Age determination and pollen profile of sediment core Trentelmoor, Peine, northern Germany

The filling up of the lake which existed in the basin of the Trentelmoor (40 km E of Hannover, Germany) - in Preboreal times was finished 2000 years ago. Since then fen vegetation has covered the former lake's surface. The postglacial development of the vegetation follows the pattern which is typical of Central Europe. However, due to the poorness of the soils around the Trentelmoor, the frequencies of some tree species differ. Beech for example never reached - for the benefit of oak - that importance which this tree species usually gains on better soils. Human impact becomes recognisable in the upper Neolithic for the first time. The area has been settled continuously, but with changing intensities, throughout the last 3000 years. When the manuscript of this paper went to press the results of two radiocarbon age determinations only were completed. An additional three determinations were completed somewhat later. See the accompanying table for results.

Contribution of woody habitat islands to the conservation of birds and their potential ecosystem services in an extensive Colombian rangeland

The conservation of birds and their habitats is essential to maintain well-functioning ecosystems including human-dominated habitats. In simplified or homogenized landscapes, patches of natural and semi-natural habitat are essential for the survival of plant and animal populations. We compared species composition and diversity of trees and birds between gallery forests, tree islands and hedges in a Colombian savanna landscape to assess how fragmented woody plant communities affect forest bird communities and how differences in habitat characteristics influenced bird species traits and their potential ecosystem function. Bird and tree diversity was higher in forests than in tree islands and hedges. Soil depth influenced woody species distribution, and canopy cover and tree height determined bird species distribution, resulting in plant and bird communities that mainly differed between forest and non-forest habitat. Bird and tree species and traits widely co-varied. Bird species in tree islands and hedges were on average smaller, less specialized to habitat and more tolerant to disturbance than in forest, but dietary differences did not emerge. Despite being less complex and diverse than forests, hedges and tree islands significantly contribute to the conservation of forest biodiversity in the savanna matrix. Forest fragments remain essential for the conservation of forest specialists, but hedges and tree islands facilitate spillover of more tolerant forest birds and their ecological functions such as seed dispersal from forest to the savanna matrix.

Two pollen profiles of the middle Pleistocene at Samerberg/Upper Bavaria, Germany

Previous pollen analytical studies on sediments from the pleistocene lake basin at Samerberg, situated on the northern edge of the Bavarian Alps (47°45' N, 12°12' E, 607 m a.s.l.) had been performed on samples taken from cores and exposures close to the southern shore of the former lake. After geoelectric and refraction-seismic measurements had shown that the lake basin had been much deeper in its northern part, another core was taken where maximum depth could be expected. The corer penetrated three moraines, two of them lying above pollen-bearing sediments, and one below them, and reached the hard rock (Kössener Kalk) at a depth of 93 m. Two forest phases could be identified by pollen analysis. The pollen record begins abruptly in a forest phase at the end of a spruce-dominated period when fir started to spread (DA 1, DA = pollen zone). Following this, Abies (fir) was the main tree species at Samerberg, Picea being second, and deciduous trees were almost non-existent. First box (Buxus) was of major importance in the fir forests (DA 2), but later on beech (Fagus) and wing-nut (Pterocarya) spread (DA 3). Finally this forest gave way to a spruce forest with pine (DA 4). The beginning and the end of this interglacial cycle are not recorded. Its vegetational development is different from the eemian one known from earlier studies at Samerberg. It is characterized by the occurrence of Abies together with Buxus, Pterocarya and Fagus. A similar association of woody species is known only from the Holsteinian age deposits in an area ranging from England to Poland, though at no other place these species were such important constituents of the vegetation as at Samerberg. Therefore zone 1 to 4 are attributed to the Holsteinian interglacial period. The younger forest phase, separated from the interglacial by a stadial with open vegetation (DA 5), seems to be completely represented, though its sediments are disturbed, apparently by sliding which caused repetition of same-age-sediments in the core (DA 7a, b, c) The vegetational development is simple. A juniper phase (DA 6) was followed by reforestation with spruce, accompanied by some fir (DA 7, 9). Finally pine became the dominant species (DA 9). The simple vegetational development of this younger forest phase does not allow a safe correlation with one of the known pre-eemian interstadials, but for stratigraphical reasons it can be related best to the Dömnitz-interglacial, which among others is also known as Wacken- or Holstein-II-interglacial. Possibly another phase of reforestation is indicated at the end of the following stadial (DA 10). But due to an erosional unconformity nothing than the rise of the juniper curve can be stated. It was only after this sequence of forest phases and periods with open vegetation that glaciers reached the Samerberg area again.

