The effects of growth phase and salinity on the hydrogen isotopic composition of alkenones produced by coastal haptophyte algae

The isotopic fractionation of hydrogen during the biosynthesis of alkenones produced by marine haptophyte algae has been shown to depend on salinity and, as such, the hydrogen isotopic composition of alkenones is emerging as a palaeosalinity proxy. The relationship between fractionation and salinity has previously only been determined during exponential growth, whilst it is not yet known in which growth phases natural haptophyte populations predominantly exist. We have therefore determined the relationship between the fractionation factor, alpha alkenones-water, and salinity for C37 alkenones produced in different growth phases of batch cultures of the major alkenone-producing coastal haptophytes Isochrysis galbana (strain CCMP 1323) and Chrysotila lamellosa (strain CCMP 1307) over a range in salinity from ca. 10 to ca. 35. alpha alkenones-water was similar in both species, ranging over 0.841-0.900 for I. galbana and 0.838-0.865 for C. lamellosa. A strong (0.85 <= R**2 <= 0.97; p < 0.0001) relationship between salinity and fractionation factor was observed in both species at all growth phases investigated. This suggests that alkenone dD has the potential to be used as a salinity proxy in coastal areas where haptophyte communities are dominated by these coastal species. However, there was a marked difference in the sensitivity of alpha alkenones-water to salinity between different growth phases: in the exponential growth phase of I. galbana, alpha alkenones-water increased by 0.0019 per salinity unit (S 1), but was less sensitive at 0.0010 S 1 and 0.0008 S 1 during the stationary and decline phases, respectively. Similarly, in C. lamellosa alpha alkenones-water increased by 0.0010 S 1 in the early stationary phase and by 0.0008 S 1 during the late stationary phase. Assuming the shift in sensitivity of alpha alkenones-water to salinity observed at the end of exponential growth in I. galbana is similar in other alkenone-producing species, the predominant growth phase of natural populations of haptophytes will affect the sensitivity of the alkenone salinity proxy. The proxy is likely to be most sensitive to salinity when alkenones are produced in a state similar to exponential growth.

Impact of salinity and growth phase on alkenone distributions in coastal haptophytes

Batch cultures of Isochrysis galbana (strain CCMP 1323) and Chrysotila lamellosa (strain CCMP 1307) were grown at salinity ca. 10 to ca. 35 and the alkenone distributions determined for different growth phases. UK'37 values decreased slightly with salinity for C. lamellosa but were largely unaffected for I. galbana except during the decline phase. The values decreased with incubation time in both species. The proportion of C37:4, used as proxy for salinity, increased in both species at 0.16-0.20% per salinity unit, except during the stationary phase for I. galbana. C37:4 was much more abundant in C. lamellosa (30-44%) than in I. galbana (4-12%). Although our results suggest that salinity has a direct effect on alkenone distributions, growth phase and species composition will also have a marked impact, complicating the use of alkenone distributions as a proxy for salinity in the marine environment.

Experimental growth characteristics, cell size, yessotoxin values and bioassay data of Protoceratium reticulatum (Dinophyceae)

Harmful algal blooms are mainly caused by marine dinoflagellates and are known to produce potent toxins that may affect the ecosystem, human activities and health. Such events have increased in frequency and intensity worldwide in the past decades. Numerous processes involved in Global Change are amplified in the Arctic, but little is known about species specific responses of arctic dinoflagellates. The aim of this work was to perform an exhaustive morphological, phylogenetical and toxinological characterization of Greenland Protoceratium reticulatum and, in addition, to test the effect of temperature on growth and production of bioactive secondary metabolites. Seven clonal isolates, the first isolates of P. reticulatum available from arctic waters, were phylogenetically characterized by analysis of the LSU rDNA. Six isolates were further characterized morphologically and were shown to produce both yessotoxins (YTX) and lytic compounds, representing the first report of allelochemical activity in P. reticulatum. As shown for one of the isolates, growth was strongly affected by temperature with a maximum growth rate at 15 °C, a significant but slow growth at 1 °C, and cell death at 25 °C, suggesting an adaptation of P. reticulatum to temperate waters. Temperature had no major effect on total YTX cell quota or lytic activity but both were affected by the growth phase with a significant increase at stationary phase. A comparison of six isolates at a fixed temperature of 10 °C showed high intraspecific variability for all three physiological parameters tested. Growth rate varied from 0.06 to 0.19 per day, and total YTX concentration ranged from 0.3 to 15.0 pg YTX/cell and from 0.5 to 31.0 pg YTX/cell at exponential and stationary phase, respectively. All six isolates performed lytic activity; however, for two isolates lytic activity was only detectable at higher cell densities in stationary phase.

