Nitrate Utilization Research Articles

Genome-wide identification of nitrate transporter 2 (NRT2) gene family and functional analysis of MeNRT2.2 in cassava (Manihot esculenta Crantz).

Nitrate transporter 2 (NRT2) proteins play an important role in nitrate uptake and utilization in plants. The NRT2 family has been identified and functionally characterized in many plants. However, no systematic identification of NRT2 family members has been reported in cassava (Manihot esculenta Crantz). In this study, six MeNRT2 genes were identified from cassava genome and named as MeNRT2.1-2.6 according to their chromosomal locations. Phylogenetic tree showed that NRT2 proteins were divided into four main subgroups, which was further supported by their gene structure and conserved motifs. All six MeNRT2 genes are randomly distributed on 4 chromosomes (LG8, LG11, LG13, and LG17), two tandem duplicated genes (MeNRT2.3/MeNRT2.4) and a pair of segmental duplicated gene (MeNRT2.1/MeNRT2.2) was detected. Subsequently, expression profiles of MeNRT2 genes in eight different tissues and in response to nitrate deficient treatment were analyzed. The results showed that the MeNRT2 genes had differential expression patterns. All of MeNRT2 genes induced by nitrate deficiency, of them the MeNRT2.2 had the highest expression level after treatment. Arabidopis transformed with MeNRT2.2 gene showed higher fresh weight than wild type plants in response to N starvation, suggesting that MeNRT2.2 play important role in adapting to low nitrogen. Taken together, our results provide the reference for further analyses of the molecular functions of the MeNRT2 gene family, but also some candidate genes for developing nitrogen efficient crops.

Utilisation of Nitrate Accumulated in Soil During Greenhouse Cultivation

Utilisation of Nitrate Accumulated in Soil During Greenhouse Cultivation

Winter soil temperature and its effect on soil nitrate Status: A Support Vector Regression-based approach on the projected impacts

Winter season is characterized by lowering of air temperature and soil temperature, and deceleration of soil nitrogen cycle. Despite low soil temperature, soil NO3– is found on the soil surface and in the streams of temperate and higher latitude regions towards the end of winter season. With the rise in global air temperature, the soil temperature is expected to warm up accordingly. Plausible effects of future soil temperature changes in affecting the production and movement of soil NO3– within a vertical profile were studied in the present paper. Four Representative Concentration Pathways (RCP) scenarios were considered to estimate the effects of change in air temperature on soil temperature at different depths within a vertical profile and on the soil nitrate status. Since winter season is known to have low to no soil nitrate utilization (due to microbial inactivity and low utilization by biomass), plausible effects of winter warming in the future decades were studied. The occurrence and movement of soil NO3– during winter, raises a concern regarding the future warming climate’s response. Long-term records from Hubbard Brook Experimental Forest, New Hampshire, USA, were used for observation, calibration, and validation purposes. Non-linear relationship between air temperature, soil moisture, soil temperature, and soil NO3– status at different soil depths, were assessed using support vector regression (SVR) models. Soil temperature and soil NO3– status at different soil depths were predicted using SVR models, under the four different RCP scenarios. The projections under different RCP scenarios suggested - significant production of soil NO3– during winter (approx. 18 mg/l by the year 2095 under a “business as usual” scenario), and movement of soil NO3– towards soil surface before the beginning of plant-growing season.

Identification and characterization of apple MdNLP7 transcription factor in the nitrate response

Nitrogen is an essential nutrient for plant growth and development. Low utilization of nitrogen fertilizer during agricultural production causes a series of environmental problems, such as water eutrophication, soil acidity, and air pollution. Investigating the patterns and mechanisms of crop NO3− absorption and utilization therefore key to fully improving crop nitrogen utilization rates and promoting sustainable agricultural development. Apple is one of the most important horticultural crops in the world. Its nitrogen demand by apple during the growth period is very high, but few studies have been performed on apple genes, that regulate the NO3− response. Here, we found that the apple transcription factor MdNLP7 promoted nitrogen absorption and assimilation by activating the expression of MdNIA2 and MdNRT1.1. MdNLP7 also regulated H2O2 content by increasing catalase activity, which may also influence nitrate utilization. Our findings provide insight into the mechanisms by which MdNLP7 controls nitrate utilization in apple.

