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Plant Physiology 57:290-296 (1976)
© 1976 American Society of Plant Biologists

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Articles

Osmotic Shrinkage as a Factor in Freezing Injury in Plant Tissue Cultures

Leigh E. Towill1 and Peter Mazur

a The University of Tennessee—Oak Ridge Graduate School of Biomedical Sciences and Biology Division, Oak Ridge National Laboratory, 2 Oak Ridge, Tennessee 37830

Haplopappus gracilis and Acer saccharum tissue culture cells are extremely sensitive to freezing injury, and exhibit a decrease in survival from 98% at –1 C to 4% at –3 C (Haplopappus) and 92% at –3 C to 13% at –5 C (Acer) when suspended in distilled H2O, seeded at –1 C, and then cooled by 0.1 C/minute. Similar results are obtained when cells are suspended in growth medium. The extent of shrinkage of cells during freezing can be duplicated by exposure of the cells to plasmolyzing solutions of nonpenetrating substances ({Delta} Tf = 1.86 {varphi}vm). Solutions of sucrose and glycerol that produce extensive plasmolysis cause a decrease in survival within 3 to 5 minutes at room temperature, and the higher the molality to which the cell is exposed the greater the injury. Also, the rate of rehydration of the plasmolyzed cell and of the frozen cell affects its survival, with the slower rate being more beneficial. The close correlation between the decrease in survival at subzero temperatures and the decrease in survival when cells are placed in solutions having osmolalities, which could produce the same extent of shrinkage as these killing temperatures, suggests that this shrinkage is related to freezing injury in tissue culture cells.

1 Postdoctoral investigator supported by Subcontract 3322 from the Biology Division of Oak Ridge National Laboratory to the University of Tennessee.

2 Operated by Union Carbide Corporation under contract with the United States Atomic Energy Commission.




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Copyright © 1976 by the American Society of Plant Biologists