Nature, origin, and petroleum source potential of organic matter at DSDP Leg 56-57 Holes

As part of a continuing program of organic-geochemistry studies of sediments recovered by the Deep Sea Drilling Project, we have analyzed the types, amounts, and thermal-alteration indices of organic matter in samples collected from the landward wall of the Japan Trench on Legs 56 and 57. The samples were canned aboard ship, enabling us to measure also their gas contents. In addition, we analyzed the heavy C15+ hydrocarbons, NSO compounds, and asphaltenes extracted from selected samples. Our samples form a transect down the trench wall, from Holes 438 and 438A (water depth 1558 m), through Holes 435 and 435A (water depth 3401 m), and 440 (water depth 4507 m), to Holes 434 and 434B (water depth 5986 m). The trench wall is the continental slope of Japan. Its sediments are Cenozoic hemipelagic diatomaceous muds that were deposited where they are found or have slumped from farther up the slope. Their terrigenous components probably were deposited from near-bottom nepheloid layers transported by bottom currents or in low density flows (Arthur et al., 1978). Our objective was to find out what types of organic matter exist in the sediment and to estimate their potential for generation of hydrocarbons.

Age determination, carbon geochemistry and palynology of section AC003

Sedimentary records from California's Northern Channel Islands and the adjacent Santa Barbara Basin (SBB) indicate intense regional biomass burning (wildfire) at the Ållerød-Younger Dryas boundary (~13.0-12.9 ka) (All age ranges in this paper are expressed in thousands of calendar years before present [ka]. Radiocarbon ages will be identified and clearly marked "14C years".). Multiproxy records in SBB Ocean Drilling Project (ODP) Site 893 indicate that these wildfires coincided with the onset of regional cooling and an abrupt vegetational shift from closed montane forest to more open habitats. Abrupt ecosystem disruption is evident on the Northern Channel Islands at the Ållerød-Younger Dryas boundary with the onset of biomass burning and resulting mass sediment wasting of the landscape. These wildfires coincide with the extinction of Mammuthus exilis [pygmy mammoth]. The earliest evidence for human presence on these islands at 13.1-12.9 ka (~11,000-10,900 14C years) is followed by an apparent 600-800 year gap in the archaeological record, which is followed by indications of a larger-scale colonization after 12.2 ka. Although a number of processes could have contributed to a post 18 ka decline in M. exilis populations (e.g., reduction of habitat due to sea-level rise and human exploitation of limited insular populations), we argue that the ultimate demise of M. exilis was more likely a result of continental scale ecosystem disruption that registered across North America at the onset of the Younger Dryas cooling episode, contemporaneous with the extinction of other megafaunal taxa. Evidence for ecosystem disruption at 13-12.9 ka on these offshore islands is consistent with the Younger Dryas boundary cosmic impact hypothesis [Firestone et al., 2007, doi:10.1073/pnas.0706977104].

Landsat derived percent vegetation cover of Israel (as of 2003)

Wildfires are part of the Mediterranean ecosystem, however, in Israel all wildfires are human caused, either intentionally or un-intentionally. In this study we aimed to develop and test a new method for mapping fire scars from MODIS imagery, to examine the temporal and spatial patterns of wildfires in Israel in the 2000s and to examine the factors controlling Israel's wildfire regime. To map the fires we used two 'off-the-shelf' MODIS fire products as our basis-the 1 km MODIS Collection 5 fire hotspots, the 500 m MCD45A1 burnt areas-and we created a new set of fire scar maps from the 250 m MOD13Q1 product. We carried out a cross comparison of the three MODIS based wildfire scar maps and evaluated them independently against the wild fire scars mapped from 30 m Landsat TM imagery. To examine the factors controlling wildfires we used GIS layers of rainfall, land use, and a Landsat-based national vegetation map. Wildfires occurred in areas where annual rainfall was above 250 mm, mostly in areas with herbaceous vegetation. Wildfire frequency was especially high in the Golan Heights and in the foothills of the Judean mountains, and a high correspondence was found between military training zones and the spatial distribution of fire scars. The use of MODIS satellite images enabled us to map wildfires at a national scale due to the high temporal resolution of the sensor. Our MOD13Q1 based mapping of fire scars adequately mapped large (>1 km**2) fires with accuracies above 80%. Such large fires account for a large proportion of all fires, and pose the greatest threats. This database can aid managers in determining wildfire risks in space and in time.

