Cenomanian-Turonian benthic foraminifera of the Kerguelen Plateau

Recent drilling on the Kerguelen Plateau (Ocean Drilling Program Leg 183) has provided a unique and exciting high latitude record of palaeoceanographic change during the Cenomanian-Turonian in the Southern Ocean. The benthic foraminiferal succession at Site 1138 records the evolution of the Kerguelen Plateau from a subaerially exposed platform in the Cenomanian to a bathyal, pelagic environment in the early Turonian, following a major transgressive pulse and increased thermal subsidence of the Kerguelen Plateau, which led to a sea-level rise of possibly several hundred metres. Diversified benthic foraminiferal assemblages indicate an upper bathyal, mesotrophic setting after the peak of the transgression. The assemblages exhibit strong similarities to temperate, shelf and slope assemblages in the Northern Hemisphere. This bimodal distribution reflects the existence of open oceanic gateways and a dynamic trans-hemispheric global circulation. Equatorial assemblages are characterized by a low-diversity, high carbon flux biofacies. Assemblages from Alaska demonstrate high organic productivity and low oxygen conditions and the prevalence of elevated temperatures on the flooded shelf of the North Slope. Our results show that the distribution of upper bathyal benthic foraminifera was strongly modulated by carbon flux and oxygenation fluctuations, and not by physical migration barriers.

Eocene to Quaternary benthic foraminifers from the Izu-Bonin Arc

The benthic foraminifer fauna at Sumisu Rift Sites 790 and 791 indicates that a deep open-ocean (>2300 m) or a basin with open-ocean access existed between 1.1 and 0.7 Ma at the time of the initiation of rifting. The appearance of a low- to medium-oxygen fauna (1600-2300 m) between 0.7 and 0.5 Ma suggests that the open-ocean access may have been terminated at this time because of the development of volcanoes and rift flank uplifts around the basin. The occurrence of low-oxygen faunas at 0.03 Ma suggests a secondary closing of the basin. The lower bathyal benthic faunas from lower Pliocene sediments of rift margin Site 788 suggest about 0.6-1.6 km of total basement uplift. This uplift may have led to the formation of the major hiatus between 2.3 and <0.3 Ma. The faunal changes of benthic foraminifers at Sites 792 and 793 in the forearc basin document a shallowing water depth from below the carbonate compensation depth (CCD) (about 3.5 km) in the late early Oligocene to the present depths of 1800 and 2975 m, respectively. These data suggest about 1 km of total basement uplift in the inner part of the forearc basin (Site 792) and about 0.6 km total basement subsidence in the central part of the forearc basin (Site 793) since about 31 Ma. The former uplift led to a thinner sediment accumulation (800 m) and the latter subsidence to a thicker sediment accumulation (1400 m) at these sites. Faunal changes of benthic foraminifers observed in Sites 782 and 786 sequences drilled at the outer-arc high document a deepening water depth from 1.3 to 2.1 km in late Eocene to the present depth of about 3 km. These data suggest about 1.1-1.9 and 1.3-2.1 km of total basement subsidence at Sites 786 and 782, respectively. These results indicate total basement uplift in the inner part of the Bonin arc-trench system since late Oligocene and total basement subsidence in the outer part of the system since late Eocene. The last occurrence (LO) of Stilostomella spp. and Pleurostomella spp. and the first occurrence (F0) of Bulimina aculeata d'Orbigny occurred consistently at 0.7 Ma at all three arc proximal sites (790,791, and 792). This fact is taken to suggest a change of water mass, from one originating from the central part of the ocean to that originating from ocean-margin areas at that time.

Radiocarbon dating on cold-water corals, grain size, Corg, and benthic foraminfera assemblage of two sediment cores from the Ionian Sea

Continuous sedimentary records from an eastern Mediterranean cold-water coral ecosystem thriving in intermediate water depths (~600 m) reveal a temporary extinction of cold-water corals during the Early to Mid Holocene from 11.4-5.9 cal kyr BP. Benthic foraminiferal assemblage analysis shows low-oxygen conditions of 2 ml l**-1 during the same period, compared to bottom-water oxygen values of 4-5 ml l**-1 before and after the coral-free interval. The timing of the corals' demise coincides with the sapropel S1 event, during which the deep eastern Mediterranean basin turned anoxic. Our results show that during the sapropel S1 event low oxygen conditions extended to the rather shallow depths of our study site in the Ionian Sea and caused the cold-water corals temporary extinction. This first evidence for the sensitivity of cold-water corals to low oceanic oxygen contents suggests that the projected expansion of tropical oxygen minimum zones resulting from global change will threaten cold-water coral ecosystems in low latitudes in the same way that ocean acidification will do in the higher latitudes.

