Thompson J, Costerton J W, MacLeon R A
J Bacteriol. 1970 Jun;102(3):843-54. doi: 10.1128/jb.102.3.843-854.1970.
When cells of a marine pseudomonad were washed with a solution consisting of 0.3 m NaCl, 0.05 m MgSO(4), and 0.01 m KCl (complete salts), they maintained their normal morphology. When washed with a solution of 0.05 m MgSO(4), they became plasmolyzed as indicated by both phase and electron microscopy. Suspensions of cells washed with 0.05 m MgSO(4) showed an increase in optical density (OD) when 0.3 m NaCl was added, and this was followed by a decrease in OD upon the further addition of 0.01 m KCl. Salts of other monovalent cations were not effective in replacing K(+) in producing the OD decrease. Phase-contrast microscopy revealed that the increase in OD was accompanied by a decrease in cell size, and the decrease in OD, by an increase in the cell size. Both phase and electron microscopy showed that the K(+)-dependent decrease in OD was accompanied by deplasmolysis of the cells. Na(+) was required in the suspending medium in addition to K(+) to obtain deplasmolysis. The intracellular K(+) concentration in cells which had been washed with complete salts and which had retained their normal morphology was found to be 0.290 m. In cells plasmolyzed by washing with 0.05 m MgSO(4), the intracellular K(+) concentration was 0.004 m. Deplasmolyzed cells contained 0.330 m K(+). The membrane profile of plasmolyzed cells was retained when protoplasts were formed. The protoplasts became spherical if incubated in a solution permitting the deplasmolysis of the parent cells. The evidence obtained indicates that plasmolysis and deplasmolysis under the conditions described was due to the loss and gain, respectively, of K(+) by the cells. The effect of Na(+) could be ascribed to its capacity to control the porosity of the cytoplasmic membrane of this organism.
当用含有0.3m NaCl、0.05m MgSO₄和0.01m KCl的溶液(完全盐溶液)洗涤海洋假单胞菌的细胞时,它们保持正常形态。当用0.05m MgSO₄溶液洗涤时,相差显微镜和电子显微镜均显示细胞发生了质壁分离。用0.05m MgSO₄洗涤的细胞悬液在加入0.3m NaCl时光密度(OD)增加,进一步加入0.01m KCl后OD降低。其他单价阳离子的盐在替代K⁺以产生OD降低方面无效。相差显微镜显示,OD增加伴随着细胞大小减小,OD降低伴随着细胞大小增加。相差显微镜和电子显微镜均显示,依赖K⁺的OD降低伴随着细胞的质壁分离复原。除了K⁺外,悬浮介质中还需要Na⁺才能实现质壁分离复原。用完全盐溶液洗涤并保持正常形态的细胞内K⁺浓度为0.290m。用0.05m MgSO₄洗涤而发生质壁分离的细胞内K⁺浓度为0.004m。质壁分离复原的细胞含有0.330m K⁺。形成原生质体时,质壁分离细胞的膜形态得以保留。如果在允许母细胞质壁分离复原的溶液中孵育,原生质体会变成球形。所获得的证据表明,在所描述的条件下,质壁分离和质壁分离复原分别是由于细胞K⁺的丢失和获得。Na⁺的作用可归因于其控制该生物体细胞质膜孔隙率的能力。