Storage and hydrolysis of seawater samples for inorganic carbon isotope analysis

Preservation of seawater samples was tested for total inorganic carbon (ΣCO2), stable carbon isotope (δ13C), and radiocarbon (14C) applications using foil bags and storage by refrigeration and freezing. The aim was to preserve representative samples with minimal storage effects but without using toxic methods such as mercuric chloride poisoning. Hydrolysis of samples to CO2 was based on existing methods. Results of IAEA-C2 standard used with deionized water stored in the foil bags showed complete reaction yields, 14C results within 2σ of the consensus value, and δ13C that were internally consistent, indicating that there were no procedural effects associated with the foil bags. 14C results were statistically indistinguishable across the storage times, for frozen and refrigerated seawater samples from a coastal site, Elie Ness, Fife, UK. The scatter of ΣCO2 concentrations and δ13C was within scatter observed in other studies for lake- and seawater samples preserved by acidification or using mercuric chloride. However, both ΣCO2 and δ13C were less variable for frozen samples compared with refrigerated samples. The foil bags are lighter, safer to transport, and similar in cost to glass bottles and allow sample collection in the field and transfer to the hydrolysis vessel without exposure of the sample to atmosphere. Storage of seawater samples in the foil bags was considered a reliable, alternative method to poisoning for ΣCO2, δ13C, and 14C, and freezing the samples is recommended for storage time beyond a week

Investigation into background levels of small organic samples at the NERC Radiocarbon Laboratory

Recent progress in preparation/combustion of submilligram organic samples at our laboratories is presented. Routine methods had to be modified/refined to achieve acceptable and consistent procedural blanks for organic samples smaller than 1000 g C. A description of the process leading to a modified combustion method for smaller organic samples is given in detail. In addition to analyzing different background materials, the influence of different chemical reagents on the overall radiocarbon background level was investigated, such as carbon contamination arising from copper oxide of different purities and from different suppliers. Using the modified combustion method, small amounts of background materials and known-age standard IAEA-C5 were individually combusted to CO2. Below 1000 g C, organic background levels follow an inverse mass dependency when combusted with the modified method, increasing from 0.13 0.05 pMC up to 1.20 0.04 pMC for 80 g C. Results for a given carbon mass were lower for combustion of etched Iceland spar calcite mineral, indicating that part of the observed background of bituminous coal was probably introduced by handling the material in atmosphere prior to combustion. Using the modified combustion method, the background-corrected activity of IAEA-C5 agreed to within 2 s of the consensus value of 23.05 pMC down to a sample mass of 55 g C

Progress in AMS target production in sub-milligram samples at the NERC Radiocarbon Laboratory

. Recent progress in graphite target production for sub-milligram environmental samples in our facility is presented. We describe an optimized hydrolysis procedure now routinely used for the preparation of CO2 from inorganic samples, a new high-vacuum line dedicated to small sample processing (combining sample distillation and graphitization units), as well as a modified graphitization procedure. Although measurements of graphite targets as small as 35 µg C have been achieved, system background and measurement uncertainties increase significantly below 150 µg C. As target lifetime can become critically short for targets <150 µg C, the facility currently only processes inorganic samples down to 150 µg C. All radiocarbon measurements are made at the Scottish Universities Environmental Research Centre (SUERC) accelerator mass spectrometry (AMS) facility. Sample processing and analysis are labor-intensive, taking approximately 3 times longer than samples ≥500 µg C. The technical details of the new system, graphitization yield, fractionation introduced during the process, and the system blank are discussed in detail

Shifting access to pools of shoot water sustains gas exchange and increases stem hydraulic safety during seasonal atmospheric drought

Understanding how plants acclimate to drought is crucial for predicting future vulnerability, yet seasonal acclimation of traits that improve drought tolerance in trees remains poorly resolved. We hypothesized that dry season acclimation of leaf and stem traits influencing shoot water storage and hydraulic capacitance would mitigate the drought-associated risks of reduced gas exchange and hydraulic failure in the mangrove Sonneratia alba. By late dry season, availability of stored water had shifted within leaves and between leaves and stems. While whole shoot capacitance remained stable, the symplastic fraction of leaf water increased 86%, leaf capacitance increased 104% and stem capacitance declined 80%. Despite declining plant water potentials, leaf and whole plant hydraulic conductance remained unchanged, and midday assimilation rates increased. Further, the available leaf water between the minimum water potential observed and that corresponding to 50% loss of stem conductance increased 111%. Shifting availability of pools of water, within and between organs, maintained leaf water available to buffer periods of increased photosynthesis and losses in stem hydraulic conductivity, mitigating risks of carbon depletion and hydraulic failure during atmospheric drought. Seasonal changes in access to tissue and organ water may have an important role in drought acclimation and avoidance.Research was conducted with financial support from Australian Research Council Discovery Grant DP180102969 awarded to M.B., L.S. and M.M. CB was supported by an Australian Government Research Training Program (RTP) Scholarship. TIF was supported by the Becas Chile PhD scholarship program granted by ANID. OB was supported by Australian Research Council Discovery Grant DP170104091. We thank Catherine Bone for outstanding support of field work conducted in mangrove forest along the Daintree River

