Plant Tissue Culture: An Upcoming Alternative To Clones

[jumpincactus]

I used the search feature for this forum. It didn't turn up a thread on this subject.

Finding a reliable source of new cannabis plant starts, or clones, that are healthy and free of pests and disease is a constant challenge for cannabis growers. The conventional way to produce cannabis clones is to take cuttings from plant “moms” and grow them out into new plants. Many growers do this themselves, and cannabis nurseries do the same process on a larger scale. This method has been successful, but scaling it is expensive and resource intensive. As an alternative to the traditional method of making clones from cuttings, many people have experimented with using a technology called plant tissue culture.

Tissue culture technology has been around for decades and is widely used to produce genetically identical plant starts for many common crops such as orchids, potatoes, flowers and fruit trees. Tissue cultured plants begin their lives as tiny shoots in a laboratory. The cultured plants go through a long process of first removing all outside pathogens, then multiplying and sprouting new shoots, finally to emerge into the outside world as beautiful, happy and vigorously growing new plants.

This technology has the potential to produce plants unlike anything produced through conventional cloning. The process of culturing cannabis rejuvenates old, mature plant tissue into a healthy, vigorously growing juvenile state, in some ways like newborn seedlings. This plant fountain of youth gives cultured plants many new advantages for growers: higher yields, stronger growth, and better resilience to environmental stress. Plus, mass production of high quality disease free clones will eventually mean greater availability and variety for growers. Tissue cultured cannabis, when it becomes widely available in the near future, could solve most if not all the typical problems growers have always had to live with when finding clones.
http://www.theganjier.com/2015/09/11/plant-tissue-culture-an-upcoming-alternative-to-clones/



Heres a link for more in depth on performing the procedure and the science to it all.

http://www.apsnet.org/edcenter/k-12...iotechnology/documents/planttissueculture.pdf

 

[We Solidarity]

Tissue culture is way way too labor intensive to ever replace conventional clone propagation, hence the reason there are so many failed labs in the U.S., and the worlds only major exporter of tissue cultured material is China, where they can afford the specialized labor.

 

[geologic]

Good call,
I knew this thread was needed;
I don't do lights,
so no overseason clones;
started looking into this 'till I got to the "sterile conditions" part--
that ain't me...

 

[jumpincactus]

Tissue culture is way way too labor intensive to ever replace conventional clone propagation, hence the reason there are so many failed labs in the U.S., and the worlds only major exporter of tissue cultured material is China, where they can afford the specialized labor.

Thanks for sharing that @We Solidarity , I am just starting to delve into the science. From what I gather it is best left alone to the mass producers of plant stock with deep pockets. ........Well, that rulez me out fo sho..... :(

None the less, still very interesting stuff.

 

[We Solidarity]

Thanks for sharing that @We Solidarity , I am just starting to delve into the science. From what I gather it is best left alone to the mass producers of plant stock with deep pockets. ........Well, that rulez me out fo sho..... :(

None the less, still very interesting stuff.

yep, tissue culture starts paying out when you need to supply millions of identical plants, which unfortunately will never be the case in our industry since you can cover an entire city's need off 12-15,000 plants, which is very manageable and affordable with traditional methods.

Another instance in which tissue culture is valuable is in maintaining one specific cultivar, (i.e. the cavendish banana) which may play a role in preserving some of the more important cannabis genetics in the future

 

[geologic]

yep, tissue culture starts paying out when you need to supply millions of identical plants, which unfortunately will never be the case in our industry since you can cover an entire city's need off 12-15,000 plants, which is very manageable and affordable with traditional methods.

Another instance in which tissue culture is valuable is in maintaining one specific cultivar, (i.e. the cavendish banana) which may play a role in preserving some of the more important cannabis genetics in the future

All I wanted to do was to overwinter ~50 plants--
an' cull 'em as they're tasted properly...

 

[Sativied]

Another obstacle that makes it less interesting for cannabis growers is that it takes too long. The plantlets have to acclimatize and adapt slowly.

 

[jumpincactus]

Another obstacle that makes it less interesting for cannabis growers is that it takes too long. The plantlets have to acclimatize and adapt slowly.

What is the typical time to prep and harden off a tissue cultured start as compared to a clone??? Curious, just getting my head wrapped around the whole process...... peace

 

[Sativied]

I don't know how long it takes for cannabis, it takes several months for some species and is on top of the time it takes to grow the plantlet. Those large tissue culture-rers have plantlets in all different stages allowing them to supply constantly.