A fractional vegetation cover remote sensing product on pan-arctic scale with link to geotiff image

The paper presents first results of a pan-boreal scale land cover harmonization and classification. A methodology is presented that combines global and regional vegetation datasets to extract percentage cover information for different vegetation physiognomy and barren for the pan-arctic region within the ESA Data User Element Permafrost. Based on the legend description of each land cover product the datasets are harmonized into four LCCS (Land Cover Classification System) classifiers which are linked to the MODIS Vegetation Continuous Field (VCF) product. Harmonized land cover and Vegetation Continuous Fields products are combined to derive a best estimate of percentage cover information for trees, shrubs, herbaceous and barren areas for Russia. Future work will concentrate on the expansion of the developed methodology to the pan-arctic scale. Since the vegetation builds an isolation layer, which protects the permafrost from heat and cold temperatures, a degradation of this layer due to fire strongly influences the frozen conditions in the soil. Fire is an important disturbance factor which affects vast processes and dynamics in ecosystems (e.g. biomass, biodiversity, hydrology, etc.). Especially in North Eurasia the fire occupancy has dramatically increased in the last 50 years and has doubled in the 1990s with respect to the last five decades. A comparison of global and regional fire products has shown discrepancies between the amounts of burn scars detected by different algorithms and satellite data.

Miocene microflora of Belorado (Spain)

Neogene deposits from Belorado (Province de Burgos, Spain) in the NE border of the Duero Basin have been analyzed. The palynologic analysis of the samples suggests the existence of arboreal landscapes associated with the herbaceous cover creating belts of vegetation around restricted aquatic areas (marshy and lacustrine environments). The characteristics of the palynomorph assemblages allow to define a warm-template climate with strong seasons. The age of these deposits could be included in the middle Miocene (Aragonian).

(Table 2) Paleoclimate reconstruction for palynology samples of Turkey

Palynofloras of the Kocaçay and Cumaovasi basins in western Turkey that belong to a time-span from the late Early to late Middle Miocene (the late Burdigalian-Serravallian) are studied and compared with published palynofloras of Europe and Turkey. Palynological data and numerical climatic results obtained by the coexistence approach indicate palaeoclimate changed from warm subtropical to temperate during the late Burdigalian-Serravallian. Moreover, the palaeoclimates of the Kocacay and Cumaovasi basins are compared with continental palaeoclimatic records of coal-bearing sediments in western Turkey and current temperatures in the Izmir region. According to this comparison, palaeoclimatic results of these basins and other localities in western Turkey show a distinct difference as a result of orographic change. The palaeovegetation in the Kocaçay and Cumaovasi basins during the studied time-span was affected by palaeotopography and palaeoclimate. In these basins mixed mesophytic, coniferous forests, and swamp palaeovegetation generally predominated during the late Early-early Middle Miocene. The role of the herbaceous taxa increased at the end of the late Middle Miocene (the Serravallian) in the Kocaçay and Cumaovasi basins. It is obvious from the palynomorph data of these basins that grassland palaeovegetation started to expand in the late Middle Miocene. Unlike in Central Europe, where late Burdigalian and Langhian represent a period of outstanding warmth, the so-called Mid-Miocene Climatic Optimum, cold month mean temperatures reconstructed in this study point to an ongoing cooling trend, already from the late Burdigalian onwards, possibly related to increasing terrestrial conditions in the study area.

Climate and total tree cover reconstructions based on the 43.7-kyr fossil pollen record from Khoe (NW Sakhalin Island) and modern climate variables of 236 modern surface pollen sampling sites

