Abundance Of Diazotrophs Research Articles

Fertilization with inorganic and organic nutrients changes diazotroph community composition and N-fixation rates

Nitrogen fixation by free-living diazotrophs from the atmosphere is an important pathway for nitrogen input into the soil. However, there is little information regarding soil diazotrophic community composition and diversity under long-term fertilization in rice paddy ecosystems. Using the 15N2-tracing method and nifH gene as a molecular marker, we investigated the abundance, structure, and activity of soil diazotrophic community in soil at a 30-year-old filed experimental site treated with four different fertilizer management practices: control (non-fertilization), chemical NPK fertilizers, NPK plus rice straw (NPK+RS), or NPK plus chicken manure (NPK+OM). Among all the treatments, the NPK+OM treatment significantly improved the soil nutritional status. Fertilization increased both bacteria and nifH gene abundances, with the highest values (p < 0.05) found in the NPK+OM treatment. The potential nitrogen fixation rate ranged from 14.6 to 118 μg kg−1 day−1, and the highest rates (p < 0.05) were also observed in the NPK+OM treatment. Long-term chemical NPK fertilization decreased the diversity of diazotrophic community, whereas NPK+RS and NPK+OM treatments maintained the diversity of diazotrophic community. Long-term fertilization changed diazotrophic community as compared to non-fertilization, but there were no significant differences among fertilized treatments. Most nifH sequences were closely linked to Alphaproteobacteria, which was dominated by the genera Bradyrhizobium. Relatively higher Cyanobacteria abundances were observed in the unfertilized soil as compared with fertilized soil. Our results suggest that long-term fertilization increased the abundance of diazotrophs and changed their community structure, and combined use of chicken manure and chemical NPK fertilizers can significantly improve the activity of diazotrophic community.

Nitrogen Fixation Aligns with nifH Abundance and Expression in Two Coral Trophic Functional Groups.

Microbial nitrogen fixation (diazotrophy) is a functional trait widely associated with tropical reef-building (scleractinian) corals. While the integral role of nitrogen fixation in coral nutrient dynamics is recognized, its ecological significance across different coral functional groups remains yet to be evaluated. Here we set out to compare molecular and physiological patterns of diazotrophy (i.e., nifH gene abundance and expression as well as nitrogen fixation rates) in two coral families with contrasting trophic strategies: highly heterotrophic, free-living members of the family Fungiidae (Pleuractis granulosa, Ctenactis echinata), and mostly autotrophic coral holobionts with low heterotrophic capacity (Pocilloporidae: Pocillopora verrucosa, Stylophora pistillata). The Fungiidae exhibited low diazotroph abundance (based on nifH gene copy numbers) and activity (based on nifH gene expression and the absence of detectable nitrogen fixation rates). In contrast, the mostly autotrophic Pocilloporidae exhibited nifH gene copy numbers and gene expression two orders of magnitude higher than in the Fungiidae, which coincided with detectable nitrogen fixation activity. Based on these data, we suggest that nitrogen fixation compensates for the low heterotrophic nitrogen uptake in autotrophic corals. Consequently, the ecological importance of diazotrophy in coral holobionts may be determined by the trophic functional group of the host.

Impact of fertilization regimes on diazotroph community compositions and N2-fixation activity in paddy soil

Abstract Nutrient status in soil is crucial for the growth and activity of resident microorganisms, which in turn regulate nitrogen transformation in the terrestrial biosphere. Biological nitrogen fixation (BNF) is an important N input in agricultural ecosystems, but the influence of fertilization on the structural and functional behavior of diazotrophs is not clear. In this study, we assessed the nutrient limitations on the abundance and N2-fixation activity of diazotrophs in a long term (20 years) fertilization experiment. Although both phosphorus (P) deficiency and potassium (K) deficiency resulted in significant decreases in nifH gene expression and N2-fixation activity, P deficiency exhibited more restrictive effects. Assessments of community structures based upon transcripts also indicated that the active diazotroph populations were more sensitive to P deficiency than K deficiency, and some diazotroph groups were detected in the P deficiency treatment. Long-term rice straw addition significantly increased diazotroph abundance, but in contrast, sharply reduced nifH gene expression and N2-fixation activity. The close relationship between N2-fixation activity and nifH gene expression rather than its copy number suggests that the nifH gene transcript level is a suitable indicator for predicting the N2-fixation activity of N2-fixing microorganisms in paddy soil of various fertility status.

