Cannabis tissue culture

burnone710 said:

Can a culture be taken from a plant that has been flowered out already during harvest or can you re-veg and take a culture? Will this save the genetics without losing the vigor and potency that you would normally after re-veg ? I have an amazing cut I misjudged and killed the mother too early, still have it flower almost a week from harvest...tissue culture kit arriving around harvest time. Really wanna get this sucker back!!

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I haven't heard confirmation of that they will take in late flowering, but it seems quite possible. Please try it and let us know!

Are you proposing to use nodes as per the most efficient current protocol from these harvestable / mature plants?

More often than not, you want relatively juvenile tissue for micropropagation (for example, if this were a leaf protocol, a common phrase you'll hear for both tissue culture and pathogen assays is 'youngest, fully expanded leaves').

When we tissue culture woody perennials we often use floral material or meristematic tissue (which becomes the same thing in the case of our plants) because juvenility is so lacking in other tissue, making it less susceptible to tissue culture treatments.

Although a poor comparison to cannabis due to the widespread differences in tissue culture, something may be learned from this example. It is probable that if the cannabis plant has however undergone extensive senescence such as at the point of harvest, you may even find these tissues are not susceptible to tissue culture. And that doesn't even remark upon the problem that good/efficient protocols for tissue culture of cannabis floral material do not exist.

There are however some reasonable leaf or possibly petiole protocols for cannabis, you could certainly give it a shot. I would maybe try a few weeks before harvest though. And perhaps try to get young 'suckers' from the bottom if there are any, and the most juvenile tissue in the meristematic region, at least that would be my guesstimate. I am not sure how such tissue will receive the current sterilization procedure commonly used, it may be injured by it if floral material or lacking lignification, you'll have to see.

I don't know if this is the appropriate forum to ask but has anyone had access to this paper(below citation) it is available upon request, but i have no luck getting any answers from my request. Commercial companies are using this Technique, it seems to produce high levels of success and is said to be much easier to apply. I have spent many many hours looking for it looked at sci hub libgen and gone through many web search results with no luck .

Cannabis sativa L. Micropropagation in Temporary Immersion Bioreactor System
Article in Planta Medica 76(05) · March 2010 with 319 Reads
DOI: 10.1055/s-0030-1251771

How long can a tissue culture be stored? Is there a lab and storage facility where I could have strains preserved and stored long term?

I have searched.. I can't find it if tissue culture plants from callus or meristem can produce a seedling like plant. Clones don't have tap roots and because they are cuttings of branches they grow staggered. Also they are sexually mature and more inclined to flower under shortish days.

My interest is with producing a hearty seedling like plant from known genetic material. Everyone seems to think seeds perform better for long season big outdoor plants like we grow in California. have searched.. I can't find it if tissue culture plants from callus or meristem can produce a seedling like plant. Clones don't have tap roots and because they are cuttings of branches they grow staggered. Also they are sexually mature and more inclined to flower in spring.

My interest is with producing a hearty seedling like plant from known genetic material. Everyone seems to think seeds perform better for long season big outdoor plants like we grow in California.
I'm not 100% sold on if seedlings really are that much more vigorous then clones, but of they were...

There are some nice videos on youtube by monster gardens that also has a kit. You can buy the parts and nutrients on ebay also.

I started a couple weeks ago with a storage bin for my hood and I already have hepa filtering fan. Tools and tubes easy.

Check out the videos guy is doing it in a warehouse with success.

Peace
AL

Here is the pamphlet provided by Bill Graham check him out on youtube too easy

Scrappy-doo said:

How long can a tissue culture be stored? Is there a lab and storage facility where I could have strains preserved and stored long term?

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callus or any stage really must be 're plated' as it were every few months or so. but I am sure if kept cool and low light with charcoal to absorb plant metabolites that could be extended somewhat. but as far as i know there is no real long term storage (6 month or year) without transfers. Dont think the labs that are doing it for cannabis offer cryogenic storage which of course may work for longer term and once frozen dont need to do anything. however I imagine that would be alot more expensive and not really practical for our purposes and also have problems of there own.

but yes there are places popping up now for cannabis to store genetics and they just do all the replating. The only issue I forsee is will those plants be able to be organic certified. and will they pass testing having PGR and antibiotics cytocides.

oh and to the guy who asked about taking tissue after flower. yes this is most certianly possible as you add all the hormones and sugars

Hi folk,
I’m looking for an anther culture and callus induction medium formulation. I’m thinking about testing:

Murashige and Skoog’s medium containing 3% (w/v) sucrose, 0.8% Type E agar supplemented with 0.5µM naphthalene acetic acid (NAA) in combination with 1.0µM of thidiazuron (TDZ)

Murashige and Skoog’s medium containing 3% (w/v) sucrose, 0.8% Type E agar supplemented with plant growth regulators: BAP (1mg/l) and NAA (0.5mg/l)

Any other suggestion would be greatly appreciated.