Protistan plankton communities in the Galápagos Archipelago respond to changes in deep water masses resulting from the 2015/16 El Niño

The Galápagos Archipelago lies within the Eastern Equatorial Pacific Ocean at the convergence of major ocean currents that are subject to changes in circulation. The nutrient-rich Equatorial Undercurrent upwells from the west onto the Galápagos platform, stimulating primary production, but this source of deep water weakens during El Niño events. Based on measurements from repeat cruises, the 2015/16 El Niño was associated with declines in phytoplankton biomass at most sites throughout the archipelago and reduced utilization of nitrate, particularly in large-sized phytoplankton in the western region. Protistan assemblages were identified by sequencing the V4 region of the 18S rRNA gene. Dinoflagellates, chlorophytes and diatoms dominated most sites. Shifts in dinoflagellate communities were most apparent between the years; parasitic dinoflagellates, Syndiniales, were highly detected during the El Niño (2015) while the dinoflagellate genus, Gyrodinium, increased at many sites during the neutral period (2016). Variations in protistan communities were most strongly correlated with changes in subthermocline water density. These findings indicate that marine protistan communities in this region are regimented by deep water mass sources and thus could be profoundly affected by altered ocean circulation.

Utilization of lithium nitrate to mitigate alkali–silica reaction of architectural glass mortar: Characteristics and mechanisms

It is low carbon footprint and low cost to use waste glass as fine aggregate in producing architectural mortar. However, glass aggregate is prone to alkali–silica reaction (ASR), which causes expansion and cracking of architectural mortar. Addition of LiNO3 is a good method to mitigate ASR but current understanding on characteristics of ASR reaction products and mitigation mechanisms of LiNO3 for glass aggregate are not sufficient. To contribute to the state of knowledge on these topics and promote the recycling of waste glass, this study systematically investigated morphologies and characteristics of ASR reaction products of glass mortar samples with and without LiNO3 by various microscopic analyses. The results showed that 1% LiNO3 reduced ASR expansion by 85.12% and map cracks disappeared. The formation location, characteristics and microstructures of ASR gel of glass aggregate were different from natural aggregate. The mitigation mechanisms of lithium were proposed: reduction in the dissolution of glass aggregate and the transformation from tobermorite-type to polymerized C–S–H, dense pozzolanic C–S–H serving as a physical barrier and strong lithium binding capacity to reduce Na/Si ratio of ASR gel.

Effect of crude oil shock load on anoxic-aerobic sequential moving bed reactors during petroleum refinery wastewater treatment

Concentrations of pollutants are suddenly increased (shock loads) in the lab-scale bioreactors to examine their stability against sudden change in feeding conditions. In the present study, sudden elevated loads of crude oil were supplied to an anoxic (A1)-aerobic (A2) sequential reactor system, as studies involving series reactors for shock analysis are scarce. The system was previously engaged in the treatment of petroleum refinery wastewater at influent crude oil of 300 mg/L along with phenol, sulfide, nitrate-N and ammonia-N. Influent crude oil was suddenly increased to 600 mg/L and 900 mg/L during first (shock 1) and second (shock 2) shock loadings, respectively, at two different times of operation. An immediate increase in the effluent organics and a decrease in efficiency was observed after each shock. The shock toxicity caused death of microbes and decrease (1–4%) in biomass concentration. Hence, biomass activity reduced, and the pollutants degradation declined by 40–50%. Nitrogen production in A1 decreased which confirmed lesser nitrate utilization during shock. Both the reactors recovered to their original stature after the withdrawal of shock 1. Shock 2 had a permanent effect on A1 as its efficiency decreased from 33% to 11% even after withdrawal of the shock. The combined A1-A2 system successfully tolerated the shock loads, and more than 99% removals of organics, sulfide, and nitrate was achieved after resumed normalcy.

