Bacterial denitrification drives elevated N2O emissions in arid southern California drylands.

Soils are the largest source of atmospheric nitrous oxide (N2O), a powerful greenhouse gas. Dry soils rarely harbor anoxic conditions to favor denitrification, the predominant N2O-producing process, yet, among the largest N2O emissions have been measured after wetting summer-dry desert soils, raising the question: Can denitrifiers endure extreme drought and produce N2O immediately after rainfall? Using isotopic and molecular approaches in a California desert, we found that denitrifiers produced N2O within 15 minutes of wetting dry soils (site preference = 12.8 ± 3.92 per mil, δ15Nbulk = 18.6 ± 11.1 per mil). Consistent with this finding, we detected nitrate-reducing transcripts in dry soils and found that inhibiting microbial activity decreased N2O emissions by 59%. Our results suggest that despite extreme environmental conditions-months without precipitation, soil temperatures of ≥40°C, and gravimetric soil water content of <1%-bacterial denitrifiers can account for most of the N2O emitted when dry soils are wetted.

Quantifying atmospheric N deposition in dryland ecosystems: A test of the Integrated Total Nitrogen Input (ITNI) method.

Estimating nitrogen (N) deposition to terrestrial ecosystems is complicated by the multiple forms and routes of N loading from the atmosphere. We used the integrated total nitrogen input (ITNI) method, which is based on the principle of isotope dilution within a plant-liquid-sand system, to quantify N inputs to coastal sage scrub ecosystems in Riverside, California. Using the ITNI method, we measured atmospheric N deposition of 29.3 kg N ha-1 yr-1 over a range of aboveground plant biomass of 228 to 424 g m-2. From 85 to 96% of the atmospheric N inputs were taken up by plants in the ITNI modules with most of the assimilation mediated by, and stored in, aboveground biomass. Parallel measurements using conventional approaches yielded deposition rates of 25.2 kg N ha-1 yr-1 when using the inferential method and 4.8 kg N ha-1 yr-1 using throughfall collectors. The relatively low throughfall estimates were attributed to canopy retention of inorganic N, low rainfall, and to the fact that the throughfall flux data did not include organic N and stomatal uptake of N gases. Also, during dry periods, frequent watering of ITNI modules may have increased stomatal conductance and led to overestimates of N deposition. Across published studies that used the ITNI method, areal N deposition rates varied by ~40-fold, were positively correlated with plant biomass and 90% of the variability in measured deposition rates can be explained by plant biomass production. The ITNI method offers a holistic approach to measuring atmospheric N deposition in arid ecosystems, although more study is needed to understand how watering rates effect N deposition measurements.

Acidity and organic matter promote abiotic nitric oxide production in drying soils.

Soils are an important source of NO, particularly in dry lands because of trade-offs that develop between biotic and abiotic NO-producing processes when soils dry out. Understanding how drier climates may offset the balance of these trade-offs as soils transition toward more arid states is, therefore, critical to estimating global NO budgets, especially because drylands are expected to increase in size. We measured NO emission pulses after wetting soils from similar lithologies along an altitudinal gradient in the Sierra Nevada, CA, where mean annual precipitation varied from 670 to 1500 mm. Along the gradient, we measured field NO emissions, and used chloroform in the laboratory to reduce microbial activity and partition between biotic and abiotic NO-producing processes (i.e., chemodenitrification). Field NO emission pulses were lowest in the acidic and SOM-rich soils (4-72 ng NO-N m-2 s-1 ), but were highest in the high-elevation barren site (~560 ng NO-N m-2 s-1 ). In the laboratory, NO emission pulses were up to 19× greater in chloroform-treated soils than in the controls, and these abiotic pulses increased with elevation as pH decreased (6.2-4.4) and soil organic matter (SOM) increased (18-157 mg C g-1 ). Drought can shift the balance between the biotic and abiotic processes that produce NO, favoring chemodenitrification during periods when biological processes become stressed. Acidic and SOM-rich soils, which typically develop under mesic conditions, are most vulnerable to N loss via NO as interactions between pH, SOM, and drought stimulate chemodenitrification.

Aridity and plant uptake interact to make dryland soils hotspots for nitric oxide (NO) emissions.

