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Cannabis tissue culture

Gray Wolf

A Posse ad Esse. From Possibility to realization.
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Here are some purdy ladies grown from meristematic cells in a special medium by our inhouse red headed geek, instead of seeds or clones. The meristematic cells are extracted form the stem nodes anywhere on the mother plant.

One of the advantages is that no diseases or systemic parasites are passed on.

Hee, hee, hee, snicker, snark, snort.....................
 

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PistilPete

Enjoying the ride
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Excellent. Never seen it done before with mj. Good luck with the trials. I'll be watching and trying to learn a thing or 3.
 
C

chefro420

Sweet! I have wanted to mess with this for a while. Any more pix and could you elaborate more bro!!
 

bloyd

Well-known member
Veteran
Awesome gray wolf and growgeek! Have you guys looked into making artificial seed at all? Would love any details you have.
 

Sheriff Bart

Deputy Spade
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plants dont have 'stem cells' so to speak. plants contain meristematic tissues, which are the tissues in the plant which are actively dividing and growing. after they divide differentiation occurs, where the cells which were once just dividing cells, start to do different things, like turn into mesophyll cells (photosynthesis) or vascular tissues(transport), etc. etc. the majority of plant cells are also totipotent, that is that they have the ability to de-differentiate from whatever tissue they were found in and can start to divide again like the meristematic tissue and grow what is known as callous tissue. you can control what organs callous tissue will differentiate out into with hormone treatments. the auxins (IAA, IBA, NAA, 2,4-D) will be used to induce root growth and the cytokinins (BAP, kinetin, zeatin, etc.) can be used to induce new plants shoots to grow off the callous tissue which can be cut and rooted into new plants (or roots can be grown in the tc and the plants can be transplanted)

it looks like though that your geek was simply tissue culturing nodes with meristems (shoots) already on them, and not doing tc from a leaf, or pith or some other single tissue culture, but a mixed tissue culture so theres probably just regular nutrients in the agar and no hormones.

and r.e. the pathogens. the theory is that viruses either cannot or do not penetrate into the meristematic tissues of the plants (because if they did they would kill the plant way faster) so you can TC the meristematic tissues and hopefully will clone off virus free growth.
 

ScrubNinja

Grow like nobody is watching
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Very cool, cheers for posting. Do you have a full blown lab, or how much trouble/expense did you go to to create a suitable environment to do this?
 

Gray Wolf

A Posse ad Esse. From Possibility to realization.
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Awesome gray wolf and growgeek! Have you guys looked into making artificial seed at all? Would love any details you have.

Yup, we do have plans for artificial seeds, but haven't tested out our theories yet, so let us get some tangible results before commenting. If it works, I will ask the geekmiester to publish a report.

These are also amongst the first two dozen strains that he cultured, and so far he has grown no roots (making these non plants) on any of them, or raised any through adult hood. I shared his tissue culture results early, because the results to date have been so encouraging.

I will continue to share pictures as they progress through adulthood and if this experiment finishes as well as it started, I will ask him to publish his notes as well.
 

Chimera

Genetic Resource Management
Veteran
are where did you get the idea for artificial seeds? hahahaha


are these from callus or micro-cuttings of meristem shoots? It's tough to see through the container.

-Chimera
 

Gray Wolf

A Posse ad Esse. From Possibility to realization.
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ICMag Donor
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plants dont have 'stem cells' so to speak. plants contain meristematic tissues, which are the tissues in the plant which are actively dividing and growing. after they divide differentiation occurs, where the cells which were once just dividing cells, start to do different things, like turn into mesophyll cells (photosynthesis) or vascular tissues(transport), etc. etc. the majority of plant cells are also totipotent, that is that they have the ability to de-differentiate from whatever tissue they were found in and can start to divide again like the meristematic tissue and grow what is known as callous tissue. you can control what organs callous tissue will differentiate out into with hormone treatments. the auxins (IAA, IBA, NAA, 2,4-D) will be used to induce root growth and the cytokinins (BAP, kinetin, zeatin, etc.) can be used to induce new plants shoots to grow off the callous tissue which can be cut and rooted into new plants (or roots can be grown in the tc and the plants can be transplanted)

it looks like though that your geek was simply tissue culturing nodes with meristems (shoots) already on them, and not doing tc from a leaf, or pith or some other single tissue culture, but a mixed tissue culture so theres probably just regular nutrients in the agar and no hormones.

