Cryo-EM structures of photocomplexes from the free-living aerobic anoxygenic phototrophic bacterium Erythrobacter sanguineus.

Aerobic anoxygenic phototrophic (AAP) bacteria are widely distributed in marine and freshwater ecosystems. The aims of this study were to investigate the diversity and abundance of AAP bacteria in cyano- bacterial bloom-forming eutrophic lakes and to study the association of AAP bacteria with the bloom- forming cyanobacteria. Analysis of pufM gene (the light-reaction center gene) clone libraries indicated that in eutrophic lakes (Lake Taihu and Lake Chaohu, China) with cyanobacterium Microcystis blooms, the AAP bacteria were related to members of Alphaproteobacteria, Betaproteobacteria, and Gammaproteobacteria. In Lake Taihu and Lake Chaohu, Alphaproteobacteria accounted for 81.5% and 75.0% of Microcystis- associated AAP bacteria, respectively, and 84.6% and 72.5% of free-living AAP bacteria, respectively. The predominance of Alphaproteobacteria in the two lakes was different from the previously reported predominance of Betaproteobacteria in freshwater lakes. Quantitative real-time PCR analysis indicated that in Lake Taihu and Lake Chaohu, AAP bacteria represented an important part of the bacterial community associated with Microcystis, and the abundance of Microcystis-associated AAP bacteria (18.3% and 11.7%, respectively) was higher than that of free-living AAP bacteria (5.1% and 7.9%, respectively). The abundance of AAP bacteria in the two bloom-forming lakes was higher than the previously reported level in other eutrophic freshwater bodies.

Aerobic anoxygenic phototrophic (AAP) bacteria are photoheterotrophs that despite their low abundances have been hypothesized to play an ecologically and biogeochemically important role in aquatic systems. Characterizing this role requires a better understanding of the in situ dynamics and activity of AAP bacteria. Here we provide the first assessment of the single-cell activity of freshwater AAP bacteria and their contribution to total bacterial production across lakes spanning a wide trophic gradient, and explore the role of light in regulating AAP activity. The proportion of cells that were active in leucine incorporation and the level of activity per cell were consistently higher for AAP than for bulk bacteria across lakes. As a result, AAP bacteria contributed disproportionately more to total bacterial production than to total bacterial abundance. Interestingly, although environmentally driven patterns in activity did not seem to differ largely between AAP and bulk bacteria, their response to light did, and exposure to light resulted in increases in the proportion of active AAP bacteria with no clear effect on their cell-specific activity. This suggests that light may play a role in the activation of AAP bacteria, enabling these photoheterotrophs to contribute more to the carbon cycle than suggested by their abundance.

Temperature is an important controlling factor in the growth activity of all microorganisms. Aerobic anoxygenic phototrophic (AAP) bacteria actively grow in the ocean and are known as one of the main driving forces in organic matter cycling in surface seawater environments. Whether temperature change affects AAP bacteria activity from an ecological viewpoint remains an open question. To date, no known studies have reported the effect of temperature change on AAP bacteria growth in the ocean. We here show that the growth rate of AAP bacteria exceeded that of other bacterial types at high water temperatures in the absence of grazers. The slope of the regression line of the net growth rate of AAP bacteria as a function of water temperature was the same as that for non‐AAP bacteria at all temperatures (10, 20, and 30°C); however, when grazers were eliminated, it was 4.7 times higher than that of non‐AAP bacteria. This result suggests that AAP bacteria are more responsive to water temperature increases than other bacteria and that AAP bacteria might become more dominant than other bacteria under elevated water temperatures.

The abundance of aerobic anoxygenic phototrophic (AAP) bacteria, cyanobacteria, and heterotrophs was examined in the Mid-Atlantic Bight and the central North Pacific Gyre using infrared fluorescence microscopy coupled with image analysis and flow cytometry. AAP bacteria comprised 5% to 16% of total prokaryotes in the Atlantic Ocean but only 5% or less in the Pacific Ocean. In the Atlantic, AAP bacterial abundance was as much as 2-fold higher than that of Prochlorococcus spp. and 10-fold higher than that of Synechococcus spp. In contrast, Prochlorococcus spp. outnumbered AAP bacteria 5- to 50-fold in the Pacific. In both oceans, subsurface abundance maxima occurred within the photic zone, and AAP bacteria were least abundant below the 1% light depth. The abundance of AAP bacteria rivaled some groups of strictly heterotrophic bacteria and was often higher than the abundance of known AAP bacterial genera (Erythrobacter and Roseobacter spp.). Concentrations of bacteriochlorophyll a (BChl a) were low ( approximately 1%) compared to those of chlorophyll a in the North Atlantic. Although the BChl a content of AAP bacteria per cell was typically 20- to 250-fold lower than the divinyl-chlorophyll a content of Prochlorococcus, the pigment content of AAP bacteria approached that of Prochlorococcus in shelf break water. Our results suggest that AAP bacteria can be quite abundant in some oceanic regimes and that their distribution in the water column is consistent with phototrophy.