Enantioselective organocatalytic synthesis of bicyclic resorcinols via an intramolecular Friedel-Crafts-type 1,4-addition: access to cannabidiol analogues

The organocatalytic transformation of resorcinols is extremely rare. In this article, we report a highly enantioselective, organocatalytic intramolecular cyclization of these systems by a Friedel–Crafts-type 1,4-addition using a Jørgensen-Hayashi-like organocatalyst with a large silyl protecting group, and show that heat improves reaction yield with virtually no detriment to enantioselectivity. A variety of bicyclic resorcinols were obtained with excellent enantioselectivities (up to 94%). To show the utility of these constructs, and as part of a wider project involving the synthesis of cannabinoid-like compounds, the resorcinol formed was used to generate both ‘normal’ and ‘abnormal’ cannabidiol (CBD) derivatives which were shown to have anticonvulsant activity

Reefs at Risk: A Map-Based Indicator of Threats to the Worlds Coral Reefs

This report presents the first-ever detailed, map-based assessment of potential threats to coral reef ecosystems around the world. "Reefs at Risk" draws on 14 data sets (including maps of land cover, ports, settle-ments, and shipping lanes), information on 800 sites known to be degraded by people, and scientific expertise to model areas where reef degradation is predicted to occur, given existing human pressures on these areas. Results are an indicator of potential threat (risk), not a measure of actual condition. In some places, particularly where good management is practiced, reefs may be at risk but remain relatively healthy. In others, this indicator underestimates the degree to which reefs are threatened and degraded.Our results indicate that:Fifty-eight percent of the world's reefs are poten-tially threatened by human activity -- ranging from coastal development and destructive fishing practices to overexploitation of resources, marine pollution, and runoff from inland deforestation and farming.Coral reefs of Asia (Southeastern); the most species-rich on earth, are the most threatened of any region. More than 80 percent are at risk (undermedium and high potential threat), and over half are at high risk, primarily from coastal development and fishing-related pressures.Overexploitation and coastal development pose the greatest potential threat of the four risk categories considered in this study. Each, individually, affects a third of all reefs.The Pacific, which houses more reef area than any other region, is also the least threatened. About 60 percent of reefs here are at low risk.Outside of the Pacific, 70 percent of all reefs are at risk.At least 11 percent of the world's coral reefs contain high levels of reef fish biodiversity and are under high threat from human activities. These "hot spot" areas include almost all Philippine reefs, and coral communities off the coasts of Asia, the Comoros, and the Lesser Antilles in the Caribbean.Almost half a billion people -- 8 percent of the total global population -- live within 100 kilometers of a coral reef.Globally, more than 400 marine parks, sanctuaries, and reserves (marine protected areas) contain coral reefs. Most of these sites are very small -- more than 150 are under one square kilometer in size. At least 40 countries lack any marine protected areas for conserving their coral reef systems

Vapour pressure deficit modulates hydraulic function and structure of tropical rainforests under nonlimiting soil water supply

Atmospheric conditions are expected to become warmer and drier in the future, but little is known about how evaporative demand influences forest structure and function independently from soil moisture availability, and how fast-response variables (such as canopy water potential and stomatal conductance) may mediate longer-term changes in forest structure and function in response to climate change. We used two tropical rainforest sites with different temperatures and vapour pressure deficits (VPD), but nonlimiting soil water supply, to assess the impact of evaporative demand on ecophysiological function and forest structure. Common species between sites allowed us to test the extent to which species composition, relative abundance and intraspecific variability contributed to site-level differences. The highest VPD site had lower midday canopy water potentials, canopy conductance (gc), annual transpiration, forest stature, and biomass, while the transpiration rate was less sensitive to changes in VPD; it also had different height–diameter allometry (accounting for 51% of the difference in biomass between sites) and higher plot-level wood density. Our findings suggest that increases in VPD, even in the absence of soil water limitation, influence fast-response variables, such as canopy water potentials and gc, potentially leading to longer-term changes in forest stature resulting in reductions in biomass

