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Phytoplankton are important drivers of aquatic ecosystem function and environmental health. Their community compositions and distributions are directly impacted by environmental processes and human activities, including in the largest estuary in North America, the Chesapeake Bay. It is crucial to uncover how planktonic eukaryotes play fundamental roles as primary producers and trophic links and sustain estuarine ecosystems. In this study, we investigated the detailed community structure and spatiotemporal variations of planktonic eukaryotes in the Chesapeake Bay across space and time for three consecutive years. A clear seasonal and spatial shift of total, abundant, and rare planktonic eukaryotes was evident, and the pattern recurred interannually. Multiple harmful algal species have been identified in the Bay with varied distribution patterns, such as Karlodinium, Heterosigma akashiwo, Protoperidinium sp., etc. Compared to abundant taxa, rare subcommunities were more sensitive to environmental disturbance in terms of richness, diversity, and distribution. The combined effects of temporal variation (13.3%), nutrient availability (10.0%), and spatial gradients (8.8%) structured the distribution of eukaryotic microbial communities in the Bay. Similar spatiotemporal patterns between planktonic prokaryotes and eukaryotes suggest common mechanisms of adjustment, replacement, and species interaction for planktonic microbiomes under strong estuarine gradients. To our best knowledge, this work represents the first systematic study on planktonic eukaryotes in the Bay. A comprehensive view of the distribution of planktonic microbiomes and their interactions with environmental processes is critical in understanding the underlying microbial mechanisms involved in maintaining the stability, function, and environmental health of estuarine ecosystems., Importance: Deep sequencing analysis of planktonic eukaryotes in the Chesapeake Bay reveals high community diversity with many newly recognized phytoplankton taxa. The Chesapeake Bay planktonic eukaryotes show distinct seasonal and spatial variability, with recurring annual patterns of total, abundant, and rare groups. Rare taxa mainly contribute to eukaryotic diversity compared to abundant groups, and they are more sensitive to spatiotemporal variations and environmental filtering. Temporal variations, nutrient availability, and spatial gradients significantly affect the distribution of eukaryotic microbial communities. Similar spatiotemporal patterns in prokaryotes and eukaryotes suggest common mechanisms of adjustment, substitution, and species interactions in planktonic microbiomes under strong estuarine gradients. Interannually recurring patterns demonstrate that diverse eukaryotic taxa have well adapted to the estuarine environment with a long residence time. Further investigations of how human activities impact estuarine planktonic eukaryotes are critical in understanding their essential ecosystem roles and in maintaining environmental safety and public health., Competing Interests: The authors declare no conflict of interest.

The microbial carbon pump (MCP) provides a mechanistic illustration of transformation of recalcitrant dissolved organic matter (DOM) in the ocean. Here, we explored and demonstrated the key roles of algae-associated microorganisms (mainly heterotrophic bacteria) in the production and transformation of carboxyl-rich alicyclic molecule (CRAM)-like DOM through a laboratory experiment involving cultures of Skeletonema dohrnii. Without the participation of the associated bacteria, CRAM-like DOM molecules were not detected via Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR MS) in algal cultures treated with antibiotics. Similarly, CRAM-like DOM were not detected in cultures of bacteria alone. Our experimental results showed that algae-associated bacteria are important in the process of converting algal-derived organic matter into CRAM-like DOM during S. dohrnii culture. Bacteroidetes (mainly Flavobacteriia) dominated the bacterial community in the stationary and degradation phases, where the predicted metabolic pathways for bacterial assemblages were mainly involved in biosynthesis, metabolism, and degradation. Facilitated by these heterotrophic bacteria, the amount and the chemodiversity of CRAM-like DOM derived from algae varied during the growth and decomposition of algal cells, and CRAM-like DOM were enriched at the later growth stage. The properties and characteristics of these CRAM-like DOM, including molecular weight, double bond equivalent, hydrogen-carbon ratio, carbon-nitrogen ratio, carbon-sulfur ratio, and modified aromaticity index increased with the growth and decay of algal cells, indicating the transformation from active to recalcitrant DOM. In contrast, the organic matter in axenic cultures of S. dohrnii mainly existed in the form of particulate organic matters (POM), and small amounts of CRAM-like DOM were detected. This study provides the first laboratory evidence to reveal and confirm the direct involvement of algae-associated microbiomes in the production and transformation of algae-derived refractory DOM, highlighting the significance of these epiphytic bacteria in marine carbon sequestration and global carbon cycling. IMPORTANCE Dissolved organic matter (DOM) serves as a major carbon and nutrient pool in oceans, and recalcitrant DOM are the primary sources for carbon sequestration in depths. Here, we demonstrate the critical roles of algae-associated microorganisms (mainly heterotrophic bacteria) in the transformation of recalcitrant dissolved organic matter through laboratory cultures of a model diatom, Skeletonema dohrnii. Our experimental results showed that in addition to affecting the growth and the physiology of S. dohrnii, algae-associated bacteria are important in processing and converting algal DOM into CRAM-like DOM. Facilitated by the associated bacteria, the amount and the chemodiversity of DOM derived from algae varied during the growth and decomposition of algal cells, and enriched recalcitrant DOM formed in the later growth stage. The properties and diversity of DOM increased with the growth and decay of algal cells, indicating the transformation from active DOM to inert organic matter. Our results confirmed that the direct involvement of algae-associated microbes in the production of CRAM-like DOM. Detailed community structure analysis of the algae-associated bacterial community and its predicted functions confirmed the involvement of certain bacterial groups (e.g., Flavobacteriia ) in biosynthesis, metabolism, and degradation.

