This study examines the genetic variation of Norway spruce (Picea abies) by analyzing ten SSR loci in 180 individuals from nine breeding populations in Northern and Central Sweden. The results reveal high genetic diversity within populations but low differentiation among them. The AMOVA showed that 86% of the total genetic variance was found within individuals, 13% among individuals, and only 1% among populations, reflecting extensive gene flow and high genetic connectivity. This pattern aligns with the species' recolonization history following the last glaciation and ongoing gene flow facilitated by Sweden's continuous landscape. Single-locus tests revealed that most loci did not show significant departures from Hardy-Weinberg Equilibrium (HWE), suggesting random mating and stable population sizes. Positive inbreeding coefficients (F) were observed across all populations, indicating some homozygosity excess, potentially due to null alleles or mating among related individuals. The findings suggest that breeding programs should prioritize controlled cross-pollination, diverse genetic source selection, and strategic planting of seed orchards to ensure long-term sustainability and resilience of spruce populations.