THE RELATION OF THE GRAM STAIN TO THE CELL WALL AND THE RIBONUCLEIC ACID CONTENT OF THE CELL

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Abstract
An attempt has been made to stain the cell wall and Gram-positive material of the same cell differentially, the purpose being to determine whether the Gram stain substrate is located within the cell wall. It was possible to apply the Gram stain and then stain for the cell wall by Dyar''s method (Jour. Bact. 53: 498. 1947.). With both bacilli and yeasts it was found that the initial presence of the Gram stain reagents prevent the visible uptake of cell wall stain. It was also possible to remove the Gram stain reagents from a stained cell and simultaneously replace the Gram stain with Congo red of the cell wall stain. These data constitute cytological evidence of the most direct sort for the presence of Gram-staining substrate as a part of the structure of the cell wall. It is probable, however, that Gram-staining material is not limited to the cell wall but also occurs within the cytoplasm. In the case of bacterial spores and ascospores of Schizo-saccharomyces pombe. Gram-staining material was not demonstrated within the spore wall but rather was limited in distribution to the interior of the spore. By shaking yeast with chloroform it is possible to remove about 1/3 of the ribonucleic acid content of the cell without destroying the Gram-positive reaction of the cell. This observation demonstrates the plural cytological nature of the ribonucleic acid of a Gram-positive cell, and raises a question as to the chemical similarity of the ribonucleic acids of the Gram-positive cell.

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