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Nitrogenase is extremely sensitive to molecular oxygen (O(2)), and unicellular diazotrophic cyanobacteria separate nitrogen (N(2))-fixation and photosynthesis to protect nitrogenase from O(2) produced by photosynthesis. When grown under 12 h light/12 h dark cycles (LD), the marine unicellular diazotrophic cyanobacterium Gloeothece sp. 68DGA expressed the nitrogenase protein and its activity (acetylene reduction activity) only during the dark phase. However, this strain was able to grow diazotrophically under continuous light (CL). To determine whether nitrogenase synthesis and N(2)-fixation are temporally separated from photosynthesis in the Gloeothece cells that have fully acclimated to CL, the proportion of cells containing nitrogenase (the Fe-protein of nitrogenase) in the culture was measured using an immunocytochemical technique. Cells were grown in a continuous-culture device to maintain constant cell density. Under LD, the cells showed diurnal oscillation of nitrogenase activity, photosynthesis, respiration and the expression and the abundance of the Fe-protein. The oscillation was gradually reduced after the transfer of the cells to CL, and was lost after 23-25 days of cultivation under CL. In CL-acclimated cultures, the Fe-protein was always detected in about 94 % of the cells, although the nitrogenase activity was about one-third of the maximum activity in LD-acclimated cultures. These results suggest that synthesis of nitrogenase proceeds without diurnal oscillation in the CL-acclimated cells of Gloeothece sp. 68DGA. As the respiration rate in CL-acclimated culture was as high as the maximum rate observed in LD-acclimated culture, O(2)-uptake mechanism(s) may have been upregulated to maintain low intracellular pO(2).

While primary productivity in the oligotrophic North Pacific Subtropical Gyre (NPSG) is changing, the micro-size plankton community has not been evaluated in the last 4 decades, prompting a re-evaluation. We collected samples over three years (2016–2018) from depths of 10 to 200 m (n = 127), and the micro-size plankton were identified and counted to understand the heterogeneity of micro-size plankton community structure. The assemblages were consistent to the those of 4 decades ago. Dinophyceae (dinoflagellates) were the most numerically abundant, followed by Cryptophyceae and Bacillariophyceae (diatoms). The other micro-size plankton classes (Cyanophyceae, Haptophyceae, Dictyochophyceae, Euglenophyceae, and Prasinophyceae) were not always detected, whereas only Trichodesmium spp. was counted in the Cyanophyceae. Other unidentified autotrophic and heterotrophic flagellates were also significantly present, and their numeric abundance was higher than or at the same level as was that of the Dinophyceae. In the Dinophyceae, Gymnodiniaceae and Peridiniales were abundant. The chlorophyll a concentration and these class-level assemblages suggested micro-size plankton is not a major primary producer in this area. We applied generalized additive models (GAMs) and principal coordination analyses (PCoAs) to evaluate the habitats of every plankton group and the heterogeneity of the assemblages. The GAMs suggested that every classified plankton abundance showed a similar response to salinity, and we observed differences in habitats in terms of temperature and nitrate concentrations. Based on the PCoAs, we observed unique communities at the 200 m depth layer compared with those at the other sampling layers. The site scores of PCoAs indicated that the micro-size plankton assemblages are most heterogeneous at the 10 m depth layer. At such depth, diazotrophic Cyanophyceae (Trichodesmium spp.) are abundant, particularly in less-saline water. Therefore, nitrogen fixation may contribute to the heterogeneity in the abundance and assemblages in the western NPSG. [ABSTRACT FROM AUTHOR]