Data files of figures 1, 2 and 3

We investigated optimal conditions for characterization of bioactivity of lytic compound(s) excreted by Alexandrium tamarense based on a cell-bioassay system. Allelochemical response of the cryptophyte Rhodomonas salina indicated the presence oflytic compound(s) in a reliable and reproducible way and allows for quantification of this lytic effect. The parameters tested were the incubation time of putatively lytic extracts or fractions with the target organism R. salina, different techniques for cell harvest from A. tamarense cultures and the optimal harvest time. A three hour incubation time was found to be optimal to yield a rapid response while accurately estimating effective concentration (ECso) values. Harvest of A. tamarense cultures by filtration resulted in loss of lytic activity in most cases and centrifugation was most efficient in terms of recovery of lytic activity. Maximum yield of extracellular lytic activity of A. tamarense cultures was achieved in the stationary phase. Such optimized bioassay guided fractionation techniques are a valuable asset in the isolation and eventual stmctural elucidation of the unknown lytic substances.

Geochemistry, microbiology and phospholipid fatty acid content of ODP Site 169-1036 sediments

Phospholipid fatty acids were measured in samples of 60°-130°C sediment taken from three holes at Site 1036 (Ocean Drilling Program Leg 169) to determine microbial community structure and possible community replacement at high temperatures. Five of six samples had similar concentrations of phospholipid fatty acids (2-6 pmol/g dry weight of sediment), and biomass estimates from these measurements compare favorably with direct microscopic counts, lending support to previous microscopic measures of deep sedimentary biomass. Very long-chain phospholipid fatty acids (21 to 30 carbons) were detected in the sediment and were up to half the total phospholipid fatty acid measured; they appear to increase in abundance with temperature, but their significance is not known. Community composition from lipid analysis showed that samples contained standard eubacterial membrane lipids but no detectable archaeal lipids, though archaea would be expected to dominate the samples at high temperatures. Cluster analysis of Middle Valley phospholipid fatty acid compositions shows that lipids in Middle Valley sediment samples are similar to each other at all temperatures, with the exception of very long-chain fatty acids. The data neither support nor deny a shift to a high-temperature microbial community in hot cores, so at the present time we cannot draw conclusions about whether the microbes observed in these hot sediments are active.

Combined effects of CO2 and temperature on carbon uptake and partitioning by the marine diatoms Thalassiosira weissflogii and Dactyliosolen fragilissimus

Carbon uptake and partitioning of two globally abundant diatom species, Thalassiosira weissflogii and Dactyliosolen fragilissimus, was investigated in batch culture experiments under four conditions: ambient (15°C, 400 µatm), high CO2 (15°C, 1000 µatm), high temperature (20°C, 400 µatm), and combined (20°C, 1000 µatm). The experiments were run from exponential growth into the stationary phase (six days after nitrogen depletion), allowing us to track biogeochemical dynamics analogous to bloom situations in the ocean. Elevated CO2 had a fertilizing effect and enhanced uptake of dissolved inorganic carbon (DIC) by about 8% for T. weissflogii and by up to 39% for D. fragilissimus. This was also reflected in higher cell numbers, build-up of particulate and dissolved organic matter, and transparent exopolymer particles. The CO2 effects were most prominent in the stationary phase when nitrogen was depleted and CO2(aq) concentrations were low. This indicates that diatoms in the high CO2 treatments could take up more DIC until CO2 concentrations in seawater became so low that carbon limitation occurs. These results suggest that, contrary to common assumptions, diatoms could be highly sensitive to ongoing changes in oceanic carbonate chemistry, particularly under nutrient limitation. Warming from 15 to 20 °C had a stimulating effect on one species but acted as a stressor on the other species, highlighting the importance of species-specific physiological optima and temperature ranges in the response to ocean warming. Overall, these sensitivities to CO2 and temperature could have profound impacts on diatoms blooms and the biological pump.