Pollen and spore deposition in different areas of the Lena Delta

Studies of the annual pollen and spore deposition in different areas of the Lena Delta were undertaken for the first time in the Asian sector of the Arctic during the Russian-German ''LENA 98'' and ''LENA 99'' expeditions in the framework of the International ''Laptev Sea System-2000'' Project. To achieve this objective, three spore-pollen traps were set up along the meridional delta profile in accordance with the European Pollen Monitoring Programme for the period July 1998 to August 1999. A comparison between the results of spore-pollen analysis of the contents of traps and the surrounding vegetation was performed. The results confirmed the current spore-pollen spectra are comprised both of pollen and spores of the local plants and of long-distance pollen and spores. The dependence of the long-distance pollen deposition on the character of the wind regime of the region was established. The prevailing southerly and southeasterly wind direction determines the main pollen influx of tree species from the areas of their growth south of the delta. The features of the morphological structure and fossilization of pollen and the features of the productive capability and plant growing conditions are of large significance in the pollen transfer and deposition.

Pollen and macroremains from four profiles from site Samerberg 1

Pollen and macrofossil analysis of lake sediments revealed the complete development of vegetation from Riss late-glacial to early Würm glacial times at Samerberg (12°12' E, 47°45' N, 600 m a.s.l) on the northern border of the Alps. The pollen bearing sediments overlie three stratigraphic units, at the base a ground-moraine, then a 13 m thick layer of pollen free silt and clay, and then a younger moraine; all the sediments including the pollen bearing sediments, lie below the Würm moraine. The lake, which had developed in an older glacial basin, became extinct, when the ice of the river Inn glacier filled its basin during Würm full-glacial time at the latest. One interglacial, three interstadials, and the interdigitating treeless periods were identified at Samerberg. Whereas the cold periods cannot be distinguished from one another pollenanalytically, the interglacial and the two older interstadials have distinctive characteristics. A shrub phase with Juniperus initiated reforestation and was followed by a pine phase during the interglacial and each of the three interstadials. The further development of the interglacial vegetation proceeded with a phase when deciduous trees (mainly Quercus, oak) and hazel (Corylus) dominated, though spruce (Picea) was present at the same time in the area. A phase with abundant yew (Taxus) led to an apparently long lasting period with dominant spruce and fir (Abies) accompanied by some hornbeam (Carpinus). The vegetational development shows the main characteristics of the Riss/Würm interglacial, though certain differences in the vegetational development in the northern alpine foreland are obvious. These differences may result from the existence of an altitudinal zonation of the vegetation in the vicinity of the site and are the expression of its position at the border of the Alps. A greater age (e.g. the Holsteinian) can be excluded by reason of the vegetational development, and is also not indicated at first sight from the geological and stratigraphical data of the site. Characteristic of the Riss/Würm vegetational development in southern Germany - at least in the region between Lake Starnberg/Samerberg/Salzach - is the conspicuous yew phase. According to absolute pollen counts, yew not only displaced the deciduous species, but also displaced spruce preferentially, thus indicating climatic conditions less favourable for spruce, caused by mild winters (Ilex spreading!) and by short-term low precipitation, indicated by the reduced sedimentation rate. The oldest interstadials is bipartite, as due to the climatic deterioration the early vegetational development, culminating in a spruce phase, had been interrupted by another expansion of pine. A younger spruce-dominated period with fir and perhaps also with hornbeam and beech (Fagus) followed. An identical climatic development has been reported from other European sites with long pollen sequences (see chapter 6.7). However, different tree species are found in the same time intervals in Middle Europe during Early Würm times. Sediments of the last interglacial (Eem or Riss/Würm) have been found in all cases below the sediments of the bipartite interstadial, and in addition one more interstadial occurs in the overlying sediments. This proves that Eem and Riss/Würm of the north-european plain resp. of the alpine foreland are contemporaneous interglacials although this has been questioned by some authors. The climax vegetation of the second interstadial was a spruce forest without fir and without more demanding deciduous tree species. The vegetational development of the third interstadial is recorded fragmentary only. But it has been established that a spruce forest was present. The oldest interstadial must correspond to the danish Brørup interstadial as it is expressed in northern Germany, the second one to the Odderade interstadial. A third Early Würm interstadial, preserved fragmentarily at Samerberg, is known from other sites. The dutch Amersfoort interstadial most likely is the equivalent to the older part of the bipartite danish Brørup interstadial.

