Boron deficiency, manifested as shoot dieback, is a problem in conifer stands growing on soils with high nitrogen availability in Fennoscandia. Earlier observations on Norway spruce (Picea abies L. Karst.) suggest that freezing tolerance is decreased by boron deficiency. Here, the effect of boron fertilization on cold acclimation of Norway spruce was studied in a young stand with initially low boron status two years after fertilization. Buds, stems, needles and roots were collected at five sampling times during cold acclimation and subsequently exposed to series of freezing temperatures. Lethal temperatures of organs were assessed by electrolyte leakage method (EL) and visual scoring of damage (VS). Freezing tolerance of buds was measured also by differential thermal analysis (DTA). The mean boron (B) concentration in needles was 4 mg kg–1 in unfertilized and 21 mg kg–1 in B-fertilized trees while critical level of B deficiency is considered to be 5 mg kg–1. The risk for increased freezing injuries in the low-B trees was not evident since all trees achieved cold hardiness that would be sufficient in central Finland. At two sampling times out of five, shoots or stem of B-fertilized trees were slightly more freezing tolerant than non-fertilized trees. However, the present study does not give strong evidence for the hypothesis that decreased freezing tolerance in B deficiency would be a triggering factor for leader dieback in Norway spruce at the B levels studied.