Cutting Edge Cloning: Tissue Culture Propagation

[pop22]

From what I've read, polyploidy rarely creates a plant worth the while, not to say gives larger harvest. Check out what Robert Conell Clark has to say on the subject.

As for tissue culture, I was thinking of using very young seedlings for taking the cuts. say 4 - 7 days at most. May not make a difference but that young, they may still have undifferentiated cells, and/or hasn't started the death clock yet. The only way to know is to try. I'm hoping to give it another shot next month. I'll start 30 or so auto seedlings to use as donors.

I've done tissue culture before ... Not 100% sure I'd it will work on autos... But I think it will considering you are capturing the plants DNA

Here's some legit information so everyone can give it a shot. View attachment 763012 View attachment 763013

Chemicals like colchicine or sulfuran can induce chromosome doubling leading to polypoid cells which have the potential to be bigger and stonger than thier parents.

You could then use silver thiosulfate to create feminized seeds from the plants and go on a poly-pheno hunt! Or do that on any plant.

I was just thinking of trying this
@HemiSync @pop22 @iampepe @SpliffScot @MaineGrower @Powerful14

 

[CTb1]

From what I've read, polyploidy rarely creates a plant worth the while, not to say gives larger harvest. Check out what Robert Conell Clark has to say on the subject.

As for tissue culture, I was thinking of using very young seedlings for taking the cuts. say 4 - 7 days at most. May not make a difference but that young, they may still have undifferentiated cells, and/or hasn't started the death clock yet. The only way to know is to try. I'm hoping to give it another shot next month. I'll start 30 or so auto seedlings to use as donors.

I've read different in a few books not just regarding cannabis. But there is always conflicting information out there.

That would make sense it would be the age of the seedling once it takes off regardless i believe.

 

[CTb1]

@pop22 @derek420colorado we should create an advanced propagation/cultivation thread where everyone inputs their research with references included. Many minds are better than one

 

[wwwillie]

Great stuff as always @pop22

 

[JanSteen]

Since I haven't seen people post results: I've been able to create callous tissue with the aid of 2,4-D and 6-bap from auto seeds. The roots work best. They grew up as I expected: after the first leaf formation, that's a few transplants after callus, the plants started flowering. Visually I'd say it looked like a revegging photo plant. The culture died not long after that with no apparent reason, no phenolic bleeding, just chlorosis and slowly fading away.
If anyone had better results, I'd love to hear how!

6-bap (4-6mg/L) + 1/2 MS alone should work well to create callus in most cannabis strains.
Older autoflower tissue seems to respond in the same matter. With photo plants it's pretty easy to establish a culture and maintain it, but it takes a lot of time and a hand full of dollars. Usually petri's for plants have a shelf life of 2 weeks max, and it's hardly worth the time and money. Especially since cuttings take 2 weeks to root, and don't need sterile agar, hormones and special nutrients with a limited shelf life.

As for cloning stock for photoplants, I'd say a bonsai is cheaper than transplanting a piece of tissue to fresh agar every 2 weeks (it takes me 45 mins to clean, 1 hour to prepare, 15 minutes of work, and 30 mins for cleaning up the mess).

I am curious about protoplast fusion with Hops and other plants. It should be pretty easy if you can get ahold of Polyethylene glycol (PEG-300 and up) and some basic plant hormones.
Has anyone ever tried to make synthetic seeds with calcium agglutinate? That might actually work, because the tissue would revert to it's original form. The process might reset some things.

I'm just shooting in the dark here, the matter isn't something I've been reading about since I gave up cannabis TC long ago.. But the clock of auto's might be influenced by telomere length, like in some other plants, which would explain why it's nearly impossible to revert them to their 'ground state'. It would explain the large difference in growth periods of AF strains as well. A simple electrophoresis of genomic dna would prove or disprove that; if older plants have shorter strands compared to seedlings, the answer would be right there. If someone would be so kind to look into that and report the findings, it would make me a happy man.

 

[JanSteen]

I've been asking around a little, and this issue seems to exist in the food crop TC as well. They don't have an answer to it either, other than reculturing fast and often.
One possible option they told me, but couldn't confirm, was to start culturing from flowers. Just like with carnivorous plants, which form new suckers from flowers if the hormone levels are right, this might be an idea.

I've been able to make callus from calyxes pretty easily in the past, but those were photo's. They already have that feature hardwired inside their genetics. I've never tried it with auto's. It might just work.

 

[Senseimillan]

But I'm going to jump in to this by starting with a photo period, a very special one. Ducksfoot. VERY hard to get in the USA, or anywhere else for that matter.

@pop22 lucky ducky! It's on my bucket list of strains to collect.

@JanSteen do you think it would be possible to culture cells from the tip of a germinating seeds taproot and multiply them? Is it possible to keep it hormonally stable at that stage of life?