Ontogenetic δ15N Trends and Multidecadal Variability in Shells of the Bivalve Mollusk, Arctica islandica

Bulk stable nitrogen isotope values of the carbonate-bound organic matrix in bivalve shells (δ15NCBOM) are increasingly used to assess past food web dynamics, track anthropogenic nitrogen pollution and reconstruct hydrographic changes. However, it remains unresolved if the δ15NCBOM values are also affected by directed ontogenetic trends which can bias ecological and environmental interpretations. This very aspect is tested here with modern and fossil specimens of the long-lived ocean quahog, Arctica islandica, collected from different sites and water depths in the NE Atlantic Ocean. As demonstrated, δ15NCBOM values from the long chronologies show a general decrease through lifetime by −0.006‰ per year. The most likely reason for the observed δ15NCBOM decline is a change in the type of proteins synthesized at different stages of life, i.e., a gradual shift from proteins rich in strongly fractionating, trophic amino acids during youth toward proteins rich in source amino acids during adulthood. Aside from this ontogenetic trend, distinct seasonal to multidecadal δ15NCBOM variations (ca. 50 to 60 years; up to 2.90‰) were identified. Presumably, the latter were governed by fluctuations in nutrient supply mediated by the Atlantic Multidecadal Variation (AMV) and Atlantic Meridional Overturning Circulation (AMOC) combined with changes in nitrate utilization by photoautotrophs and associated Rayleigh fractionation processes. Findings underline the outstanding potential of bivalve shells in studies of trophic ecology, oceanography and pollution, but also highlight the need for compound-specific isotope analyses.

Alternate Partial Root-Zone Drip Nitrogen Fertigation Reduces Residual Nitrate Loss While Improving the Water Use but Not Nitrogen Use Efficiency.

The efficient utilization of irrigation water and nitrogen is of great importance for sustainable agricultural production. Alternate partial root-zone drip irrigation (APRD) is an innovative water-saving drip irrigation technology. However, the coupling effects of water and nitrogen (N) supply under APRD on crop growth, water and N use efficiency, as well as the utilization and fate of residual nitrates accumulated in the soil profile are not clear. A simulated soil column experiment where 30–40 cm soil layer was 15NO3-labeled as residual nitrate was conducted to investigate the coupling effects of different water [sufficient irrigation (W1), two-thirds of the W1(W2)] and N [high level (N1), 50% of N1 (N2)] supplies under different irrigation modes [conventional irrigation (C), APRD (A)] on tomato growth, irrigation water (IWUE) and N use efficiencies (NUE), and the fate of residual N. The results showed that, compared with CW1N1, AW1N1 promoted root growth and nitrogen absorption, and increased tomato yield, while the N absorption and yield did not vary significantly in AW2N1. The N absorption in AW2N2 decreased by 16.1%, while the tomato yield decreased by only 8.8% compared with CW1N1. The highest IWUE appeared in AW2N1, whereas the highest NUE was observed in AW2N2, with no significant difference in NUE between AW2N1 and CW1N1 at the same N supply level. The 15N accumulation peak layer was almost the same as the originally labeled layer under APRD, whereas it moved 10–20 cm downwards under CW1N1. The amount of 15N accumulated in the 0-40 cm layer increased with the decreasing irrigation water and nitrogen supply, with an increase of 82.9–141.1% in APRD compared with that in CW1N1. The utilization of the 15N labeled soil profile by the tomato plants increased by 9–20.5%, whereas the loss rate of 15N from the plant-soil column system decreased by 21.3–50.1% in APRD compared with the CW1N1 treatment. Thus, APRD has great potential in saving irrigation water, facilitating water use while reducing the loss of residual nitrate accumulated in the soil profile, but has no significant effect on the NUE absorbed.

Comprehensive Comparative Genomics and Phenotyping of Methylobacterium Species.