Mesocosm bloom experiment results with the coccolithophore Emiliania huxleyi

We investigated the effect of CO2 and primary production on the carbon isotopic fractionation of alkenones and particulate organic matter (POC) during a natural phytoplankton bloom dominated by the coccolithophore Emiliania huxleyi. In nine semi-closed mesocosms (~11 m**3 each), three different CO2 partial pressures (pCO2) in triplicate represented glacial (~180 ppmv CO2), present (380 ppmv CO2), and year 2100 (~710 ppmv CO2) CO2 conditions. The largest shift in alkenone isotopic composition (4-5 per mil) occurred during the exponential growth phase, regardless of the CO2 concentration in the respective treatment. Despite the difference of ~500 ppmv, the influence of pCO2 on isotopic fractionation was marginal (1-2 per mil). During the stationary phase, E. huxleyi continued to produce alkenones, accumulating cellular concentrations almost four times higher than those of exponentially dividing cells. Our isotope data indicate that, while alkenone production was maintained, the interaction of carbon source and cellular uptake dynamics by E. huxleyi reached a steady state. During stationary phase, we further observed a remarkable increase in the difference between d13C of bulk organic matter and of alkenones spanning 7-12 per mil. We suggest that this phenomenon is caused mainly by a combination of extracellular release of 13C-enriched polysaccharides and subsequent particle aggregation induced by the production of transparent exopolymer particles (TEP).

Geochemistry on the Kekesai composite intrusion

The late Carboniferous to Permian is a critical period for final amalgamation of the Central Asian Orogenic Belt (CAOB), which is characterized by voluminous igneous rocks, particularly granitoids. The Kekesai composite granitoid porphyry intrusion, situated in the Chinese western Tianshan (southwest part of CAOB) includes two intrusive phases, a monzogranite phase, intruded by a granodiorite phase. LA-ICPMS U-Pb zircon analyses suggest that the monzogranitic rocks formed at 305.5±1.1 Ma, with a wide age range of inherited zircons (358-488 Ma and 1208-1391 Ma), whereas the granodioritic rocks formed at 288.7±1.5 Ma. The monzogranitic and granodioritic phases have similar geochemical features and Sr-Nd-Hf isotopic compositions. They exhibit high and variable SiO2 (66-71 wt.%) and MgO (0.41-2.14 wt.%) contents with some arc-like geochemical characteristics (e.g., enrichment of large ion lithophile elements and negative anomalies of Nb, Ta and Ti) and relatively high initial 87Sr/86Sr ratios (ISr=0.7055-0.7059), low positive eNd(t) (+0.84 to +1.03) as well as a large variation in Hf isotopic compositions with eHf(t) between +3.43 to +14.8, implying both of them were derived from similar source materials. These geochemical characteristics suggest that they might be mainly derived from the partial melting of arc-derived Mesoproterozoic mafic lower crust with involvement of a mantle-derived component in variable proportions by mantle-derived magma underplating. The presence of late-Ordovician to earliest early Carboniferous inherited zircons and the Hf isotopic compositions in the monzogranitic sample, similar to that of the widespread juvenile arc rocks, indicates some crust contamination during magma emplacement. Our new data, combined with previous studies, imply that extensive post-collisional magmatism due to underplating of mantle-derived magma, could plausibly be explained by slab break-off regime.

Total nitrogen from solid phase in the Jena Experiment (Main Experiment up to 30cm depth, year 2008)

This data set contains measurements of total nitrogen from the main experiment plots of a large grassland biodiversity experiment (the Jena Experiment; see further details below). In the main experiment, 82 grassland plots of 20 x 20 m were established from a pool of 60 species belonging to four functional groups (grasses, legumes, tall and small herbs). In May 2002, varying numbers of plant species from this species pool were sown into the plots to create a gradient of plant species richness (1, 2, 4, 8, 16 and 60 species) and functional richness (1, 2, 3, 4 functional groups). Plots were maintained by bi-annual weeding and mowing. Soil sampling and analysis: Stratified soil sampling was performed in April 2008 to a depth of 30 cm. Three independent samples per plot were taken using a split tube sampler with an inner diameter of 4.8 cm (Eijkelkamp Agrisearch Equipment, Giesbeek, the Netherlands). Soil samples were segmented to a depth resolution of 5 cm in the field, giving six depth subsamples per core, and made into composite samples per depth. Sampling locations were less than 30 cm apart from sampling locations in other years. Samples were dried at 40°C. All soil samples were passed through a sieve with a mesh size of 2 mm. Because of much higher proportions of roots in the soil, the samples were further sieved to 1 mm according to common root removal methods. No additional mineral particles were removed by this procedure. Total nitrogen concentration was analyzed on ball-milled subsamples (time 4 min, frequency 30 s-1) by an elemental analyzer at 1150°C (Elementaranalysator vario Max CN; Elementar Analysensysteme GmbH, Hanau, Germany).

