Development and Germination of the Azotobacter Cyst

The fine structure of Azotobacter vinelandii has been studied by means of electron microscopy of ultrathin sections made of the encysting and germinating cells. The organisms were fixed with KMnO(4) and embedded in epoxy resin. On an encystment medium the rod-shaped bacteria begin to assume an almost spherical form and then bark-like exine appears in 1(1/2) to 2 days. The exine thickens and an electron permeable intine forms between it and the shrinking cell body. In 5 days the intine makes up more than half of the cyst volume and begins to show a definite two-layered structure. Meanwhile the peripheral bodies, which may be extensions of the cell membrane of the vegetative cell, disappear as the encystment progresses. The cell wall and membrane of the vegetative cell remain demonstrable as the confining structure of the shrinking central body of the mature cyst. In this central body lipoidal globules appear together with aggregations of nuclear material. Cyst germination begins with an increase in the size of the central body at the expense of the intine. The nuclear aggregations become more diffuse and the lipoidal globules disappear. The exine may be pushed outward and the bark-like fragments separate as the emerging vegetative cell develops. Invagination of the cell wall and membrane may occur at this stage leading to cell division. Empty exines remain as horseshoe-shaped structures.