In this activity, we will conduct in-depth microbial community analyses using key sites identified from the 16S rRNA amplicon-based profiling of microbial communities in Activity 2. The DNA extracted in Activity 2 will be shotgun sequenced to generate metagenomes that will be assembled and binned into metagenome assembled genomes (MAGs). These MAGs represent draft genomes for microbes present in the landfill cover soils, allowing for an in-depth metabolic prediction for abundant populations. We will use proteomics to map activities back to specific community members to clarify carbon flux and key players in methane cycling. We will use the community information as a time zero for enrichment experiments in Activity 6.
We anticipate that we will be able to identify keystone organisms active in methane cycling (methanogenesis as well as methanotrophy) in landfill cover soils, including detailed taxonomic placement, metabolic prediction, and proteomic confirmation of these critical roles.