The relationship between the abundance of nitrogenase and its activity was studied in the marine unicellular cyanobacterium Gloeothece sp. 68DGA cultured under different light/dark regimens. The Fe- and MoFe-protein of nitrogenase and nitrogen (N2)-fixing (acetylene reduction) activity were detected only during the dark phase when the cells were grown under a 12 h light/12 h dark cycle (12L/12D). Nitrogenase activity appeared about 4 h after entering the dark phase. Maximum nitrogenase activity occurred at around the middle of the dark phase, and the activity rapidly decreased to zero before the start of the light phase. The rapid decrease of nitrogenase activity and the Fe-protein of nitrogenase near the end of the dark phase in 12L/12D were partly recovered by the addition ofl-methionine-sulfoximine, an inhibitor of glutamine synthetase. Diurnal oscillation of the abundance of nitrogenase was maintained in the first subjective dark phase (i.e. the period corresponding to the dark phase) after the cells were transferred from 12L/12D to continuous illumination. However, enzyme activity was detected only when photosynthetic oxygen (O2) evolution was completely suppressed by reducing the light intensity or by the addition of 3-(3,4-dichlorophenyl)-1,1-dimethylurea. Nitrogenase always appeared in the cells about 16 h after starting the light phase, even when the 12L/12D cycle was modified by the addition or subtraction of a single 6 h period of light or dark. These results suggest the following: (i) N2-fixation by Gloeothece sp. 68DGA is primarily regulated by an endogenous circadian oscillator at the level of nitrogenase synthesis. (ii) The endogenous circadian rhythm resets on a shift of the timing of the light phase. (iii) Nitrogenase activity is not always reflected in the presence of nitrogenase. (iv) The activity of nitrogenase is negatively regulated by fixed nitrogen and the concentration of ambient O2. [ABSTRACT FROM AUTHOR]

Abstract: Seasonal changes in abundances of Trichodesmium spp. and unicellular cyanobacterial phylotype groups A and B in the northern South China Sea (SCS) and upstream Kuroshio Current were investigated using qPCR techniques targeting the nifH gene that encodes nitrogenase for N2 fixation. The aim was to understand the community structures and factors associated with the abundances of the diazotrophs (nitrogen-fixers) in this important marginal sea. The results from 3 cruises covering 9 sampling stations revealed a close correlation between seawater temperature and gene abundances of the three nifH phylotypes. Trichodesmium spp. and group B were more abundant in the Kuroshio than the SCS, whereas group A was equally distributed. The abundances of Trichodesmium spp. and group B were positively correlated with nitracline depths. As nitracline is consistently deeper in the Kuroshio than in the SCS, nitracline depth could be used to track the abundance dynamics of the two phylotypes. In the station with prevailing internal waves, we observed unusually high abundances of Trichodesmium spp. but markedly low abundances of group B. Distributions of Trichodesmium spp. and group B were not only influenced by temperature and nitracline depth, but also occurrence of internal waves. [Copyright &y& Elsevier]

We studied the seasonal, diel, and vertical distribution of phytoplankton N2 fixation to understand the relative contributions of unicellular and filamentous nitrogen fixers (diazotrophs) to N2 fixation and nitrogen recycling in the northern South China Sea (SCS) and the neighboring upstream Kuroshio. N2-fixation rates were measured by the 15N2 tracer technique (addition by bubble) on unicellular (<10 or 20µm) and the filamentous diazotrophs (>10 or 20µm, mostly Trichodesmium and Richelia) fractionated by 10- or 20-µm mesh sizes. The mean depth-integrated total (unicellular+filamentous) N2-fixation rates in the SCS (51.7±6.2µmolNm−2 d−1) averaged 1/3 of that in the Kuroshio (142.7±29.6µmolNm−2 d−1), with higher rates in the winter than in other seasons in the SCS and the opposite seasonal pattern in the Kuroshio. Unicellular diazotrophs contributed 65% of the total N2 fixation in the SCS, which were negatively correlated with surface temperature and, as for total N2 fixation, were higher in the winter when Trichodesmium spp. were scarce. In comparison, the unicellular diazotrophs contributed 50% of total N2 fixation in the Kuroshio, and their contributions were not significantly correlated with surface temperature. In both the SCS and the Kuroshio, the unicellular N2 fixation was more important during the night than during the day, and in the deep euphotic layer than in the surface layer, even in the daytime. Our results show that the unicellular diazotrophs were important N2 fixers and contributed significantly to N2 fixation in the tropical marginal seas, more so in the SCS than the Kuroshio. [ABSTRACT FROM AUTHOR]