DIURNAL-VARIATION IN HEAT TOLERANCE AND HEAT-SHOCK PROTEIN EXPRESSION IN BLACK SPRUCE (PICEA-MARIANA)

Experiments were conducted using black spruce (Picea mariana (Mill.) B.S.P.) seedlings to evaluate diurnal variability in heat tolerance of roots and shoots and in levels of root heat shock proteins (HSPs). Shoot heat tolerance was significantly lower in the morning than in the afternoon. Similarly, root systems exposed to high temperatures in the morning suffered significantly more damage than those exposed in the afternoon. Root HSPs were identified using an antibody specific for the constitutive HSP73 and inducible HSP72. Both the constitutive levels of root HSPs and the levels of root HSPs following heat shock varied diurnally, and in different patterns for different protein fractions. The levels of soluble and nuclear root HSP72/73 were enhanced by heat shock, both in the morning and afternoon. In comparison, HSP levels in the root mitochondrial protein fractions increased after heat shock in the morning, but decreased after heat shock in the afternoon. HSPs in the root microsomal protein fraction declined following both morning and afternoon heat shock. The higher afternoon levels of root heat tolerance were associated with the greater constitutive levels of HSP73 in the mitochondrial and microsomal root protein fractions in the afternoon. To examine the effects of the shoot on diurnal variation in root system heat tolerance, root heat damage was compared between seedlings with intact shoots versus seedlings with shoots excised either 4-6 h prior to, or immediately before, heat treatment. Regardless of time of day, root systems died when shoots were excised and root systems were treated with high temperatures. In contrast, shoot excision or root heating alone resulted in no root mortality.