Nitrification is a central process of the aquatic nitrogen cycle that controls the supply of nitrate used in other key processes, such as phytoplankton growth and denitrification. Through time series observation and modeling of a seasonally stratified, eutrophic coastal basin, we demonstrate that physical dilution of nitrifying microorganisms by water column mixing can delay and decouple nitrification. The findings are based on a 4-y, weekly time series in the subsurface water of Bedford Basin, Nova Scotia, Canada, that included measurement of functional (amoA) and phylogenetic (16S rRNA) marker genes. In years with colder winters, more intense winter mixing resulted in strong dilution of resident nitrifiers in subsurface water, delaying nitrification for weeks to months despite availability of ammonium and oxygen. Delayed regrowth of nitrifiers also led to transient accumulation of nitrite (3 to 8 μmol · kgsw−1) due to decoupling of ammonia and nitrite oxidation. Nitrite accumulation was enhanced by ammonia-oxidizing bacteria (Nitrosomonadaceae) with fast enzyme kinetics, which temporarily outcompeted the ammonia-oxidizing archaea (Nitrosopumilus) that dominated under more stable conditions. The study reveals how physical mixing can drive seasonal and interannual variations in nitrification through control of microbial biomass and diversity. Variable, mixing-induced effects on functionally specialized microbial communities are likely relevant to biogeochemical transformation rates in other seasonally stratified water columns. The detailed study reveals a complex mechanism through which weather and climate variability impacts nitrogen speciation, with implications for coastal ecosystem productivity. It also emphasizes the value of high-frequency, multiparameter time series for identifying complex controls of biogeochemical processes in aquatic systems.
This paper proposes an integration and modular organization of the complex regulatory networks involved in the mammalian cell cycle, apoptosis, and related intracellular signaling cascades. A common node linking the cell cycle and apoptosis permits the possibility of coordinate control between the initiation of these two cellular processes. From this node, pathways emanate that lead to the activation of cyclin-dependent kinases (in the cell cycle) and caspases (in apoptosis). Computer simulations are carried out to demonstrate that the proposed network architecture and certain module-module interactions can account for the experimentally observed sequence of cellular events (quiescence, cell cycle, and apoptosis) as the transcriptional activities of E2F-1 and c-Myc are increased. Despite the lack of quantitative kinetic data on most of the pathways, it is demonstrated that there can be meaningful conclusions regarding system stability that arise from the topology of the network. It is shown that only cycles in the network graph determine stability. Thus, several positive and negative feedback loops are identified from a literature review of the major pathways involved in the initiation of the cell cycle and of apoptosis.
Depressurization and sample processing delays may impact the outcome of shipboard microbial incubations of samples collected from the deep sea. To address this knowledge gap, we developed an ROV-powered incubator instrument to carry out and compare results from in situ and shipboard RNA Stable Isotope Probing (RNA-SIP) experiments to identify the key chemolithoautotrophic microbes and metabolisms in diffuse, low-temperature venting fluids from Axial Seamount. All the incubations showed microbial uptake of labeled bicarbonate primarily by thermophilic autotrophic Epsilonbacteraeota that oxidized hydrogen coupled with nitrate reduction. However, the in situ seafloor incubations showed higher abundances of transcripts annotated for aerobic processes suggesting that oxygen was lost from the hydrothermal fluid samples prior to shipboard analysis. Furthermore, transcripts for thermal stress proteins such as heat shock chaperones and proteases were significantly more abundant in the shipboard incubations suggesting that depressurization induced thermal stress in the metabolically active microbes in these incubations. Together, results indicate that while the autotrophic microbial communities in the shipboard and seafloor experiments behaved similarly, there were distinct differences that provide new insight into the activities of natural microbial assemblages under near-native conditions in the ocean. Importance: Diverse microbial communities drive biogeochemical cycles in Earth’s ocean, yet studying these organisms and processes is often limited by technological capabilities, especially in the deep ocean. In this study, we used a novel marine microbial incubator instrument capable of in situ experimentation to investigate microbial primary producers at deep-sea hydrothermal vents. We carried out identical stable isotope probing experiments coupled to RNA sequencing both on the seafloor and on the ship to examine thermophilic, microbial autotrophs in venting fluids from an active submarine volcano. Our results indicate that microbial communities were significantly impacted by the effects of depressurization and sample processing delay, with shipboard microbial communities more stressed compared to seafloor incubations. Differences in metabolism were also apparent and are likely linked to the chemistry of the fluid at the beginning of the experiment. Microbial experimentation in the natural habitat provides new insights into understanding microbial activities in the ocean.
Isotopic enrichment factors are key to using stable isotope signatures in biogeochemical studies. However, these are typically determined in laboratory experiments and their applicability to environmental conditions is difficult to test. Here, we analyzed nitrogen stable isotope changes associated with nitrification in a coastal basin using weekly time-series measurements of δ 15 N in particulate nitrogen, ammonium, nitrite, and nitrate. Two year-long time series were selected as contrasting natural experiments in the ammonium-rich, aphotic bottom water of Bedford Basin, Nova Scotia, Canada. In 2014, ammonia oxidation (AO) was associated with Thaumarchaeota and nitrite concentrations remained low (< 0.5 μmol kg À1 ). In contrast, transient nitrite accumulation (~8 μmol kg À1 ) and a more rapid δ 15 N NH4 increase in the fall of 2017 were likely caused by ammoniaoxidizing bacteria, associated with higher AO rates and, possibly, stronger nitrogen-isotope enrichment ( 15 ε AO ). Estimates of 15 ε AO (21.8 AE 2.2‰, 24.1 AE 1.1‰) were derived empirically using Rayleigh models applied to field data from restricted periods during which the bottom waters approximated a closed system and influence on 15 ε AO from other processes was demonstrably insignificant. Using a numerical reactive-transport model, we
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