Vallisneria natans decreased CH4 fluxes in wetlands: Interactions among plant physiological status, nutrients and epiphytic bacterial community.

Submerged macrophytes provide niches for epiphytic microbes (including aerobic methanotrophs) growth. However, little is known about the impacts of submerged macrophytes growth status and nutrients loadings on methanotroph community and methane release in wetlands. In the present study, methane fluxes, bacterial and methanotroph community in epiphytic biofilm, and environmental parameters were investigated during Vallisneria natans senescence in wetlands under low (VnL) and high (VnH) nutrients for seven weeks. Relative conductivity and concentration of H2O2, total chlorophyll and malondialdehyde were higher in leaves of V. natans in VnH than VnL at the same sampling time. Nutrients loading increased methane fluxes in treatments with or without (Control) macrophytes, while healthy V. natans plants reduced the methane flux and nutrients concentration in water columns. CH4 fluxes were positively correlated to temperature and COD (p < 0.05). Methane oxidation rates were 3.04-31.68 µmol methane mg-1 fresh weight of V. natans leaves - epiphytic biofilm within 1 h. Proteobacteria, Cyanobacteria, Bacteroidetes, Verrucomicrobia, Planctomycetes, Actinobacteria and Acidobacteria were dominant phylum in all epiphytic biofilms. The mean abundances of pmoA/16S rRNA were higher in VnL than VnH. According to Illumina sequencing results of pmoA gene, γ-proteobacteria and α-proteobacteria were the dominant methanotroph class in epiphytic biofilm from VnH and VnL, respectively. Among seven detected methanotrophic genera, Methylomonas was significantly higher in VnH than VnL. Network analysis revealed that there were much closer relationships between the environmental parameters and epiphytic bacterial community in VnH than in VnL. COD and MDA were negatively correlated with Methyloglobulus, Methylosarcina, Methylobacter and Methylocystis, but positively correlated with Methylomonas and Methylosinus. This study highlights that methanotrophs in epiphytic biofilm play important roles in methane-oxidizing, which can be affected by plant physiological status and environmental parameters.

Characterization and co-occurrence of microbial community in epiphytic biofilms and surface sediments of wetlands with submersed macrophytes.

Microbes in epiphytic biofilms and surface sediments play crucial roles in the biogeochemical cycles in wetlands. However, little is known about the compositions of microbial community in wetlands dominated with submersed macrophytes. In this study, bacterial and eukaryotic community in epiphytic biofilms and surface sediments were investigated in wetlands with artificial plants and Myriophyllum verticillatum from September (~27 °C) to January (~9 °C). A total of 30 (including 13 bacterial and 17 eukaryotic) and 34 (including 14 bacterial and 20 eukaryotic) phyla were detected in epiphytic biofilms and sediments, respectively. Microbial community in epiphytic biofilms shifted with decreasing temperature, and biofilms on M. verticillatum were generally similar to those on artificial plants. Though the OTUs and Shannon values were significantly higher in sediments than epiphytic biofilms (p < 0.05), numbers of strongly correlated edges detected in biofilms (64 nodes with 182 edges) were at least three times of those in sediments (40 nodes with 57 edges) as revealed by co-occurrence networks analysis (|r| > 0.7, p < 0.05). These data suggest that there were complex interactions among microbes in epiphytic biofilms than sediments. Positive relationships among microbes revealed the predation, symbiosis, parasitism relationships and the collective degradation of organic matter, while negative ones may be ascribed to their different lifestyles. These results highlight that artificial plants play a similar role as submersed macrophytes as microbial carriers and can be potentially used an alternative substitutes to submersed macrophytes in wetlands.