Characterization of the heat-shock response in spinach (Spinacia oleracea L.)

Abstract

Spinach (Spinacia oleracea L. cultivar Longstanding Bloomsdale) grown at 20 °C was subjected to a range of rapid thermal shifts as high as 42 °C. There was a decrease in the level of protein synthesis following heat-shock treatments above 34 °C as indicated by the level of incorporation of L-[35S]methionine. In vivo labelled polypeptides and in vitro translation products of RNA isolated from leaf tissue and analyzed using one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis and fluorography, indicated that the temperature of induction of all 15 heat-shock proteins in the 20 °C grown plants was 36 °C. In addition, heat-shock RNA was coordinately expressed and the translation of heat-shock proteins was noncoordinate with respect to temperature. Treatment with cycloheximide and with chloramphenicol demonstrated that heat-shock protein synthesis in spinach was restricted to cytosolic ribosomes. Synthesis of some low molecular weight heat-shock proteins were insensitive to actinomycin D, suggesting greater stability of these heat-shock RNAs. The heat-shock polypeptide profile of plants grown at 10 °C was similar to that of plants grown at 20 °C, with 14 heat-shock proteins being induced at 36 °C. The growth temperature did not influence the final array of heat-shock proteins synthesized nor alter the temperature of induction of the heat-shock response.Key words: heat-shock response, heat-shock proteins, Spinacia oleracea.