The measurement of water relations of plant cell culture. I. Water release in response to centrifuge-induced water potentials

Water loss by cell suspensions during centrifugation is well defined by simple physical principles. The major factors affecting water release during centrifugation are: duration of centrifogation, depth of the cell mass, density of cells, relative centripetal acceleration and centripetal force. Water release during centrifugation was best described by an exponential decay process with a decay constant that increases with acceleration from 0.31 ± 0.01 to 0.66 ± 0.12 min^?1 (mean ± SE) between 4 825 and 19 300 m s^?2, respectively. The cell mass relative water content (RWC) at equilibrium was not a function of rate of water loss and was constant for each acceleration. A centripetal force was generated by the mass of the cells being accelerated away from the axis of rotation. This force generated a pressure that removed some of the cell wall and symplast water, by compression at contact points between the cells and by compression of the cytoplasm. Pressure induced by centripetal forces ranging from ?0.02 to ?0.23 MPa gave a linear relationship (r² > 0.99) between force and RWC. The slope (0.900 MPa) was proportional to the cell wall modulus of elasticity (±). and the intercept was interpreted to give the mass of the cells at full turgor without interstitial water (RWC=1). This interpretation is supported by the findings, of two independent experiments. Centrifuged cells suspended at 100% relative humidity for over 48 h reached the same water content as predicted by the intercept. Interstitial water was labelled with solutions of polyethylene glycol (PEG. M_r 8 000), the diameter of which was too large to enter the pores of plant cell walls. Centripetal accelerations greater than 10 900 m s^?2 removed PEG-labelled water to levels below 0.9% of cell water content. Removal of interstitial water and other loosely bound water provided a convenient method for determination of growth, RWC and ±. The centrifugal methods provide the foundation for new quantitative methods for cell culture water relations analyses.