Stable isotope ratios and foraminiferal abundance of sediment cores from the Western Iberian Margin

Rapid climate changes at the onset of the last deglaciation and during Heinrich Event H4 were studied in detail at IMAGES cores MD95-2039 and MD95-2040 from the Western Iberian margin. A major reorganisation of surface water hydrography, benthic foraminiferal community structure, and deepwater isotopic composition commenced already 540 years before the Last Isotopic Maximum (LIM) at 17.43 cal. ka and within 670 years affected all environments. Changes were initiated by meltwater spill in the Nordic Seas and northern North Atlantic that commenced 100 years before concomitant changes were felt off western Iberia. Benthic foraminiferal associations record the drawdown of deepwater oxygenation during meltwater and subsequent Heinrich Events H1 and H4 with a bloom of dysoxic species. At a water depth of 3380 m, benthic oxygen isotopes depict the influence of brines from sea ice formation during ice-rafting pulses and meltwater spill. The brines conceivably were a source of ventilation and provided oxygen to the deeper water masses. Some if not most of the lower deep water came from the South Atlantic. Benthic foraminiferal assemblages display a multi-centennial, approximately 300-year periodicity of oxygen supply at 2470-m water depth. This pattern suggests a probable influence of atmospheric oscillations on the thermohaline convection with frequencies similar to Holocene climate variations. For Heinrich Events H1 and H4, response times of surface water properties off western Iberia to meltwater injection to the Nordic Seas were extremely short, in the range of a few decades only. The ensuing reduction of deepwater ventilation commenced within 500-600 years after the first onset of meltwater spill. These fast temporal responses lend credence to numerical simulations that indicate ocean-climate responses on similar and even faster time scales.

Barium in seawater and sediments from the Artic Ocean

As part of ongoing circulation studies in the Arctic, seawater samples for dissolved Ba concentrations were obtained during Sep.-Oct., 1992 at several locations in the Bering Strait, Eastern Chukchi and Southern Beaufort Seas. The results reveal a dynamic rang (10 to 150 nmol/kg) for this element in the Arctic equal to or greater than that in combined Atlantic, Indian and Pacific oceans. Lowest levels are observed in surface waters, with values tending to decrease northwards in the direction of currents generally flowing frorn the Bering Strait along the Alaskan coast. Low surfacc concentrations tend to be accompanied by relatively enriched near bottom levels. On the basis of these spatial distributions, hydrographic observations and a knowledge of its behavior in other marine settings, it appears that Ba can be significantly depleted from surface waters as a result of the highly seasonal biological aclivities over Arctic marginal shelves. Removal at the surface is counteracted to some extent by regeneration at depth or in the sediments and by riverine inputs. The biologically related drawdown is likely to enhance the contrast between 'background' surface Ba levels in the Arctic and waters imprinted by regeneration and/or rivers, These preliminary findings suggest that Ba holds particular promise for tracing river waters and the ventilation of halodine waters hy laterally sinking brines produced during ice formation over the shelves.

