Arabidopsis protoplasts - can't regenerate plants from protoplasts (Aug/12/2008 )

Hi everyone

I am having huge problems with regenerating plants from Arabidopsis mesophyll protoplasts. I have searched far and wide for a recent protocol to no avail, the method I am currently using is Mathur and Koncz method from the Arabidopsis protocols book. I can isolate excellent quality protoplasts, but embedding them in alginate and regenerating plants has so far proved to be impossible. Does anyone have any experience and tips to save my rapidly departing sanity?

Cheers
Mel

See:

A. Dovzhenko, C. Dal Bosco, J. Meurer, and H. U. Koop (2003) Efficient regeneration from cotyledon protoplasts in Arabidopsis thaliana. Protoplasma 222: 107–111.

..... Culture conditions and media compositions were optimised for the development of protoplasts embedded in thin alginate
layers....................... (from the abstract)

Obviously cotyledon protoplasts are different from mesophyll protoplasts, but you might check the paper anyway.

Good luck,

Christian