Biochar has the potential to remediate heavy metals in agricultural soil and mitigate nitrous oxide (N₂O) emissions; however, the effects of biochar on heavy metal remediation, the soil microbial community and N₂O emissions are not completely understood. In this study, we conducted a pot experiment in which Glycine max L. (soybean) was cultivated in two cadmium (Cd)-contaminated soils (low, 3.14 mg kg⁻¹; high, 10.80 mg kg⁻¹) to investigate the effects of biochar on the bioremediation of Cd, N₂O emissions and the rhizosphere microbial community structure. The bioaccumulation of Cd in the plant shoots and roots increased with all biochar addition rates (0%, 1%, 5% and 10%); unexpectedly, the translocation capacity of Cd to the edible parts of the plant significantly decreased to 0.58 mg kg⁻¹, which was close to the edible threshold (0.4 mg kg⁻¹). The abundance and activities of functional marker genes of microbial nitrification (amoA) and denitrification (nirK, nirS and nosZ) were quantified with quantitative PCR, and we found that biochar addition reduced the precursor production of rhizoshpere N₂O by inhibiting the transcription of the nirK gene. In addition, the nitrogenase activity during anthesis (S) was significantly (P < 0.05) increased with 1% (v/v) biochar addition. Noticeably, biochar addition only changed the microbial community structure in the very first stage before eventually stabilize. This study highlighted that biochar has the potential ability to maintain the quality of agricultural crops, remediate Cd-contaminated soils and may help reduce N₂O emissions without disturbing the microbial community.

We investigated the ability of the aquatic fern Azolla to take up ciprofloxacin (Cipro), as well as the effects of that antibiotic on the N-fixing process in plants grown in medium deprived (-N) or provided (+N) with nitrogen (N). Azolla was seen to accumulate Cipro at concentrations greater than 160 μg g⁻¹ dry weight when cultivated in 3.05 mg Cipro l⁻¹, indicating it as a candidate for Cipro recovery from water. Although Cipro was not seen to interfere with the heterocyst/vegetative cell ratios, the antibiotic promoted changes with carbon and nitrogen metabolism in plants. Decreased photosynthesis and nitrogenase activity, and altered plant's amino acid profile, with decreases in cell N concentrations, were observed. The removal of N from the growth medium accentuated the deleterious effects of Cipro, resulting in lower photosynthesis, N-fixation, and assimilation rates, and increased hydrogen peroxide accumulation. Our results shown that Cipro may constrain the use of Azolla as a biofertilizer species due to its interference with nitrogen fixation processes.

Nitrogen fixation and its ecological regulation are poorly understood in the tropical estuaries, which are highly influenced by anthropogenic disturbances. In this study, we investigated the role of nutrient stoichiometry in the diversity, abundance and activity of N₂-fixing bacterial community and their seasonal variations in the water column of a tropical eutrophic estuary (Cochin estuary). The N₂ fixation rates in the estuary ranged from 0.1 to 2.0 nmol N₂ l⁻¹ h⁻¹, with higher activity during post-monsoon and lower during monsoon. The rates are appeared to be primarily controlled by dissolved inorganic nitrogen and phosphorous (N:P) ratio. Clone library analysis of nitrogenase (nifH) gene revealed that the major N₂ fixing phylotypes belong to Cluster I and Cluster III diazotrophs. The overall findings of this study suggest that monsoon induced seasonal changes in nutrient stoichiometry control the distribution and activity of diazotrophs in a tropical estuary.

Few studies focus on the effects of aluminum (Al) on marine nitrogen-fixing cyanobacteria, which play important roles in the ocean nitrogen cycling. To examine the effects of Al on the nitrogen-fixing cyanobacteria, bioassay experiments in the oligotrophic South China Sea (SCS) and culture of Crocosphaera watsonii in the laboratory were conducted. Field data showed that 200 nM Al stimulated the growth and the nitrogenase gene expression of Trichodesmium and unicellular diazotrophic cyanobacterium group A, and the nitrogen fixation rates of the whole community. Laboratory experiments demonstrated that Al stimulated the growth and nitrogen fixation of C. watsonii under phosphorus limited conditions. Both field and laboratory results indicated that Al could stimulate the growth of diazotrophs and nitrogen fixation in oligotrophic oceans such as the SCS, which is likely related to the utilization of phosphorus, implying that Al plays an important role in the ocean nitrogen and carbon cycles by influencing nitrogen fixation.