And to add to that China comment someone made. Many companies in the Netherlands have tc facilities in lower wages countries because it's so labor intensive. When it's done here it more often for scientific purposes, to improve the process, for breeding (like tc pollen to create a haploid and double it with colchicine into a fully homozygous plant, or create virus free breeding stock) and rejuvanation (strip it from the effects of nurture and rebuild it from only its dna) and not as a direct alternative for large scale cloning.

 

[jumpincactus]

The more I learn and hear about this process the more inclined I am to not even messing with it due to the cost and time constraints. I will say this tho. I always pay my help a rightful/livable wage and would never sub/sellout to another country just cause I can take advantage of a lower wage base. This is a lot of the NEW WORLD/global economy and personally I think it suxxxx. Nuff said

 

[shadowshell]

Would tissue culture make sense for long term genetic preservation projects?

Have you all seen the Lablets offerings from Dark Hearts in the SF Bay area? They are currently available from a few Bay area vendors. DHN makes some interesting and seemingly reasonable claims about the improved vigor of clones generated this way. I get the impression that they generate a mother from culture for the vigor and offer clones from that. It doesn't seem like they would generate every clone from culture. Though I've been wrong before and am only guessing now.

http://darkheartnursery.com/news/introducing-lablets/
http://www.theganjier.com/2015/09/11/plant-tissue-culture-an-upcoming-alternative-to-clones/

 

[jumpincactus]

Do you have a link for the information concerning DHN's claims t more vigor with a TC plant as I was told they lose some vigor compared to the hybrid vigor we are all accustomed to. Not saying either camp is right. Would like to read up on it. @shadowshell :D

 

[shadowshell]

@jumpincactus The only mention I can find are in the links I posted and in some instagram comments. I've emailed them for more info, but I don't know what will result of that. I may try to scope them out at the next Bay Area event and get the lowdown in person. Let me know if you find anything definitive.

 

[shadowshell]

"Lablets are produced using a new type of stock plants which are created in our tissue culture laboratory. These new stock plants, which we call “producers,” grow much differently than traditionally produced clones. By taking cuttings from producers, we are able to create clones that develop similarly to those growers are used to, but with added vigor. What we are hearing from our beta-test partners, and seeing in our in-house trials, is that the clones have the health and vigor more characteristic of very juvenile plants; such as seedlings. From our in-house Fire OG trial, we’re seeing that Lablets produce more branching with tighter internodal distances. Ironically, we’ve received some anecdotal feedback that the opposite may be true in some Indica varietals, where growers are actually seeing longer branches."
http://darkheartnursery.com/news/introducing-lablets/ @jumpincactus

 

[jumpincactus]

@jumpincactus The only mention I can find are in the links I posted and in some instagram comments. I've emailed them for more info, but I don't know what will result of that. I may try to scope them out at the next Bay Area event and get the lowdown in person. Let me know if you find anything definitive.

you got it, and likewise if you find more info please share!!

 

[ArcticOrange]

Thanks for sharing that @We Solidarity , I am just starting to delve into the science. From what I gather it is best left alone to the mass producers of plant stock with deep pockets. ........Well, that rulez me out fo sho..... :(

None the less, still very interesting stuff.

Ive got a friend at university propogating berry plants from tissue culture, hoping to learn some mad science from them lol

 

[GrwrGoneWild]

you got it, and likewise if you find more info please share!!

Look closely at TC lablets vs. Premium Heart cuts, they have thin stems. I'm finding the same thing with the TC''s I've run, thin stems. My work is based off of the directions in Melinda's work in cannabis TC. Seems to be a common problem. I think its something to do with CO2 exhange, most of these TC's are in a closed container. You have sucrose in the mix but I still think you need CO2 for real bulk,..in photosynthesis co2 is the main producer of glucose. My theory anyways. I got some more agar for another run. It was kinda cool to have TC grown plantlets but just not impressive next to a regular clone. Not to mention explant hardening.. etc etc.. yaddda.. yadda.

One thing I found out when messing around with a cutting just sitting in liquid Murashige and Skoog medium plus hormones (IBA/IAA) but no agar The stem doubled in thickness, that was in contact with the solution. And had the nubs for roots ~7 days, I still had to ditch the solution because of a mold infection, thats even with PPM (Plant Preservative Mixture) in the solution in open air in a tray. Interesting stuff :). I should make a container that could have sterile gas exchange.