Physical and chemical oceanography in the Arctic Ocean

Data from sections across the Eurasian Basin of the Arctic Ocean occupied by the German Research Vessel Polarstern in 1987 and by the Swedish icebreaker Oden in 1991 are used to derive information on the freshwater balance of the Arctic Ocean halocline and on the sources of the deep waters of the Nansen, Amundsen and Makarov basins. Salinity, d18O and mass balances allow separation of the river-runoff and the sea-ice meltwater fractions contained in the Arctic halocline. This provides the basis for tracking the river-runoff signal from the shelf seas across the central Arctic Ocean to Fram Strait. The halocline has to be divided into at least three lateral regimes: the southern Nansen Basin with net sea-ice melting, the northern Nansen Basin and Amundsen Basin with net sea-ice formation and increasing river-runoff fractions, and the Canadian Basin with minimum sea-ice meltwater and maximum river-runoff fractions and water of Pacific origin. In the Canadian Basin, silicate is used as a tracer to identify Pacific water entering through Bering Strait and an attempt is made to quantify its influence on the halocline waters of the Canadian Basin. For this purpose literature data from the CESAR and LOREX ice camps are used. Based on mass balances and depending on the value of precipitation over the area of the Arctic Ocean the average mean residence time of the river-runoff fraction contained in the Arctic Ocean halocline is determined to be about 14 or 11 years. Water column inventories of river-runoff and sea-ice meltwater are calculated for a section just north of Fram Strait and implications for the ice export rate through Fram Strait are discussed. Salinity, tritium, 3He and the d18O ratio of halocline waters sampled during the 1987 Polarstern cruise to the Nansen Basin are used to estimate the mean residence time of the river-runoff component in the halocline and on the shelves of the Arctic Ocean. These estimates are done by comparing ages of the halocline waters based on a combination of tracers yielding different time information: the tritium 'vintage' age which records the time that has passed since the river-runoff entered the shelf and the tritium/3He age which reflects the time since the shelf waters left the shelf. The difference between the ages determined by these two methods is about 3 to 6 years. Correction for the initial tritium/3He age of the shelf waters (about 0.5 to 1.5 years) yields a mean residence time of the river-runoff on the shelves of about 3.5 ± 2 years. Comparison of the 18O/16O ratios of shelf water, Atlantic water and the deep waters of the Arctic Ocean indicate that the sources of the deep and bottom waters of the Eurasian Basin are located in the Barents and Kara seas.

Physicochemical characteristics and protist occurrence at stations sampled in 2007/2008 in the eastern Beaufort Sea

This study documents, for the first time, the abundance and species composition of protist assemblages in Arctic sea ice during the dark winter period. Lack of knowledge of sea-ice assemblages during the dark period has left questions about the retention and survival of protist species that initiate the ice algal bloom. Sea-ice and surface water samples were collected between December 27, 2007 and January 31, 2008 within the Cape Bathurst flaw lead, Canadian Beaufort Sea. Samples were analyzed for protist identification and counts, chlorophyll (chl) a, and total particulate carbon and nitrogen concentrations. Sea-ice chl a concentrations (max. 0.27 µg/l) and total protist abundances (max. 4 x 10**3 cells/l) were very low, indicating minimal retention of protists in the ice during winter. The diversity of winter ice protists (134 taxa) was comparable to spring ice assemblages. Pennate diatoms dominated the winter protist assemblage numerically (averaging 77% of total protist abundances), with Nitzschia frigida being the most abundant species. Only 56 taxa were identified in surface waters, where dinoflagellates were the dominant group. Our results indicate that differences in the timing of ice formation may have a greater impact on the abundance than structure of protist assemblages present in winter sea ice and at the onset of the spring ice algal bloom.

Mercury species concentrations near Casey station, Antarctica and along the SR3 CASO-GEOTRACES transect