Kudzu (Pueraria lobata) is an important medicinal plant, which can associate with rhizobia for nitrogen fixation. The mutualistic symbiosis between rhizobium and kudzu is not well understood, but it is necessary to fully utilize kudzu. Nodules and rhizosphere soils collected from 16 sampling sites were characterized based on phylogenetic analyses of the rpoB gene; 16S rRNA gene; the housekeeping genes SMc00019, truA, and thrA; and the symbiotic genes nodA and nifH. The relationships between biogeographic pattern, nitrogenase activity, and environmental factors were studied. Results indicated that a clear biogeographic pattern of rhizobial communities in the kudzu rhizosphere existed in southern China; latitude and soil pH were found to be the most important factors affecting the biogeographic pattern. Bradyrhizobium diazoefficiens and Bradyrhizobium erythrophlei were the dominant species in kudzu rhizosphere. The symbiotic rhizobia in kudzu nodules mainly belonged to B. lablabi, B. elkanii, B. pachyrhizi, and B. japonicum. Nitrogenase activities in the nodules of kudzu in the Jiangxi sampling region were significantly higher than those in the Guangxi and Hunan sampling regions, and they were significantly negatively correlated to pH and exchangeable Ca. These results constitute the first report of the existence of symbiotic genes in kudzu bradyrhizobia, which are similar to those in B. elkanii and B. pachyrhizi. Our findings could improve the understanding of kudzu–rhizobium symbiosis and could advance the application of rhizobial inoculation in medicinal legumes in terms of increasing the content of active ingredients.

Diazotrophy is considered as one of the most crucial and dynamic phenomena in the rice field and also a major source of nitrogen input. The objective of this study was to elucidate possible interactions between diverse and dominant diazotrophic bacterial community and organic carbon composition of the paddy soil. Our results suggest that most abundantly found diazotrophs belong to a proteobacteria group and uncultured bacterial forms. A gene abundance study clearly showed significantly higher diazotrophic abundance (P < 0.01) at Chandauli (CHN) as compared to Varanasi (VNS) and Ghazipur (GHJ) districts of Eastern Uttar Pradesh, India, with nitrogenase reductase (nifH) copy number between 1.44 × 10³ and 3.34 × 10³ copy g⁻¹ soil. Fourier-transform infrared (FT-IR) spectroscopy data identified –CO–, C=O ([Formula: see text] and –NH–), [Formula: see text], and OH– as dominant organic functional groups in the paddy soil. Multivariate analysis was performed to get a clear and more accurate picture of interactions between free-living diazotrophs and abiotic soil factors. Regression analysis suggested a similar trend of distribution of different functional groups along each site. Relative abundance and diversity of diazotrophic population increased in response to FT-IR-based soil organic fractions. Maximum number of FT-IR spectral peak at sites in the Chandauli district augmented its bacterial diazotrophic diversity and abundance. Taken together, the present study sheds light on the substrate-driven composition of the microbial population of selected paddy areas.

Nitrogen-fixing microorganisms (diazotrophs) play important roles in aquatic biogeochemistry and ecosystem functioning. However, little is known about the spatiotemporal variation of diazotrophic microbial communities in deep subtropical reservoirs. In this study, denaturing gradient gel electrophoresis (DGGE), clone libraries, quantitative PCR, and quantitative reverse transcription (RT)-PCR were used together to examine the vertical and seasonal patterns of diazotrophic microbial communities based on nitrogenase (nifH) gene sequences in the Dongzhen Reservoir, China, across time (every 3 months for 1 year) and space (five different water depths). In general, the numbers of DGGE bands increased with water depth during the stratification seasons (spring, summer, and autumn), with the clone-library-based operational taxonomic unit (OTU) number and nifH gene diversity being highest in autumn (6 OTUs at depth 0 m; 15 OTUs at 33 m) and winter (12 OTUs at 0 m, 13 OTUs at 33 m) but decreasing drastically in spring (2 OTUs at 0 m, 3 OTUs at 33 m) and summer (3 OTUs at 0 m, 2 OTUs at 33 m). The nifH gene abundance was lowest in the water mixing season (winter average, 5.17 × 10⁷ copies/L) but increased in the three other seasons (9.03 × 10⁹ copies/L). Cyanobacteria (dominated by filamentous thermophilic cyanobacteria and Cylindrospermopsis raciborskii) were the most dominant diazotrophic group at all depths and seasons, while both alphaproteobacteria and gammaproteobacteria were co-dominant in the bottom waters in autumn and winter. The distinct seasonal and spatial patterns in diazotrophic communities were significantly related to total nitrogen (TN) and ammonium nitrogen (NH₄-N) in the reservoir (P < 0.01). Further, TN showed a significant positive correlation with nifH RNA copy number (P < 0.05) and DGGE band number (P < 0.01), whereas the NH₄-N was negatively correlated with nifH DNA copy number (P < 0.01) and positively with both RNA/DNA ratio (P < 0.01) and DGGE band number (P < 0.01). Our data indicated that water stratification, mixing, and nitrogen might drive the diazotrophic community structure and activity in complex ways, thereby influencing the aquatic nitrogen cycle. Therefore, adaptive reservoir management strategies should carefully consider the effects of water stratification for protecting drinking water quality and for controlling the potential for diazotrophic cyanobacteria blooms.