CELL DIVISION IN ISOLATED SINGLE PLANT CELLS IN VITRO

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Abstract
A method has been devised for culturing single isolated cells derived from a friable pea-root callus tissue, under conditions which allow cell division and the initial stages of callus tissue development to occur. An agar drop of nutrient medium on a double cover slip is inoculated with a small piece of callus tissue, sealed over a hollow-ground slide, and incubated for 2-4 days at 25[degree] C. The culture is opened, and single cells, isolated in a micropipette with a deFonbrune micro-manipulator from a cell suspension, are placed on the agar medium in the peripheral area of the drop surrounding the callus "nurse." The slide is resealed and incubated in the dark at 25[degree]C. Microscopic observation of these cells can be readily made at a magnification of 400X. Isolated mature vacuolated single cells can divide under these conditions. Of the cells which were viable 1 week after isolation, about 1% had divided; in some, division was not followed by enlargement; in others, division was followed by cell enlargement and then by further divisions. Although no independent callus-tissue clones have yet been established by this method of culture, it provides conclusive evidence that isolated single mature plant cells are capable of dividing in vitro.