We present here the first mercury speciation study in the water column of the Southern Ocean, using a high-resolution south-to-north section (27 stations from 65.50°S to 44.00°S) with up to 15 depths (0-4440 m) between Antarctica and Tasmania (Australia) along the 140°E meridian. In addition, in order to explore the role of sea ice in Hg cycling, a study of mercury speciation in the 'snow-sea ice-seawater' continuum was conducted at a coastal site, near the Australian Casey station (66.40°S; 101.14°E). In the open ocean waters, total Hg (Hg(T)) concentrations varied from 0.63 to 2.76 pmol/L with 'transient-type' vertical profiles and a latitudinal distribution suggesting an atmospheric mercury source south of the Southern Polar Front (SPF) and a surface removal north of the Subantartic Front (SAF). Slightly higher mean Hg(T) concentrations (1.35 ± 0.39 pmol/L) were measured in Antarctic Bottom Water (AABW) compared to Antarctic Intermediate water (AAIW) (1.15 ± 0.22 pmol/L). Labile Hg (Hg(R)) concentrations varied from 0.01 to 2.28 pmol/L, with a distribution showing that the Hg(T) enrichment south of the SPF consisted mainly of Hg(R) (67 ± 23%), whereas, in contrast, the percentage was half that in surface waters north of PFZ (33 ± 23%). Methylated mercury species (MeHg(T)) concentrations ranged from 0.02 to 0.86 pmol/L. All vertical MeHg(T) profiles exhibited roughly the same pattern, with low concentrations observed in the surface layer and increasing concentrations with depth up to an intermediate depth maximum. As for Hg(T), low mean MeHg(T) concentrations were associated with AAIW, and higher ones with AABW. The maximum of MeHg(T) concentration at each station was systematically observed within the oxygen minimum zone, with a statistically significant MeHg(T) vs Apparent Oxygen Utilization (AOU) relationship (p <0.001). The proportion of Hg(T) as methylated species was lower than 5% in the surface waters, around 50% in deep waters below 1000 m, reaching a maximum of 78% south of the SPF. At Casey coastal station Hg(T) and Hg(R) concentrations found in the 'snow-sea ice-seawater' continuum were one order of magnitude higher than those measured in open ocean waters. The distribution of Hg(T) there suggests an atmospheric Hg deposition with snow and a fractionation process during sea ice formation, which excludes Hg from the ice with a parallel Hg enrichment of brine, probably concurring with the Hg enrichment of AABW observed in the open ocean waters. Contrastingly, MeHg(T) concentrations in the sea ice environment were in the same range as in the open ocean waters, remaining below 0.45 pmol/L. The MeHg(T) vertical profile through the continuum suggests different sources, including atmosphere, seawater and methylation in basal ice. Whereas Hg(T) concentrations in the water samples collected between the Antarctic continent and Tasmania are comparable to recent measurements made in the other parts of the World Ocean (e.g., Soerensen et al., 2010; doi:10.1021/es903839n), the Hg species distribution suggests distinct features in the Southern Ocean Hg cycle: (i) a net atmospheric Hg deposition on surface water near the ice edge, (ii) the Hg enrichment in brine during sea ice formation, and (iii) a net methylation of Hg south of the SPF.

Geochemistry of gas hydrates of DSDP Hole 84-570

During Deep Sea Drilling Project (DSDP) Leg 84 a core 1 m long and 6 cm in diameter of massive gas hydrate was unexpectedly recovered at Site 570 in upper slope sediment of the Middle America Trench offshore of Guatemala. This core contained only 5-7% sediment, the remainder being the solid hydrate composed of gas and water. Samples of the gas hydrate were decomposed under controlled conditions in a closed container maintained at 4°C. Gas pressure increased and asymptotically approached the equilibrium decomposition pressure for an ideal methane hydrate, CH4.5-3/4H2O, of 3930 kPa and approached to this pressure after each time gas was released, until the gas hydrate was completely decomposed. The gas evolved during hydrate decomposition was 99.4% methane, ~0.2% ethane, and ~0.4% CO2. Hydrocarbons from propane to heptane were also present, but in concentrations of less than 100 p.p.m. The carbon-isotopic composition of methane was -41 to -44 per mil, relative to PDB standard. The observed volumetric methane/water ratio was 64 or 67, which indicates that before it was stored and analyzed, the gas hydrate probably had lost methane. The sample material used in the experiments was likely a mixture of methane hydrate and water ice. Formation of this massive gas hydrate probably involved the following processes: (i) upward migration of gas and its accumulation in a zone where conditions favored the growth of gas hydrates, (ii) continued, unusually rapid biological generation of methane, and (iii) release of gas from water solution as pressure decreased due to sea level lowering and tectonic uplift.

Middle Miocene stable carbon and oxygen isotope record of foraminifera of ODP Hole 120-747A