Aerobic anoxygenic phototrophic (AAP) bacteria are well known to be abundant in estuaries, coastal regions and in the open ocean, but little is known about their activity in any aquatic ecosystem. To explore the activity of AAP bacteria in the Delaware estuary and coastal waters, single-cell (3)H-leucine incorporation by these bacteria was examined with a new approach that combines infrared epifluorescence microscopy and microautoradiography. The approach was used on samples from the Delaware coast from August through December and on transects through the Delaware estuary in August and November 2011. The percent of active AAP bacteria was up to twofold higher than the percentage of active cells in the rest of the bacterial community in the estuary. Likewise, the silver grain area around active AAP bacteria in microautoradiography preparations was larger than the area around cells in the rest of the bacterial community, indicating higher rates of leucine consumption by AAP bacteria. The cell size of AAP bacteria was 50% bigger than the size of other bacteria, about the same difference on average as measured for activity. The abundance of AAP bacteria was negatively correlated and their activity positively correlated with light availability in the water column, although light did not affect (3)H-leucine incorporation in light-dark experiments. Our results suggest that AAP bacteria are bigger and more active than other bacteria, and likely contribute more to organic carbon fluxes than indicated by their abundance.

Abundance and diversity of aerobic anoxygenic phototrophic bacteria in saline lakes on the Tibetan plateau

Little is known about the abundance, distribution, and ecology of aerobic anoxygenic phototrophic (AAP) bacteria, particularly in oligotrophic environments, which represent 60% of the ocean. We investigated the abundance of AAP bacteria across the South Pacific Ocean, including the center of the gyre, the most oligotrophic water body of the world ocean. AAP bacteria, Prochlorococcus, and total prokaryotic abundances, as well as bacteriochlorophyll a (BChl a) and divinyl-chlorophyll a concentrations, were measured at several depths in the photic zone along a gradient of oligotrophic conditions. The abundances of AAP bacteria and Prochlorococcus were high, together accounting for up to 58% of the total prokaryotic community. The abundance of AAP bacteria alone was up to 1.94 x 10(5) cells ml(-1) and as high as 24% of the overall community. These measurements were consistent with the high BChl a concentrations (up to 3.32 x 10(-3) microg liter(-1)) found at all stations. However, the BChl a content per AAP bacterial cell was low, suggesting that AAP bacteria are mostly heterotrophic organisms. Interestingly, the biovolume and therefore biomass of AAP bacteria was on average twofold higher than that of other prokaryotic cells. This study demonstrates that AAP bacteria can be abundant in various oligotrophic conditions, including the most oligotrophic regime of the world ocean, and can account for a large part of the bacterioplanktonic carbon stock.

Photoheterotrophic microbes use organic substrates and light energy to satisfy their demand for carbon and energy and seem to be well adapted to eutrophic estuarine and oligotrophic oceanic environments. One type of photoheterotroph, aerobic anoxygenic phototrophic (AAP) bacteria, is especially abundant in particle-rich, turbid estuaries. To explore questions regarding the controls of these photoheterotrophic bacteria, we examined their abundance by epifluorescence microscopy, concentrations of the light-harvesting pigment, bacteriochlorophyll a (BChl a) and the diversity of pufM and 16S ribosomal RNA (rRNA) genes in the Chesapeake Bay. Concentrations of BChl a varied substantially, much more so than AAP bacterial abundance, along the estuarine salinity gradient. The BChl a concentration was correlated with turbidity only when oceanic and estuarine waters were considered together. Concentrations of BChl a and BChl a quotas were higher in particle-associated than in free-living AAP bacterial communities and appear to reflect physiological adaptation, not different AAP bacterial communities; pufM genes did not differ between particle-associated and free-living communities. In contrast, particle-associated and free-living bacterial communities were significantly different, on the basis of the analysis of 16S rRNA genes. The BChl a quota of AAP bacteria was not correlated with turbidity, suggesting that pigment synthesis varies in direct response to particles, not light attenuation. The AAP bacteria seem to synthesize more BChl a when dissolved and particulate substrates are available than when only dissolved materials are accessible, which has implications for understanding the impact of substrates on the level of photoheterotrophy compared with heterotrophy in AAP bacteria.

