I am interested in what resources you have on the topic, what input you have with regards to the topic, and if you believe a particular method/mix/approach will work better than others for not simply regular cannabis/hemp embryo culture, but also for embryo rescue with older genetics in mind.
I am developing SOP’s and would appreciate input past ScienceDirect, Springer, JSTOR, and Wiley.
I am thinking on running a MT or MS medium with agar, balancing cytokinins and auxins, with other added aminos, vitamins, and carbon sources contributing to successful embryo rescue .
I am asking at the excision of the embryo. let’s say these extremely old seeds did not germinate post-sterilization, gibberellic and moderate brix content failed, the same methods opened, “hemp seeds,” 1-5 years old without issue, but these are let’s say stubborn, or that they weren’t kept in the correct conditions for decades
I assume you have a few seeds from a large number of strains.
Dissecting out the embryo was a non-starter with tobacco, I doubt it will help with cannabis, but at least the seed is big enough that you can attempt that.
in short, yes this is one method that works, but I am interested in other’s experience, tips, tricks, methods, and never assume a thing about seed game, could be just one or two seeds of gorilla glue
I believe organogenesis is best for PTC, but this is a question about Embryo rescue. I’m trying this out with as many trials as necessary, goal is under 4000 trials.
And I’m trying to understand how many seed you can afford to lose….4000 trials suggests more that a couple of GG#4
You’re certain to get strain specific germination rates with old and abused seeds…the problem is different if you have two seeds of 2000 strains vs 2000 seeds of two strains….by all means continue to obsfucate, and I’ll wander off.
Yes, I get that you’re looking at embryo rescue.
AND those guiding me 30 years ago suggested that IF I could not get “germination” on hormone free media, then trying callus inducing media to get any cell division was the next step…
Have you optimized your sterilization tech? You want to be as gentle as possible without losing sterilization.
What are you using for osmoticum? Something less edible than sucrose can help when you’ve dialed sterilization down to a knifes edge.
If you’ve got hundreds or thousands of seeds from each strain, you’re at least in a place where you can actually TRY things.
First things I would try are 1/2 strength versions of published meristem culture media. MS and ??
Then I’d play with osmoticum. Both level and constituents.
The gelling agent can make a difference. Try a couple.
Then hormones. Shoot inducing/root inducing/callus inducing/gibberellin/none.
Without knowing what you’ve tried and how successful/unsuccessful you’ve been, or how far you are from your goal, it’s difficult to go beyond that.
For tissue culture in general, making progress is often about the right person wielding the scalpel & tweezers. Either they do less damage, or they recognize subtle differences in cell types/organization. If you’re dissecting your seeds, or even just removing the seed coat, this may apply.
I get why you’re obsfucating, but why then would you expect anyone who’s actually solved it to share?
There have been enough studies published to know my way about this and sterilization isn’t an issue here. I have honed studies from protoplast to callus, and I am simply seeking input on straight from embryo
thank you for your input.
We’ll see if there is another interested contributor
When embryo culture fails, getting ANY cells to play is the next trick…”rescuing” the embryo requires that most of it be viable.
Culturing any viable cells from older material that doesn’t respond well to “rescue” is probably closer to going from protoplasts (which sucks more, being alone without a cell wall, or being surrounded by your dead buddies?!?)
Although you should have the remains of two meristems with pluripotent cells…primed to respond differently…which is where playing with the hormone balance (shoot vs root vs callus first) comes in.
If you’ve got the various organogenic media at your fingertips, dissecting some embryos or even cellulase treating and gently making “blobs” may help with the recalcitrant line(s).
I made some seeds in 2005 and they are popping near 100 percent but do come up weak and stunted. im not taking proper care either but most are still alive. Store your seeds in the fridge.
