Impact of grazing on shaping abundance and composition of active methanotrophs and methane oxidation activity in a grassland soil

The effect of grazing on the abundance, composition, and methane (CH4) uptake of methanotrophs in grasslands has been well documented in the past few decades, but the dominant communities of active methanotrophs responsible for CH4 oxidation activity in grazed soils are still poorly understood. In this study, we characterized the metabolically active, aerobic methanotrophs in grasslands with three different levels of grazing (light, medium, and heavy) by combining DNA-stable isotope probing (SIP) and quantitative PCR (qPCR) for methane monooxygenase (pmoA) gene– and 16S rRNA gene–based amplicon sequencing. The CH4 oxidation potential was as low as 0.51 μmol g dry weight−1 day−1 in the ungrazed control, while it decreased as grazing intensity increased in grazed fields, ranging from 2.25 μmol g dry weight−1 day−1 in light grazed fields to 1.59 in heavily grazed fields. Increased CH4 oxidation activity was paralleled by twofold increases in abundance of pmoA genes and relative abundance of methanotroph-affiliated 16S rRNA genes in the total microbial community in grazed soils. SIP and sequencing revealed that the genera Methylobacter and Methylosarcina (type I; Gammaproteobacteria) and Methylocystis (type II; Alphaproteobacteria) were active methanotrophs responsible for CH4 oxidation in grazed soils. Light and intermediate grazing stimulated the growth and activity of methanotrophs, while heavy grazing decreased the abundance and diversity of the active methanotrophs in the typical steppe. Redundancy and correlation analysis further indicated that the variation of bulk density and soil C and N induced by grazing determined the abundance, diversity of active methanotrophs, and methane oxidation activity in the long-term grazed grassland soil.