Protoplast Fusion in Streptomyces: Conditions for Efficient Genetic Recombination and Cell Regeneration

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Abstract
Protoplasts from 4 different species of Streptomyces regenerated cells efficiently in hypertonic soft agar medium overlaid on partially dehydrated medium. The efficiencies of regeneration were strongly dependent upon the incubation temperatures for cell growth and for protoplast regeneration. Cell growth temperatures (before protoplast formation) required for efficient protoplast regeneration varied from species to species and did not necessarily correlate with the optimum temperatures for protoplast regeneration. Under the best conditions, protoplasts from all 4 spp. were able to regenerate viable cells at nearly 100% efficiency and also formed confluent lawns of mycelia when plated in high concentrations. The temperatures for cell growth and protoplast regeneration also affected the frequencies of genetic recombinants obtained by protoplast fusion in S. fradiae and highest recombinant frequencies were obtained under conditions which favored efficient protoplast regeneration. With the modified procedure described, maximum frequencies of genetic recombinants were obtained by treating parental protoplasts with 40-60% polyethylene glycol 1000.

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