Urbanization reduces resource use efficiency of phytoplankton community by altering the environment and decreasing biodiversity.

Urbanization often exerts multiple effects on aquatic and terrestrial organisms, including changes in biodiversity, species composition and ecosystem functions. However, the impacts of urbanization on river phytoplankton in subtropical urbanizing watersheds remain largely unknown. Here, we explored the effects of urbanization on phytoplankton community structure (i.e., biomass, community composition and diversity) and function (i.e., resource use efficiency) in a subtropical river at watershed scale in southeast China over 6 years. A total of 318 phytoplankton species belonging into 120 genera and 7 phyla were identified from 108 samples. Bacillariophyta biomass showed an increasing trend with increasing urbanization level. The phytoplankton community shifted from Chlorophyta dominance in rural upstream waters to Bacillariophyta dominance in urbanized downstream waters. Furthermore, phytoplankton diversity and resource use efficiency (RUE = phytoplankton biomass/total phosphorus) were significantly decreased with increasing urbanization level from upstream to downstream. Phytoplankton RUE exhibited a significant positive correlation with species richness, but a negative correlation with phytoplankton evenness. The variation in environmental factors (turbidity, total nitrogen, NH4+-N, total phosphorus, PO43--P and percentage urbanized area) was significantly correlated with phytoplankton diversity and RUE. Overall, our results revealed the influence of urbanization on phytoplankton community structure and ecosystem function was due to its altering the environmental conditions. Therefore, human-driven urbanization may play crucial roles in shaping the structure and function of phytoplankton communities in subtropical rivers, and the mechanism of this process can provide important information for freshwater sustainable uses, watershed management and conservation.

Structural and functional variations of phytoplankton communities in the face of multiple disturbances.

The global decline of freshwater biodiversity caused by climate change and human activities are supposed to disrupt ecosystem services related to water quality and alter the structure and function of aquatic communities across space and time, yet the effects of the combination of these factors on plankton community ecosystem has received relatively little attention. This study aimed to explore the impacts of disturbances (e.g. human activity, temperature, precipitation, and water level) on phytoplankton community structure (i.e. community evenness and community composition) and function (i.e. resource use efficiency) in four subtropical reservoirs over 7 years from 2010 to 2016. Our results showed that community turnover (measured as community dissimilarity) was positively related to disturbance frequency, but no significant correlation was found between phytoplankton biodiversity (i.e. evenness) and disturbance frequency. Phytoplankton resource use efficiency (RUE = phytoplankton biomass/ total phosphorus) was increased with a higher frequency of disturbance with an exception of cyanobacteria. The RUE of Cyanobacteria and diatoms showed significantly negative correlations with their community evenness, while the RUE of Chlorophyta exhibited a positive correlation with their community turnover. We suggest that multiple environmental disturbances may play crucial roles in shaping the structure and functioning of plankton communities in subtropical reservoirs, and mechanism of this process can provide key information for freshwater uses, management and conservation.

Microbial one­carbon and nitrogen metabolisms are beneficial to the reservoir recovery after cyanobacterial bloom.

Cyanobacterial blooms have profound effects on the structure and function of plankton communities in inland waters, but few studies have focused on the effects of microbial-based processes in one­carbon and nitrogen cycling on water quality improvement following the bloom. Here, we compared the structure and function of the bacterial community, focusing on microbial one­carbon and nitrogen metabolisms during and after a cyanobacterial Microcystis bloom in a deep subtropical reservoir. Our data showed that microbial one­carbon and nitrogen cycles were closely related to different periods of the bloom, and the changes of functional genes in microbial carbon and nitrogen cycling showed the same consistent trend as that of Methylomonas sp. With the receding of the bloom, the abundance of Methylomonas as well as the functional genes of microbial one­carbon and nitrogen cycling reached the peak and then recovered. Our results indicate that microbial one­carbon and nitrogen metabolisms were beneficial to the recovery of water quality from the cyanobacterial bloom. This study lays a foundation for a deep understanding of the cyanobacterial decomposition mediated by microbes in one­carbon and nitrogen cycles in inland freshwaters.