Darpa

Darpa said:

Hi folk,
I’m looking for an anther culture and callus induction medium formulation. I’m thinking about testing:

Murashige and Skoog’s medium containing 3% (w/v) sucrose, 0.8% Type E agar supplemented with 0.5µM naphthalene acetic acid (NAA) in combination with 1.0µM of thidiazuron (TDZ)

Murashige and Skoog’s medium containing 3% (w/v) sucrose, 0.8% Type E agar supplemented with plant growth regulators: BAP (1mg/l) and NAA (0.5mg/l)

Any other suggestion would be greatly appreciated.

Darpa

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I'd try equal amounts of cytokinins and auxins to initiate a callus.

Darpa said:

Hi folk,
I’m looking for an anther culture and callus induction medium formulation. I’m thinking about testing:

Murashige and Skoog’s medium containing 3% (w/v) sucrose, 0.8% Type E agar supplemented with 0.5µM naphthalene acetic acid (NAA) in combination with 1.0µM of thidiazuron (TDZ)

Murashige and Skoog’s medium containing 3% (w/v) sucrose, 0.8% Type E agar supplemented with plant growth regulators: BAP (1mg/l) and NAA (0.5mg/l)

Any other suggestion would be greatly appreciated.

Darpa

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Ebay has a product medium made in germany for sativa plants pre mixed

with DNA and bubbleman behind the tissue culture company here in canada. anyone know if tissue cultured plants could be considered organic since grown with salts and PGRs for part of there life.. esp TDZ and the cytocides to prevent contam maybe not alot makes it into the plant but likely not super awesome for concentrates? think certified organic would def be off the table?

clearheaded said:

with DNA and bubbleman behind the tissue culture company here in canada. anyone know if tissue cultured plants could be considered organic since grown with salts and PGRs for part of there life.. esp TDZ and the cytocides to prevent contam maybe not alot makes it into the plant but likely not super awesome for concentrates? think certified organic would def be off the table?

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Hi Clearheaded, I haven't do an extensive check up on the subject, but it seem that tissue cultures plants wouldn't be an issue for a biologic certification. (at least under Canadian regulation).


Natural PGR are autorized (sorry it's in french)
Régulateurs de croissance des plantes: Les hormones végétales naturelles, comme l’acide gibberellique, l’acide indole-acétique (AIC) et les cytokinines, sont permises. Voir aussi Acide gibberellique.

Therefore, TDZ, wich is a citokinin, is autorized...

Ref: List of autorized substances for biologic production
https://publications.gc.ca/collections/collection_2014/ongc-cgsb/P29-32-311-2006-fra.pdf

there are a few of the needed PGRs that are considered safe and organic. your right. however several/many are not. TDZ is has a small amount of toxicity(mostly to fish) and is infact banned in the EU. while of course biological mechanisms are different in fish and humans to me it means it should be avoided as something that will be inhaled. like pyrethrin. "ok" to eat however inhaling is a little different and also toxi to fish. also TDZ is not organic and 100% synthetic compund. just because a chemical falls into a class that is naturally occuring does not make it organic or safe...

Also because 90% of PGR are synthetic and infact not organic I do find it hard to believe. As PGR are used for cotton and one of the reasons people shy away from non organic cotton. does organic cotton still use the PGRs before harvest??

1 other point which would prevent them from being organic is of course they are grown on salt based mediums. how could a TC plant become organic because its grown on slat based nutes? antiiotics antifungals in the medium is common aswell to produce in mass. are those something you want to smoke? are they considered organic even if antibiotics may be gotten from fungus?? organic beef use organic antibiotics???


Pushkar Kale, Agricultural and Food Engineering, IIT Kgp
Answered Mar 17, 2016 · Author has 206 answers and 322.7k answer views
"In case of Tissue Culture, due to use of artificial media, it is prohibited."

So my sense is while seed potatos are started in tissue culture they are grown out and then those potatoes are then the seed potato ie generation 1year away. they can be organic because are not grown inorganic medium. however a cannabis plant started in inorganic salts would not be considered organic..