Impact of 25 years of inorganic fertilization on diazotrophic abundance and community structure in an acidic soil in southern China

Although diazotrophs are important in the nitrogen (N)-cycle and contribute to the pool of plant available N, the population response to long-term inorganic fertilization is largely unknown. Here, we investigated the diazotrophic populations in both the bulk and rhizosphere soils of maize grown in an acidic farmland soil that experienced 25 years of inorganic fertilization. The fertilization regimes included unfertilized control, N fertilizer alone, N fertilizer with quicklime, phosphorus (P) and potassium (K) fertilizers, N + P + K fertilizers, and N + P + K fertilizers with quicklime. Quantitative PCR and high-throughput pyrosequencing of the nifH gene were used to analyze diazotrophic abundance and community composition. All of the fertilizer treatments improved soil nutrient availability, but those without quicklime caused soil acidification. Maize biomasses and nifH copy numbers were significantly lower under N and N + P + K treatments but increased under P + K fertilization. Quicklime applications effectively alleviated the inhibitory effect of N input. Fertilization led to decreases in operational taxonomic unit richness and shifts in diazotrophic community composition. Soil pH and nutrient availability had a cooperative effect on diazotrophic abundance, while soil nutrient availability appeared to be the main factor shaping diazotrophic community structure. Rhizosphere effects increased the nifH gene copy number but did not obviously change the diazotrophic community composition on the current research scale. Overall, the long-term inorganic fertilization affected both diazotrophic abundance and community composition, and the fertilizer treatment had a greater influence than quicklime remediation or crop cultivation on community composition.

Diazotrophs: Overlooked Key Players within the Coral Symbiosis and Tropical Reef Ecosystems?

Coral reefs are highly productive ecosystems thriving in nutrient-poor waters. Their productivity depends largely on the availability of nitrogen, the proximate limiting nutrient for primary production. In reefs, the major nitrogen pathways include regeneration, nitrification, ammonification and dinitrogen (N2) fixation. N2 fixation is performed by prokaryotes called ‘diazotrophs’ that abound in coral rubbles, sandy bottoms, microbial mats or seagrass meadows. Corals, which are the main reef builders, have developed a partnership with dinoflagellates which transform dissolved inorganic nitrogen into amino acids and protein, and with diazotrophs to gain diazotrophically-derived nitrogen (DDN). Pioneering studies found active diazotrophic cyanobacteria in the corals’ mucus and/or tissue, and later high throughput sequencing efforts have described diverse communities of non-cyanobacterial diazotrophs associated with scleractinian corals. Yet, the metabolic processes behind these associations and how they benefit corals is currently not well understood. While genomic studies describe the diversity of diazotrophs and isotopic tracer experiments quantify N2 fixation rates, combined advanced methods are needed to elucidate the mechanisms behind the transfer of DDN to the coral holobiont and whether these mechanisms change according to the identity of the diazotrophs or coral species. Here we review our current knowledge on N2 fixation in corals: the diversity and localization of diazotrophs in the coral holobiont, the environmental factors controlling N2 fixation, the fate of DDN within the coral symbiosis as well as its potential role in coral resilience. We finally summarize the unknowns: are the diversity, abundance and localization of diazotrophs within the holobiont species- and/or site-specific? Do they have an impact on DDN production? What are the metabolic mechanisms implicated? Do they change spatially, temporally or according to environmental factors? We encourage scientists to undertake research efforts to tackle these questions in order to shed light on nitrogen cycling in reef ecosystems and to understand if coral-associated N2 fixation can improve coral’s resilience in the face of climate change.

The Assimilation of Diazotroph-Derived Nitrogen by Scleractinian Corals Depends on Their Metabolic Status.