Lithology, age determination and pollen records of five profiles for nothern Bavaria, Germany

Palynological investigations in northeastern Bavaria (Bavarian Vogtland, Fichtelgebirge, Steinwald) reveal the Late Glacial and Postglacial history of the regional vegetation. Radiocarbon data in comparison with those from the neighbouring regions (Rhön, Oberpfälzer Wald, Bavarian Forests) show a time lag in the development of the arboreal vegetation due to migration processes. The Fichtelgebirge is the southernmost part ofnortheastern Bavaria where the early Alleröd period (pollen zone IIa) is characterised by a dominance of birch forests. Hazel reached maximal values around 8000 BP in the area from the Fichtelgebirge to the Bavarian Forests, e.g. about 600 years earlier than in the more northern Rhön mountains. For spruce there is a considerable time lag between the Bavarian Forests and the Fichtelgebirge. Spruce spreading started in the Fichtelgebirge during the older part of the Atlantic period (pollen zone VI). At the same time, spruce already was the dominant tree in the Bavarian Forests. During the younger part of the Atlantic period (pollen zone VII) spruce and mixed oak forest tree species frequently occurred in the Fichtelgebirge. At the end of pollen zone VI, spruce came to dominance. At the same time, the immigration of beech started. During the Subboreal period (pollen zone VIII), spruce remained being a dominant member in the forests and at the end of pollen zone VIII, fir began to spread rapidly. During the first part of the Subatlantic period (pollen zone IX) spruce, beech, fir and pine formed the mountainous forests in the Fichtelgebirge. In the area of the Bavarian Vogtland, however, fir was a dominant forest tree during pollen zone IX, while spruce and beech played a less important role. During the 12th century, human colonisation started in the area of the Fichtelgebirge. This is 400 years later as in the area of the Rhön mountains. Indicators for earlier forest clearances are rare or absent.

Drought risk assessment as changes in the potential stand yield classes and potential forest production for Sitka spruce, Scots pine, and pedunculate oak over the next 80 years in the Great Britain

Probabilistic climate data have become available for the first time through the UK Climate Projections 2009, so that the risk of tree growth change can be quantified. We assess the drought risk spatially and temporally using drought probabilities and tree species vulnerabilities across Britain. We assessed the drought impact on the potential yield class of three major tree species (Picea sitchensis, Pinus sylvestris, and Quercus robur) which presently cover around 59% (400,700 ha) of state-managed forests, across lowland and upland sites. Here we show that drought impacts result mostly in reduced tree growth over the next 80 years when using b1, a1b and a1fi IPCC emissions scenarios. We found a maximum reduction of 94% but also a maximum increase of 56% in potential stand yield class in the 2080s from the baseline climate (1961-1990). Furthermore, potential production over the national forest estate for all three species in the 2080s may decrease due to drought by 42% in the lowlands and 32% in the uplands in comparison to the baseline climate. Our results reveal that potential tree growth and forest production on the national forest estate in Britain is likely to reduce, and indicate where and when adaptation measures are required. Moreover, this paper demonstrates the value of probabilistic climate projections for an important economic and environmental sector.

Urban forest health survey on Roznik in Ljubljana in 2013 by two methods

Urban forest health was surveyed on Roznik in Ljubljana (46.05141 N, 14.47797 E) in 2013 by two methods: ICP Forests and UFMO. ICP Forests is most commonly used monitoring programme in Europe - the International Co-operative Programme on the Assessment and Monitoring of Air Pollution Effects on Forests, which is based on systematic grid. UFMO method - Urban Forests Management Oriented method was developed in the frame of EMoNFUr Project - Establishing a monitoring network to assess lowland forest and urban plantations in Lombardy and urban forest in Slovenia (LIFE10 ENV/IT/000399). UFMO is based on non-linear transects (GPS tracks). ICP forests monitoring plots were established in July 2013 in the urban forest Roznik in Ljubljana .The 32 plots are located on sampling grid 500 × 500 m. The grid was down-scaled from the National Forest Monitoring survey, which bases on national sample grid 4 × 4 km. With the ICP forests method the following parameters for each tree within the 15 plots were gathered according to the ICP forests manual for Visual assessment of crown condition and damaging agents: tree species, percentage of defoliation, affected part of the tree, specification of affected part, location in crown, symptom, symptom specification, causal agents / factors, age of damage, damage extent, and damage extent on the trunk. With the UFMO method, the following parameters for each tree that needed sylviculture measure (felling, pruning, sanitary felling, thinning, etc.) were recorded: tree species, breast diameter, causal agent / damaging factor, GPS waypoint and GPS track. For overall picture in the urban forest health problems, also other biotic and abiotic damaging factors that did not require management action were recorded.