Also if use your logic of PGR OK because bsed on organic structures but synthetically produced or tottaly new chamical. Can plants then be sprayed with PGRs and be considered organic??

love to hear chimera chime in.

Hi Clearheaded,

After further verification, you are wright. It would be very difficult to get a biological certification if you are starting with tissue culture material. Synthetic growth regulator are not authorized under Canadian Organic certification (I missed that note regarding section 4.1.3: Substances listed in Tables 4.2 and 4.3 shall comply with prohibitions in 1.4 of CAN/CGSB-32.310.):

CAN/CGSB-32.310, Organic Production Systems – General Principles and Management Standards, describes the principles and management standard of organic production systems.

CAN/CGSB-32.311, Organic Production Systems – Permitted Substances Lists, provides lists of substances that are allowed for use in organic production systems.
General Principles and Management Standards.

CAN/CGSB-32.310-2015 Amended March 2018

1.4 Prohibited substances, materials or techniques in organic production and preparation If producing or preparing organic products, the following substances, materials or techniques are prohibited since they are incompatible with the general principles of organic production:

a) all products of and materials from genetic engineering (GE), as defined in this standard, and as specified in 4.1.3, 5.1.2 and 6.2.1 of CAN/CGSB-32.311;

b) all products, materials or processes intentionally using nanotechnology, as defined in this standard, with the following exceptions:

1) naturally occurring nano-sized particles or those produced incidentally through processes such as grinding flour;

2) contact surfaces, such as equipment, work surfaces, or packaging, where transference of nano-sized particles to organic crops, livestock or products is unintended and unlikely to occur;

c) irradiation, as defined in this standard, for the treatment of organic products and inputs used in the production of organic products, except as specified in CAN/CGSB-32.311;

d) soil amendments, such as fertilizer or composted plant and animal material, that contain a substance not listed in CAN/CGSB-32.311;

e) sewage sludge;

f) synthetic crop production aids and materials, except as specified in CAN/CGSB-32.311;

g) synthetic growth regulators;

h) cloned livestock and its descendants;

i) synthetic allopathic veterinary drugs, including antibiotics and parasiticides, except as permitted by this standard;

j) synthetic substances used in organic product preparation, such as ingredients, food additives and processing aids, including sulphates, nitrates and nitrites, except as specified in CAN/CGSB-32.311;

k) equipment, harvest and storage containers, storage facilities and packaging materials treated with synthetic fungicides, preservatives, fumigants and pesticides;

l) substances that are not listed in CAN/CGSB-32.311, except as permitted in this standard.

That been said, Certified Organic or non-certified organic products, it’s just a marketing tool in order to differentiate your product on today market. There is no health concern about consuming non certified organic product… But if you prefer bio, just go with it….

It exists for a reason, some reason. To say it is just a marketing tool and there is no health concern does not sound entirely comprehensive/accurate. Tissue culture may be ok to use right away or after 1-2 generations but your post is dismissive of organic standards.

Thanks for clearing that up Darp.

speaking of marketing Bman is convinced it is organic buut think he just doesnt know enough, but thats on him, everyone is free to go through this world as they like, just a little let down he didnt read more as they will be pushin this potentially laced stuff to potentially all/most large LPs (at least if is found to be viable, which is yet to be seen). whats worse is for it to be viable it isnt going to be for moms it will be 1000s of plants to be flowered. So much talk on hash church about clean products and issue with even using clones of clones that are sprayed failing testing...

Well while "organic" has a life of its own pun intended, as far as marketing and meanings to people. may not cover all we would like or what BIO is in euro. there is certainly more then marketing in organic herb or organic tomato, once you experience the difference its hard to deny and more then sugar content. Also it is making a choice to reduce chemicals in environment not only leakage but manufacture/transport and also those chems in your body. once you get older it seems the folks who sprayed there lawns and didnt bother washing things seem to be the ones getting cancer 100% anectdote, but after being exposed to multiple things over the years that we are told is "OK" then being told its not 15-20 years later and taken off the market, makes sense it would add up. What were we talking about again?? lol anyway. long story short unfortunete TC isnt organic..

tried to get chimera to chime in hopefully he does, not sure if he invested or not. feel like he would of held off as knows there is no real barrier to entry.

Bump for 2020. Thinkin about getting into tissue culture, just to preserve my genetics so I can bring things out and pht um away for certain breeding projects..