Studies on frost hardiness in Chlorella ellipsoidea II. Effects of inhibitors of RNA and protein synthesis and surfactants on the process of hardening

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Abstract
Chlorella ellipsoidea cells at an intermediate stage in the ripening phase of the cell cycle were brought into contact with inhibitors of RNA and protein synthesis, and surfactants during or after hardening at 3°C. Cycloheximide, actinomycin D and 5-fluorouracil inhibited the development of frost hardiness in the algal cells. In contrast with cycloheximide, chloramphenicol did not cause complete inhibition of hardiness increase, even at a high concentration. These results suggest that protein synthesis on cytoplasmic ribosomes is indispensable for the development of frost hardiness, but protein synthesis on chloroplast ribosomes has little or no involvement in the hardening process. Triton X-100 and sodium dodecyl sulfate inhibited the development of frost hardiness, but sodium cholate and sodium deoxycholate did not. Hardened cells were more susceptible to sodium dodecyl sulfate, in contrast to sodium cholate, sodium deoxycholate and Triton X-100, than unhardened cells. From the results, considerable alterations in both lipids and proteins constituting cellular membranes appear to be involved in the process of hardening.

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