RESUMEN
Grazing exclusion has been implemented worldwide as a nature-based solution for restoring degraded grassland ecosystems that arise from overgrazing. However, the effect of grazing exclusion on soil nitrogen cycle processes, subsequent greenhouse gas emissions and underlying mechanisms remain unclear. Here, we investigated the effect of four-year grazing exclusion on plant communities, soil properties, and soil nitrogen cycle-related functional gene abundance in an alpine meadow on the Qinghai-Tibet Plateau. Using an automated continuous-flow incubation system, we performed an incubation experiment and measured soil-borne N2O, N2, and CO2 fluxes to three successive "hot moment" events (precipitation, N deposition, and oxic-to-anoxic transition) between grazing-excluded and grazing soil. Higher soil N contents (total nitrogen, NH4+, NO3-) and extracellular enzyme activities (ß-1,4-glucosidase, ß-1,4-N-acetyl-glucosaminidase, cellobiohydrolase) are observed under grazing exclusion. The aboveground and litter biomass of plant community was significantly increased by grazing exclusion, but grazing exclusion decreased the average number of plant species and microbial diversity. The N2O + N2 fluxes observed under grazing exclusion were higher than those observed under free grazing. The N2 emissions and N2O/(N2O + N2) ratios observed under grazing exclusion were higher than those observed under free grazing in oxic conditions. Instead, higher N2O fluxes and lower denitrification functional gene abundances (nirS, nirK, nosZ, and nirK + nirS) under anoxia were found under grazing exclusion than under free grazing. The N2O site-preference value indicates that under grazing exclusion, bacterial denitrification contributes more to higher N2O production compared with under free grazing (81.6 % vs. 59.9 %). We conclude that grazing exclusion could improve soil fertility and plant biomass, nevertheless it may lower plant and microbial diversity and increase potential N2O emission risk via the alteration of the denitrification end-product ratio. This indicates that not all grassland management options result in a mutually beneficial situation among wider environmental goals such as greenhouse gas mitigation, biodiversity, and social welfare.
Asunto(s)
Desnitrificación , Gases de Efecto Invernadero , Tibet , Ecosistema , Pradera , Suelo , Microbiología del Suelo , Óxido Nitroso/análisisRESUMEN
Estuarine and coastal environments are assumed to contribute to nitrous oxide (N2O) emissions under increasing nitrogen loading. However, isotopic and molecular mechanisms underlying N2O production pathways under elevated nitrogen concentration remain poorly understood. Here we used microbial inhibition, isotope mass balance, and molecular approaches to investigate N2O production mechanisms in estuarine and coastal sediments through a series of anoxic incubations. Site preference of the N2O molecule increased due to increasing nitrate concentration, suggesting the changes in N2O production pathways. Enhanced N2O production under high nitrate concentration was not mediated by bacterial denitrification, but instead was mainly regulated by fungal denitrification. Elevated nitrate concentration increased the contribution of fungal denitrification to N2O production by 11-25%, whereas it decreased bacterial N2O production by 16-33%. Chemodenitrification was also enhanced by high nitrate concentration, contributing to 13-28% of N2O production. Elevated nitrate concentration significantly mediated nirK-type denitrifiers structure and abundance, which are the keystone taxa driving N2O production. Collectively, these results suggest that increasing nitrate concentration can shift N2O production pathways from bacterial to fungal and chemodenitrification, which are mainly responsible for the enhanced N2O production and have widespread implications for N2O projections under ongoing nitrogen pollution in estuarine and coastal ecosystems.
Asunto(s)
Nitrógeno , Óxido Nitroso , Nitrógeno/metabolismo , Desnitrificación , Nitratos/química , EcosistemaRESUMEN
Ocean acidification in nitrogen-enriched estuaries has raised global concerns. For decades, biotic and abiotic denitrification in estuarine sediments has been regarded as the major ways to remove reactive nitrogen, but they occur at the expense of releasing greenhouse gas nitrous oxide (N2 O). However, how these pathways respond to acidification remains poorly understood. Here we performed a N2 O isotopocules analysis coupled with respiration inhibition and molecular approaches to investigate the impacts of acidification on bacterial, fungal, and chemo-denitrification, as well as N2 O emission, in estuarine sediments through a series of anoxic incubations. Results showed that acidification stimulated N2 O release from sediments, which was mainly mediated by the activity of bacterial denitrifiers, whereas in neutral environments, N2 O production was dominated by fungi. We also found that the contribution of chemo-denitrification to N2 O production cannot be ignored, but was not significantly affected by acidification. The mechanistic investigation further demonstrated that acidification changed the keystone taxa of sedimentary denitrifiers from N2 O-reducing to N2 O-producing ones and reduced microbial electron-transfer efficiency during denitrification. These findings provide novel insights into how acidification stimulates N2 O emission and modulates its pathways in estuarine sediments, and how it may contribute to the acceleration of global climate change in the Anthropocene.