Foliar water uptake via cork warts in mangroves of the Sonneratia genus

Foliar water uptake (FWU) occurs in plants of diverse ecosystems; however, the diversity of pathways and their associated FWU kinetics remain poorly resolved. We characterized a novel FWU pathway in two mangrove species of the Sonneratia genus, S. alba and S. caseolaris. Further, we assessed the influence of leaf wetting duration, wet-dry seasonality and leaf dehydration on leaf conductance to surface water (Ksurf ). The symplastic tracer dye, disodium fluorescein, revealed living cells subtending and encircling leaf epidermal structures known as cork warts as a pathway of FWU entry into the leaf. Rehydration kinetics experiments revealed a novel mode of FWU, with slow and steady rates of water uptake persistent over a duration of 12 h. Ksurf increased with longer durations of leaf wetting and was greater in leaves with more negative water potentials at the initiation of leaf wetting. Ksurf declined by 68% between wet and dry seasons. Our results suggest that FWU via cork warts in Sonneratia sp. may be rate limited and under active regulation. We conclude that FWU pathways in halophytes may require ion exclusion to avoid uptake of salt when inundated, paralleling the capacity of halophyte roots for ion selectivity during water acquisition. This article is protected by copyright. All rights reserved.The project was funded by the Australian Research Council Discovery Grant Projects DP150104437 and DP180102969 awarded to MCB. TIF was supported by the Becas Chile PhD scholarship program granted by ANID. CB, HAB and RJH were supported by Australian Government Research Training Program (RTP) Scholarships. HIH was supported by an NSF Graduate Research Fellowship and a NSF GROW Fellowship. OB was supported by Australian Research Council Discovery Grant DP17010409

Predicting dark respiration rates of wheat leaves from hyperspectral reflectance

Greater availability of leaf dark respiration (R dark) data could facilitate breeding efforts to raise crop yield and improve global carbon cycle modelling. However, the availability of R dark data is limited because it is cumbersome, time consuming, or destructive to measure. We report a non‐destructive and high‐throughput method of estimating R dark from leaf hyperspectral reflectance data that was derived from leaf R dark measured by a destructive high‐throughput oxygen consumption technique. We generated a large dataset of leaf R dark for wheat (1380 samples) from 90 genotypes, multiple growth stages, and growth conditions to generate models for R dark. Leaf R dark (per unit leaf area, fresh mass, dry mass or nitrogen, N) varied 7‐ to 15‐fold among individual plants, whereas traits known to scale with R dark, leaf N, and leaf mass per area (LMA) only varied twofold to fivefold. Our models predicted leaf R dark, N, and LMA with r 2 values of 0.50–0.63, 0.91, and 0.75, respectively, and relative bias of 17–18% for R dark and 7–12% for N and LMA. Our results suggest that hyperspectral model prediction of wheat leaf R dark is largely independent of leaf N and LMA. Potential drivers of hyperspectral signatures of R dark are discussed

MAP Kinase Phosphatase-2 Plays a Critical Role in Response to Infection by Leishmania mexicana

In this study we generated a novel dual specific phosphatase 4 (DUSP4) deletion mouse using a targeted deletion strategy in order to examine the role of MAP kinase phosphatase-2 (MKP-2) in immune responses. Lipopolysaccharide (LPS) induced a rapid, time and concentration-dependent increase in MKP-2 protein expression in bone marrow-derived macrophages from MKP-2+/+ but not from MKP-2−/− mice. LPS-induced JNK and p38 MAP kinase phosphorylation was significantly increased and prolonged in MKP-2−/− macrophages whilst ERK phosphorylation was unaffected. MKP-2 deletion also potentiated LPS-stimulated induction of the inflammatory cytokines, IL-6, IL-12p40, TNF-α, and also COX-2 derived PGE2 production. However surprisingly, in MKP-2−/− macrophages, there was a marked reduction in LPS or IFNγ-induced iNOS and nitric oxide release and enhanced basal expression of arginase-1, suggesting that MKP-2 may have an additional regulatory function significant in pathogen-mediated immunity. Indeed, following infection with the intracellular parasite Leishmania mexicana, MKP-2−/− mice displayed increased lesion size and parasite burden, and a significantly modified Th1/Th2 bias compared with wild-type counterparts. However, there was no intrinsic defect in MKP-2−/− T cell function as measured by anti-CD3 induced IFN-γ production. Rather, MKP-2−/− bone marrow-derived macrophages were found to be inherently more susceptible to infection with Leishmania mexicana, an effect reversed following treatment with the arginase inhibitor nor-NOHA. These findings show for the first time a role for MKP-2 in vivo and demonstrate that MKP-2 may be essential in orchestrating protection against intracellular infection at the level of the macrophage