Nitric oxide (NO) is an important trace gas and regulator of atmospheric photochemistry. Theory suggests moist soils optimize NO emissions, whereas wet or dry soils constrain them. In drylands, however, NO emissions can be greatest in dry soils and when dry soils are rewet. To understand how aridity and vegetation interact to generate this pattern, we measured NO fluxes in a California grassland, where we manipulated vegetation cover and the length of the dry season and measured [δ(15)-N]NO and [δ(18)-O]NO following rewetting with (15)N-labeled substrates. Plant N uptake reduced NO emissions by limiting N availability. In the absence of plants, soil N pools increased and NO emissions more than doubled. In dry soils, NO-producing substrates concentrated in hydrologically disconnected microsites. Upon rewetting, these concentrated N pools underwent rapid abiotic reaction, producing large NO pulses. Biological processes did not substantially contribute to the initial NO pulse but governed NO emissions within 24 h postwetting. Plants acted as an N sink, limiting NO emissions under optimal soil moisture. When soils were dry, however, the shutdown in plant N uptake, along with the activation of chemical mechanisms and the resuscitation of soil microbial processes upon rewetting, governed N loss. Aridity and vegetation interact to maintain a leaky N cycle during periods when plant N uptake is low, and hydrologically disconnected soils favor both microbial and abiotic NO-producing mechanisms. Under increasing rates of atmospheric N deposition and intensifying droughts, NO gas evasion may become an increasingly important pathway for ecosystem N loss in drylands.

20th century atmospheric deposition and acidification trends in lakes of the Sierra Nevada, California, USA.

We investigated multiple lines of evidence to determine if observed and paleo-reconstructed changes in acid neutralizing capacity (ANC) in Sierra Nevada lakes were the result of changes in 20th century atmospheric deposition. Spheroidal carbonaceous particles (SCPs) (indicator of anthropogenic atmospheric deposition) and biogenic silica and δ(13)C (productivity proxies) in lake sediments, nitrogen and sulfur emission inventories, climate variables, and long-term hydrochemistry records were compared to reconstructed ANC trends in Moat Lake. The initial decline in ANC at Moat Lake occurred between 1920 and 1930, when hydrogen ion deposition was approximately 74 eq ha(-1) yr(-1), and ANC recovered between 1970 and 2005. Reconstructed ANC in Moat Lake was negatively correlated with SCPs and sulfur dioxide emissions (p = 0.031 and p = 0.009). Reconstructed ANC patterns were not correlated with climate, productivity, or nitrogen oxide emissions. Late 20th century recovery of ANC at Moat Lake is supported by increasing ANC and decreasing sulfate in Emerald Lake between 1983 and 2011 (p < 0.0001). We conclude that ANC depletion at Moat and Emerald lakes was principally caused by acid deposition, and recovery in ANC after 1970 can be attributed to the United States Clean Air Act.

Correcting for background nitrate contamination in KCl-extracted samples during isotopic analysis of oxygen and nitrogen by the denitrifier method.

RATIONALE: Previous research has shown that the denitrifying bacteria Pseudomonas chlororaphis ssp. aureofaciens (P. aureofaciens) can be used to measure the δ(15)N and δ(18)O values of extracted soil nitrate (NO3(-)) by isotope ratio mass spectrometry. We discovered that N2O production from reference blanks made in 1 M KCl increased relative to blanks made of deionized water (DIW). Further investigation showed that isotopic standards made in KCl yielded δ(15)N and δ(18)O values different from the standards prepared in DIW. METHODS: Three grades of crystalline KCl were dissolved in DIW to create solutions of increasing molarity (0.1 M to 2 M), which were added to P. aureofaciens broth and measured as blanks. Reference standards USGS-32, USGS-34, and USGS-35 were then dissolved in a range of KCl concentrations to measure isotopic responses to changing KCl molarity. Reference blanks and standards created in DIW were analyzed as controls to measure the impact of KCl on the δ(15)N and δ(18)O values. RESULTS: The amount of N2O in the KCl blanks increased linearly with increasing molarity, but at different rates for each KCl grade. The isotopic values of the reference standards measured in KCl were systematically different from those measured in DIW, suggesting contamination by background NO3(-) in the KCl reagents. However, we also noted reduced conversion of NO3(-) into N2O as the KCl molarity increased, suggesting there is a physiological response of P. aureofaciens to KCl. CONCLUSIONS: There is a small amount of NO3(-) present in crystalline KCl, which can bias isotopic measurement of NO3(-) at low sample concentrations. This can be minimized by making standards and blanks in the same KCl as is used in samples, diluting all samples and standards to the appropriate NO3(-) concentration using matched KCl solutions, and adding samples and standards to the broth at a constant volume to standardize the KCl molarity in the reaction vial.