and r.e. the pathogens. the theory is that viruses either cannot or do not penetrate into the meristematic tissues of the plants (because if they did they would kill the plant way faster) so you can TC the meristematic tissues and hopefully will clone off virus free growth.

Thanks for drawing my attention to my error and the enlightenment bro!

The geek tells me you are correct on my technically incorrect use of the word stem cell instead of meristematic cell.

You are incorrect however in your assumption that these were grown from shoots, or that there were no hormones used.
 

Bullfrog44

Active member
Veteran
Not sure what company you are using, but I know the guy that makes these out of Mountain View California. This guy really knows his shit and makes a great product.
 

Sheriff Bart

Deputy Spade
Veteran
TC is really quite simple in practice and theory. the difficulties lie in 1st determining optimum hormone levels, and 2nd sterile practice and 3rd right equipment. generally a flow hood or glove box is necessary (anybody else do any mycology work and clone shrooms? same deal)
bleach and peroxide are used to sterilize tissues, and you can work with standard Hoaglands solutions in agarose but there are a lot more complicated nutrient broths that are worked with. it all depends what you want to do, what you are propagating, and why.

and sorry i was just assuming based on what i could see. i would love to hear more about how its done, but i know a lot of people get pissy about that sort of shit thinking its trade secret details of theirs that they have to protect so i wasnt expecting to hear much about the nutrients and hormone levels.

here is a simple lab in a plant phsyiology class I had a while back, workin on tobacco leaves and pith tissue and doing tc
quite simple
just keeping it all sterile is a bitch
View attachment tissculturemanual.pdf
 
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420247

Plant Whisperer
Veteran
TC is really quite simple in practice and theory. the difficulties lie in 1st determining optimum hormone levels, and 2nd sterile practice and 3rd right equipment. generally a flow hood or glove box is necessary (anybody else do any mycology work and clone shrooms? same deal)
bleach and peroxide are used to sterilize tissues, and you can work with standard Hoaglands solutions in agarose but there are a lot more complicated nutrient broths that are worked with. it all depends what you want to do, what you are propagating, and why.

and sorry i was just assuming based on what i could see. i would love to hear more about how its done, but i know a lot of people get pissy about that sort of shit thinking its trade secret details of theirs that they have to protect so i wasnt expecting to hear much about the nutrients and hormone levels.

Yes TC and growing mushrooms are very similar :wave:

Cloning shrooms??? I thought it was just cutting a piece of mycelium and culturing it... LOL what do I know... :wave:
 

Chimera

Genetic Resource Management
Veteran
Shoot Culture is neat and all but pretty basic TC wise.

The real fun comes with callous culture and shoot regenation. Callous is the real 'stem cells' or totipotent cells in plants... using callous and regenerating shoots opens up all sorts of possibilities.

Coming soon to a med state/country near you!

-Chimera
 

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Microbeman

The Logical Gardener
ICMag Donor
Veteran
plants dont have 'stem cells' so to speak. plants contain meristematic tissues, which are the tissues in the plant which are actively dividing and growing. after they divide differentiation occurs, where the cells which were once just dividing cells, start to do different things, like turn into mesophyll cells (photosynthesis) or vascular tissues(transport), etc. etc. the majority of plant cells are also totipotent, that is that they have the ability to de-differentiate from whatever tissue they were found in and can start to divide again like the meristematic tissue and grow what is known as callous tissue. you can control what organs callous tissue will differentiate out into with hormone treatments. the auxins (IAA, IBA, NAA, 2,4-D) will be used to induce root growth and the cytokinins (BAP, kinetin, zeatin, etc.) can be used to induce new plants shoots to grow off the callous tissue which can be cut and rooted into new plants (or roots can be grown in the tc and the plants can be transplanted)

it looks like though that your geek was simply tissue culturing nodes with meristems (shoots) already on them, and not doing tc from a leaf, or pith or some other single tissue culture, but a mixed tissue culture so theres probably just regular nutrients in the agar and no hormones.