Total nitrogen from solid phase in the Jena Experiment (Main Experiment up to 30cm depth, year 2004)

This data set contains measurements of total nitrogen from the main experiment plots of a large grassland biodiversity experiment (the Jena Experiment; see further details below). In the main experiment, 82 grassland plots of 20 x 20 m were established from a pool of 60 species belonging to four functional groups (grasses, legumes, tall and small herbs). In May 2002, varying numbers of plant species from this species pool were sown into the plots to create a gradient of plant species richness (1, 2, 4, 8, 16 and 60 species) and functional richness (1, 2, 3, 4 functional groups). Plots were maintained by bi-annual weeding and mowing. Soil sampling and analysis: Stratified soil sampling was performed in April 2004 to a depth of 30 cm. Three independent samples per plot were taken using a split tube sampler with an inner diameter of 4.8 cm (Eijkelkamp Agrisearch Equipment, Giesbeek, the Netherlands). Soil samples were segmented to a depth resolution of 5 cm in the field, giving six depth subsamples per core, and made into composite samples per depth. Sampling locations were less than 30 cm apart from sampling locations in other years. Samples were dried at 40°C. All soil samples were passed through a sieve with a mesh size of 2 mm. Because of much higher proportions of roots in the soil, the samples were further sieved to 1 mm according to common root removal methods. No additional mineral particles were removed by this procedure. Total nitrogen concentration was analyzed on ball-milled subsamples (time 4 min, frequency 30 s-1) by an elemental analyzer at 1150°C (Elementaranalysator vario Max CN; Elementar Analysensysteme GmbH, Hanau, Germany).

Fossil pollen record of composite sediment core TMD from Tso Moriri, analysis of modern surface pollen samples, mean annual precipitation reconstruction, and digitisation and recalibration of different discussed palaeoclimate proxy records

This paper presents a new fossil pollen record from Tso Moriri (32°54'N, 78°19'E, 4512 m a.s.l.) and seeks to reconstruct changes in mean annual precipitation (MAP) during the last 12,000 years. This high-alpine lake occupies an area of 140 km**2 in a glacial-tectonic valley in the northwestern Himalaya. The region has a cold climate, with a MAP <300 mm, and open vegetation. The hydrology is controlled by the Indian Summer Monsoon (ISM), but winter westerly-associated precipitation also affects the regional water balance. Results indicate that precipitation levels varied significantly during the Holocene. After a rapid increase in MAP, a phase of maximum humidity was reached between ca. 11 to 9.6 cal ka BP, followed by a gradual decline in MAP. This trend parallels the reduction in the Northern Hemisphere summer insolation. Comparison of different palaeoclimate proxy records reveal evidence for a stronger Holocene decrease in precipitation in the northern versus the southern parts of the ISM domain. The long-term trend of ISM weakening is overlaid with several short periods of greater dryness, which are broadly synchronous with the North Atlantic cold spells, suggesting reduced amounts of westerly-associated winter precipitation. Compared to the mid and late Holocene, it appears that westerlies had a greater influence on the western parts of the ISM domain during the early Holocene. During this period, the westerly-associated summer precipitation belt was positioned at Mediterranean latitudes and amplified the ISM-derived precipitation. The Tso Moriri pollen record and moisture reconstructions also suggest that changes in climatic conditions affected the ancient Harappan Civilisation, which flourished in the greater Indus Valley from approximately 5.2 to 3 cal ka BP. The prolonged Holocene trend towards aridity, punctuated by an interval of increased dryness (between ca. 4.5 to 4.3 cal ka BP), may have pushed the Mature Harappan urban settlements (between ca. 4.5 to 3.9 cal ka BP) to develop more efficient agricultural practices to deal with the increasingly acute water shortages. The amplified aridity associated with North Atlantic cooling between ca. 4 to 3.6 and around 3.2 cal ka BP further hindered local agriculture, possibly causing the deurbanisation that occurred from ca. 3.9 cal ka BP and eventual collapse of the Harappan Civilisation between ca. 3.5 to 3 cal ka BP.

