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Biddle, Jennifer F (Ed.)ABSTRACT Aerobic anoxygenic phototrophic (AAP) bacteria harvest light energy using bacteriochlorophyll-containing reaction centers to supplement their mostly heterotrophic metabolism. While their abundance and growth have been intensively studied in coastal environments, much less is known about their activity in oligotrophic open ocean regions. Therefore, we combinedin situsampling in the North Pacific Subtropical Gyre, north of O'ahu island, Hawaii, with two manipulation experiments. Infra-red epifluorescence microscopy documented that AAP bacteria represented approximately 2% of total bacteria in the euphotic zone with the maximum abundance in the upper 50 m. They conducted active photosynthetic electron transport with maximum rates up to 50 electrons per reaction center per second. Thein situdecline of bacteriochlorophyll concentration over the daylight period, an estimate of loss rates due to predation, indicated that the AAP bacteria in the upper 50 m of the water column turned over at rates of 0.75–0.90 d−1. This corresponded well with the specific growth rate determined in dilution experiments where AAP bacteria grew at a rate 1.05 ± 0.09 d−1. An amendment of inorganic nitrogen to obtain N:P = 32 resulted in a more than 10 times increase in AAP abundance over 6 days. The presented data document that AAP bacteria are an active part of the bacterioplankton community in the oligotrophic North Pacific Subtropical Gyre and that their growth was mostly controlled by nitrogen availability and grazing pressure.IMPORTANCEMarine bacteria represent a complex assembly of species with different physiology, metabolism, and substrate preferences. We focus on a specific functional group of marine bacteria called aerobic anoxygenic phototrophs. These photoheterotrophic organisms require organic carbon substrates for growth, but they can also supplement their metabolic needs with light energy captured by bacteriochlorophyll. These bacteria have been intensively studied in coastal regions, but rather less is known about their distribution, growth, and mortality in the oligotrophic open ocean. Therefore, we conducted a suite of measurements in the North Pacific Subtropical Gyre to determine the distribution of these organisms in the water column and their growth and mortality rates. A nutrient amendment experiment showed that aerobic anoxygenic phototrophs were limited by inorganic nitrogen. Despite this, they grew more rapidly than average heterotrophic bacteria, but their growth was balanced by intense grazing pressure.more » « less
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Abstract We optimized a high throughput method to quantify turnover rates of adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) in marine microbes from simultaneous measures of the respective stocks and phosphorylation rates. We combined a microbial adenylate extraction method using boiling 20 mM Tris buffer with purification and analysis by high pressure liquid chromatography optimized to quantify these intracellular adenylate concentrations in marine microbes. Additionally, we incorporated radiolabeled phosphate (32Pi) incubations to quantify phosphorus (P) uptake rates and the phosphorylation rates for these adenylate compounds in microbial cells. With this method, we can directly assess the variations in microbial growth rates, metabolic turnover rates, energy charge, and adenylate storage. We applied and validated this method application with environmental samples from Biscayne Bay, Florida, and quantified adenylate turnover times of 12, 15, and 73 min, for ATP, ADP, and AMP, respectively. Future incorporation of this method into experiments and geographic surveys across marine environments will allow for direct assessments of changes in microbial metabolic activity in relation to other ecological variables.more » « less
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Abstract Heterotrophic bacteria in the surface ocean play a critical role in the global carbon cycle and the magnitude of this role depends on their growth rates. Although methods for determining bacterial community growth rates based on incorporation of radiolabeled thymidine and leucine are widely accepted, they are based on a number of assumptions and simplifications. We sought to independently assess these methods by comparing bacterial growth rates to turnover rates of bacterial membranes using previously published methods in a range of open‐ocean settings. We found that turnover rates for heterotrophic bacterial phospholipids averaged 0.80 ± 0.35 d−1. This was supported by independent measurements of turnover rates of a membrane‐bound pigment in photoheterotrophic bacteria, bacteriochlorophyll a(0.85 ± 0.09 d−1). By contrast, bacterial growth rates measured by uptake of radiolabeled thymidine and leucine were 0.12 ± 0.08 d−1, well within the range expected from the literature. We explored whether the discrepancies between phospholipid turnover rates and bacterial growth rate could be explained by membrane recycling/remodeling and other factors, but were left to conclude that the radiolabeled thymidine and leucine incorporation methods substantially underestimated actual bacterial growth rates. We use a simple model to show that the faster bacterial growth rates we observed can be accommodated within the constraints of the microbial carbon budget if bacteria are smaller than currently thought, grow with greater efficiency, or some combination of these two factors.more » « less
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