ABSTRACTTropical corals are associated with a diverse community of dinitrogen (N2)-fixing prokaryotes (diazotrophs) providing the coral an additional source of bioavailable nitrogen (N) in oligotrophic waters. The overall activity of these diazotrophs changes depending on the current environmental conditions, but to what extent it affects the assimilation of diazotroph-derived N (DDN) by corals is still unknown. Here, in a series of 15N2 tracer experiments, we directly quantified DDN assimilation by scleractinian corals from the Red Sea exposed to different environmental conditions. We show that DDN assimilation strongly varied with the corals’ metabolic status or with phosphate availability in the water. The very autotrophic shallow-water (~5 m) corals showed low or no DDN assimilation, which significantly increased under elevated phosphate availability (3 µM). Corals that depended more on heterotrophy (i.e., bleached and deep-water [~45 m] corals) assimilated significantly more DDN, which contributed up to 15% of the corals’ N demand (compared to 1% in shallow corals). Furthermore, we demonstrate that a substantial part of the DDN assimilated by deep corals was likely obtained from heterotrophic feeding on fixed N compounds and/or diazotrophic cells in the mucus. Conversely, in shallow corals, the net release of mucus, rich in organic carbon compounds, likely enhanced diazotroph abundance and activity and thereby the release of fixed N to the pelagic and benthic reef community. Overall, our results suggest that DDN assimilation by corals varies according to the environmental conditions and is likely linked to the capacity of the coral to acquire nutrients from seawater.

Evidence for polyploidy in the globally important diazotroph Trichodesmium.

Polyploidy is a well-described trait in some prokaryotic organisms; however, it is unusual in marine microbes from oligotrophic environments, which typically display a tendency towards genome streamlining. The biogeochemically significant diazotrophic cyanobacterium Trichodesmium is a potential exception. With a relatively large genome and a comparatively high proportion of non-protein-coding DNA, Trichodesmium appears to allocate relatively more resources to genetic material than closely related organisms and microbes within the same environment. Through simultaneous analysis of gene abundance and direct cell counts, we show for the first time that Trichodesmium spp. can also be highly polyploid, containing as many as 100 genome copies per cell in field-collected samples and >600 copies per cell in laboratory cultures. These findings have implications for the widespread use of the abundance of the nifH gene (encoding a subunit of the N2-fixing enzyme nitrogenase) as an approach for quantifying the abundance and distribution of marine diazotrophs. Moreover, polyploidy may combine with the unusual genomic characteristics of this genus both in reflecting evolutionary dynamics and influencing phenotypic plasticity and ecological resilience.

Effects of herbivores on nitrogen fixation by grass endophytes, legume symbionts and free-living soil surface bacteria in the Serengeti

Grass roots can harbor abundant endophytic N2-fixing microbes (diazotrophs), but their abundance and activity compared to those on legumes and in soil crusts is still unknown. Here, in a natural ecosystem, the Serengeti of East Africa, we explored whether herbivores and soil nutrients limited grass root endophyte diazotroph abundance and their root mass-specific and area-specific N2-fixation, as they often do for diazotrophs symbiotic with legumes and those free-living in soil. N2-fixation and copy number of the nitrogenase gene nifH was measured with stable isotope and molecular methods, respectively, for the dominant grass Themeda triandra, and legume, Indigofera volkensii, and in the top 5cm of soil in a 16-year herbivore exclosure experiment across four sites that varied in mean annual rainfall and soil N, P, and moisture. T. triandra nifH gene copy number was highly variable across sites and individuals but often approached or exceeded that of I. volkensii roots and soils. T. triandra roots generally exhibited lower root mass-specific N2-fixation (activity), which was not reduced by herbivores and increased in drier soils. In contrast, I. volkensii activity was only reduced by herbivores and soil diazotrophs were mostly inactive. T. triandra exhibited greater area-specific N2-fixation than I. volkensii, due to its much greater root biomass, but this difference was reduced by herbivores. Grass-associated endophytic diazotrophs may fix far more N2 in natural systems than previously realized, and may be limited by different factors those affecting symbiotic legume and free-living soil diazotrophs.

Contribution of mono and polysaccharides to heterotrophic N2 fixation at the eastern Mediterranean coastline.