Calcium isotope ratios of pore fluids and solid phase of organic rich sediments

Pore fluid calcium isotope, calcium concentration and strontium concentration data are used to measure the rates of diagenetic dissolution and precipitation of calcite in deep-sea sediments containing abundant clay and organic material. This type of study of deep-sea sediment diagenesis provides unique information about the ultra-slow chemical reactions that occur in natural marine sediments that affect global geochemical cycles and the preservation of paleo-environmental information in carbonate fossils. For this study, calcium isotope ratios (d44/40Ca) of pore fluid calcium from Ocean Drilling Program (ODP) Sites 984 (North Atlantic) and 1082 (off the coast of West Africa) were measured to augment available pore fluid measurements of calcium and strontium concentration. Both study sites have high sedimentation rates and support quantitative sulfate reduction, methanogenesis and anaerobic methane oxidation. The pattern of change of d44/40Ca of pore fluid calcium versus depth at Sites 984 and 1082 differs markedly from that of previously studied deep-sea Sites like 590B and 807, which are composed of nearly pure carbonate sediment. In the 984 and 1082 pore fluids, d44/40Ca remains elevated near seawater values deep in the sediments, rather than shifting rapidly toward the d44/40Ca of carbonate solids. This observation indicates that the rate of calcite dissolution is far lower than at previously studied carbonate-rich sites. The data are fit using a numerical model, as well as more approximate analytical models, to estimate the rates of carbonate dissolution and precipitation and the relationship of these rates to the abundance of clay and organic material. Our models give mutually consistent results and indicate that calcite dissolution rates at Sites 984 and 1082 are roughly two orders of magnitude lower than at previously studied carbonate-rich sites, and the rate correlates with the abundance of clay. Our calculated rates are conservative for these sites (the actual rates could be significantly slower) because other processes that impact the calcium isotope composition of sedimentary pore fluid have not been included. The results provide direct geochemical evidence for the anecdotal observation that the best-preserved carbonate fossils are often found in clay or organic-rich sedimentary horizons. The results also suggest that the presence of clay minerals has a strong passivating effect on the surfaces of biogenic carbonate minerals, slowing dissolution dramatically even in relation to the already-slow rates typical of carbonate-rich sediments.

Total nitrogen from solid phase in the Jena Experiment (Main Experiment up to 30cm depth, year 2006)

This data set contains measurements of total nitrogen from the main experiment plots of a large grassland biodiversity experiment (the Jena Experiment; see further details below). In the main experiment, 82 grassland plots of 20 x 20 m were established from a pool of 60 species belonging to four functional groups (grasses, legumes, tall and small herbs). In May 2002, varying numbers of plant species from this species pool were sown into the plots to create a gradient of plant species richness (1, 2, 4, 8, 16 and 60 species) and functional richness (1, 2, 3, 4 functional groups). Plots were maintained by bi-annual weeding and mowing. Soil sampling and analysis: Stratified soil sampling was performed in April 2006 to a depth of 30 cm. Three independent samples per plot were taken using a split tube sampler with an inner diameter of 4.8 cm (Eijkelkamp Agrisearch Equipment, Giesbeek, the Netherlands). Soil samples were segmented to a depth resolution of 5 cm in the field, giving six depth subsamples per core, and made into composite samples per depth. Sampling locations were less than 30 cm apart from sampling locations in other years. Samples were dried at 40°C. All soil samples were passed through a sieve with a mesh size of 2 mm. Because of much higher proportions of roots in the soil, the samples were further sieved to 1 mm according to common root removal methods. No additional mineral particles were removed by this procedure. Total nitrogen concentration was analyzed on ball-milled subsamples (time 4 min, frequency 30 s-1) by an elemental analyzer at 1150°C (Elementaranalysator vario Max CN; Elementar Analysensysteme GmbH, Hanau, Germany).

(Table 1) Age control points of ODP Site 160-968

The astronomical timescale of the Eastern Mediterranean Plio-Pleistocene builds on tuning of sapropel layers to Northern Hemisphere summer insolation maxima. A 3000-year precession lag has become instrumental in the tuning procedure as radiocarbon dating revealed that the midpoint of the youngest sapropel, S1, in the early Holocene occurred approximately 3000 years after the insolation maximum. The origin of the time lag remains elusive, however, because sapropels are generally linked to maximum African monsoon intensities and transient climate modeling results indicate an in-phase behavior of the African monsoon relative to precession forcing. Here we present new high-resolution records of bulk sediment geochemistry and benthic foraminiferal oxygen isotopes from ODP Site 968 in the Eastern Mediterranean. We show that the 3000-year precession time lag of the sapropel midpoints is consistent with (1) the global marine isotope chronology, (2) maximum (monsoonal) precipitation conditions in the Mediterranean region and China derived from radiometrically dated speleothem records, and (3) maximum atmospheric methane concentrations in Antarctica ice cores. We show that the time lag relates to the occurrence of precession-paced North Atlantic cold events, which systematically delayed the onset of strong boreal summer monsoon intensity. Our findings may also explain a non-stationary behavior of the African monsoon over the past 3 million years due to more frequent and intensive cold events in the Late Pleistocene.