
https://imgur.com/a/t0bbTIU
https://i.imgur.com/f01Gb0E.jpg
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cultivando en la miseria
SuctoSpore® Pictorial Tek
 
x7x_x7x@shroomery.org
carl_jung_in_lsd@yahoo.com
koh samui and oak ridge are my favourite strains
Edited by x7x_x7x (05/19/23 07:02 PM)
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 the_chosen_one
On the Darkslide
Registered: 09/11/06
Posts: 3,133
Loc: 1984
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 Re: Cannabis micropropagation project 2023 [Re: x7x_x7x]
#28327527 - 05/20/23 12:29 PM (10 months, 21 days ago) |
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How did you prepare and select your samples? They look very mature. Where I live the samples need to be taken from an invitro grown source and treated with Tween. Otherwise I just can't beat the Fusarium as it embeds deep into the plant tissue.
Is there a particular reason for starting with the Tomato agar? Are you introducing hormones at various stages or going for a one in all?
So many questions.  I've had some experience with this myself and would love to compare notes. Thanks for posting it as a focused topic. 
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"Luck favors the observant." - Workman
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x7x_x7x
x7x, my problem child.
Registered: 05/05/07
Posts: 3,817
Loc: buenos aires
Last seen: 7 days, 19 hours
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Re: Cannabis micropropagation project 2023 [Re: the_chosen_one] 1
#28328722 - 05/21/23 12:08 PM (10 months, 20 days ago) |
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Hi! Tomatoes have antimicrobial properties. I'm been using tomatoe agar for years for my mushroom cultures.
In this case, the red colour comes from an colouring food paste made whit amaranths extract, that is venom for the molds (and basidiomycete too  )
I have no experience in plant tissue grow. Never read about the maturity of the plant sample, it's a good point I been missing.
I suppose the whole tomatoe is plenty of vitamins, cytokinins, and all those things the plant needs. Let's see what happen. Is a cheap experiment. The only limitation is lack of space for storing jars
Let's see what happens with Fusarium, I treated the explants with tap water and 70% alcohol, then clean it in sterile water.
A very potent white LED light stimulate very well the growth of my Mary Janes, probably the explants are enjoying the light too
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cultivando en la miseria
SuctoSpore® Pictorial Tek
x7x_x7x@shroomery.org
carl_jung_in_lsd@yahoo.com
koh samui and oak ridge are my favourite strains
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x7x_x7x
x7x, my problem child.
Registered: 05/05/07
Posts: 3,817
Loc: buenos aires
Last seen: 7 days, 19 hours
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 Re: Cannabis micropropagation project 2023 [Re: x7x_x7x] 2
#28328885 - 05/21/23 02:22 PM (10 months, 20 days ago) |
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--------------------
cultivando en la miseria
SuctoSpore® Pictorial Tek
x7x_x7x@shroomery.org
carl_jung_in_lsd@yahoo.com
koh samui and oak ridge are my favourite strains
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greenladel
Registered: 05/27/20
Posts: 636
Last seen: 6 months, 29 days
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Re: Cannabis micropropagation project 2023 [Re: x7x_x7x]
#28329418 - 05/22/23 01:20 AM (10 months, 20 days ago) |
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watching this.
i have recently been thinking about getting started with tissue culture but haven't gotten around to it yet. for now i am taking things in from the back of the room.
thanks for sharing 
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x7x_x7x
x7x, my problem child.
Registered: 05/05/07
Posts: 3,817
Loc: buenos aires
Last seen: 7 days, 19 hours
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Re: Cannabis micropropagation project 2023 [Re: greenladel]
#28329605 - 05/22/23 07:13 AM (10 months, 19 days ago) |
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thanks! seeing the evidence, looks very easy.
yesterday I discarded one culture heavily contaminated with God knows what. Other shows some molds in one side, probably Fusarium as @the_chosen_one says; in the other side some growth is visible, let's see if are contams or tissue.
I find here a local vendor of vitamins, culture media and citikinins.
Quote:
greenladel said:
watching this.
i have recently been thinking about getting started with tissue culture but haven't gotten around to it yet. for now i am taking things in from the back of the room.
thanks for sharing
--------------------
cultivando en la miseria
SuctoSpore® Pictorial Tek
x7x_x7x@shroomery.org
carl_jung_in_lsd@yahoo.com
koh samui and oak ridge are my favourite strains
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x7x_x7x
x7x, my problem child.
Registered: 05/05/07
Posts: 3,817
Loc: buenos aires
Last seen: 7 days, 19 hours
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Re: Cannabis micropropagation project 2023 [Re: x7x_x7x]
#28334527 - 05/25/23 05:47 PM (10 months, 16 days ago) |
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Discarded all the plates except one showing root grow. It's a rose stem in water agar + red colouring paste.
Will post pics tomorrow.
--------------------
cultivando en la miseria
SuctoSpore® Pictorial Tek
x7x_x7x@shroomery.org
carl_jung_in_lsd@yahoo.com
koh samui and oak ridge are my favourite strains
|
x7x_x7x
x7x, my problem child.
