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Plant Physiology 66:835-837 (1980)
© 1980 American Society of Plant Biologists

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Effects of Divalent Cations and Polyethylene Glycol on the Membrane Fluidity of Protoplast 1

Wendy F. Boss and Ralph L. Mott

Department of Botany, North Carolina State University, Raleigh, North Carolina 27650

Calcium is often used to stabilize membranes and enhance membrane fusion. We have used the fatty acid spin label, 5-nitroxy stearic acid to measure fluidity changes in the plasma membrane of carrot suspension culture cell protoplasts in response to divalent cations. Electron spin resonance spectra from spin-labeled protoplasts showed no membrane fluidity changes (as determined by the hyperfine splitting constant, 2Amax) in the presence of Mg from 0 to 10 millimolar or Ca from 0 to 5 millimolar. Protoplasts in 10 millimolar Ca, however, showed a dramatic increase of 5 gauss in 2Amax and evidence of exchange-broadening. The original (control) spectrum was regained by removing bound Ca with a Ca chelator. Polyethylene glycol, which enhances protoplast fusion, did not alter the membrane fluidity in the region of the 5-nitroxy stearic acid probe if added simultaneously with or following 10 millimolar Ca. Pretreatment with polyethylene glycol did, however, inhibit the Ca-induced phase separation. These data on a living system describe membrane structural changes under conditions similar to those used for protoplast fusion.

1 This work was funded under Project No. 03585 of the North Carolina Agricultural Research Service. This paper is No. 6187 in the Journal Series of the North Carolina Agricultural Research Service.




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Copyright © 1980 by the American Society of Plant Biologists