Geochemical composition and atomic formulas of saponites, celadonites and clays from ODP Leg 168 sites

A drilling transect across the sedimented eastern flank of the Juan de Fuca Ridge, conducted during Leg 168 of the Ocean Drilling Program, resulted in the recovery of samples of volcanic basement rocks (pillow basalts, massive basalts, and volcanic glass breccias) that exhibit the effects of low-temperature hydrothermal alteration. Secondary clays are ubiquitous, with Mg-rich and Fe-rich saponite and celadonitic clays commonly accounting for several percent, and up to 10%-20% by volume. Present-day temperatures of the basement sites vary from 15° to 64°C, with the coolest site being about 0.8 Ma, and the warmest site being about 3.5 Ma. Whereas clays are abundant at sites that have been heated to present temperatures of 23°C and higher, the youngest site at 15°C has only a small trace of secondary clay alteration. Alteration increases as temperatures increase and as the volcanic basement ages. The chemical compositions of secondary clays were determined by electron microprobe, and additional trace element data were determined by both conventional nebulization inductively coupled plasma-mass spectroscopy (ICP-MS) and laser-ablation ICP-MS. Trioctahedral saponite and pyrite are characteristic of the interior of altered rock pieces, forming under conditions of low-oxygen fugacity. Dioctahedral celadonite-like clays along with iron oxyhydroxide and Mg-saponite are characteristic of oxidized haloes surrounding the nonoxidized rock interiors. Chemical compositions of the clays are very similar to those determined from other deep-sea basalts altered at low temperature. The variable Mg:Fe of saponite appears to be a systematic function both of the Mg:Fe of the host rock and the oxidation state during water-rock interaction.

Oliogocene to Pleistocene benthic foraminifera in ODP Leg 121 sites

Oligocene to Pleistocene bathyal benthic foraminifers at Broken Ridge (Site 754) and Ninetyeast Ridge (Site 756), eastern Indian Ocean, were investigated for then- stratigraphic distribution and their response to paleoceanographic changes. Q-mode factor analysis was applied to relative abundance data of the most abundant benthic foraminifers. At Site 754, seven varimax assemblages were recognized from the upper Oligocene to the Pleistocene: the Gyroidina orbicularis-Rectuvigerina striata Assemblage in the uppermost Oligocene; the Lenticulina spp. Assemblage in the upper Oligocene to lower Miocene, and in lower Miocene to lowermost middle Miocene; the Burseolina cf. pacifica-Cibicidoides mundulus Assemblage in the lower Miocene; the Planulina wuellerstorfi Assemblage in the upper middle Miocene; the Globocassidulina spp. Assemblage in the upper Miocene; the Gavelinopsis lobatulus-Uvigerina proboscidea Assemblage in the Pliocene; and the Ehrenbergina spp. Assemblage in the Pleistocene. The major faunal changes are complex, but exist between the Lenticulina spp. Assemblage and the P. wuellerstorfi Assemblage at ~13.8 Ma, and between the Ehrenbergina spp. Assemblage and the G. lobatulus Assemblage at ~5 Ma. The development of the P. wuellerstorfi and Globocassidulina spp. Assemblages after 13.8 Ma is correlated with the decrease in temperature of the intermediate waters of the ocean, in turn related to Antarctic glacial expansion. The faunal changes at ~5 Ma are related to the development of low oxygen intermediate water, formed in the presence of a strong thermocline. At Site 756, six varimax assemblages are distributed as follows: the Cibicidoides cf. mundulus-Oridorsalis umbonatus Assemblage in the lower Oligocene; the Epistominella umbonifera-Cibicidoides mundulus Assemblage from the upper Oligocene to the lower Miocene; the Cibicidoides mundulus-Burseolinapacifica Assemblage from lower Miocene to the lower middle Miocene; the Globocassidulina spp. Assemblage from the upper lower Miocene to the Pliocene; the Uvigerina proboscidea Assemblage in the upper Miocene and the Pliocene; and the Globocassidulina sp. D Assemblage in the Pliocene. The main faunal change at this site is between the E. umbonifera Assemblage and the Globocassidulina spp. Assemblage, at ~17.1 Ma. The timing of this faunal change is coeval with faunal changes in the North Atlantic and the Pacific. The change is related to a change in bottom water characteristics caused by an increased influence of carbonate corrosive water from the Antarctic source region, and a change in surface productivity. A low oxygen event at Site 756, which started at about 7.3 Ma, occurred about 2.3 m.y. before that at Site 754. The different response to global paleoceanographic changes is not yet explained, but may be due to the difference of marine topography and the degree of upwelling