Registered: 05/05/07
Posts: 3,817
Loc: buenos aires
Last seen: 7 days, 19 hours
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Re: Cannabis micropropagation project 2023 [Re: x7x_x7x]
#28334528 - 05/25/23 05:49 PM (10 months, 16 days ago) |
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I responded a WA from the local vendor of MS culture media asking for prices and the guy dissapeared.
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cultivando en la miseria
SuctoSpore® Pictorial Tek
x7x_x7x@shroomery.org
carl_jung_in_lsd@yahoo.com
koh samui and oak ridge are my favourite strains
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the_chosen_one
On the Darkslide
Registered: 09/11/06
Posts: 3,133
Loc: 1984
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Re: Cannabis micropropagation project 2023 [Re: x7x_x7x] 1
#28335699 - 05/26/23 02:48 PM (10 months, 15 days ago) |
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Here's where I get most of my supplies..
https://phytotechlab.com/
For my first attempt I went with mostly home made medium and naturally sourced hormones. I was able to achieve cell division but not much more before contamination set in. Eventually I broke down and began to purchase some of the stuff the pros use.
The Tween is a mild solvent that kills most invaders when used with alcohol and peroxide pre-treatments. Unfortunately, it hasn't done much against embedded Fusarium. BUT! I did figure out how to beat it (to be continued).
The best results I've had have been with the Murashige & Skoog based nutrients.
and an arsenal of hormones.
With these I was able to go beyond just cell division and make it to the rooting and branching stage before the dreaded fusarium set in.
I had one make it all the way to a final pot where it promptly died. 
Those trials were a great learning experience and the final failure really had me thinking about what it was that I was trying to accomplish. I believe there is some confusion in the terms. The Cannabis thread gang knows how anal I am about those.
Now I see this a couple of different ways based on what it is you want to do. "Micropropagation".. not to be confused with "Tissue Culture" although it's pretty easy to do. To me microprop is more like tiny cloning and has little to do with tissue storage or clean up from virus's etc. Microprop is more taking large node samples or exoplants and making it clone when normally it wouldn't. Where as TC is more from smaller tissue samples and making them storable for long periods of time. There's also the benefit of being able to clean up the culture much like we do with mushrooms. The ultimate dream is SynthSeeds.  I'm aiming more for those aspects more so than tiny cloning. Shipping and space are not big concerns for me, but long term storage of tissue would be a huge break through. Especially if one could bypass microprop with synthseed. *insert dreamy happy music here.
Continued.. so how to beat the Fusarium and most molds. Thinking of MP and TC as two different things for different purposes and getting picked on by Miss Botany Degree opened my eyes.. and it was so damn simple. I'm attempting TC. I don't need fully mature plants to sample from. As long as there is nodal regions it's all good. So how can I gain reasonably clean samples? The answer is usually written right before Tissue Culture.. Micropropagation. No body said one can't germinate a seed in a clean invitro environment. Kills me because we did this in grade school, but there's how the chain usually starts. If you want clean samples you have to grow clean samples. That rarely happens with cannabis in normal growing conditions. Invitro gives much more control and is typically pretty clean if not sterile initially.
My next attempt will be samples taken from an invitro grown donator.
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"Luck favors the observant." - Workman
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Muffin_Man
Sunshine Muffins
Registered: 05/24/23
Posts: 1,066
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Re: Cannabis micropropagation project 2023 [Re: x7x_x7x] 1
#28336327 - 05/26/23 11:16 PM (10 months, 15 days ago) |
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Wow this is some pretty amazing and good info. I had no idea about tomato agar and the amaranthus extract. Love how you said "venom" for the molds, that just cracked me up haha. Following this!
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x7x_x7x
x7x, my problem child.
Registered: 05/05/07
Posts: 3,817
Loc: buenos aires
Last seen: 7 days, 19 hours
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Re: Cannabis micropropagation project 2023 [Re: the_chosen_one]
#28337082 - 05/27/23 02:19 PM (10 months, 14 days ago) |
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nice! here in Argentina none of these supplies are available, at least until i know, thats why I',m trying with DIY media hahahah
great job!
--------------------
cultivando en la miseria
SuctoSpore® Pictorial Tek
x7x_x7x@shroomery.org
carl_jung_in_lsd@yahoo.com
koh samui and oak ridge are my favourite strains
|
x7x_x7x
x7x, my problem child.
Registered: 05/05/07
Posts: 3,817
Loc: buenos aires
Last seen: 7 days, 19 hours
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Re: Cannabis micropropagation project 2023 [Re: the_chosen_one]
#28337083 - 05/27/23 02:21 PM (10 months, 14 days ago) |
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one of my roses cuttings still clean and growing roots. let's see what happen with cannabis cuttings.
--------------------
cultivando en la miseria
SuctoSpore® Pictorial Tek
x7x_x7x@shroomery.org
carl_jung_in_lsd@yahoo.com
koh samui and oak ridge are my favourite strains
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