Investigations on the cell volumes of Azotobacter vinelandii by scanning electron microscopy

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Abstract
Previous experiments by other investigators on the DNA content of Azotobacter vinelandii have demonstrated that the DNA content in these cells is several folds higher than that of E. coli. On the basis of this observation, it was hypothesized that A. vinelandii has at least 40 to 80 identical chromosomes per cell. However, the gene dosage analysis in A. vinelandii cells suggested that many genetic operations can be performed in these cells without the constraints expected in a polyploid bacterium. In an attempt to explain this apparent discrepancy, we have done systematic analysis of the relationship between the DNA content and the cell volume of this bacterium. Since a linear correlation is observed between the DNA content and the cell size in many other cell types, we hypothesized that if A. vinelandii is polyploid in nature, it should have a much larger cell volume to accommodate such a large amount of DNA. Our scanning electron microscopic analysis revealed that the cell volume of the vegetative cells of A. vinelandii is about 16 times larger than the cell volume of E. coli. This result is apparently consistent with the concept that the A. vinelandii is a polyploid bacterium. It was also reported that the encysted cells of A. vinelandii contain about 25% of the DNA content of the vegetative cells. This would mean that an encysted cell of A. vinelandii could contain about 10 copies of its chromosome. Since the estimated molecular weight of A. vinelandii chromosome is very similar to that of E. coli chromosome, the DNA content of the encysted cells also should be about 10 times higher than that of E. coli cells. If we assume that the relationship between the DNA content and the cell size is linear, then the encysted cells should have a cell volume larger than that of E. coli and smaller than that of the vegetative cells of A. vinelandii. However our scanning electron microscopic analysis showed that the cell volume of the encysted cells of A. vinelandii is in fact very similar to the cell volume of E. coli.