Abstract Moss‐associated cyanobacteria nitrogen (N2‐) fixation can contribute to support moss growth and constitutes a major source of new N in boreal forest ecosystems. The biomass of moss‐colonizing cyanobacteria and their N2‐fixation are usually considered linearly correlated. Yet, recent evidence has shown that cyanobacterial biomass and N2‐fixation can be decoupled, suggesting that they are not necessary affected by the same environmental and ecological drivers. Climate and nutrients were reported as affecting moss‐associated N2‐fixation, with equivocal results, whereas drivers of moss cyanobacterial biomass remain unclear. In addition, these drivers are often determined through manipulative experiments (e.g. fertilization and incubation) and remain to be validated with complementary observational studies to help us better understand future impacts of global change on the moss–cyanobacteria symbiosis. We hypothesized that moss‐associated cyanobacterial biomass is controlled in situ by factors affecting bacterial growth, whereas N2‐fixation is controlled by factors affecting enzymatic reactions. Using random forests, Spearman correlations and linear mixed‐effects models, we determined the main drivers of cyanobacterial biomass and N2‐fixation of two feather moss species, which were collected over 3 years along a 1000‐km latitudinal transect in the eastern Canadian boreal forest. We found that temperature, precipitation and phosphorus were the main positive drivers of moss cyanobacterial biomass and that temperature and molybdenum were the main positive drivers of N2‐fixation. Vanadium was a negative driver of N2‐fixation, suggesting the use of alternative nitrogenases by cyanobacteria. Both cyanobacterial biomass and N2‐fixation were strongly influenced by the moss species and were negatively correlated with moss C:N stoichiometry, highlighting the role of N2‐fixation in moss N enrichment. Synthesis. We identified for the first time some environmental drivers of moss‐associated cyanobacterial biomass and showed that they contrast with the drivers of N2‐fixation, which should be considered in further research and confirmed in other experimental settings. This is an important advance in our knowledge of moss–cyanobacteria associations, which would greatly help in better predicting the impacts of global change on this symbiosis and on nitrogen inputs in boreal forest ecosystems.
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