The pink-pigmented facultative methylotrophs (PPFMs), a major bacterial group found in the plant phyllosphere, comprise two genera: Methylobacterium and Methylorubrum. They have been separated into three major clades: A, B (Methylorubrum), and C. Within these genera, however, some species lack either pigmentation or methylotrophy, which raises the question of what actually defines the PPFMs. The present study employed a comprehensive comparative genomics approach to reveal the phylogenetic relationship among the PPFMs and to explain the genotypic differences that confer their different phenotypes. We newly sequenced the genomes of 29 relevant-type strains to complete a dataset for almost all validly published species in the genera. Through comparative analysis, we revealed that methylotrophy, nitrate utilization, and anoxygenic photosynthesis are hallmarks differentiating the PPFMs from the other Methylobacteriaceae. The Methylobacterium species in clade A, including the type species Methylobacterium organophilum, were phylogenetically classified into six subclades, each possessing relatively high genomic homology and shared phenotypic characteristics. One of these subclades is phylogenetically close to Methylorubrum species; this finding led us to reunite the two genera into a single genus Methylobacterium. Clade C, meanwhile, is composed of phylogenetically distinct species that share relatively higher percent G+C content and larger genome sizes, including larger numbers of secondary metabolite clusters. Most species of clade C and some of clade A have the glutathione-dependent pathway for formaldehyde oxidation in addition to the H4MPT pathway. Some species cannot utilize methanol due to their lack of MxaF-type methanol dehydrogenase (MDH), but most harbor an XoxF-type MDH that enables growth on methanol in the presence of lanthanum. The genomes of PPFMs encode between two and seven (average 3.7) genes for pyrroloquinoline quinone-dependent alcohol dehydrogenases, and their phylogeny is distinctly correlated with their genomic phylogeny. All PPFMs were capable of synthesizing auxin and did not induce any immune response in rice cells. Other phenotypes including sugar utilization, antibiotic resistance, and antifungal activity correlated with their phylogenetic relationship. This study provides the first inclusive genotypic insight into the phylogeny and phenotypes of PPFMs.

Denitrification Process Enhancement and Diversity of the Denitrifying Community in the Full Scale Activated Sludge System after Adaptation to Fusel Oil

Implementation of anaerobic digestion of primary sludge in modern wastewater treatment plants (WWTPs) limits the availability of organic carbon for denitrification in conventional nitrification-denitrification (N/DN) systems. In order to ensure efficient denitrification, dosage of the external carbon source is commonly undertaken. However, application of commercial products, such us ethanol or acetate, greatly increases operational costs. As such, inexpensive and efficient alternative carbon sources are strongly desirable. In this study, the use of the fusel oil, a by-product from the distillery industry, was validated in terms of the denitrification process enhancement and impact on the activated sludge bacterial community structure. The experiment was conducted at a full scale biological nutrient removal facility (210,000 PE), in the set of the two technological lines: the experimental line (where fusel oil was introduced at 45 cm3/m3 dose) and the reference line (without an external carbon source addition). During the experimental period of 98 days, conventional nitrate utilization rate (NUR) measurements were carried out on a regular basis in order to assess the biomass adaptation to the fusel oil addition and denitrification process enhancement. While the NURs remained at a stable level in the reference line (1.4 ± 0.1 mg NO3-N/g VSS·h) throughout the entire duration of the experiment, the addition of fusel oil gradually enhanced the denitrification process rate up to 2.7 mg NO3-N/g VSS·h. Moreover, fusel oil contributed to the mitigation of the variability of NO3-N concentrations in the effluent from the anoxic zone. The bacterial community structure, characterized by 16S rRNA PCR—DGGE and the clone libraries of the genes involved in the denitrification process (nirS and nirK), was comparable between the reference and the experimental line during the entire experimental period. In both analyzed lines, the most frequent occurrence of denitrifiers belonging to the genera Acidovorax, Alcaligenes, Azoarcus, Paracoccus and Thauera was noticed. Our results proved that fusel oil would a valuable substrate for denitrification. The addition of fusel oil enhances the process rate and does not reflect a severe selection pressure on the bacterial community at applicable doses. Practical application of fusel oil generates opportunities for the WWTPs to meet effluent standards and reduce operational costs, as well as optimizing waste management for the distillery industry.