Aerobic anoxygenic phototrophic (AAP) bacteria are bacteriochlorophyll a-containing prokaryotes which can use both light and organic compounds as energy sources. This functional group is ubiquitous in the euphotic zone of the oceans. Nevertheless, life strategies, distribution pat- terns and physiology of AAP bacteria remain largely unknown. We combined infrared fluorometry, microscopic counts and HPLC pigment analysis to characterize free-living and particle-attached AAP bacterial populations. Using a size-fractionation approach, we found that the size distribution of AAP bacteria and the fraction of particle-attached cells varied greatly among different marine environ- ments. In the open sea environments (Atlantic Ocean, offshore Mediterranean Sea), the main portion of AAP bacterial fluorescence was in the 50% of AAP bacteria were free living. In a coastal lagoon and in the deep chlorophyll a maximum at an offshore Mediterranean station, parti- cle-attached AAP bacteria formed up to half of the AAP bacterial community. The results presented here suggest that AAP bacteria can take on either free-living or particle-attached lifestyles depend- ing on environmental conditions.

With the ability of photoheterotrophic metabolism, aerobic anoxygenic phototrophic (AAP) bacteria are an important bacterial group in the marine microbial community. We investigated the diel variations of AAP bacteria in the coral reef water of the South China Sea, and report, for the first time, the frequency of dividing cells (FDC) of AAP bacteria. Our results showed that AAP bacte- rial abundance ranged from 1.54 × 10 4 ± 3.21 × 10 2 to 3.59 × 10 4 ± 3.04 × 10 3 cells ml -1 . The diel pat- tern of AAP bacterial abundance had 2 peak values, one at night around 21:00 h and the other in the daytime at around 9:00 h. Tidal current and light were the primary factors controlling the diel varia- tions of AAP bacteria in this area during our study period. FDC values in AAP and total bacteria were both higher during the night than during the day. The FDC values in AAP bacteria (3.53 ± 1.09 to 9.43 ± 0.80%) were significantly higher than those in total bacteria (1.06 ± 0.06 to 3.14 ± 1.38%), indicat- ing higher growth rate of AAP bacteria than total bacteria in the study area.

Phytoplankton is a key component of aquatic microbial communities, and metabolic coupling between phytoplankton and bacteria determines the fate of dissolved organic carbon (DOC). Yet, the impact of primary production on bacterial activity and community composition remains largely unknown, as, for example, in the case of aerobic anoxygenic phototrophic (AAP) bacteria that utilize both phytoplankton-derived DOC and light as energy sources. Here, we studied how reduction of primary production in a natural freshwater community affects the bacterial community composition and its activity, focusing primarily on AAP bacteria. The bacterial respiration rate was the lowest when photosynthesis was reduced by direct inhibition of photosystem II and the highest in ambient light condition with no photosynthesis inhibition, suggesting that it was limited by carbon availability. However, bacterial assimilation rates of leucine and glucose were unaffected, indicating that increased bacterial growth efficiency (e.g., due to photoheterotrophy) can help to maintain overall bacterial production when low primary production limits DOC availability. Bacterial community composition was tightly linked to light intensity, mainly due to the increased relative abundance of light-dependent AAP bacteria. This notion shows that changes in bacterial community composition are not necessarily reflected by changes in bacterial production or growth and vice versa. Moreover, we demonstrated for the first time that light can directly affect bacterial community composition, a topic which has been neglected in studies of phytoplankton-bacteria interactions.IMPORTANCE Metabolic coupling between phytoplankton and bacteria determines the fate of dissolved organic carbon in aquatic environments, and yet how changes in the rate of primary production affect the bacterial activity and community composition remains understudied. Here, we experimentally limited the rate of primary production either by lowering light intensity or by adding a photosynthesis inhibitor. The induced decrease had a greater influence on bacterial respiration than on bacterial production and growth rate, especially at an optimal light intensity. This suggests that changes in primary production drive bacterial activity, but the effect on carbon flow may be mitigated by increased bacterial growth efficiencies, especially of light-dependent AAP bacteria. Bacterial activities were independent of changes in bacterial community composition, which were driven by light availability and AAP bacteria. This direct effect of light on composition of bacterial communities has not been documented previously.

