;Moss chloroplasts should prove useful for studying the cyanobacteria-derived system in chloroplasts. To determine the effects of antibiotics that inhibit bacterial peptidoglycan synthesis, the numbers of chloroplasts in treated Physcomitrella patens cells were counted. Ampicillin and D-cycloserine caused a rapid decrease in the number of chloroplasts per cell. Fosfomycin affected half of the cells, while vancomycin affected a few cells. Conversely, bacitracin had no effect. With the decrease in chloroplast number, macrochloroplasts appeared in antibiotic-treated cells. Removal of the antibiotics resulted in the recovery of chloroplast number, suggesting that the decrease in number was directly dependent on the antibiotic treatment. Microscopic observations showed that the decrease in the number of chloroplasts resulted from cell division without chloroplast division. These results suggest that enzymes derived from the bacterial peptidoglycan synthesis pathway are related to moss chloroplast division.

Eleven mutant lines exhibiting decreased numbers of chloroplasts per cell were isolated from 8 800 tagged mutant lines of Physcomitrella patens by microscopic observations. Chloronema subapical cells in wild-type plants had a mean of 48 chloroplasts, whereas chloroplast numbers in subapical cells in mutant lines 215 and 222 decreased to 75 % of that in the wild type. Seven mutant lines - 473, 122, 221, 129, 492, 207, and 138 - had about half as many chloroplasts as the wild type. Mutant line 11 had a few remarkably enlarged chloroplasts, and mutant line 347 had chloroplasts of various sizes. Whereas the cell volume was the same as in the wild type in mutant lines 222, 473, 221, 129, 492, and 207, the cell volume of the other mutants increased. The chloroplast number of leaf cells was the same as that of chloronema cells in each mutant line when gametophores could be formed. Treatment with ampicillin decreased the number of chloroplasts in all mutant lines. Southern hybridization using DNA in tags as probes showed that only one insertion occurred in mutant lines 473 and 221. To determine whether the tagged DNA inserted into the known genes for plastid division, we isolated the PpMinD1, PpMinD2, and PpMinE1 genes. Genomic polymerase chain reaction analysis showed that the PpFtsZ and PpMinD/E genes were not disrupted by the insertion of the tags in mutant lines 11 and 347, respectively.

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