I have been looking for a way to transform common vetch and regenerate shoot from transformed cells.
So far, I have observed no de-novo shoot regeneration from vetch tissue (callus, hypocotyl, epicotyl, and root) under the effect of cytokinin or auxin+cytokinin treatment.
I also noticed that If the initial tissue not containing shoot meristem, it barely responded to cytokinin treatment. However, with addition auxin treatment, follow by cytokinin treatment, tissue would eventually develop into a callus (green callus if cultured under light).
If the initial tissue containing shoot meristem, under the cytokinin treatment, shoot multiplication happened vigourously.
Through this observation, I concluded that shoot meristem is the best material to be transformed. However, I need the opinion of other researchers who might have encountered the same issue as I am aware of the transformation efficiency of shoot meristem is extremely low.
Thank you.
Whenever I see regeneration challenges at my lab, I suggest moving more natural as supplements than defined media. I would suggest if coconut milk can be supplemented to vetch shoot meristem. If I get you correctly, Why are you looking for transforming the meristematic cells particularly (DID YOU MEAN GENETIC TRANSFROMATION?).
Vy Nguyen,
My suggestions,
I think shoot meristems are better explants for genetic transformation and regeneration studies as it contains meristematic cells. Shoot meristem can be pre-cultured in cytokinin containing medium (your standardized composition) for few days or can be used directly as explants for transformation studies.
Please find the reference links for more details.
Article Involvement of exogenous polyamines enhances regeneration an...
https://link.springer.com/article/10.1007%2Fs12010-014-1360-x
https://crec.ifas.ufl.edu/media/crecifasufledu/faculty/dutt/Transgenic_plants_from_shoot.pdf
Best wishes.
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