The responses of denitrifiers and denitrification ability to dissolved oxygen (DO) concent in different layers of surface lake sediments are still poorly understood. Here, the optimal denitrification condition was constructed based on response surface methodology (RSM) to analyze the denitrification characteristics of surface sediments. The aerobic zone (AEZ), hypoxic zone (HYZ), up-anoxic zone (ANZ-1) and sub-anoxic zone (ANZ-2) were partitioned based on the oxygen contents, and sediments were collected using a customized-designed sub-millimeter scale sampling device. Integrated real-time quantitative PCR, Illumina Miseq-based sequencing and denitrifying enzyme activities analysis revealed that denitrification characteristics varied among different DO layers. Among the four layers, the DNA abundance and RNA expression levels of norB, nirS and nosZ were the highest at the aerobic layer, hypoxic layer and up-axoic layer, respectively. The hypoxia and up-anaerobic layer were the active nitrogen removal layers, since these two layers displayed the highest DNA abundance, RNA expression level and enzyme activities of denitrification functional genes. The abundance of major denitrifying bacteria showed significant differences among layers, with Azoarcus, Pseudogulbenkiania and Rhizobium identified as the main nirS, nirK and nosZ-based denitrifiers. Pearson’s correlation revealed that the response of denitrifiers to environmental factors differed greatly among DO layers. Furthermore, napA showed higher DNA abundance and RNA expression level in the aerobic and hypoxic layers than anaerobic layers, indicating that aerobic denitrifiers might play important roles at these layers.

Microcystis is one of the most common bloom-forming cyanobacteria in freshwater ecosystems throughout the world. However, the underlying life history mechanism and distinct temporal dynamics (inter- and intra-annual) of Microcystis populations in different geographical locations and lakes remain unclear but is critical information needed for the development of robust prediction, prevention, and management strategies. Perennial observations indicate that temperature may be the key factor driving differences in the overwintering strategy. This study quantitatively compared the overwintering abilities of Microcystis aeruginosa (Ma) in both the water column and sediments under a gradient of overwintering water temperatures (i.e., 4, 8, and 12 °C) using the death and proliferation rates of Ma. The results show that the dynamics of the Microcystis overwintering strategy were significantly affected by water temperatures. At 4 and 8 °C, Ma mainly overwintered in sediments and disappeared from the water column after exposure to low temperatures for a long duration, although some Microcystis cells can overwinter in the water column for short durations at low temperatures. At 12 °C, most Ma can overwinter in the water column. Rising temperatures promoted the proliferation of pelagic Ma but accelerated the death of benthic Ma. With warmer winter temperatures, pelagic Microcystis might become the primary inoculum sources in the spring. Our study highlights the overwintering strategy flexibility in explaining temporal dynamics differences of Microcystis among in geographical locations and should be considered in the context of global warming.