Arabidopsis thaliana: protocol for plant regeneration from protoplasts

We report a protocol for plant regeneration from leaf protoplasts of Arabidopsis thaliana ecotype Zürich. The protocol has been established in 1988 and has since been in routine use in our laboratory. Whereas recovery of proliferating protoplast-derived clones is routine, the success in plant regeneration from protoplast-derived clones is highly variable. In the hands of one of us (H.K.) average shoot regeneration frequency (% of calli regenerating at least one shoot) was ca. 60% and average plant regeneration frequency (% of calli yielding fertile plants in soil) was ca. 40%.