Excessive ammonium assimilation by plastidic glutamine synthetase causes ammonium toxicity in Arabidopsis thaliana

Plants use nitrate, ammonium, and organic nitrogen in the soil as nitrogen sources. Since the elevated CO2 environment predicted for the near future will reduce nitrate utilization by C3 species, ammonium is attracting great interest. However, abundant ammonium nutrition impairs growth, i.e., ammonium toxicity, the primary cause of which remains to be determined. Here, we show that ammonium assimilation by GLUTAMINE SYNTHETASE 2 (GLN2) localized in the plastid rather than ammonium accumulation is a primary cause for toxicity, which challenges the textbook knowledge. With exposure to toxic levels of ammonium, the shoot GLN2 reaction produced an abundance of protons within cells, thereby elevating shoot acidity and stimulating expression of acidic stress-responsive genes. Application of an alkaline ammonia solution to the ammonium medium efficiently alleviated the ammonium toxicity with a concomitant reduction in shoot acidity. Consequently, we conclude that a primary cause of ammonium toxicity is acidic stress.

Simultaneous reduction of perchlorate and nitrate using fast-settling anoxic sludge

Fast-settling, anoxic sludge (FAS) was cultivated and utilized in this study to simultaneously reduce elevated levels of perchlorate and nitrate in an anaerobic sequencing batch reactor (AnSBR). Average perchlorate and nitrate removal efficiencies of 96.5 ± 8.44 % and 99.8 ± 0.32 %, respectively, were achieved from an average perchlorate and nitrate loading rate of 159 ± 101 g ClO4−/m3·d and 10.8 ± 7.25 g NO3−-N/m3·d, respectively, throughout long-term operation (>500-d). Batch activity tests revealed a preferential utilization of nitrate over perchlorate, where significant perchlorate reduction inhibition occurred when nitrate was present as a competing electron acceptor under carbon-limiting conditions. Specific perchlorate and nitrate reduction rates were shown to increase as the hydraulic retention time (HRT) of the AnSBR was step-wise decreased and subsequently the perchlorate and nitrate loading rates were step-wise increased. Functional, mRNA-based expression of the nitrite reductase (nirS and nirK), nitrous oxide reductase (nosZ), perchlorate reductase subunit A (pcrA), and the chlorite dismutase (cld) genes illustrated the simultaneous activity of heterotrophic denitrification and perchlorate reduction occurring throughout a complete standard reactor operational cycle, and allowed for expression trends to be documented as the HRT of the AnSBR was reduced from 5-d to 1.25-d. Nitrous oxide (N2O) production was detected as a result of incomplete denitrification, where the largest N2O production occurred at the highest nitrate loading rates investigated in this study. Thauera species were heavily enriched at a longer HRT of 5-d, but were out-competed by Dechloromonas species as the HRT of the AnSBR was step-wise reduced to 1.25-d.

Transcriptome Variations in Verticillium dahliae in Response to Two Different Inorganic Nitrogen Sources.

The fungus Verticillium dahliae causes vascular wilt disease on hundreds of plant species. The main focus of the research to control this fungus has been aimed at infection processes such as penetration peg formation and effector secretion, but the ability of the fungus to acquire and utilize nutrients are often overlooked and may hold additional potential to formulate new disease control approaches. Little is known about the molecular mechanisms of nitrogen acquisition and assimilation processes in V. dahliae. In this present study, RNA sequencing and gene expression analysis were used to examine differentially expressed genes in response to the different nitrogen sources, nitrate and ammonium, in V. dahliae. A total of 3244 and 2528 differentially expressed genes were identified in response to nitrate and ammonium treatments, respectively. The data indicated nitrate metabolism requires additional energy input while ammonium metabolism is accompanied by reductions in particular cellular processes. Gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses of DEGs during nitrate metabolism revealed that many of the genes encoded those involved in protein biosynthetic and metabolic processes, especially ribosome and RNA polymerase biosynthesis, but also other processes including transport and organonitrogen compound metabolism. Analysis of DEGs in the ammonium treatment indicated that cell cycle, oxidoreductase, and certain metabolic activities were reduced. In addition, DEGs participating in the utilization of both nitrate and ammonium were related to L-serine biosynthesis, energy-dependent multidrug efflux pump activity, and glycerol transport. We further showed that the mutants of three differentially expressed transcription factors (VdMcm1, VdHapX, and VDAG_08640) exhibited abnormal phenotypes under nitrate and ammonium treatment compared with the wild type strain. Deletion of VdMcm1 displayed slower growth when utilizing both nitrogen sources, while deletion of VdHapX and VDAG_08640 only affected nitrate metabolism, inferring that nitrogen assimilation required regulation of bZIP transcription factor family and participation of cell cycle. Taken together, our findings illustrate the convergent and distinctive regulatory mechanisms between preferred (ammonium) and alternative nitrogen (nitrate) metabolism at the transcriptome level, leading to better understanding of inorganic nitrogen metabolism in V. dahliae.

