Methanogenesis and Salt Tolerance Genes of a Novel Halophilic Methanosarcinaceae Metagenome-Assembled Genome from a Former Solar Saltern

Anaerobic archaeal methanogens are key players in the global carbon cycle due to their role in the final stages of organic matter decomposition in anaerobic environments such as wetland sediments. Here we present the first draft metagenome-assembled genome (MAG) sequence of an unclassified Methanosarcinaceae methanogen phylogenetically placed adjacent to the Methanolobus and Methanomethylovorans genera that appears to be a distinct genus and species. The genome is derived from sediments of a hypersaline (97–148 ppt chloride) unrestored industrial saltern that has been observed to be a significant methane source. The source sediment is more saline than previous sources of Methanolobus and Methanomethylovorans. We propose a new genus name, Methanosalis, to house this genome, which we designate with the strain name SBSPR1A. The MAG was binned with CONCOCT and then improved via scaffold extension and reassembly. The genome contains pathways for methylotrophic methanogenesis from trimethylamine and dimethylamine, as well as genes for the synthesis and transport of compatible solutes. Some genes involved in acetoclastic and hydrogenotrophic methanogenesis are present, but those pathways appear incomplete in the genome. The MAG was more abundant in two former industrial salterns than in a nearby reference wetland and a restored wetland, both of which have much lower salinity levels, as well as significantly lower methane emissions than the salterns.

Quantifying functional increases across a large-scale wetland restoration chronosequence

Over 300,000 ha of forested wetlands have undergone restoration within the Mississippi Alluvial Valley region. Restored forest successional stage varies, providing opportunities to document wetland functional increases across a large-scale restoration chronosequence using the Hydrogeomorphic (HGM) approach. Results from >600 restored study sites spanning a 25-year chronosequence indicate that: 1) wetland functional assessment variables increased toward reference conditions; 2) restored wetlands generally follow expected recovery trajectories; and 3) wetland functions display significant improvements across the restoration chronosequence. A functional lag between restored areas and mature reference wetlands persists in most instances. However, a subset of restored sites have attained mature reference wetland conditions in areas approaching or exceeding tree diameter and canopy closure thresholds. Study results highlight the importance of site selection and the benefits of evaluating a suite of wetland functions in order to identify appropriate restoration success milestones and design monitoring programs. For example, wetland functions associated with detention of precipitation (a largely physical process) rapidly increased under post restoration conditions, while improvements in wetland habitat functions (associated with forest establishment and maturation) required additional time. As the wetland science community transitions towards larger scale restoration efforts, effectively quantifying restoration functional improvements will become increasingly important.

Microbial drivers of methane emissions from unrestored industrial salt ponds

Wetlands are important carbon (C) sinks, yet many have been destroyed and converted to other uses over the past few centuries, including industrial salt making. A renewed focus on wetland ecosystem services (e.g., flood control, and habitat) has resulted in numerous restoration efforts whose effect on microbial communities is largely unexplored. We investigated the impact of restoration on microbial community composition, metabolic functional potential, and methane flux by analyzing sediment cores from two unrestored former industrial salt ponds, a restored former industrial salt pond, and a reference wetland. We observed elevated methane emissions from unrestored salt ponds compared to the restored and reference wetlands, which was positively correlated with salinity and sulfate across all samples. 16S rRNA gene amplicon and shotgun metagenomic data revealed that the restored salt pond harbored communities more phylogenetically and functionally similar to the reference wetland than to unrestored ponds. Archaeal methanogenesis genes were positively correlated with methane flux, as were genes encoding enzymes for bacterial methylphosphonate degradation, suggesting methane is generated both from bacterial methylphosphonate degradation and archaeal methanogenesis in these sites. These observations demonstrate that restoration effectively converted industrial salt pond microbial communities back to compositions more similar to reference wetlands and lowered salinities, sulfate concentrations, and methane emissions.