On the use of chemotaxonomy, a phytoplankton identification and quantification method based on pigment for quick surveys of subtropical reservoirs.

Traditionally, composition and biomass of phytoplankton communities are determined by microscopy, but this method is time-consuming and so does not allow for high-frequency data acquisition across space and time. Pigment-based chemotaxonomy (CHEMTAX) is now widely applied to study of phytoplankton community structure on broader spatial and temporal scales of oceans, but the ability of this approach to provide estimates of phytoplankton assemblage in freshwater ecosystems is yet underdeveloped. To investigate the efficiency of the high-performance liquid chromatography (HPLC)-CHEMTAX in quantifying the different phytoplankton groups in inland freshwater, we present a comparison between phytoplankton pigment analyses by HPLC with CHEMTAX and microscopic counting of phytoplankton samples from four subtropical reservoirs in January and July 2014, respectively. The correlation between pigment and phytoplankton abundance detected by microscopy was stronger than that between pigment and phytoplankton biomass. The published marker pigments and their revised ratios can be used to describe phytoplankton abundances in a mixed community of freshwater phytoplankton, and pigment-based CHEMTAX can successfully describe the overall pattern of phytoplankton community dynamics during different seasons. The use of pigment-based CHEMTAX for quick surveys of phytoplankton communities can be recommended as a useful supplement or alternative tool to microscopy for freshwater ecosystem management.

Precision early detection of invasive and toxic cyanobacteria: A case study of Raphidiopsis raciborskii.

Blooms of the toxic cyanobacterium, Raphidiopsis raciborskii (basionym Cylindrospermopsis raciborskii), are becoming a major environmental issue in freshwater ecosystems globally. Our precision prevention and early detection of R. raciborskii blooms rely upon the accuracy and speed of the monitoring method. A duplex digital PCR (dPCR) monitoring approach was developed and validated to detect the abundance and toxin-producing potential of R. raciborskii simultaneously in both laboratory spiked and environmental samples. Results of dPCR were strongly correlated with traditional real time quantitative PCR (qPCR) and microscopy for both laboratory and environmental samples. However, discrepancies between methods were observed when measuring R. raciborskii at low abundance (1 - 105 cells L - 1), with dPCR showing a higher precision compared to qPCR at low cell concentration. Furthermore, the dPCR assay had the highest detection rate for over two hundred environmental samples especially under low abundance conditions, followed by microscopy and qPCR. dPCR assay had the advantages of simple operation, time-saving, high sensitivity and excellent reproducibility. Therefore, dPCR would be a fast and precise monitoring method for the early warning of toxic bloom-forming cyanobacterial species and assessment of water quality risks, which can improve prediction and prevention of the impacts of harmful cyanobacterial bloom events in inland waters.

Community dynamics of free-living and particle-attached bacteria following a reservoir Microcystis bloom.

The composition of microbial communities can vary at the microspatial scale between free-living (FL) and particle-attached (PA) niches. However, it remains unclear how FL and PA bacterial communities respond to cyanobacterial blooms across water depths. Here, we examined the community dynamics of the FL (0.2-3 µm) and PA (>3 µm) bacterioplankton based on 16S rRNA gene high-throughput sequencing in a subtropical stratified reservoir under Microcystis aeruginosa bloom and non-bloom conditions. Both FL and PA bacterioplankton communities showed different responses in alpha- and beta-diversities to the bloom, suggesting the idea that the responses of bacterial community could depend on lifestyle. Specifically, abundant PA subcommunities showed a greater variation between bloom and non-bloom groups than abundant FL ones. In contrast, rare FL subcommunities exhibited a stronger response to water depth than rare PA ones. Furthermore, the rare taxa exhibited a preference for PA status, shaped and stimulated by the M. aeruginosa bloom. Our analyses also showed that PA bacterial communities were generally more diverse and appeared to be more responsive to routinely measured environmental variables than FL bacteria. Microcystis blooms had a facilitative influence on specific bacteria by mediating the transitions from free-living to particle-attached lifestyles. Altogether, these findings highlight the importance of bacterial lifestyle and abundance in understanding the dynamics of microbial community in cyanobacterial bloom aquatic ecosystem.