Nitrate-responsive NIN-like protein transcription factors perform unique and redundant roles in Arabidopsis.

Upon sensing nitrate, NODULE INCEPTION (NIN)-like protein (NLP) transcription factors alter gene expression to promote nitrate uptake and utilization. Of the nine NLPs in Arabidopsis, the physiological roles of only three NLPs (NLP6-NLP8) have been characterized to date. To evaluate the unique and redundant roles of Arabidopsis NLPs, we assessed the phenotypes of single and higher order nlp mutants. Unlike other nlp single mutants, nlp2 and nlp7 single mutants showed a reduction in shoot fresh weight when grown in the presence of nitrate as the sole nitrogen source, indicating that NLP2, like NLP7, plays a major role in vegetative growth. Interestingly, the growth defect of nlp7 recovered upon the supply of ammonium or glutamine, whereas that of nlp2 did not. Furthermore, complementation assays using chimeric constructs revealed that the coding sequence, but not the promoter region, of NLP genes was responsible for the differences between nlp2 and nlp7 single mutant phenotypes, suggesting differences in protein function. Importantly, nitrate utilization was almost completely abolished in the nlp septuple mutant (nlp2 nlp4 nlp5 nlp6 nlp7 nlp8 nlp9), suggesting that NLPs other than NLP2 and NLP7 also assist in the regulation of nitrate-inducible gene expression and nitrate-dependent promotion of vegetative growth in Arabidopsis.

Effects of ocean acidification on the growth, photosynthetic performance, and domoic acid production of the diatom Pseudo-nitzschia australis from the California Current System

Pseudo-nitzschia australis (Frenguelli), a toxigenic pennate diatom capable of producing the neurotoxin domoic acid (DA), was examined in unialgal laboratory cultures to quantify its physiological response to ocean acidification (OA) - the decline in pH resulting from increasing partial pressure of CO2 (pCO2) in the oceans. Toxic blooms of P. australis are common in the coastal waters of eastern boundary upwelling systems (EBUS), including those of the California Current System (CCS) off the west coast of the United States where increased pCO2 and decreased seawater pH are well-known. This study determined the production of dissolved (dDA) and particulate DA (pDA), the rates of growth and nutrient (nitrate, silicate and phosphate) utilization, cellular elemental ratios of carbon and nitrogen, and the photosynthetic response to declining pH during the exponential and stationary growth phases of a strain of P. australis isolated during a massive toxic bloom that persisted for months along much of the U.S. west coast during 2015. Our controlled lab studies showed that DA production significantly increased as pCO2 increased, and total DA (pDA + dDA) normalized to cell density was 2.7 fold greater at pH 7.8 compared to pH 8.1 (control) during nutrient-limited stationary growth. However, exponential growth rates did not increase with declining pH, but remained constant until pH of 7.8 was reached, and then specific growth rates declined by ca. 30%. The toxin results demonstrate that despite minimal effects of OA observed during the nutrient-replete exponential growth phase, the enhancement of DA production, notably the 3-fold increase in particulate DA per cell, with declining pH from 8.1 to 7.8 during the nutrient-depleted stationary phase, supports the hypothesis that increasing pCO2 will result in greater toxic risk to coastal ecosystems from elevated ambient concentrations of particulate DA. The ecological consequences of decreasing silicate uptake rates and increasing cellular carbon quotas with declining pH may potentially ameliorate some negative impacts of OA on Pseudo-nitzschia growth in natural systems.

