Ammonia-oxidizing archaea and comammox Nitrospira clade B as freeze–thaw resistant nitrifiers in wetland soils

Wetland ecosystems are hotspots for nitrogen cycling, while the community dynamics of nitrification-driving microbes include ammonia-oxidizing archaea (AOA), ammonia-oxidizing bacteria (AOB), complete ammonia oxidizer (comammox) Nitrospira, and nitrite-oxidizing bacteria were not well-studied. In this study, we investigated the nitrification process in typical inland and coastal wetlands in northern China and found a high diversity of ammonia oxidizers. To simulate the freezing-thawing state under natural environment, the soil samples were first frozen at −20 °C for two months, and then gradually thawed at 4 °C. Further microcosm incubation combined with ¹³C-DNA-stable isotope probing analyses showed that after the freeze–thaw cycle, only the activities of AOA and comammox Nitrospira clade B recovered. The results indicated that these two groups of nitrifiers were more freeze–thaw resistant, which is in line with literature showing that AOA and comammox Nitrospira clade B were relatively more abundant than their counterparts in low-temperature environments. We tested, for the first time, the cold-shock resilience of nitrifying microbes in wetland soils and spotted candidates driving nitrification under fluctuating temperatures.