Creation of novel nitrogen-fixing actinomycetes by protoplast fusion of Frankia with streptomyces.

Protoplast fusion was used for the creation of a novel actinomycete capable of fixing atmospheric nitrogen. Protoplasts of Streptomyces griseofuscus, a fast-growing actinomycete, and Frankia, a slow-growing actinomycete which fixes atmospheric nitrogen in culture and in symbiotic association with alders, were allowed to fuse and regenerate on media without supplied nitrogen. Colonies which regenerated acquired the fast-growing characteristic of Streptomyces and the ability to grow on nitrogen-deficient media from Frankia. These colonies resembled Streptomyces in their morphology and fixed atmospheric nitrogen in culture. They contained both the parent Streptomyces DNA sequences and the Frankia DNA sequences homologous to nif structural genes HDK of K. pneumoniae. In addition to in vitro nitrogen-fixing capacity, one out of 20 colonies also formed nitrogen-fixing root nodules on Alnus rubra, the host plant for the Frankia strain. Examination of the root nodules induced by the hybrids showed only the presence of hyphae-like structures. The typical vesicle-like structures present in Frankia were absent.