Response of the eukaryotic plankton community to the cyanobacterial biomass cycle over 6 years in two subtropical reservoirs.

Although it is widely recognized that cyanobacterial blooms have substantial influence on the plankton community in general, their correlations with the whole community of eukaryotic plankton at longer time scales remain largely unknown. Here, we investigated the temporal dynamics of eukaryotic plankton communities in two subtropical reservoirs over a 6-year period (2010-2015) following one cyanobacterial biomass cycle-the cyanobacterial bloom (middle 2010), cyanobacteria decrease (late 2010-early 2011), non-bloom (2011-2014), cyanobacteria increase, and second bloom (late 2014-2015). The eukaryotic community succession that strongly correlated with this cyanobacterial biomass cycle was divided into four periods, and each period had distinct characteristics in cyanobacterial biomass and environments in both reservoirs. Integrated co-occurrence networks of eukaryotic plankton based on the whole study period revealed that the cyanobacterial biomass had remarkably high network centralities, and the eukaryotic OTUs that had stronger correlations with the cyanobacterial biomass exhibited higher centralities. The integrated networks were also modularly responded to different eukaryotic succession periods, and therefore correlated with the cyanobacterial biomass cycle. Moreover, sub-networks based on the different eukaryotic succession periods indicated that the eukaryotic co-occurrence patterns were not constant but varied largely associating with the cyanobacterial biomass. Based on these long-term observations, our results reveal that the cyanobacterial biomass cycle created distinct niches between persistent bloom, non-bloom, decrease and increase of cyanobacteria, and therefore associated with distinct eukaryotic plankton patterns. Our results have important implications for understanding how complex aquatic plankton communities respond to cyanobacterial blooms under the changing environments.

Distinct patterns and processes of abundant and rare eukaryotic plankton communities following a reservoir cyanobacterial bloom.

Plankton communities normally consist of few abundant and many rare species, yet little is known about the ecological role of rare planktonic eukaryotes. Here we used a 18S ribosomal DNA sequencing approach to investigate the dynamics of rare planktonic eukaryotes, and to explore the co-occurrence patterns of abundant and rare eukaryotic plankton in a subtropical reservoir following a cyanobacterial bloom event. Our results showed that the bloom event significantly altered the eukaryotic plankton community composition and rare plankton diversity without affecting the diversity of abundant plankton. The similarities of both abundant and rare eukaryotic plankton subcommunities significantly declined with the increase in time-lag, but stronger temporal turnover was observed in rare taxa. Further, species turnover of both subcommunities explained a higher percentage of the community variation than species richness. Both deterministic and stochastic processes significantly influenced eukaryotic plankton community assembly, and the stochastic pattern (e.g., ecological drift) was particularly pronounced for rare taxa. Co-occurrence network analysis revealed that keystone taxa mainly belonged to rare species, which may play fundamental roles in network persistence. Importantly, covariations between rare and non-rare taxa were predominantly positive, implying multispecies cooperation might contribute to the stability and resilience of the microbial community. Overall, these findings expand current understanding of the ecological mechanisms and microbial interactions underlying plankton dynamics in changing aquatic ecosystems.

Community assembly processes underlying phytoplankton and bacterioplankton across a hydrologic change in a human-impacted river.

