How litter at various stages of decomposition reacts to C and N additions is unclear. Here we used five substrates (litter, fermentation [Oe] layer, bulk soil, and the light fraction [LF] and heavy fraction [HF] of SOM) obtained from sites supporting five plant monocultures (Alnus glutinosa , Quercus robur , Salix caprea , Calamagrostis epigejos , or Picea omorica) with foliage C:N ratios ranging from 17 to 48. These plant-specific communities were experimentally planted on a post-mining heap and had affected the substrates used in this study for 40 years. Soils and other environmental factors were similar among the sites. Substrates were incubated for 3 weeks without nutrient addition or with C (glucose) or N (ammonium nitrate) addition, and microbial respiration was determined weekly. Substrate C:N ratios were determined at the start of the incubation and were highest for litter followed by Oe layer > LF > bulk soil and HF. Foliage C:N ratio was a better indicator of microbial respiration than the substrate C:N ratio, suggesting that the foliage C:N ratio reflected unmeasured leaf properties that determined microbial respiration. Respiration was highest in the litter followed by Oe layer > bulk soil > LF > HF. C addition increased respiration of the bulk soil (+39%), LF (+48%), and HF (+72%). Priming of SOM respiration was therefore higher in substrates with less available C. N significantly increased respiration of litter (+19%) but decreased respiration of bulk soil (−18%). The difference in respiration of HF vs. bulk soil following N addition suggested that, in addition to the stage of decomposition, environmental properties present in bulk soil but absent in HF may cause the reduction in respiration after N addition to bulk soil. Overall, the results indicate that differences in the contents of SOM fractions among soils will affect the responses of those soils to C and N additions. • Initial foliage C:N was a better indicator of microbial respiration than substrate C:N. • C addition increased respiration of the more-decomposed organic matter. • N addition increased respiration of litter but decreased respiration of bulk soil. • Different responses to N addition may be caused by interactions with the soil matrix. [ABSTRACT FROM AUTHOR]