TC could be used to clone ruderalis. But I only mess with photoperiod strains. There hasn't been a huge demand for clones, I've been able to keep up with regular cloning techniques. So there hasn't been alot of motivation for me to TC but I'm revisiting it, see if I can improve on my last TC run.

I think DHN is a model for TC to show it works. TC just needs a little imagination to get it to work, the other day I couldn't take a cutting from a buddys plant, If I had a flask and a way to do a sterile transfer I could have got a cutting :). I dont do singles when working a TC, I drop about 4-6 to a wide mouth container. PP5 plastics I get from the deli :), I got those plastics pretty hot, 225F @ regular pressures in a microwave a little distortion from keeping the lid on too tight when the steam cools and the container returns to normal pressure, something to think about instead of using fancy TC tubes.

What keeps people out of TC is the cost it takes about 100 bucks to get all the materials, and the sterile transfer. Its just alot easier to make clones with rockwool that costs like 10 bucks a slab. TC is just more complicated, and technical but if I can learn it you can learn it.

 

[GrwrGoneWild]

The more I learn and hear about this process the more inclined I am to not even messing with it due to the cost and time constraints.

Its not that bad, if you've ever done any work with mushroom cultures its about the same. Or even canning food at home, not too technical. The technical part is the hormones, and agar formulas.

-Make your Agar, or buy a preformulated one. I went the hard way and made mine from Murashige and Skoog and added hormones, following Melinda's TC work that is cannabis specific.

-Cook/sterlilize your agar in a container. You can use this one antibiotic called PPM, its for Tissue culture to keep the agar free from contamination. You can either microwave or pressure cook the agar.
Hey, plastic works! to for TC.

-Flask your plantlets after sterilizing. Bleach +water dip and rinse. One last dip in PPM + water if you water. Work needs to be done in a makeshift hood or laminar flow hood if you just so happen to have one.

-Put under or near lights, you dont need alot, remember that sucrose is the fuel. And wait about 21-30 days, transfer to rockwool in humidity dome under light for hardening.

It took me a few weeks to wrap my mind around the TC process. I think its the hormone balance is the hard part.

I think if you had the same plant, the TC will give you lots of little plants in the same amount of time. But if you cut clones the old fashioned way, you'll have less clones but bigger healthier looking clones..
Observe how different the clones from the Dark Heart lablets, vs the premium hearts.

 

[CBDCURES]

The process of Plant Tissue culture really is not that difficult, But you do have to spend some time
studying the process, basic chemistry , and the ability to follow procedures are outlined
I have my lab setup complete with Laminar flow table and sterilization of media Pressure cooker or microwave
serves as a Autoclave, Here again you do not really need a laminar flow table, Best source for starting out
is to go to , they seel a kit for $188.14, They are currently out of stock
however you can download the protocol, and instructions, and just purchase individual components.

This is a protocol for hemp that works well for MJ.
Plant tissue culturing preserves the exact DNA, whereas cloning you will lose the original genetics.
And the best reason for Plant Tissue Culture is, if you find a mother plant that has been tested for ideal THC/CBD ratios
Desirable Terpene profiles, you can keep these genetics going, and not have to worry about that mother plant
and having to keep it from overgrowing, and continuous cloning, after so many generations of clones
you will lose that original profile.

 

[ArcticOrange]

The process of Plant Tissue culture really is not that difficult, But you do have to spend some time
studying the process, basic chemistry , and the ability to follow procedures are outlined
I have my lab setup complete with Laminar flow table and sterilization of media Pressure cooker or microwave
serves as a Autoclave, Here again you do not really need a laminar flow table, Best source for starting out
is to go to , they seel a kit for $188.14, They are currently out of stock
however you can download the protocol, and instructions, and just purchase individual components.

This is a protocol for hemp that works well for MJ.
Plant tissue culturing preserves the exact DNA, whereas cloning you will lose the original genetics.
And the best reason for Plant Tissue Culture is, if you find a mother plant that has been tested for ideal THC/CBD ratios
Desirable Terpene profiles, you can keep these genetics going, and not have to worry about that mother plant
and having to keep it from overgrowing, and continuous cloning, after so many generations of clones
you will lose that original profile.

Thanks for this info I've been waiting around for someone to do that part of the work for me XD