Although the influence of microbial community assembly processes on aquatic ecosystem function and biodiversity is well known, the processes that govern planktonic communities in human-impacted rivers remain largely unstudied. Here, we used multivariate statistics and a null model approach to test the hypothesis that environmental conditions and obstructed dispersal opportunities, dictate a deterministic community assembly for phytoplankton and bacterioplankton across contrasting hydrographic conditions in a subtropical mid-sized river (Jiulong River, southeast China). Variation partitioning analysis showed that the explanatory power of local environmental variables was larger than that of the spatial variables for both plankton communities during the dry season. During the wet season, phytoplankton community variation was mainly explained by local environmental variables, whereas the variance in bacterioplankton was explained by both environmental and spatial predictors. The null model based on Raup-Crick coefficients for both planktonic groups suggested little evidences of the stochastic processes involving dispersal and random distribution. Our results showed that hydrological change and landscape structure act together to cause divergence in communities along the river channel, thereby dictating a deterministic assembly and that selection exceeds dispersal limitation during the dry season. Therefore, to protect the ecological integrity of human-impacted rivers, watershed managers should not only consider local environmental conditions but also dispersal routes to account for the effect of regional species pool on local communities.

Disturbance-induced phytoplankton regime shifts and recovery of cyanobacteria dominance in two subtropical reservoirs.

Many countries in the world still suffer from high toxic cyanobacterial blooms in inland waters used for human consumption. Regional climate change and human activities within watersheds exert a complex and diverse influence on aquatic ecosystem structure and function across space and time. However, the degree to which these factors may contribute to the long-term dynamics of plankton communities is still not well understood. Here, we explore the impacts of multiple disturbance events (e.g. human-resettlement, temperature change, rainfall, water level fluctuations), including six combined disturbances, on phytoplankton and cyanobacteria in two subtropical reservoirs over six years. Our data showed that combined environmental disturbances triggered two apparent and abrupt switches between cyanobacteria-dominated state and non-cyanobacterial taxa-dominated state. In late 2010, the combined effect of human-resettlement (emigration) and natural disturbances (e.g. cooling, rainfall, water level fluctuations) lead to a 60-90% decrease in cyanobacteria biomass accompanied by the disappearance of cyanobacterial blooms, in tandem with an abrupt and persistent shift in phytoplankton community. After summer 2014, however, combined weather and hydrological disturbances (e.g. warming, rainfall, water level fluctuations) occurred leading to an abrupt and marked increase of cyanobacteria biomass, associated with a return to cyanobacteria dominance. These changes in phytoplankton community were strongly related to the nutrient concentrations and water level fluctuations, as well as water temperature and rainfall. As both extreme weather events and human disturbances are predicted to become more frequent and severe during the twenty-first century, prudent sustainable management will require consideration of the background limnologic conditions and the frequency of disturbance events when assessing the potential impacts on reservoir biodiversity and ecosystem functioning and services.

Cyanobacterial bloom significantly boosts hypolimnelic anammox bacterial abundance in a subtropical stratified reservoir.

The degradation of freshwater quality induced by cyanobacterial blooms is a major global environmental concern. Microbially driven nitrogen removal could alleviate eutrophication to some degree in freshwater ecosystems. However, the response of anaerobic ammonium oxidizing (anammox) bacterial communities to cyanobacterial blooms remains poorly understood, especially in reservoir ecosystems. Here we compared the dynamics of anammox bacterial communities during and after a cyanobacterial bloom in a subtropical reservoir. Our data showed that a cyanobacterial bloom triggered a significant increase in bottom anammox bacterial abundance. During the bloom period, anammox bacterial abundance in bottom waters was 9-fold and 52-fold higher compared with non-bloom stratification and mixing periods, respectively. The community composition of anammox bacteria in surface waters changed substantially accompanied by the disappearance of the cyanobacterial bloom, and a shift of dominance from unidentified anammox genera to Ca. Brocadia was observed. Although Ca. Brocadia was always predominant in both middle and bottom waters, the non-bloom period had more unique taxa than the bloom period. Cyanobacterial bloom-related changes in environmental conditions (e.g. NH4-N and total organic carbon) and water stratification together influenced the distribution and dynamics of anammox bacteria. Altogether, our study lays the basis for a better understanding of the breakdown of cyanobacterial blooms in a stratified reservoir.