The study focused on the effects of gap size on natural regeneration of European beech (Fagus sylvatica L.) and Norway spruce (Picea abies L.) and micro-environmental soil conditions in gaps of different sizes under temperate mixed forest in the Czech Republic. Six gaps comprising two for small (≥ 200 m<sup>2</sup>), medium (≥ 500 m<sup>2</sup>) and big (≥ 900 m<sup>2</sup>) each were selected. Ten circular 1 m<sup>2</sup> subsampling plots were established at 2 m intervals along individual North-South-East-West transects, including one at the gap centre. Regeneration was monitored in 2014 and repeatedly in 2019. Soil conditions were only measured in 2019. Gap size was found to be a significant parameter for European beech natural regeneration in 2014. Besides, the quick occupation of European beech in gaps at natural beech zone provoked its prolific regeneration compared to Norway spruce in 2014. However, in 2019 the recent threat of weather variabilities was responsible for the general abysmal growth performance of natural regeneration. Division of gap microsites into different within-gap positions based on prevailing light or shade conditions was helpful in assessing the significant variations of soil conditions within-gap positions and among gap sizes. Soil temperature and moisture significantly influenced the regeneration of European beech and Norway spruce, respectively.
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