Paleoceanographic variability at southern high latitude Ocean Drilling Program (ODP) Site 747 was investigated in this study through the interval which spans the Middle Miocene Climate Transition (MMCT). Between 15.0 and 12.2 million years ago (Ma), foraminiferal d18O records derived from both benthic (Cibicidoides spp.) and planktonic taxa (Globorotalia praescitula and Globigerina bulloides) reveal a history of changes in water column thermal and salinity structure and a strong imprint of seasonality. Prior to the MMCT, in the interval between 14.35 and 13.9 Ma, G. bulloides displays relatively high d18O values similar to those of G. praescitula, interpreted to indicate weakening of the thermocline and/or increased seasonality with cooler early-spring and/or late-fall temperatures. Following this interval, G. bulloidesd18O values diverge significantly from benthic and G. praescitula values, with G. bulloides values remaining relatively low for at least 600 kyr following the benthic foraminiferal d18O shift during the MMCT at ~13.9 Ma. This divergence in d18O records occurs in direct association with the Mi3 cooling and glaciation event and may suggest: (1) a strengthening of the vertical temperature gradient, with greater cooling of deep waters than surface waters, (2) changes in the depth habitat of G. bulloides, (3) changes in the dominant season of G. bulloides calcification, (4) modification of surface-water d18O values in association with enhanced sea-ice formation, (5) increased surface-water carbonate ion concentration, and/or (6) a significant decrease in surface-water salinity across the MMCT. The first of these possible scenarios is not likely, particularly in light of recent Mg/Ca evidence for significant surface-water cooling in the Southern Ocean associated with the MMCT. Of the remaining possibilities, we favor a change in surface salinity to explain the observed trends in d18O values and hypothesize that surface salinity may have decreased by up to 2 salinity units at ~13.9 Ma. In this scenario, the development of a lower-salinity Antarctic surface layer coincided with regional cooling of both surface and deep waters of the Southern Ocean during the Mi3 glaciation of East Antarctica, and contributed into the dominance of Neogloboquadrina spp. between 13.8 and 13.2 Ma. Additionally, the distinct patterns observed in planktonic foraminiferal d18O records spanning the MMCT correspond with changes in the vertical d13C gradient between planktonic and benthic foraminiferal records and major changes in planktonic foraminiferal assemblages at Site 747, providing further evidence of the environmental significance of this climatic transition.

Tab. 3: Concentrations of selected chemical species at various depths in Ace Lake

This article reviews the history, chemical stratification, biology and biogeochemistry of Ace Lake, which is one of the many marine-derived meromictic (permanently stratified) lakes in the Vestfold Hills, Eastern Antarctica. The lake has an area of 18 ha, a maximum depth of 25 m, and a salinity range from 7 to 43 g l**-1. The lake mixes to a depth of 7 m in late winter as a result of brine freeze out during ice formation. Deeper mixing is precluded by a sharp halocline. The water beneath 12 m is permanently anoxic, The lake was formed approximately 10,800 yr BP as the polar ice cap melted. Sea level rise 7,800 yr BP resulted in invasion of seawater into the initially freshwater lake. Subsequently, sea level dropped, and the now saline lake became isolated from the ocean. The biota of the lake was derived from species trapped when the connection between the lake and the ocean was cut off. The oxic zone above 12 m supports a relatively simple community which includes microbial mats, four major species of phytoplankton (including a picocyanobacterium), two copepod species, and a variety of heterotrophic flagellates and ciliates. The anoxic zone contains populations of photosynthetic sulfur, sulfate reducing, fermentative and methanogenic bacteria, which combine to remineralise organic carbon which sediments from the upper waters. Research on the physics, biology and chemistry of Ace Lake has contributed significantly to knowledge of Antarctic meromictic lakes.

Stable isotope record of foraminifera from the subarctic Pacific

Benthic (Uvigerina spp., Cibicidoides spp., Gyroidinoides spp.) and planktonic (N. pachyderma sinistral, G. bulloides) stable isotope records from three core sites in the central Gulf of Alaska are used to infer mixed-layer and deepwater properties of the late glacial Subarctic Pacific. Glacial-interglacial amplitudes of the planktonic delta18O records are 1.1-1.3 per mil, less than half the amplitude observed at core sites at similar latitudes in the North Atlantic; these data imply that a strong, negative deltaw anomaly existed in the glacial Subarctic mixed layer during the summer, which points to a much stronger low-salinity anomaly than exists today. If true, the upper water column in the North Pacific would have been statically more stable than today, thus suppressing convection even more efficiently. This scenario is further supported by vertical (i.e., planktic versus benthic) delta18O and delta13C gradients of >1 per mil, which suggest that a thermohaline link between Pacific deep waters and the Subarctic Pacific mixed layer did not exist during the late glacial. Epibenthic delta13C in the Subarctic Pacific is more negative than at tropical-subtropical Pacific sites but similar to that recorded at Southern Ocean sites, suggesting ventilation of the deep central Pacific from mid-latitude sources, e.g., from the Sea of Japan and Sea of Okhotsk. Still, convection to intermediate depths could have occurred in the Subarctic during the winter months when heat loss to the atmosphere, sea ice formation, and wind-driven upwelling of saline deep waters would have been most intense. This would be beyond the grasp of our planktonic records which only document mixed-layer temperature-salinity fields extant during the warmer seasons. Also we do not have benthic isotope records from true intermediate water depths of the Subarctic Pacific.