A method to incorporate the effect of taxonomic uncertainty on multivariate analyses of ecological data

Researchers in ecology commonly use multivariate analyses (e.g. redundancy analysis, canonical correspondence analysis, Mantel correlation, multivariate analysis of variance) to interpret patterns in biological data and relate these patterns to environmental predictors. There has been, however, little recognition of the errors associated with biological data and the influence that these may have on predictions derived from ecological hypotheses. We present a permutational method that assesses the effects of taxonomic uncertainty on the multivariate analyses typically used in the analysis of ecological data. The procedure is based on iterative randomizations that randomly re-assign non identified species in each site to any of the other species found in the remaining sites. After each re-assignment of species identities, the multivariate method at stake is run and a parameter of interest is calculated. Consequently, one can estimate a range of plausible values for the parameter of interest under different scenarios of re-assigned species identities. We demonstrate the use of our approach in the calculation of two parameters with an example involving tropical tree species from western Amazonia: 1) the Mantel correlation between compositional similarity and environmental distances between pairs of sites, and; 2) the variance explained by environmental predictors in redundancy analysis (RDA). We also investigated the effects of increasing taxonomic uncertainty (i.e. number of unidentified species), and the taxonomic resolution at which morphospecies are determined (genus-resolution, family-resolution, or fully undetermined species) on the uncertainty range of these parameters. To achieve this, we performed simulations on a tree dataset from southern Mexico by randomly selecting a portion of the species contained in the dataset and classifying them as unidentified at each level of decreasing taxonomic resolution. An analysis of covariance showed that both taxonomic uncertainty and resolution significantly influence the uncertainty range of the resulting parameters. Increasing taxonomic uncertainty expands our uncertainty of the parameters estimated both in the Mantel test and RDA. The effects of increasing taxonomic resolution, however, are not as evident. The method presented in this study improves the traditional approaches to study compositional change in ecological communities by accounting for some of the uncertainty inherent to biological data. We hope that this approach can be routinely used to estimate any parameter of interest obtained from compositional data tables when faced with taxonomic uncertainty.

Comparison of different stomatal conductance algorithms for ozone flux modelling

A multiplicative and a semi-mechanistic, BWB-type [Ball, J.T., Woodrow, I.E., Berry, J.A., 1987. A model predicting stomatalconductance and its contribution to the control of photosynthesis under different environmental conditions. In: Biggens, J. (Ed.), Progress in Photosynthesis Research, vol. IV. Martinus Nijhoff, Dordrecht, pp. 221–224.] algorithm for calculating stomatalconductance (gs) at the leaf level have been parameterised for two crop and two tree species to test their use in regional scale ozone deposition modelling. The algorithms were tested against measured, site-specific data for durum wheat, grapevine, beech and birch of different European provenances. A direct comparison of both algorithms showed a similar performance in predicting hourly means and daily time-courses of gs, whereas the multiplicative algorithm outperformed the BWB-type algorithm in modelling seasonal time-courses due to the inclusion of a phenology function. The re-parameterisation of the algorithms for local conditions in order to validate ozone deposition modelling on a European scale reveals the higher input requirements of the BWB-type algorithm as compared to the multiplicative algorithm because of the need of the former to model net photosynthesis (An)

Promoting mixed stands through conversion treatments. Effect of holm oak coppices thinning on black pine regeneration

The effects of conversion treatments, depending on ecological factors and silvicultural parameters (thinning intensity, thinning type and rotation, among others) have been studied during the last fifteen years in an experimental trial in Central Spain. The general climate is continental Mediterranean; soils are low depth and limy; vegetation is an homogeneous dense coppices of Quercus ilex with isolated Pinus nigra trees. The experimental design (three locations) includes different thinning intensities (from 0 to 100% of extracted basal area). Inventories have been carried out in 1994 and 2010; thinning treatments were done in 1995 and 2011. Analysis of the effects of the conversion treatment show the increment of diameter and height growth rates, the canopy recovery and the stand resprouting, finding differences in these effects between thinning treatments. Besides the induced changes at holm oak stand, the application of conversion treatment clearly changed the woodland dynamics. Fifteen years after the thinnings, floristic composition varied and an abundant pine regeneration was installed in the woodland. In this work we describe the changes between inventories in tree species composition and diameter distribution, specially in the case of black pine. The conversion treatment caused changes in forest dynamics in the short term, increasing biodiversity and diversifying the forest structure. The fast installation of Pinus regeneration suggests the potential of the zone for the establishment of multipurpose mixed Quercus-Pinus stands in wide areas where Quercus species were favoured by human populations for firewood production. Conversion treatment of coppices, with the creation of mixed stands, constitutes a good management alternative for extensive areas and an interesting technique to adaptation to global change.