Characterization of dead-zone eddies in the tropical Northeast Atlantic

Localized open-ocean low-oxygen dead-zones in the tropical Northeast Atlantic are recently discovered ocean features that can develop in dynamically isolated water masses within cyclonic eddies (CE) and anticyclonic modewater eddies (ACME). Analysis of a comprehensive oxygen dataset obtained from gliders, moorings, research vessels and Argo floats revealed that eddies with low oxygen concentrations at 50-150 m depths can be found in surprisingly high numbers and in a large area (from about 4°N to 22°N, from the shelf at the eastern boundary to 38°W). Minimum oxygen concentrations of about 9 µmol kg-1 in CEs and severely suboxic concentrations (< 1 µmol kg-1) in ACMEs were observed. In total, 173 profiles with oxygen concentrations below the minimum background concentration of 40 µmol kg-1 could be associated with 27 independent "dead-zone" eddies (10 CEs; 17 ACMEs) over a period of 10 years. The eddies' oxygen minimum is located in the eddy core beneath the mixed layer at a mean depth of 80 m. Compared to the surrounding waters, the mean oxygen anomaly between 50 and 150 m depth for CEs (ACMEs) is -38 (-79) µmol kg-1. The low oxygen concentration right beneath the mixed layer has been attributed to the combination of high productivity in the eddies' surface waters and the isolation of their cores with respect to lateral oxygen supply. Indeed, eddies of both types feature a cold sea surface temperature anomaly and enhanced chlorophyll concentrations in their center. The locally increased consumption within these eddies represents an essential part of the total consumption in the open tropical Northeast Atlantic Ocean and might be partly responsible for the formation of the shallow oxygen minimum zone. Eddies south of 12°N carry weak hydrographic anomalies in their cores and seem to be generated in the open ocean away from the boundary. North of 12°N, eddies of both types carry anomalously low salinity water of South Atlantic Central Water origin from the eastern boundary upwelling region into the open ocean. Water mass properties and satellite eddy tracking both point to an eddy generation near the eastern boundary.

Dual role of HupF in the biosynthesis of [NiFe] hydrogenase in Rhizobium leguminosarum

Background: [NiFe] hydrogenases are enzymes that catalyze the oxidation of hydrogen into protons and electrons, to use H2 as energy source, or the production of hydrogen through proton reduction, as an escape valve for the excess of reduction equivalents in anaerobic metabolism. Biosynthesis of [NiFe] hydrogenases is a complex process that occurs in the cytoplasm, where a number of auxiliary proteins are required to synthesize and insert the metal cofactors into the enzyme structural units. The endosymbiotic bacterium Rhizobium leguminosarum requires the products of eighteen genes (hupSLCDEFGHIJKhypABFCDEX) to synthesize an active hydrogenase. hupF and hupK genes are found only in hydrogenase clusters from bacteria expressing hydrogenase in the presence of oxygen. Results: HupF is a HypC paralogue with a similar predicted structure, except for the C-terminal domain present only in HupF. Deletion of hupF results in the inability to process the hydrogenase large subunit HupL, and also in reduced stability of this subunit when cells are exposed to high oxygen tensions. A ?hupF mutant was fully complemented for hydrogenase activity by a C-terminal deletion derivative under symbiotic, ultra low-oxygen tensions, but only partial complementation was observed in free living cells under higher oxygen tensions (1% or 3%). Co-purification experiments using StrepTag-labelled HupF derivatives and mass spectrometry analysis indicate the existence of a major complex involving HupL and HupF, and a less abundant HupF-HupK complex. Conclusions: The results indicate that HupF has a dual role during hydrogenase biosynthesis: it is required for hydrogenase large subunit processing and it also acts as a chaperone to stabilize HupL when hydrogenase is synthesized in the presence of oxygen.