The Mid-Atlantic Bight Dissolved Inorganic Carbon System Observed in the March 1996 DOE Ocean Margins Program (OMP)—A Baseline Study

The United States Department of Energy (DOE)’s Ocean Margins Program (OMP) cruise EN279 in March 1996 provides an important baseline for assessing long-term changes in the carbon cycle and biogeochemistry in the Mid-Atlantic Bight (MAB) as climate and anthropogenic changes have been substantial in this region over the past two decades. The distributions of O2, nutrients, and marine inorganic carbon system parameters are influenced by coastal currents, temperature gradients, and biological production and respiration. On the cross-shelf direction, pH decreases seaward, but carbonate saturation state (ΩArag) does not exhibit a clear trend. In contrast, ΩArag increases from north to south, while pH has no clear spatial patterns in the along-shelf direction. In order to distinguish between the effects of physical mixing of various water masses and those of biological activities on the marine inorganic carbon system, we use the potential temperature-salinity diagram to identify water masses, and differences between observations and theoretical mixing concentrations to measure the non-conservative (primarily biological) effects. Our analysis clearly shows the degree to which ocean margin pH and ΩArag are regulated by biological activities in addition to water mass mixing, gas exchange, and temperature. The correlations among anomalies in dissolved inorganic carbon, phosphate, nitrate, and apparent oxygen utilization agree with known biological stoichiometry. Biological uptake is substantial in nearshore waters and in shelf-slope mixing areas. This work provides valuable baseline information to assess the more recent changes in the marine inorganic carbon system and the status of coastal ocean acidification.

Climate‐Modulated Nutrient Conditions Along the Labrador Shelf: Evidence From Nitrogen Isotopes in a Six‐Hundred‐Year‐Old Crustose Coralline Alga

AbstractThe impacts of climate change on north Atlantic nutrient chemistry remain poorly understood, as there exist a multitude of rapidly changing biological and physical drivers of nutrient conditions throughout the region. Here, we present nitrogen isotope measurements derived from a six‐hundred‐year‐old crustose coralline alga (δ15Nalgal) to elucidate historical and contemporary trends in nitrate utilization and circulation patterns along the Labrador Shelf. Prior to the early 1900s, we argue that intervals during which utilization approached completion were controlled by reduced nitrate advection linked to an increased proportion of nitrate‐poor polar waters and subdued Atlantic influence, as expected from concurrent negative modes of the Atlantic multidecadal oscillation. While nitrate conditions should have recovered in recent years, our record suggests that high utilization persisted since ∼1870, which we also attribute to reduced Atlantic advection, likely associated with the twentieth‐century anthropogenic weakening of the Atlantic meridional overturning circulation. These results highlight the role of ongoing climate‐induced circulation changes in modulating nutrient distributions throughout the subpolar north Atlantic, which may have implications for other environmental phenomena such as fisheries and oceanic carbon storage.

Summertime Biogenic Silica Production and Silicon Limitation in the Pacific Arctic Region From 2006 to 2016

AbstractWe present a decade of dissolved and particulate silica concentrations within five biological “hot spots” in the Pacific Arctic Region (PAR) and the first measurements of both biogenic silica production rates (ρSi) and the kinetics of silicon utilization from a period of four years at the same sites. The “hot spots” were located within the Bering and Chukchi Seas and identified as part of the Distributed Biological Observatory (DBO). Across all hot spots, the highest concentrations of silicic acid (Si(OH)4) and biogenic silica were found near the bottom of the euphotic zone and often correlated with increased ρSi. For the entire region, the average ρSi was 19 mmol m−2 day−1 and siliceous microplankton (i.e., diatoms) contributed an average of 62% to primary productivity and 82% to nitrate utilization. Irradiance and [Si(OH)4] had separate and interactive effects on ρSi. Irradiance modulated both the magnitude of ρSi and the response of diatoms to changes in Si(OH)4. Availability of Si(OH)4 limited ρSi in all hot spots in at least one of the four years. Kinetic experiments conducted in all hot spots demonstrated that the half‐saturation constant (Ks) for ρSi was 4–8 times higher than ever reported in the literature. In the southeastern Chukchi Sea, an east to west gradient in [bSiO2] and ρSi may have been driven by differences in the availability of NO3− rather than Si(OH)4. Despite strong interannual variability, we suggest that phytoplankton phenology responds to short‐term climatic changes, which can have far‐reaching effects on Arctic regions influenced by the Pacific‐origin waters flowing through the PAR.