Small-scale variation of vegetation in a mixed forest understorey is partly controlled by the effect of overstory composition on litter accumulation

We investigated how richness and composition of vascular plant species in the understory of a mixed hardwood forest stand varied with respect to the abundance and composition of the overstory. The stand is in central Spain and represents the southernmost range of distribution of several tree and herbaceous species in Europe. Understory species were identified in 46 quadrats (0.25 m2) where variables litter depth and light availability were measured. In addition, we estimated tree density, basal area, and percent basal area by tree species within 6-m-radius areas around each plot. Species richness and composition were studied using path analysis and scale-dependent geostatistical methods, respectively. We found that the relative abundance of certain trees species in the overstory was more important than total overstory abundance in explaining understory species richness. Richness decreased as soil litter depth increased, and soil litter increased as the relative proportion of Fagus sylvatica in the overstory increased, which accounted for a negative, indirect effect of Fagus sylvatica on richness. Regarding understory species composition, we found that some species distributed preferentially below certain tree species. For example, Melica uniflora was most frequent below Fagus sylvatica and Quercus petraea while the increasing proportion of Q. pyrenaica in the overstory favored the presence of Cruciata glabra, Arenaria montana, Prunus avium, Conopodium bourgaei, Holcus mollis, Stellaria media and Galium aparine in the understory. Overall, these results emphasize the importance of individual tree species in controlling the assemblage and richness of understory species in mixed stands. We conclude that soil litter accumulation is one way through which overstory composition shapes the understory community.

An integral model for natural regeneration of Pinus pinea L. in the Northern Plateau (Spain) = Modelo integral de regeneración natural para Pinus pinea L. en la Meseta Norte (España)

Natural regeneration is an ecological key-process that makes plant persistence possible and, consequently, it constitutes an essential element of sustainable forest management. In this respect, natural regeneration in even-aged stands of Pinus pinea L. located in the Spanish Northern Plateau has not always been successfully achieved despite over a century of pine nut-based management. As a result, natural regeneration has recently become a major concern for forest managers when we are living a moment of rationalization of investment in silviculture. The present dissertation is addressed to provide answers to forest managers on this topic through the development of an integral regeneration multistage model for P. pinea stands in the region. From this model, recommendations for natural regeneration-based silviculture can be derived under present and future climate scenarios. Also, the model structure makes it possible to detect the likely bottlenecks affecting the process. The integral model consists of five submodels corresponding to each of the subprocesses linking the stages involved in natural regeneration (seed production, seed dispersal, seed germination, seed predation and seedling survival). The outputs of the submodels represent the transitional probabilities between these stages as a function of climatic and stand variables, which in turn are representative of the ecological factors driving regeneration. At subprocess level, the findings of this dissertation should be interpreted as follows. The scheduling of the shelterwood system currently conducted over low density stands leads to situations of dispersal limitation since the initial stages of the regeneration period. Concerning predation, predator activity appears to be only limited by the occurrence of severe summer droughts and masting events, the summer resulting in a favourable period for seed survival. Out of this time interval, predators were found to almost totally deplete seed crops. Given that P. pinea dissemination occurs in summer (i.e. the safe period against predation), the likelihood of a seed to not be destroyed is conditional to germination occurrence prior to the intensification of predator activity. However, the optimal conditions for germination seldom take place, restraining emergence to few days during the fall. Thus, the window to reach the seedling stage is narrow. In addition, the seedling survival submodel predicts extremely high seedling mortality rates and therefore only some individuals from large cohorts will be able to persist. These facts, along with the strong climate-mediated masting habit exhibited by P. pinea, reveal that viii the overall probability of establishment is low. Given this background, current management –low final stand densities resulting from intense thinning and strict felling schedules– conditions the occurrence of enough favourable events to achieve natural regeneration during the current rotation time. Stochastic simulation and optimisation computed through the integral model confirm this circumstance, suggesting that more flexible and progressive regeneration fellings should be conducted. From an ecological standpoint, these results inform a reproductive strategy leading to uneven-aged stand structures, in full accordance with the medium shade-tolerant behaviour of the species. As a final remark, stochastic simulations performed under a climate-change scenario show that regeneration in the species will not be strongly hampered in the future. This resilient behaviour highlights the fundamental ecological role played by P. pinea in demanding areas where other tree species fail to persist.

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.