Summary: Regeneration of protoplasts to bacilli was attempted in several strains of closely related to 168. On DM3 and similar media using succinate as osmotic support, only 168 and ATCC 15245 were able to regenerate. Media containing mannitol as osmotic support, and agar as gelling agent gave rise to L-form colonies with NCTC 6346. Many of the L-form colonies were able to regenerate to the bacillary form when plated on the mannitol medium solidified with gelatin. All of the species tested were able to regenerate on the latter medium at rates sufficient to allow protoplast transformation and fusion experiments.


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