Extensive stalk elongation in Caulobacter and Asticcacaulis can be obtained in a defined medium by limiting the concentration of phosphate. Caulobacter cells which were initiating stalk formation were labeled with tritiated glucose. After removal of exogenous tritiated material, the cells were subjected to phosphate limitation while stalk elongation occurred. The location of tritiated material in the elongated stalks as detected by radioautographic techniques allowed identification of the site of stalk development. The labeling pattern obtained was consistent with the hypothesis that the materials of the stalk are synthesized at the juncture of the stalk with the cell. Complementary labeling experiments with Caulobacter and Asticcacaulis confirmed this result. In spheroplasts of C. crescentus prepared by treatment with lysozyme, the stalks lost their normal rigid outline after several minutes of exposure to the enzyme, indicating that the rigid layer of the cell wall attacked by lysozyme is present in the stalk. In spheroplasts of growing cells induced with penicillin, the stalks did not appear to be affected, indicating that the stalk wall is a relatively inert, nongrowing structure. The morphogenetic implications of these findings are discussed.