Aerobic anoxygenic phototrophic (AAnP) bacteria are bacteriochlorophyll a (BChl a)- containing prokaryotes that can use both dissolved organic matter and light as energy sources. AAnP bacteria are widely distributed in aquatic environments where they are expected to play an important role in carbon cycling. However, little is known about their spatio-temporal distribution in marine ecosystems. In this study we examined the dynamics of AAnP bacteria in a coastal saline lagoon from November 2007 to September 2008. AAnP cells were enumerated by infrared (IR) microscopy, and BChl a concentrations were measured by both IR kinetic fluorometry and high-performance liquid chromatography (HPLC). The distribution of AAnP bacteria varied seasonally, but no clear spatial pat- tern emerged. The abundance of these bacteria ranged from 1.0 to 13.5 × 10 4 cells ml -1 from winter to summer, comprising 0.1 to 3% of total bacterial abundance. Size fractionation of the BChl a fluores- cence signal showed that AAnP bacteria were mainly particle-attached in winter and free-living in spring and summer. BChl a concentrations (up to 108.7 ng l -1 ), BChl a content per cell (up to 1.7 fg cell -1 ) and the ratios of BChl a to chlorophyll a (chl a) (up to 15%) were high in spring and summer, suggesting that AAnP bacteria contributed significantly at this time to photosynthetically driven en- ergy production in the lagoon. Temperature and light were the main factors driving seasonal varia- tions in the abundance of AAnP bacteria, while total bacterial abundance was closely related to varia- tions in the concentration of dissolved organic carbon. These results highlight for the first time the numerical importance and the dynamics of AAnP bacteria in a coastal lagoon.

Marine microbes are major drivers of marine biogeochemical cycles and play critical roles in the ecosystems. Aerobic anoxygenic phototrophic bacteria (AAPB) are an important bacterial functional group with capability of harvesting light energy and wide distribution, and appear to have a particular role in the ocean’s carbon cycling. Yet the global pattern of AAPB distribution was controversial at the beginning of the 21st century due to the defects of the AAPB enumeration methods. An advanced time-series observation-based infrared epifluorescence microscopy (TIREM) approach was established to amend the existing AAPB quantitative deviation and led to the accurate enumeration of AAPB in marine environments. The abundance of AAPB and AAPB% were higher in coastal and continental shelf waters than in oceanic waters, which does not support the idea that AAPB are specifically adapted to oligotrophic conditions due to photosynthesis in AAPB acting a supplement to their organic carbon respiration. Further investigation revealed that dependence of AAPB on dissolved organic carbon produced by phytoplankton (PDOC) may limit their competition and control AAPB distribution. So, the selection of carbon sources by AAPB indicated that they can effectively fractionate the carbon flow in the sea. Enlightened by these findings, the following studies on the interactions between marine microbes and DOC led to the discovery of a new mechanism of marine carbon sequestration— the Microbial Carbon Pump (MCP). The conceptual framework of MCP addresses the sources and mechanism of the vast DOC reservoir in the ocean and represents a breakthrough in the theory of ocean carbon sequestration.

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 combined in situ sampling 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. The in situ decline 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.

The study aims to reveal phylogenetic and evolutionary relationship between aerobic anoxygenic phototrophic bacteria (AAnPB) and their relatives, anaerobic anoxygenic phototrophic bacteria (AnAnPB) and nonphototrophic bacteria (NPB, which had high homology of 16S rDNA gene with AAnPB and fell into the same genus), and validate reliability and usefulness of farnesyl pyrophosphate synthase (FPPS) gene for the phylogenetic determination. FPPS genes with our modified primers and 16S rDNA genes with general primers, were amplified and sequenced or retrieved from GenBank database. In contrast to 16S rDNA gene phylogenetic tree, AAnPB were grouped into two clusters and one branch alone with no intermingling with NPB and AnAnPB in the tree constructed on FPPS. One branch of AAnPB, in both trees, was located closer to outgroup species than AnAnPB, which implicated that some AAnPB would be diverged earlier in FPPS evolutionary history than AnAnPB and NPB. Some AAnPB and NPB were closer located in both trees and this suggested that they were the closer relatives than AnAnPB. Combination codon usage in FPPS with phylogenetic analysis, the results indicates that FPPS gene and 16S rRNA gene have similar evolutionary pattern but the former seems to be more reliable and useful in determining the phylogenic and evolutionary relationship between AAnPB and their relatives. This is the first attempt to use a molecular marker beside 16S rRNA gene for studying the phylogeny of AAnPB, and the study may also be helpful in understanding the evolutionary relationship among phototrophic microbes and the trends of photosynthetic genes transfer.