Simple Summary: The Pearl River Estuary (PRE) in China, a highly urbanized coastal area, presents a unique opportunity to study the effects of environmental changes on the phytoplankton com-munity. In September 2018, a field study was conducted to examine how the photosynthetic status and spatial distribution of these organisms varied from freshwater to oceanic waters. Dinophyta and Haptophyta were prevalent in seawater, while Chlorophyta and Cryptophyta dominated from freshwater to estuarine water. Bacillariophyta were found across all regions. Phytoplankton in the mixing zone showed signs of stress due to fluctuating environmental conditions, whereas those in freshwater and oceanic areas appeared more photosynthetically active. Human activities have in-creased nutrient levels in the estuary, leading to higher chlorophyll concentrations and more diverse phytoplankton communities upstream. Understanding these patterns helps us assess the health of coastal ecosystems, which is crucial for managing the impacts of climate change and human development on marine environments. The Pearl River Estuary (PRE) is one of the world's most urbanized subtropical coastal systems. It presents a typical environmental gradient suitable for studying estuarine phytoplankton communities' dynamics and photosynthetic physiology. In September 2018, the maximum photochemical quantum yield (Fv/Fm) of phytoplankton in different salinity habitats of PRE (oceanic, estuarine, and freshwater zones) was studied, revealing a complex correlation with the environment. Fv/Fm of phytoplankton ranged from 0.16 to 0.45, with taxa in the upper Lingdingyang found to be more stressed. Community composition and structure were analyzed using 18S rRNA, accompanied by a pigment analysis utilized as a supplementary method. Nonmetric multidimensional scaling analysis indicated differences in the phytoplankton spatial distribution along the estuarine gradients. Specificity-occupancy plots identified different specialist taxa for each salinity habitat. Dinophyta and Haptophyta were the predominant taxa in oceanic areas, while Chlorophyta and Cryptophyta dominated freshwater. Bacillariophyta prevailed across all salinity gradients. Canonical correlation analysis and Mantel tests revealed that temperature, salinity, and elevated nutrient levels (i.e., NO3−-N, PO43−-P, and SiO32−-Si) associated with anthropogenic activities significantly influenced the heterogeneity of community structure. The spatial distribution of phytoplankton, along with in situ photosynthetic characteristics, serves as a foundational basis to access estuarine primary productivity, as well as community function and ecosystem health. [ABSTRACT FROM AUTHOR]

Marine chromophytic phytoplankton are a diverse group of algae and contribute significantly to the total oceanic primary production. However, the spatial distribution of chromophytic phytoplankton is understudied in the West Pacific Ocean (WPO). In this study, we have investigated the community structure and spatial distribution of chromophytic phytoplankton using RuBisCO genes (Form ID rbc L). Our results showed that Haptophyceae, Pelagophyceae, Cyanophyceae, Xanthophyceae, and Bacillariophyceae were the dominant groups. Further, chromophytic phytoplankton can be distinguished between upwelling and non-upwelling zones of the WPO. Surface and 75 m depths of a non-upwelling area were dominated by Prochlorococcus strains, whereas chromophytic phytoplankton were homogenously distributed at the surface layer in the upwelling zone. Meanwhile, Pelagomonas -like sequences were dominant at DCM (75 m) and 150 m depths of the upwelling zone. Non-metric multidimensional scaling (NMDS) analysis did not differentiate between chromophytic phytoplankton in the upwelling and non-upwelling areas, however, it showed clear trends of them at different depths. Further, redundancy analysis (RDA) showed the influence of physicochemical parameters on the distribution of chromophytic phytoplankton. Along with phosphate (p < 0.01), temperature and other dissolved nutrients were important in driving community structure. The upwelling zone was impacted by a decrease in temperature, salinity, and re-supplement of nutrients, where Pelagomonas -like sequences outnumbered other chromophytic groups presented. [ABSTRACT FROM AUTHOR]