and r.e. the pathogens. the theory is that viruses either cannot or do not penetrate into the meristematic tissues of the plants (because if they did they would kill the plant way faster) so you can TC the meristematic tissues and hopefully will clone off virus free growth.

Thanks Bart for pointing this out. I was verging before I saw your post. One can actually purchase plant tissue cloning kits from science supply houses if so connected. I'm hoping to grow filet mignon one day and dispense with the damn cow.
 

GrowGeek

Member
Howdy all. GW told me he shared a pic and I wanted to chime in on a few details. First off, TC is easier than you think. If I can do it, anyone can. My 6 year old Goddaughter cultures African Violets and Orchids with me... Cannabis is no different as others have noted.

You need about $50.00 worth of supplies and some sort of clean work area. A laminar flow hood is the wet dream of any micropropagator (is that a word? One who micropropagates)... But more practical are things you already have around your house.

All info is on the net. No trade secrets here Sheriff. Google and Youtube are fine resources for all information from culturing basics to media requirements and what parts of the plant work best with what growth regulators / hormones.

Not too up on the details of the seed making. I know people way smarter than me that are dabbling in that and I would love to try a homemade bean or two myself! :)

I do believe the pic GW posted is one that I didn't trim as much as I "should" have. I was expecting callus and got it, but also forced the node to put up new foliage. Still working the kinks out of where to slice and such. My brain thinks electronics for 99% of it's day, gotta give me a break. :) It is still a excellent and viable subject for micropropagation, just going to have to get into a taller culture vessel sooner than the others or be divided.

This is simply a science project that took on a mind of it's own. I've been enjoying it quite a bit, and do culture more than just cannabis.. It has become almost a plant nerd obsession thing... And now all I do is look at different plants in my travels and wonder... "How am I going to culture you!?!".

Go Chimera! Looking good! You got it right about the possibilities!
 
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Gray Wolf

A Posse ad Esse. From Possibility to realization.
Mentor
ICMag Donor
Veteran
Howdy all. GW told me he shared a pic and I wanted to chime in on a few details. First off, TC is easier than you think. If I can do it, anyone can. My 6 year old Goddaughter cultures African Violets and Orchids with me... Cannabis is no different as others have noted.

You need about $50.00 worth of supplies and some sort of clean work area. A laminar flow hood is the wet dream of any micropropagator (is that a word? One who micropropagates)... But more practical are things you already have around your house.

All info is on the net. No trade secrets here Sheriff. Google and Youtube are fine resources for all information from culturing basics to media requirements and what parts of the plant work best with what growth regulators / hormones.

Not too up on the details of the seed making. I know people way smarter than me that are dabbling in that and I would love to try a homemade bean or two myself! :)

I do believe the pic GW posted is one that I didn't trim as much as I "should" have. I was expecting callus and got it, but also forced the node to put up new foliage. Still working the kinks out of where to slice and such. My brain thinks electronics for 99% of it's day, gotta give me a break. :) It is still a excellent and viable subject for micropropagation, just going to have to get into a taller culture vessel sooner than the others or be divided.

This is simply a science project that took on a mind of it's own. I've been enjoying it quite a bit, and do culture more than just cannabis.. It has become almost a plant nerd obsession thing... And now all I do is look at different plants in my travels and wonder... "How am I going to culture you!?!".

Go Chimera! Looking good! You got it right about the possibilities!

Thanks or sharing brother Growgeek!

I'll ask brother Joe to share when we do the bean making.
 
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