N2 fixation should be a critical process in the nitrogen-poor surface water of the eastern Mediterranean Sea. Despite favorable conditions, diazotroph abundance and N2 fixation rates remains low for reasons yet explained. The main goal of this study was to investigate the limiting nutrients for diazotrophy in this oligotrophic environment. Hence, we conducted dedicated bottle-microcosms with eastern Mediterranean Sea water that were supplemented with mono and polysaccharides as well as inorganic nitrogen and phosphorous. Our results indicate that the diazotrophic community expressing nifH was primarily represented by heterotrophic Proteobacteria. N2 fixation and heterotrophic bacterial activity increased up-to tenfold following two days of dark incubations, once seawater was supplemented with organic carbon substrate in the form of glucose (monosaccharides) or gum-xanthan (polysaccharide surrogate). Furthermore, our results point that carbon-rich polysaccharides, such as transparent exopolymer particles, enhance heterotrophic N2 fixation, by forming microenvironments of intense metabolic activity, high carbon: nitrogen ratio, and possibly low O2 levels. The conclusions of this study indicate that diazotrophs in the eastern Mediterranean coast are primarily limited by organic carbon substrates, as possibly in many other marine regions.

High levels of heterogeneity in diazotroph diversity and activity within a putative hotspot for marine nitrogen fixation.

Australia's tropical waters represent predicted 'hotspots' for nitrogen (N2) fixation based on empirical and modelled data. However, the identity, activity and ecology of diazotrophs within this region are virtually unknown. By coupling DNA and cDNA sequencing of nitrogenase genes (nifH) with size-fractionated N2 fixation rate measurements, we elucidated diazotroph dynamics across the shelf region of the Arafura and Timor Seas (ATS) and oceanic Coral Sea during Austral spring and winter. During spring, Trichodesmium dominated ATS assemblages, comprising 60% of nifH DNA sequences, while Candidatus Atelocyanobacterium thalassa (UCYN-A) comprised 42% in the Coral Sea. In contrast, during winter the relative abundance of heterotrophic unicellular diazotrophs (δ-proteobacteria and γ-24774A11) increased in both regions, concomitant with a marked decline in UCYN-A sequences, whereby this clade effectively disappeared in the Coral Sea. Conservative estimates of N2 fixation rates ranged from <1 to 91 nmol l(-1) day(-1), and size fractionation indicated that unicellular organisms dominated N2 fixation during both spring and winter, but average unicellular rates were up to 10-fold higher in winter than in spring. Relative abundances of UCYN-A1 and γ-24774A11 nifH transcripts negatively correlated to silicate and phosphate, suggesting an affinity for oligotrophy. Our results indicate that Australia's tropical waters are indeed hotspots for N2 fixation and that regional physicochemical characteristics drive differential contributions of cyanobacterial and heterotrophic phylotypes to N2 fixation.

Fungal endophyte Phomopsis liquidambari affects nitrogen transformation processes and related microorganisms in the rice rhizosphere.

The endophytic fungus Phomopsis liquidambari performs an important ecosystem service by assisting its host with acquiring soil nitrogen (N), but little is known regarding how this fungus influences soil N nutrient properties and microbial communities. In this study, we investigated the impact of P. liquidambari on N dynamics, the abundance and composition of N cycling genes in rhizosphere soil treated with three levels of N (urea). Ammonia-oxidizing archaea (AOA), ammonia-oxidizing bacteria (AOB) and diazotrophs were assayed using quantitative real-time polymerase chain reaction and denaturing gradient gel electrophoresis at four rice growing stages (S0: before planting, S1: tillering stage, S2: grain filling stage, and S3: ripening stage). A significant increase in the available nitrate and ammonium contents was found in the rhizosphere soil of endophyte-infected rice under low N conditions. Moreover, P. liquidambari significantly increased the potential nitrification rates, affected the abundance and community structure of AOA, AOB, and diazotrophs under low N conditions in the S1 and S2 stages. The root exudates were determined due to their important role in rhizosphere interactions. P. liquidambari colonization altered the exudation of organic compounds by rice roots and P. liquidambari increased the concentration of soluble saccharides, total free amino acids and organic acids in root exudates. Plant-soil feedback mechanisms may be mediated by the rice-endophyte interaction, especially in nutrient-limited soil.