Obtención de clones de alcornoque (Quercus suber L.) mediante embriogénesis somática: aplicación de medios líquidos y biorreactores en la sincronización de los procesos de desarrollo y maduración de los embriones

El alcornoque tiene un gran valor ambiental, como integrante de los ecosistemas forestales mediterráneos, e interés comercial por el valor de la bellota (alimentación del cerdo ibérico), el carbón, la madera y sobre todo por las aplicaciones industriales del corcho. Las posibilidades de mejora genética del alcornoque, como las de otras especies forestales, están limitadas por sus largos ciclos reproductivos y porque su propagación vegetativa mediante estaquillado solo es posible en estados muy juveniles. Por ello este sistema de propagación tiene muy poca, o ninguna, utilidad práctica en la mejora genética. La embriogénesis somática es la vía más apropiada para la clonación de muchas especies forestales y ha hecho posible el desarrollo a gran escala de plantaciones multivarietales de coníferas. En alcornoque es posible la regeneración completa de árboles adultos mediante embriogénesis somática. Con los protocolos actuales (en medio semisólido), los embriones se generan formando acúmulos y en la fase de multiplicación conviven embriones en distintos estados de desarrollo. Es un sistema asincrónico, con baja eficacia para la propagación en masa, que no elimina completamente las dificultades para el desarrollo de programas de mejora genética del alcornoque. En otras especies la utilización de medios líquidos ha mejorado: la sincronización, productividad de los cultivos, el manejo y reducido los costes de producción. Por ello el desarrollo de suspensiones embriogénicas de alcornoque se plantea como una vía para aumentar la eficacia de la propagación clonal a gran escala. En la presente tesis se desarrollan cultivos embriogénicos de alcornoque en medio líquido. El capítulo 3 aborda el establecimiento y mantenimiento de suspensiones, el capítulo 4 el desarrollo de una fase de proliferación en medio líquido y el capítulo 5 la utilización de sistemas de cultivo en medio líquido, estacionarios y de inmersión temporal, como vía para favorecer la maduración de los embriones somáticos. Para iniciar los cultivos en medio líquido se emplearon agregados de embriones tomados de la fase de proliferación en medio semisólido. Cuando estos agregados se inocularon directamente en medio líquido no se logró el establecimiento de las suspensiones. El establecimiento se consiguió empleando como inóculo las células y Resumen pequeños agregados embriogénicos, de tamaño comprendido entre 41 y 800 μm, desprendidas por agitación breve de los agregados de embriones. El mantenimiento se logró inoculando en baja densidad masas embriogénicas compactas de tamaño comprendido entre 0,8 y 1,2 mm. Estas suspensiones, muy heterogéneas, mantuvieron su capacidad de proliferación y de regeneración de embriones al menos durante diez subcultivos consecutivos. El protocolo de iniciación y mantenimiento, desarrollado inicialmente con un solo genotipo, fue eficaz cuando se probó sobre otros 11 genotipos de alcornoque. En la fase de proliferación se ensayaron tres tipos de envase y tres velocidades de agitación. La combinación envase × velocidad determinó el intercambio gaseoso, la disponibilidad de oxígeno y el estrés hidrodinámico. Los agregados embriogénicos de alcornoque crecieron incluso en condiciones de hipoxia no siendo la disponibilidad de oxígeno un factor limitante del crecimiento para tasas de trasferencia de oxígeno comprendidas entre 0,11 h-1 y 1,47 h-1. Por otra parte la producción de biomasa creció con el estrés hidrodinámico para valores de índice de cizalladura inferiores a 5 x 10-3 cm min-1. La mayor producción de biomasa se obtuvo con matraces Erlenmeyer de 100 ml y alta velocidad de agitación (160 rpm) mientras que la diferenciación de embriones se vio favorecida por bajas velocidades de agitación (60 rpm) asociadas con bajas disponibilidades de oxígeno. La posibilidad de madurar embriones de alcornoque en medio líquido se estudió utilizando sistemas de inmersión permanente y sistemas de inmersión temporal. En inmersión permanente no se diferenciaron embriones cotiledonares (posiblemente por hiperhidricidad). Los sistemas de inmersión temporal permitieron obtener embriones maduros en estado cotiledonar y capaces de regenerar plantas in vitro. Concentraciones de sacarosa superiores a 60 g l-1 y frecuencias de inmersión iguales o inferiores a una diaria, tuvieron efectos negativos para el desarrollo de los embriones somáticos. En los sistemas de inmersión temporal los parámetros físico-químicos del