Age models of northern North Atlantic sediment cores

Benthic foraminiferal stable isotope records from four high-resolution sediment cores, forming a depth transect between 1237 m and 2303 m on the South Iceland Rise, have been used to reconstruct intermediate and deep water paleoceanographic changes in the northern North Atlantic during the last 21 ka (spanning Termination I and the Holocene). Typically, a sampling resolution of ~100 years is attained. Deglacial core chronologies are accurately tied to North Greenland Ice Core Project (NGRIP) ice core records through the correlation of tephra layers and changes in the percent abundance of Neogloboquadrina pachyderma (sinistral) with transitions in NGRIP. The evolution from the glacial mode of circulation to the present regime is punctuated by two periods with low benthic d13C and d18O values, which do not lie on glacial or Holocene water mass mixing lines. These periods correlate with the late Younger Dryas/Early Holocene (11.5-12.2 ka) and Heinrich Stadial 1 (14.7-16.8 ka) during which time freshwater input and sea-ice formation led to brine rejection both locally and as an overflow exported from the Nordic seas into the northern North Atlantic, as earlier reported by Meland et al. (2008). The export of brine with low ?13C values from the Nordic seas complicates traditional interpretations of low d13C values during the deglaciation as incursions of southern sourced water, although the spatial extent of this brine is uncertain. The records also reveal that the onset of the Younger Dryas was accompanied by an abrupt and transient (~200-300 year duration) decrease in the ventilation of the northern North Atlantic. During the Holocene, Iceland-Scotland Overflow Water only reached its modern flow strength and/or depth over the South Iceland Rise by 7-8 ka, in parallel with surface ocean reorganizations and a cessation in deglacial meltwater input to the North Atlantic.

Physical oceanography and hydrochemistry from the Laptev Sea, Arctic Ocean

Combined d18O/salinity data reveal a distinctive water mass generated during winter sea ice formation which is found predominantly in the coastal polynya region of the southern Laptev Sea. Export of the brine-enriched bottom water shows interannual variability in correlation with atmospheric conditions. Summer anticyclonic circulation is favoring an offshore transport of river water at the surface as well as a pronounced signal of brine-enriched waters at about 50 m water depth at the shelf break. Summer cyclonic atmospheric circulation favors onshore or an eastward, alongshore water transport, and at the shelf break the river water fraction is reduced and the pronounced brine signal is missing, while on the middle Laptev Sea shelf, brine-enriched waters are found in high proportions. Residence times of bottom and subsurface waters on the shelf may thereby vary considerably: an export of shelf waters to the Arctic Ocean halocline might be shut down or strongly reduced during "onshore" cyclonic atmospheric circulation, while with "offshore" anticyclonic atmospheric circulation, brine waters are exported and residence times may be as short as 1 year only.

Sedimentology and geochemistry of core GIK23258-2 in the Barents Sea

At the western continental margin of the Barents Sea, 75°N, hemipelagic sediments provide a record of Holocene climate change with a time resolution of 10-70 years. Planktic foraminifera counts reveal a very early Holocene thermal optimum 10.7-7.7 kyr BP, with summer sea surface temperatures (SST) of 8°C and a much enhanced West Spitsbergen Current. There was a short cooling between 8.8 and 8.2 kyr BP. In the middle and late Holocene summer, SST dropped to 2.5°-5.0°C, indicative of reduced Atlantic heat advection, except for two short warmings near 2.2 and 1.6 kyr BP. Distinct quasi-periodic spikes of coarse sediment fraction (with large portions of lithic grains, benthic and planktic foraminifera) record cascades of cold, dense winter water down the continental slope as a result of enhanced seasonal sea ice formation and storminess on the Barents shelf over the entire Holocene. The spikes primarily cluster near recurrence intervals of 400-650 and 1000-1350 years, when traced over the entire Holocene, but follow significant 885-/840- and 505-/605-year periodicities in the early Holocene. These non-stationary periodicities mimic the Greenland-[Formula: See Text]Be variability, which is a tracer of solar forcing. Further significant Holocene periodicities of 230, (145) and 93 years come close to the deVries and Gleissberg solar cycles.

