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At maximum inhibition chloramphenicol reduced [35S] methionine incorporation into acid-insoluble materials of sterile protoplasts from young tobacco leaves 5-7 cm in length by 30% compared to 70% by cycloheximide, indicating that 30% of the [35S] methionine became incorporated into chloroplast proteins and 70% into cytoplasmic proteins. [35S] Methionine became incorporated into both the large and small subunits of Fraction I protein, the major soluble protein of chloroplasts. Rifampicin and streptolydigin inhibited [3H] uridine incorporation into the 23 and 16 S rRNAs of chloroplasts to a much greater extent than into the 25 and 18 S cytoplasmic rRNAs. Rifampicin inhibited [35S] metionine incorporation into Fraction I protein after the third hour of incubation; streptolydigin after 2 h, the former evidently preventing initiation of mRNA for the large subunit of Fraction I protein and the latter its elongation. About 2.5 h was required between initiation of the large subunit mRNA synthesis, and appearance of the protein. It was estimated that 45 min is required for transcription of the mRNA which has a half-life of 2 h and that 105 min is required for its translation into approximately 350 amino acids constituting the large subunit monomeric polypeptide. The effect of chloramphenicol, cycloheximide and 2-(4-methyl-2,6-dinitroanaline)-N-methyl propionamide, the latter an inhibitor of protein initiation by 80 S ribosomes, on kinetics of Fraction I protein synthesis indicate that protoplasts contain a pool of small subunit polypeptides and that 30 min is required to polymerize the approximately 100 amino acids constituting the primary structure.

Ferredoxin was purified from 10 species of Nicotiana and spinach leaves. Fingerprints showed all to contain five major tryptic peptides. Some of the spinach peptides were different in RF and mobility from the Nicotiana peptides, but none of the Nicotiana ferredoxins had peptides which could distinguish one species of ferredoxin from another. Electrofocusing S-carbaminomethylcysteinyl ferredoxins showed spinach ferredoxin to have a more acidic and N. glutinosa ferredoxin a slightly more acidic isoelectric point than the other 9 Nicotiana species which were alike. Electro-focusing ferredoxin from the hybrid N. glutinosa female times N. glauca male resolved two bands or isozymes of ferredoxin, one corresponding to N. glutinosa, the other to N. glauca, the code for the latter having come from the DNA in the N. glauca pollen used to form the hybrid plant. N. glutinosa ferredoxin does not contain methionine and is different from N. tabacum and N. glauca ferredoxins which contain methionine. The N. glutinosa female times N. glauca male ferredoxin contained one-half the methionine found in N. glauca ferredoxin, thus confirming that some of the genetic information for ferredoxin in the hybrid was originally contained in the nuclear DNA of N. glauca.