medio de cultivo se mantuvieron estables y no se observó ninguna limitación de nutrientes. No obstante, estos sistemas se vieron afectados por la evaporación que generó el flujo de aire necesario para desplazar el líquido en cada periodo de inmersión. Abstract ABSTRACT Cork oak is one of the most important tree species of the Mediterranean ecosystem. Besides its high environmental value has a great economic interest due to the sustainable production of acorns (to feed the Iberian pig) charcoal, timber and cork, which is a renewable natural product with various technological applications. As happens with other forest species, cork oak genetic improvement programs are limited by their long life cycles and because vegetative propagation by cuttings it´s only possible in very juvenile plants. Hence this propagation system is useless or has little practical use for breeding cork oak. Plant regeneration by somatic embryogenesis is the most suitable way for cloning many forest species, and it is the enabling technology which has allowed the establishment of large-scale conifer multi-varietal plantations. Clonal plant regeneration of mature cork oak trees can be achieved through somatic embryogenesis. Somatic embryos at different stages of development and forming clusters are produced during the multiplication phase with current protocols (using semisolid medium). This is an asynchronous low-efficient process not suitable for mass propagation, and therefore it does not solve the difficulties presented by cork oak breeding programs. Culture in liquid medium has been used with other species to improve: synchronization, yield, handling, and to reduce production costs. Thus the development of cork oak embryogenic suspension cultures is envisaged as a way to increase the efficiency of large scale clonal propagation. The thesis herein develops cork oak embryogenic cultures in liquid medium. In chapter 3 establishment and maintenance of suspension cultures are developed, chapter 4 studies proliferation phase in liquid medium and chapter 5 considers the use of different systems of culture in liquid medium, both stationary and temporary immersion, as a way to promote somatic embryos maturation. Clusters of embryos taken from proliferating cultures on semisolid medium were used to initiate the cultures in liquid medium. When these clusters were inoculated directly in liquid medium establishment of suspension cultures was not executed. However using, as initial inoculum, cells and cell aggregates with a size between 41 and 800 μm detached from these clusters of embryos, subjected to a brief shaking, suspension cultures could be established. Suspension maintenance was achieved by inoculating compact embryogenic Abstract clumps with a size between 0.8 and 1.2 mm at low density. The suspension cultures, very heterogeneous, retained both their proliferation and embryo regeneration capacity for at least ten consecutive subcultures. The initiation and maintenance protocol, initially developed with a single genotype, was effective when tested on 11 additional genotypes of cork oak. In proliferation phase three types of vessels and three different levels of agitation were assayed. The combination vessel × orbiting speed determined gas exchange, oxygen availability and hydrodynamic stress. Cork oak embryogenic aggregates grew even under hypoxia conditions; oxygen availability at transfer rates between 0.11 and 1.47 h-1 was not a limiting factor for growth. Furthermore the biomass production was increased with hydrodynamic stress when shear rate values were of less than 5 x 10-3 cm min-1. The highest biomass production was obtained with 100 ml Erlenmeyer flask and high stirring speed (160 rpm) while the differentiation of embryos was favored by low agitation speeds (60 rpm) associated with low oxygen availability. The possibility to mature cork oak somatic embryos in liquid medium was studied using both permanent immersion systems and temporary immersion systems. Cotyledonary embryos did not differentiate in permanent immersion conditions (probably due to hyperhydricity). Temporary immersion systems allowed obtaining mature cotyledonary embryos, which were able to regenerate plants in vitro. Sucrose concentrations above 60 g l-1 and immersion frequencies equal to or lower than one each 24 h had negative effects on somatic embryo development. Physicochemical parameters of the culture medium in temporary immersion systems were stable and showed no limitation of nutrients. However, these systems were affected by the evaporation generated by the airflow necessary to relocate the medium at each immersion period.