First Report of Bacterial Leaf Spot of Cucurbita pepo Caused by Erwinia persicina in China.

In February 2020, the common symptoms of water-soaked spots on Cucurbita pepo L. cotyledon were observed in Guangrao county in Shandong province, China. Field investigation showed that 40% of the Cucurbita pepo cotyledons in an area of approximately 0.8 ha were infected. The disease resulted in a severe loss in seedling production. Samples of C. pepo with water-soaked leaf spots were collected and prepared for pathogen analysis. Symptomatic cotyledon tissue was surface disinfested in 75% ethanol for 30 sec, then rinsed three times in sterilized water. Bacteria were released in sterile water in Petri dish for 2 min by cutting symptomatic tissue into small sections and stirring the plant tissue mixture fully. The diffusate was streaked onto plates containing nutrient agar (NA) and plates were incubated at 28℃ for 2 days. Three representative isolates were purified eventually from each of the plates. Colonies on NA were small, round and with smooth margins. All bacterial isolates characterized as gram-negative, white to cream color, and pink pigment was formed on the plates over long-term culture. The isolates were positive for catalase, Voges-Proskauer, potato rot, methyl red, acetoin production, nitrate utilization and citrate utilization, and acid production from maltose, glucose, melezitose, sucrose, D-arabinose, D-trehalose, cellobiose, lactose, raffinose, mannitol, D-sorbitol, melibiose and xylitol. KOH production was demonstrated according to strand formation within the potassium hydroxide test (Suslow et al. 1982). Isolates were negative for oxidase, arginine dihydrolase, phenylalanine deaminase, gelatinase, esculine, indole production and H2S production. Total genomic DNA was extracted from isolate XHL2002230201 with TIANamp Bacteria DNA Kit (TIANGEN). Universal primers 27F and 1492R (Monciardini et al. 2002) were used in PCR to amplify a 1,307-bp DNA fragment of the 16S rRNA region for molecular identification. Furthermore, four additional housekeeping genes (gyrB, atpD, rho, and rpoS) were selected and amplified using specially designed primers. The amplification products of 16S rRNA were sequenced and submitted to GenBank under accession number (MT568607.1). Sequence analysis showed 99% similarity to Erwinia persicina strains B57 (LM651373.1) and B64 (CI789_17875) by BLAST search in GenBank database (Gálvez et al. 2015; Cho et al. 2019). A phylogenetic tree was constructed, and the taxonomic position of strain XHL2002230201 was determined from the multilocus sequence analysis (MLSA) on 16S rRNA and other four housekeeping genes with E. persicina and not with other closely related Erwinia species. Pathogenicity tests and re-isolation and re-identification of the bacteria were performed to confirm the isolate and fulfill the Koch' postulates. The strain XHL2002230201 suspensions (108 CFU ml-1) were spray inoculated onto fifteen Cucurbita pepo seedlings with two true leaves, and the same number of control plants were inoculated with water. Experiments were repeated three times. All inoculated plants were kept in a moist chamber placed in a greenhouse at 28℃. Initial symptoms were observed on leaves of inoculated plants at 5 days post-inoculation, whereas no symptoms appeared on the plants inoculated with sterile distilled water. Based on morphological and biochemical characteristics, phylogenetic analysis, and Koch's postulates, the bacterial isolates were identified as E. persicina. To our knowledge, this is the first report of E. persicina causing leaf spot disease on Cucurbita pepo in China.