Water mass evolution of the Greenland Sea since late glacial times

Four sediment cores from the central and northern Greenland Sea basin, a crucial area for the renewal of North Atlantic deep water, were analyzed for planktic foraminiferal fauna, planktic and benthic stable oxygen and carbon iso- topes as well as ice-rafted debris to reconstruct the environ- mental variability in the last 23 kyr. During the Last Glacial Maximum, the Greenland Sea was dominated by cold and sea-ice bearing surface water masses. Meltwater discharges from the surrounding ice sheets affected the area during the deglaciation, influencing the water mass circulation. During the Younger Dryas interval the last major freshwater event occurred in the region. The onset of the Holocene interglacial was marked by an increase in the advection of Atlantic Wa- ter and a rise in sea surface temperatures (SST). Although the thermal maximum was not reached simultaneously across the basin, benthic isotope data indicate that the rate of overturn- ing circulation reached a maximum in the central Greenland Sea around 7ka. After 6-5ka a SST cooling and increas- ing sea-ice cover is noted. Conditions during this so-called "Neoglacial" cooling, however, changed after 3 ka, probably due to enhanced sea-ice expansion, which limited the deep convection. As a result, a well stratified upper water column amplified the warming of the subsurface waters in the central Greenland Sea, which were fed by increased inflow of At- lantic Water from the eastern Nordic Seas. Our data reveal that the Holocene oceanographic conditions in the Green- land Sea did not develop uniformly. These variations were a response to a complex interplay between the Atlantic and Polar water masses, the rate of sea-ice formation and melting and its effect on vertical convection intensity during times of Northern Hemisphere insolation changes.

Glassy State and Cryopreservation of Mint Shoot Tips

Vitrification refers to the physical process by which a liquid supercools to very low tem- peratures and finally solidifies into a metastable glass, without undergoing crystallization at a practical cooling rate. Thus, vitrification is an effective freeze-avoidance mechanism and living tissue cryopreservation is, in most cases, relying on it. As a glass is exceedingly vis- cous and stops all chemical reactions that require molecular diffusion, its formation leads to metabolic inactivity and stability over time. To investigate glassy state in cryopreserved plant material, mint shoot tips were submitted to the different stages of a frequently used cryopreservation protocol (droplet-vitrification) and evaluated for water content reduction and sucrose content, as determined by ion chromatography, frozen water fraction and glass transitions occurrence by differential scanning calorimetry, and investigated by low-tempera- ture scanning electron microscopy, as a way to ascertain if their cellular content was vitri- fied. Results show how tissues at intermediate treatment steps develop ice crystals during liquid nitrogen cooling, while specimens whose treatment was completed become vitrified, with no evidence of ice formation. The agreement between calorimetric and microscopic observations was perfect. Besides finding a higher sucrose concentration in tissues at the more advanced protocol steps, this level was also higher in plants precultured at 25/21?C than in plants cultivated at 25?C.

Estudios termodinámicos de los procesos de vitrificación en la crioconservación de germoplasma vegetal=Thermodynamic studies of vitrification processes in plant germplasm cryopreservation