The core root microbiome of sugarcanes cultivated under varying nitrogen fertilizer application.

Diazotrophic bacteria potentially supply substantial amounts of biologically fixed nitrogen to crops, but their occurrence may be suppressed by high nitrogen fertilizer application. Here, we explored the impact of high nitrogen fertilizer rates on the presence of diazotrophs in field-grown sugarcane with industry-standard or reduced nitrogen fertilizer application. Despite large differences in soil microbial communities between test sites, a core sugarcane root microbiome was identified. The sugarcane root-enriched core taxa overlap with those of Arabidopsis thaliana raising the possibility that certain bacterial families have had long association with plants. Reduced nitrogen fertilizer application had remarkably little effect on the core root microbiome and did not increase the relative abundance of root-associated diazotrophs or nif gene counts. Correspondingly, low nitrogen fertilizer crops had lower biomass and nitrogen content, reflecting a lack of major input of biologically fixed nitrogen, indicating that manipulating nitrogen fertilizer rates does not improve sugarcane yields by enriching diazotrophic populations under the test conditions. Standard nitrogen fertilizer crops had improved biomass and nitrogen content, and corresponding soils had higher abundances of nitrification and denitrification genes. These findings highlight that achieving a balance in maximizing crop yields and minimizing nutrient pollution associated with nitrogen fertilizer application requires understanding of how microbial communities respond to fertilizer use.

Non-cyanobacterial diazotrophs mediate dinitrogen fixation in biological soil crusts during early crust formation.

Biological soil crusts (BSCs) are key components of ecosystem productivity in arid lands and they cover a substantial fraction of the terrestrial surface. In particular, BSC N2-fixation contributes significantly to the nitrogen (N) budget of arid land ecosystems. In mature crusts, N2-fixation is largely attributed to heterocystous cyanobacteria; however, early successional crusts possess few N2-fixing cyanobacteria and this suggests that microorganisms other than cyanobacteria mediate N2-fixation during the critical early stages of BSC development. DNA stable isotope probing with (15)N2 revealed that Clostridiaceae and Proteobacteria are the most common microorganisms that assimilate (15)N2 in early successional crusts. The Clostridiaceae identified are divergent from previously characterized isolates, though N2-fixation has previously been observed in this family. The Proteobacteria identified share >98.5% small subunit rRNA gene sequence identity with isolates from genera known to possess diazotrophs (for example, Pseudomonas, Klebsiella, Shigella and Ideonella). The low abundance of these heterotrophic diazotrophs in BSCs may explain why they have not been characterized previously. Diazotrophs have a critical role in BSC formation and characterization of these organisms represents a crucial step towards understanding how anthropogenic change will affect the formation and ecological function of BSCs in arid ecosystems.

The Impact of Mineral Nitrogen Fertilization on the Occurrence of Native Diazotrophic Bacteria in Kohlrabi (Brassica oleracea) Shoots and Roots

Biological Nitrogen Fixation (BNF) is a process of great importance in crop production systems, as it provides additional natural sources of mineral nitrogen. BNF is catalyzed by diazotrophs that are identified by the nif operon presence comprising the nifH gene that encodes for enzyme nitrogenase synthesis. Thoroughly understanding of factors that influence diazotrophic abundance is crucial for their utilization to enhance sustainability and prevent land degradation in modern agriculture. In this study the impacts of nitrogen fertilization on diazotrophic abundance in Brassica oleracea roots and leaves was investigated in greenhouse experiments by real-time qPCR. One way ANOVA was used to compare means and bivariate Pearson correlation tested for relationships between variables. Increased nitrogen fertilization significantly increased the nitrogen content in leaves but not in roots. No significant changes in nifH gene copy numbers nor in proportion of nifH gene copy numbers were detectable. This indicates no effect of mineral N fertilization on the abundance of total native diazotrophic bacterial numbers in Brassica oleracea plants.