Phytoplankton are the basis of primary production and play important roles in regulating energy export in marine ecosystems. Compared to other regions, chromophytic phytoplankton are considerably understudied in the Bay of Bengal (BOB). Here, we investigated community structure and spatial distribution of chromophytic phytoplankton in the BOB by using RuBisCO genes (Form ID rbc L). High throughput sequencing of rbc L genes revealed that diatoms, cyanobacteria (Cyanophyceae), Pelagophyceae, Haptophyceae, Chrysophyceae, Eustigamatophyceae, Xanthophyceae, Cryptophyceae, Dictyochophyceae, and Pinguiophyceae were the most abundant groups recovered in the BOB. Abundances and distribution of diatoms and Pelagophyceae were further verified using quantitative PCR analyses which showed the dominance of these groups near the Equator region (p < 0.01) where upwelling was likely the source of nutrients. Further, redundancy analysis demonstrated that temperature was an important environmental driver in structuring distributions of Cyanophyceae and dominant chromophytic phytoplankton. Morphological identification and quantification confirmed the dominance of diatoms, and also detected other cyanobacteria and dinoflagellates that were missing in our molecular characterizations. Pearson's correlations of these morphologically identified phytoplankton with environmental gradients also indicated that nutrients and temperature were key variables shaping community structure. Combination of molecular characterization and morphological identification provided a comprehensive overview of chromophytic phytoplankton. This is the first molecular study of chromophytic phytoplankton accomplished in the BOB, and our results highlight a combination of molecular analysis targeting rbc L genes and microscopic detection in examining phytoplankton composition and diversity. [ABSTRACT FROM AUTHOR]

High-biomass red tides occur frequently in some semi-enclosed bays of Hong Kong where ambient nutrients are not high enough to support such a high phytoplankton biomass. These high-biomass red tides release massive inorganic nutrients into local waters during their collapse. We hypothesized that the massive inorganic nutrients released from the collapse of red tides would fuel growth of other phytoplankton species. This could influence phytoplankton species composition. We tested the hypothesis using a red tide event caused by Mesodinium rubrum (M. rubrum) in a semi-enclosed bay, Port Shelter. The red tide patch had a cell density as high as 5.0×105 cells L−1, and high chlorophyll a (63.71 μg L−1). Ambient inorganic nutrients (nitrate: NO3−, ammonium: NH4+, phosphate: PO43−, silicate: SiO43−) were low both in the red tide patch and the non-red-tide patch (clear waters outside the red tide patch). Nutrient addition experiments were conducted by adding all the inorganic nutrients to water samples from the two patches followed by incubation for 9 days. The results showed that the addition of inorganic nutrients did not sustain high M. rubrum cell density, which collapsed after day 1, and did not drive M. rubrum in the non-red-tide patch sample to the same high-cell density in the red tide patch sample. This confirmed that nutrients were not the driving factor for the formation of this red tide event, or for its collapse. The death of M. rubrum after day 1 released high concentrations of NO3−, PO43−, SiO43−, NH4+, and urea. Bacterial abundance and heterotrophic activity increased, reaching the highest on day 3 or 4, and decreased as cell density of M. rubrum declined. The released nutrients stimulated growth of diatoms, such as Chaetoceros affinis var. circinalis, Thalassiothrix frauenfeldii, and Nitzschia sp., particularly with additions of SiO43− treatments, and other species. These results demonstrated that initiation of M. rubrum red tides in the bay was not directly driven by nutrients. However, the massive inorganic nutrients released from the collapse of the red tide could induce a second bloom in low-ambient nutrient water, influencing phytoplankton species composition. [ABSTRACT FROM AUTHOR]