Procedures for forming and regenerating protoplasts of four Frankia strains are described. Cells obtained from growth medium containing 0.1% glycine were digested with lysozyme (250 μg/ml) in a medium containing 0.5 M sucrose, 5.0 mM CaCl2, and 5.0 mM MgCl2. Protoplasts were formed during 15 to 120 min of digestion at 25°C. Optimum conditions for protoplast regeneration involved placing protoplasts on a layer of complex growth medium containing 0.3 M sucrose, 5.0 mM CaCl2, and 5.0 mM MgCl2 which was overlaid with a layer of 0.8% low-melting-point agarose containing 0.5 M sucrose, 5.0 mM MgCl2, and 5.0 mM CaCl2. The maximum regeneration efficiency was 36.9% for strain CpI1, 1.3% for strain ACN1AG, 27% for strain EAN1pec, and 20% for strain EuI1c.
Molecular, Cellular and Biomedical Sciences
Applied and Environmental Microbiology
American Society for Microbiology
Tisa, L.S. and J.C. Ensign. 1987. Formation and regeneration of protoplasts of the actinorhizal nitrogen-fixing actinomycete Frankia. Appl. Environ. Microbiol. 53:53-56.
© 1987, American Society for Microbiology