Protein synthesis has been implicated in the cold-hardening process. Ribosomes from cold hardy and nonhardy black locust (Robinia pseudoacacia L.) seedlings were compared to determine if cold acclimation is related to alteration of ribosomal structure. Ribosomal structure, as indicated by thermal melting profiles, appears to be altered during induction of hardiness. Two-dimensional polyacrylamide gel electrophoresis of ribosomal proteins indicates at least 17 proteins from hardy seedlings that are different from those of nonhardy seedlings. These different proteins may be partially responsible for the different thermal melting profiles observed.Plants capable of developing freezing resistance demonstrate altered metabolism during low temperature acclimation (cold hardening) (23). There have been many suggestions that protein synthesis plays a role in cold hardening (1, 4, 9, 13, 23). Among the many changes found during induction of cold hardiness are those which occur in RNA (13, 26) and protein (3, 4,9,13,26) metabolism. This study was undertaken to examine the ribosome as related to induction of cold hardiness. The objective was to determine if ribosomal structure is altered during induction of hardiness.
MATERLALS AND METHODSPlant Material. Black locust seedlings (Robinia pseudoacacia L.) were grown in heat-sterilized vermiculite. Moisture was maintained relatively constant. Two-month seedlings were cold hardened as outlined in Table I. At specific points in the coldhardening regime, seedlings were placed into a programmed freezer and frozen at a rate of 6 C/hr. Samples were removed every 4 C starting at -4 and allowed to thaw overnight at 5 C. Stems were then harvested, placed into a mist chamber with a 15-hr photoperiod, allowed to recover for 7 days and visually evaluated for survival. Seedlings were considered uninjured if their bark remained green and tightly bound to the stem. The bark of injured seedlings became brown and loose. Stems were assigned a value of 1 if uninjured, 0.5 if injured with 50% or more of the stem alive and 0 if dead or more than 50% injured. Values were totaled and divided by the total number of stems to determine percent survival (31).