Effects of stock use and backpackers on water quality in wilderness in Sequoia and Kings Canyon National Parks, USA.

During 2010-2011, a study was conducted in Sequoia and Kings Canyon National Parks (SEKI) to evaluate the influence of pack animals (stock) and backpackers on water quality in wilderness lakes and streams. The study had three main components: (1) a synoptic survey of water quality in wilderness areas of the parks, (2) paired water quality sampling above and below several areas with differing types and amounts of visitor use, and (3) intensive monitoring at six sites to document temporal variations in water quality. Data from the synoptic water quality survey indicated that wilderness lakes and streams are dilute and have low nutrient and Escherichia coli concentrations. The synoptic survey sites were categorized as minimal use, backpacker-use, or mixed use (stock and backpackers), depending on the most prevalent type of use upstream from the sampling locations. Sites with mixed use tended to have higher concentrations of most constituents (including E. coli) than those categorized as minimal-use (P ≤ 0.05); concentrations at backpacker-use sites were intermediate. Data from paired-site sampling indicated that E. coli, total coliform, and particulate phosphorus concentrations were greater in streams downstream from mixed-use areas than upstream from those areas (P ≤ 0.05). Paired-site data also indicated few statistically significant differences in nutrient, E. coli, or total coliform concentrations in streams upstream and downstream from backpacker-use areas. The intensive-monitoring data indicated that nutrient and E. coli concentrations normally were low, except during storms, when notable increases in concentrations of E. coli, nutrients, dissolved organic carbon, and turbidity occurred. In summary, results from this study indicate that water quality in SEKI wilderness generally is good, except during storms; and visitor use appears to have a small, but statistically significant influence on stream water quality.

The effect of permafrost thaw on old carbon release and net carbon exchange from tundra.

Permafrost soils in boreal and Arctic ecosystems store almost twice as much carbon as is currently present in the atmosphere. Permafrost thaw and the microbial decomposition of previously frozen organic carbon is considered one of the most likely positive climate feedbacks from terrestrial ecosystems to the atmosphere in a warmer world. The rate of carbon release from permafrost soils is highly uncertain, but it is crucial for predicting the strength and timing of this carbon-cycle feedback effect, and thus how important permafrost thaw will be for climate change this century and beyond. Sustained transfers of carbon to the atmosphere that could cause a significant positive feedback to climate change must come from old carbon, which forms the bulk of the permafrost carbon pool that accumulated over thousands of years. Here we measure net ecosystem carbon exchange and the radiocarbon age of ecosystem respiration in a tundra landscape undergoing permafrost thaw to determine the influence of old carbon loss on ecosystem carbon balance. We find that areas that thawed over the past 15 years had 40 per cent more annual losses of old carbon than minimally thawed areas, but had overall net ecosystem carbon uptake as increased plant growth offset these losses. In contrast, areas that thawed decades earlier lost even more old carbon, a 78 per cent increase over minimally thawed areas; this old carbon loss contributed to overall net ecosystem carbon release despite increased plant growth. Our data document significant losses of soil carbon with permafrost thaw that, over decadal timescales, overwhelms increased plant carbon uptake at rates that could make permafrost a large biospheric carbon source in a warmer world.

Sterols of the syndinian dinoflagellate Amoebophrya sp., a parasite of the dinoflagellate Alexandrium tamarense (Dinophyceae).

Several harmful photosynthetic dinoflagellates have been examined over past decades for unique chemical biomarker sterols. Little emphasis has been placed on important heterotrophic genera, such as Amoebophrya, an obligate, intracellular parasite of other, often harmful, dinoflagellates with the ability to control host populations naturally. Therefore, the sterol composition of Amoebophrya was examined throughout the course of an infective cycle within its host dinoflagellate, Alexandrium tamarense, with the primary intent of identifying potential sterol biomarkers. Amoebophrya possessed two primary C(27) sterols, cholesterol and cholesta-5,22Z-dien-3beta-ol (cis-22-dehydrocholesterol), which are not unique to this genus, but were found in high relative percentages that are uncommon to other genera of dinoflagellates. Because the host also possesses cholesterol as one of its major sterols, carbon-stable isotope ratio characterization of cholesterol was performed in order to determine whether it was produced by Amoebophrya or derived intact from the host. Results indicated that cholesterol was not derived intact from the host. A comparison of the sterol profile of Amoebophrya to published sterol profiles of phylogenetic relatives revealed that its sterol profile most closely resembles that of the (proto)dinoflagellate Oxyrrhis marina rather than other extant genera.