16S rRNA gene amplicon sequencing of simplified methanotrophic consortium and its initial consortium
Samples for enrichment were collected from rice rhizosphere soil (Shanghai, China), sludge from a wastewater treatment plant (Shanghai Chengtou Sewage Treatment, China), and coalmine soil (0-2 m depth, Liangshuijing Coal Mine, Sichuan, China). Primary enrichment was conducted using CH4 as the sole carbon source in NMS medium to obtain initial methanotrophic consortia (RF, ST, CM). Dilution-to-extinction culturing yielded simplified consortia (SRF, SST, SCM). One replicate of each simplified methanotrophic consortium and its initial consortium collected after 6 days of cultivation under enrichment conditions, was used for 16S rRNA gene amplicon sequencing. Moreover, Six cell suspension samples of SRF and SST consortia (24, 72, 144 h) and SCM (24, 48, 144 h) under typical terrestrial CH4 hotspot concentrations were used for Illumina MiSeq sequencing of 16S rRNA.