Digital recordings of gas-vesicle collapse used to measure turgor pressure and cell-water relations of cyanobacterial cells.
The gas vesicles of the cyanobacterium Microcystis sp. collapse under pressures ranging from 0.65-1.10 MPa, determined from turbidity changes in a pressure nephelometer. In turgid cells, collapse occurs at a lower range of pressures; the difference is equal to the cell turgor pressure. The turgor pressure decreases, however, as gas vesicles collapse; this decrease is minimised by calculating the turgor pressure in samples with few of their gas vesicles collapsed. Previously, pressure and turbidity were measured in discrete steps, using analogue meters, or continuously, using chart recorders: turgor pressure was calculated from the mean or median collapse pressures. We describe modifications allowing continuous digital recording; the output was modelled with polynomial or sigmoid functions, the latter providing the best fit over the full collapse-pressure curve; turgor pressure could then be calculated for any point on the collapse-pressure curve. The shape of the collapse-pressure curve was affected by the rate of pressure rise; curves were similar to those from step-wise methods when the pressure was raised at approximately 4 kPa s(-1). Under a rapid, almost instantaneous, rise in pressure there was a larger initial decrease in turgor and from the subsequent recovery the hydraulic conductivity of the cell surface could be calculated; the new method gave improved measurements of the cell volumetric elastic modulus. Following collapse of half the gas vesicles, cells recovered their full turgor pressure after 3 h. This suggests turgor homeostasis. These methods are applicable to other bacteria with gas vesicles, including Escherichia coli, if it could be genetically modified to express transgenic gas vesicles.
Holland
Daryl P
DP
School of Biological Sciences, University of Bristol, Bristol BS8 1UG, UK. daryl.holland@sci.monash.edu.au
Walsby
Anthony E
AE
D000465:Algorithms; D055119:Elastic Modulus; D004867:Equipment Design; D005740:Gases; D046931:Microcystis / Q000502:physiology*; D008954:Models, Biological; D009391:Nephelometry and Turbidimetry; D009997:Osmotic Pressure; D010539:Permeability; D011312:Pressure; D014617:Vacuoles / Q000502:physiology*
D005740:Gases