Macroalgal blooms favor heterotrophic diazotrophic bacteria in nitrogen-rich and phosphorus-limited coastal surface waters in the Yellow Sea

Macroalgal blooms may lead to dramatic changes in physicochemical variables and biogeochemical cycling in affected waters. However, little is known about the effects of macroalgal blooms on marine bacteria, especially those functioning in nutrient cycles. We measured environmental factors and investigated bacterial diazotrophs in two niches, surface waters that were covered (CC) and non-covered (CF) with massive macroalgal canopies of Ulva prolifera, in the Yellow Sea in the summer of 2011 using real-time PCR and clone library analysis of nifH genes. We found that heterotrophic diazotrophs (Gammaproteobacteria) dominated the communities and were mostly represented by Vibrio-related phylotypes in both CC and CF. Desulfovibrio-related phylotypes were only detected in CC. There were significant differences in community composition in these two environments (p < 0.001) and a much higher abundance of nifH in CC (4.55 × 106 copies l−1) than in CF (2.49 × 106 copies l−1). The nifH copy number was inversely related to concentrations of ammonium and dissolved inorganic nitrogen and to the stoichiometric ratios of N:P and N:Si. This indicates that macroalgal blooms significantly affect diazotrophic abundance and community composition and that vibrios and Desulfovibrio-related heterotrophic diazotrophs adapt well to the (N-rich but P-limited) environment during blooming. Potential ecological and microbiological mechanisms behind this scenario are discussed.

Nitrogen fixation in shallow‐water sediments: Spatial distribution and controlling factors

Nitrogenase activity (NA) in shallow‐water (&lt; 1 m) sediments was investigated at 60 randomly selected sites along a 150 km stretch on the brackish‐water Swedish west coast, without targeting any specific type of sediments, such as microbial mats. Benthic nitrogen (N) fixation and diazotrophs (nifH genes) were found at all sites, regardless of the presence of cyanobacterial or microbial mats. The majority of sites showed N fixation rates between 0.03 and 1 mmol N m−2 d−1. These rates were similar to those of benthic denitrification previously measured in the area. Maximum rates up to 3.4 mmol N m−2 d−1 were measured. A structural equation model was used to investigate direct and indirect effects of biogeochemical and physical factors on NA. Number of nifH genes had the largest direct positive influence on NA, whereas increasing wave exposure had an indirect negative effect on NA through its influence on the diazotrophic abundance. Increased salinity, previously been shown to suppress NA in coastal waters, was found to directly stimulate benthic N fixation, likely by generating favorable conditions for diazotrophic sulfate‐reducing bacteria. Our field data confirmed previously observed negative effects of dissolved inorganic nitrogen on NA, which have so far mainly been experimentally studied. Both NA rates and the number of nifH genes correlated positively with pore‐water dissolved inorganic phosphorus concentrations. These findings show that the potential for N fixation in illuminated sediments can be considerable, stretching beyond cyanobacterial mats, being controlled by complex interactions between biotic and abiotic factors.

Production of dissolved organic carbon enriched in deoxy sugars representing an additional sink for biological C drawdown in the Amazon River plume

AbstractIn North Atlantic waters impacted by discharges from the Amazon and Orinoco Rivers, where planktonic diatom‐diazotroph associations (DDA) were active, we observed that an average (± standard deviation) of 61 ± 12% of the biological drawdown of dissolved inorganic carbon (DIC) was partitioned into the accumulating total organic carbon pool, representing a flux of up to 9 ± 4 Tg C yr−1. This drawdown corresponded with chemical alteration of ultrafiltered dissolved organic matter (UDOM), including increases in stable C isotopic composition (δ13C) and C:N. The dissolved carbohydrate component of UDOM also increased with biological DIC drawdown and diatom‐associated diazotroph (i.e., Richelia) abundance. New carbohydrates could be distinguished by distinctively high relative abundances of deoxy sugars (up to 55% of monosaccharides), which may promote aggregate formation and enhance vertical carbon export. The identified production of non‐Redfieldian, C‐enriched UDOM thus suggests a mechanism to explain enhanced C sequestration associated with DDA N2 fixation, which may be widespread in mesohaline environments.