En la actualidad, las técnicas de crioconservación poseen una importancia creciente para el almacenamiento a largo plazo de germoplasma vegetal. En las dos últimas décadas, estos métodos experimentaron un gran desarrollo y se han elaborado protocolos adecuados a diferentes sistemas vegetales, utilizando diversas estrategias como la vitrificación, la encapsulación-desecación con cuentas de alginato y el método de “droplet”-vitrificación. La presente tesis doctoral tiene como objetivo aumentar el conocimiento sobre los procesos implicados en los distintos pasos de un protocolo de crioconservación, en relación con el estado del agua presente en los tejidos y sus cambios, abordado mediante diversas técnicas biofísicas, principalmente calorimetría diferencial de barrido (DSC) y microscopía electrónica de barrido a baja temperatura (crio-SEM). En un primer estudio sobre estos métodos de crioconservación, se describen las fases de enfriamiento hasta la temperatura del nitrógeno líquido y de calentamiento hasta temperatura ambiente, al final del periodo de almacenamiento, que son críticas para la supervivencia del material crioconservado. Tanto enfriamiento como calentamiento deben ser realizados lo más rápidamente posible pues, aunque los bajos contenidos en agua logrados en etapas previas de los protocolos reducen significativamente las probabilidades de formación de hielo, éstas no son del todo nulas. En ese contexto, se analiza también la influencia de las velocidades de enfriamiento y calentamiento de las soluciones de crioconservación de plantas en sus parámetros termofísicos referente a la vitrificación, en relación su composición y concentración de compuestos. Estas soluciones son empleadas en la mayor parte de los protocolos actualmente utilizados para la crioconservación de material vegetal. Además, se estudia la influencia de otros factores que pueden determinar la estabilidad del material vitrificado, tales como en envejecimiento del vidrio. Se ha llevado a cabo una investigación experimental en el empleo del crio-SEM como una herramienta para visualizar el estado vítreo de las células y tejidos sometidos a los procesos de crioconservación. Se ha comparado con la más conocida técnica de calorimetría diferencial de barrido, obteniéndose resultados muy concordantes y complementarios. Se exploró también por estas técnicas el efecto sobre tejidos vegetales de la adaptación a bajas temperaturas y de la deshidratación inducida por los diferentes tratamientos utilizados en los protocolos. Este estudio permite observar la evolución biofísica de los sistemas en el proceso de crioconservación. Por último, se estudió la aplicación de películas de quitosano en las cuentas de alginato utilizadas en el protocolo de encapsulación. No se observaron cambios significativos en su comportamiento frente a la deshidratación, en sus parámetros calorimétricos y en la superficie de las cuentas. Su aplicación puede conferir propiedades adicionales prometedoras. ABSTRACT Currently, cryopreservation techniques have a growing importance for long term plant germplasm storage. These methods have undergone great progress during the last two decades, and adequate protocols for different plant systems have been developed, making use of diverse strategies, such as vitrification, encapsulation-dehydration with alginate beads and the dropletvitrification method. This PhD thesis has the goal of increasing the knowledge on the processes underlying the different steps of cryopreservation protocols, in relation with the state of water on tissues and its changes, approached through diverse biophysical techniques, especially differential scanning calorimetry (DSC) and low-temperature scanning electron microscopy (cryo-SEM). The processes of cooling to liquid nitrogen temperature and warming to room temperature, at the end of the storage period, critical for the survival of the cryopreserved material, are described in a first study on these cryopreservation methods. Both cooling and warming must be carried out as quickly as possible because, although the low water content achieved during previous protocol steps significantly reduces ice formation probability, it does not completely disappear. Within this context, the influence of plant vitrification solutions cooling and warming rate on their vitrification related thermophysical parameters is also analyzed, in relation to its composition and component concentration. These solutions are used in most of the currently employed plant material cryopreservation protocols. Additionally, the influence of other factors determining the stability of vitrified material is studied, such as glass aging. An experimental research work has been carried out on the use of cryo-SEM as a tool for visualizing the glassy state in cells and tissues, submitted to cryopreservation processes. It has been compared with the better known differential scanning calorimetry technique, and results in good agreement and complementary have been obtained. The effect on plant tissues of adaptation to low temperature and of the dehydration induced by the different treatments used in the protocols was explored also by these techniques. This study allows observation of the system biophysical evolution in the cryopreservation process. Lastly, the potential use of an additional chitosan film over the alginate beads used in encapsulation protocols was examined. No significant changes could be observed in its dehydration and calorimetric behavior, as well as in its surface aspect; its application for conferring additional properties to gel beads is promising.