regeneration from protoplasts

[Giovanni Sena]

Hello,
I am trying to induce calli from root-derived protoplasts and 
subsequently to regenerate whole plants from such calli.
I am following the protocol by Mathur J. and Koncz C. published in 
the vol. 82 ("Arabidopsis Protocols") of the Methods in Molecular 
Biology series (1998).
Unfortunately, I am getting very low efficiency in callus formation 
from such protoplasts liquid culture (not embedded).

Can anybody suggest me how to improve this protocol, or what might be 
common mistakes I could be doing?

Any comment/advice on regeneration from protoplasts is more than 
welcome and appreciated !

Thank you so much for your help!



Giovanni Sena