Experimental assessment of diazotroph responses to elevated seawater pCO2 in the North Pacific Subtropical Gyre

AbstractWe examined short‐term (24–72 h) responses of naturally occurring marine N2 fixing microorganisms (termed diazotrophs) to abrupt increases in the partial pressure of carbon dioxide (pCO2) in seawater during nine incubation experiments conducted between May 2010 and September 2012 at Station ALOHA (A Long‐term Oligotrophic Habitat Assessment) (22°45′N, 158°W) in the North Pacific Subtropical Gyre (NPSG). Rates of N2 fixation, nitrogenase (nifH) gene abundances and transcripts of six major groups of cyanobacterial diazotrophs (including both unicellular and filamentous phylotypes), and rates of primary productivity (as measured by 14C‐bicarbonate assimilation into plankton biomass) were determined under contemporary (~390 ppm) and elevated pCO2 conditions (~1100 ppm). Quantitative polymerase chain reaction (QPCR) amplification of planktonic nifH genes revealed that unicellular cyanobacteria phylotypes dominated gene abundances during these experiments. In the majority of experiments (seven out of nine), elevated pCO2 did not significantly influence rates of dinitrogen (N2) fixation or primary productivity (two‐way analysis of variance (ANOVA), P &gt; 0.05). During two experiments, rates of N2 fixation and primary productivity were significantly lower (by 79 to 82% and 52 to 72%, respectively) in the elevated pCO2 treatments relative to the ambient controls (two‐way ANOVA, P &lt; 0.05). QPCR amplification of nifH genes and gene transcripts revealed that diazotroph abundances and nifH gene expression were largely unchanged by the perturbation of the seawater pCO2. Our results suggest that naturally occurring N2 fixing plankton assemblages in the NPSG are relatively resilient to large, short‐term increases in pCO2.

Climate change affects key nitrogen-fixing bacterial populations on coral reefs

Coral reefs are at serious risk due to events associated with global climate change. Elevated ocean temperatures have unpredictable consequences for the ocean's biogeochemical cycles. The nitrogen cycle is driven by complex microbial transformations, including nitrogen fixation. This study investigated the effects of increased seawater temperature on bacteria able to fix nitrogen (diazotrophs) that live in association with the mussid coral Mussismilia harttii. Consistent increases in diazotroph abundances and diversities were found at increased temperatures. Moreover, gradual shifts in the dominance of particular diazotroph populations occurred as temperature increased, indicating a potential future scenario of climate change. The temperature-sensitive diazotrophs may provide useful bioindicators of the effects of thermal stress on coral reef health, allowing the impact of thermal anomalies to be monitored. In addition, our findings support the development of research on different strategies to improve the fitness of corals during events of thermal stress, such as augmentation with specific diazotrophs.

Analysis of the occurrence and activity of diazotrophic communities in organic and conventional horticultural soils

Nitrogen (N)-fixing microorganisms are an important soil component as they help replenish the pool of available N. Organic management can influence the N-fixing community; however, diazotroph community structure and activity in horticultural systems and the impacts of specific cultivation methods (i.e., greenhouse and open field) are unclear. Using the nifH gene, a marker gene for the microbial community involved in N fixation, we investigated the occurrence (DNA) and activity (RNA) of the diazotrophic community in organically and conventionally managed soils in a horticultural system over the course of 1 year. Ordination analysis of DGGE profiles revealed organic management affected the community structure in the greenhouse but not the open field; fertilization intensity may explain this divergent response, as indicated by the relevance of total C content to community structure. Quantitative PCR revealed that organic management increased the abundance and activity of diazotrophs. The soluble organic N concentration was higher in organically managed soils and during warmer months, and correlated with diazotroph abundance. Most identified sequences were from known diazotrophs, predominantly β-, γ- and α-proteobacteria. Twenty-four bands resembled Pseudomonas stutzeri and eight resembled Azoarcus sp. Our results show that the cultivation method controls the extent of the effects of season and organic management on diazotrophs, and that greenhouse cultivation can boost the effects of organic management on this community. Organic management intensified the positive effect of seasonal temperature on diazotroph abundance and activity, which may increase biological nitrogen fixation rates. In tandem, soil DNA and RNA analyses provide a comprehensive picture of the community.