Tissue Culture/Micro Propagation Grow from Scratch

[canndo]

These are NOT part of the experiment, they are different strains and are older than the Lemon Skunk. I figured I'd put them in here just to keep interest up (if there is much). I suppose you could call them my "moms" but all I am doing is fattening them up so I can see if they will flower in vitro. These are a number of generations old.

 

[JealousGreen]

How do you get new plants that can be planted? Do you have to clone into another jar, wait for roots, then transplant? Is it that simple? Will shoots ever be big enough to put into an aero-cloner?

 

[canndo]

Usually yes, you move shoots into a rooting medium. In some cases you do that but not to the point of rooting and then place them in some more natural medium to harden off as they root. I suppose you could aero-clone, the plants can get large enough.

 

[JealousGreen]

Are you only limited by the size of the jar then?

 

[canndo]

Correct. Although the media could get expensive if you wanted to use really big jars.

 

[vh13]

This is the first time I've ever seen anything like this.

+ Subscribed, Rep, and very curious.

 

[canndo]

I am getting ready to do several things, although some are not a part of this journal, I'm motivated by my chia pet to do something today.

I want to demonstrate the potential to flower in vitro - this should have a real functionality as I could sex plants that have been entirely grown in vitro from seed (when I get that working). I am anticipating trying to flower without roots and with roots. I don't have enough plantlets that are big enough to try this both ways, nor do I have enough jars. So, I cleared out some duplicates and took pictures of some of them that are more clear than anything I could take through the glass of the jar.




As you can see, there are healthy shoots, they are almost big enough to place into new jars and new media, no vitrification and not much callus production. I'm thinking I will do my first operation on the established Lemon Skunk plantlets - there are, I believe, 9 left and I think 5 are productive enough to continue with.

 

[canndo]

More pictures of the Lemon Skunk plantlets today.

 

[canndo]

 

[DarthD3vl]

what are you going to do with these? Im not sure i understand why your doing this?

 

[canndo]

Darth, I intend to keep a strain library. Furthermore I intend to create large numbers of clones in a small space. There is a market for commonly sized, robust and pre-packaged clones in my state that I would like to take advantage of. In the months that I have finally garnered some sucess I have gone from about a dozen jars each with a single cutting to 50 (well, not now since I dumped a lot of duplicates) - some of the 50 have multiple plantlets. From here I have larger containers which will each hold 8 to 10 plantlets all the way up to rooting stage.

 

[vh13]

So this is a kind of hyper-bonsai-ism, except instead of focusing on aesthetics you're interest is in the preservation and production of genetics?

Very fascinating.

What's the cycling rate? I imagine you have to cut new specimens and enter them in at the lowest levels periodically, the medium will only last so long, right?

 

[canndo]

VH13, I suppose you could call it that but it is just a natural result of feeding the plants sucrose. Each plantlet will produce between 2 and 5 transplantable shoots a month - I am still tweeking the Cytokinin/Auxin/Ga balance so 3 is the norm. Now figure I can put 8 plantlets in a tiny tub. In one month I have 24 "clones" - transplant all of them and you do the math, I can potentialy do hundreds of thousands in a year. As for my cycling rate, the medium seems to last about 3 weeks and then the plantlets start exhibiting callus formation and vitrification. I suspect however that this may be caused by a lack of air exchange or too high a humidity in the jars.

Regardless, I either transplant the original plantlet every three to four weeks, or I slice off the transplantable new shoots, put them in new media and, well, generaly tidy up the plants for their next month of growing. I don't use new cuttings from a "real" plant unless I want a new genetic lineage - like the Lemon skunk I am doing now.

 

[DarthD3vl]

OH thats awsome man! I've never seen any one do this. very cool

 

[LucienMD]

Do you use a glove box or a laminar flow hood? Do you know how results vary between those two approaches?

 

[canndo]

Lucien, I've used both, I like the hood better of course but I have never seen a difference. I think the secret is in the heat tolerant biocide I use in the media. About the only contamination I ever get is from direct contact with an improperly sanitized plantlet.

 

[canndo]

Today's pictures, the best 5 jars. Growth is slowing a bit, time for that transplant and clean up I've been putting off. As you can see I have a number of useable shoots although in order to have this all work I may wait a few more weeks for that process.

 

[canndo]

View attachment 1495328View attachment 1495327View attachment 1495326View attachment 1495325View attachment 1495324

 

[Wolverine97]

Subbed. As I said in the other thread, I'm intensely interested. I want to see some roots though, why wouldn't regular old Clonex work for that? Sterility issue? Too strong?

 

[haole420]

3 weeks and no roots? not sure what benefits are to be had with tissue culture over cloning.

kudos to canndo for actually doing this experiment, though.

if you used tiny tissue samples (hole punched leaf) and ramped this up to commercial production scale (i.e., starting a new batch of thousands of tissue samples every day), then it might make sense. i think that's why they use tissue culture for large-scale crops like bamboo, strawberries, asparagus, etc.

unless you're growing tens of thousands of square feet of weed, i'm not sure tissue culture is worth all the trouble when the same can be accomplished by cloning. plus with cloning, you can take bigger clippings that'll put you at least a month ahead of this kind of propagation in terms of growth.

keep it going though: i want to see how they end up!