Asunto(s)
Desnitrificación , Agua de Mar , Bacterias/genética , Concentración de Iones de Hidrógeno , Nitrógeno , Óxido NitrosoRESUMEN
We measured the δ values of N2O using gas chromatography isotope ratio mass spectrometry with a preconcentrator (precon-GC-IRMS). The instrumental precision of the mass spectrometer was restricted to below the shot noise limit, which agreed with the theoretical and experimental results of 0.02 (δ15N) and 0.04 (δ18O), respectively. The precision of the measured δ values was significantly improved by the temperature regulation protocol of the LN2 preconcentrator, which was monitored by various temperature sensors placed along the U-trap. The reproducibility of the He-diluted N2O gas measurements resulted in 0.063 (δ15N) and 0.075 (δ18O) due to additional sources of uncertainty in the vials used for autosampling and in the general preconcentration process. Multipoint normalization of the dual δ values of the measured N2O samples was conducted using United States Geological Survey reference materials denitrified by Pseudomonas aureofaciens. Kaiser's ion correction method, based on International Atomic Energy Agency parameters, exhibited low bias for the atomic isotope ratio reduction of the nitrate reference material, for which the oxygen anomaly was considerably high. Dedicated corrections for net isotope fractionation and water exchange were important in improving uncertainties in the procedure for normalizing the oxygen isotope ratio. Blank measurements for correcting biases in isotope ratios caused by pre-dissolved nitrate and nitrite ions in the water solvent led to further improvements, i.e. beyond unevenly controlled net isotope fractionation, throughout the bacterial denitrification process. The uncertainty evaluation revealed that three-point normalization can significantly improve the normalization accuracy compared with two-point normalization. In addition, an alternative strategy was suggested for assigning δ18O using a CO2 lab tank, allowing its use as a reference material for N2O gas tanks.
Asunto(s)
Desnitrificación , Óxido Nitroso , Pseudomonas , Reproducibilidad de los ResultadosRESUMEN
The copper-containing enzyme nitrous oxide reductase (N2OR) catalyzes the transformation of nitrous oxide (N2O) to dinitrogen (N2) in microbial denitrification. Several accessory factors are essential for assembling the two copper sites CuA and CuZ, and for maintaining the activity. In particular, the deletion of either the transmembrane iron-sulfur flavoprotein NosR or the periplasmic protein NosX, a member of the ApbE family, abolishes N2O respiration. Here we demonstrate through biochemical and structural studies that the ApbE protein from Pseudomonas stutzeri, where the nosX gene is absent, is a monomeric FAD-binding protein that can serve as the flavin donor for NosR maturation via covalent flavinylation of a threonine residue. The flavin transfer reaction proceeds both in vivo and in vitro to generate post-translationally modified NosR with covalently bound FMN. Only FAD can act as substrate and the reaction requires a divalent cation, preferably Mg2+ that was also present in the crystal structure. In addition, the reaction is species-specific to a certain extent.
Asunto(s)
Proteínas Bacterianas/metabolismo , Flavinas/metabolismo , Flavoproteínas/metabolismo , Óxido Nitroso/metabolismo , Oxidorreductasas/metabolismo , Pseudomonas stutzeri/metabolismo , Transferasas/metabolismo , Catálisis , Cobre/metabolismo , Magnesio/metabolismo , Proteínas de la Membrana/metabolismo , Procesamiento Proteico-Postraduccional/fisiologíaRESUMEN
Rhizobia are recognized to establish N2-fixing symbiotic interactions with legume plants. Bradyrhizobium japonicum, the symbiont of soybeans, can denitrify and grow under free-living conditions with nitrate (NO3 (-)) or nitrite (NO2 (-)) as sole nitrogen source. Unlike related bacteria that assimilate NO3 (-), genes encoding the assimilatory NO3 (-) reductase (nasC) and NO2 (-) reductase (nirA) in B. japonicum are located at distinct chromosomal loci. The nasC gene is located with genes encoding an ABC-type NO3 (-) transporter, a major facilitator family NO3 (-)/NO2 (-) transporter (NarK), flavoprotein (Flp) and single-domain haemoglobin (termed Bjgb). However, nirA clusters with genes for a NO3 (-)/NO2 (-)-responsive regulator (NasS-NasT). In the present study, we demonstrate NasC and NirA are both key for NO3 (-) assimilation and that growth with NO3 (-), but not NO2 (-) requires flp, implying Flp may function as electron donor to NasC. In addition, bjgb and flp encode a nitric oxide (NO) detoxification system that functions to mitigate cytotoxic NO formed as a by-product of NO3 (-) assimilation. Additional experiments reveal NasT is required for NO3 (-)-responsive expression of the narK-bjgb-flp-nasC transcriptional unit and the nirA gene and that NasS is also involved in the regulatory control of this novel bipartite assimilatory NO3 (-)/NO2 (-) reductase pathway.