Should I plate them to plain media to let the shoot develop and grow regardless of the callus? Or if I plate it to a cytokinin, will the callus produce additional shoots despite already being on a plant?
If I understand correctly you have a seedling with callus growing from the hypocotyl. I would cut the callus from the hypocotyl and plate it by itself. Then you can keep growing the seedling in soil. I would grow the callus on plain media. MS basal salts, phytoagar, 3% sucrose, no hormones.
It may be tempting to grow callus on "callus media" but that is for inducing new callus from differentiated tissue. Typically contains 2,4-D and some other growth regulators like 6-benzylaminopurine, indole-3-acetic-acid, etc depending on the species. This will often inhibit growth of callus even though it induces the formation of callus in the first place.
Once you have as much callus as your heart desires, transfer to plates containing shoot inducing hormones, then to plates with root inducing hormones. The exact recipe will vary by species. Some guess work may be required for less well studied species.
> Or if I plate it to a cytokinin, will the callus produce additional
> shoots despite already being on a plant?
> Or if I plate it to a cytokinin, will the callus produce additional
> shoots despite already being on a plant?
That *might* work but it will also mess with the seedling. So I would grow the seedling and callus separately if you want the seedling to continue growling normally.
-Cory
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