A late Quaternary pollen record from northern Sakhalin Island (51.34°N, 142.14°E, 15 m a.s.l.) spanning the last 43.7 ka was used to reconstruct regional climate dynamics and vegetation distribution by using the modern analogue technique (MAT). The long-term trends of the reconstructed mean annual temperature (TANN) and precipitation (PANN), and total tree cover are generally in line with key palaeoclimate records from the North Atlantic region and the Asian monsoon domain. TANN largely follows the fluctuations in solar summer insolation at 55°N. During Marine Isotope Stage (MIS) 3, TANN and PANN were on average 0.2 °C and 700 mm, respectively, thus very similar to late Holocene/modern conditions. Full glacial climate deterioration (TANN = -3.3 °C, PANN = 550 mm) was relatively weak as suggested by the MAT-inferred average climate parameters and tree cover densities. However, error ranges of the climate reconstructions during this interval are relatively large and the last glacial environments in northern Sakhalin could be much colder and drier than suggested by the weighted average values. An anti-phase relationship between mean temperature of the coldest (MTCO) and warmest (MTWA) month is documented during the last glacial period, i.e. MIS 2 and 3, suggesting more continental climate due to sea levels that were lower than present. Warmest and wettest climate conditions have prevailed since the end of the last glaciation with an optimum (TANN = 1.5 °C, PANN = 800 mm) in the middle Holocene interval (ca 8.7-5.2 cal. ka BP). This lags behind the solar insolation peak during the early Holocene. We propose that this is due to continuous Holocene sea level transgression and regional influence of the Tsushima Warm Current, which reached maximum intensity during the middle Holocene. Several short-term climate oscillations are suggested by our reconstruction results and correspond to Northern Hemisphere Heinrich and Dansgaard-Oeschger events, the Bølling-Allerød and the Younger Dryas. The most prominent fluctuation is registered during Heinrich 4 event, which is marked by noticeably colder and drier conditions and the spread of herbaceous taxa.

Palynofacies analysis of sediments in the eastern Equatorial Atlantic

Analyses of the palynofacies and sporomorph thermal alteration indices (TAI) of sediments from Ocean Drilling Program (ODP) Sites 959 to 962 in the Cote d'Ivoire-Ghana Transform Margin, West Africa were undertaken to (1) determine the source and depositional conditions of the organic matter in the sediments, (2) refine a paleobathymetric curve derived from other data for Site 959, which drilled the most continuous sedimentary sequence from Pleistocene to Albian and (3) interpret the paleothermal history of the area. Twelve types of dispersed organic matter were identified: amorphous organic matter (AOM), marine palynomorphs, algae, resins, black debris, yellow-brown fragments, black-brown fragments, cuticles, plant tissue, wood, sporomorphs and fungi, The relative abundances of these organic matter components at each site were analyzed using cluster analysis, resulting in the identification of seven palynofacies assemblages at Site 959, five each at sites 960 and 961, and four at Site 962. Amorphous organic matter (which is chiefly marine derived), black debris and wood have played the most significant role in defining palynofacies assemblages. The palynofacies assemblages show some correlation with lithologic units, sediment sources and depositional environments. Previous palynofacies studies in passive margins have demonstrated that changes in the ratio of AOM to terrestrial organic matter are related primarily to proximal-distal positions of depositional environments relative to the shoreline. However, this assumption does not always hold true for a transform margin where tectonic factors play an important role in the organic matter distribution, at least in the early stages of evolution. Lithofacies, CCD paleodepths for the North Atlantic, trace fossil association, benthic foraminifera and palynofacies data were the criteria used for reconstructing a paleobathymetric curve for Site 959. A cyclicity in the organic matter distribution of the Upper Miocene to Lower Pliocene pelagic sediments could be related to fluctuations in productivity of biosiliceous and calcareous organisms, and sedimentation rates. A drastic increase in the amount of AOM and a decrease in black debris and wood in the carbonate and clastic rocks (Lithologic Unit IV) overlying the tectonized Albian sediments (Lithologic Unit V) at Sites 959 and 960 coincide with the presence of an unconformity. Qualitative color analysis of palynomorphs was undertaken for all sites, although the main focus was on Site 959 where detailed organic geochemical data were available. At Site 959, TAI values indicate an immature stage of organic maturation (<2) down to the black claystones of Lithologic Unit III at about 918.47 mbsf. Below this, samples show an increase with depth to a moderately mature stage (>2 except for the claystone samples between 1012.52 and 1036.5 mbsf, and one limestone sample at 1043.4 mbsf), reaching peak levels of 2.58 to 3.0 in the tectonized Albian sediments below the unconformity. These TAI values show a positive correlation with the Tma x values derived from Rock-Eval pyrolysis data. The highest values recorded in the basal tectonized units at all the sites (Sites 960-962 have mean values between 2.25 and 3.13) may be related to high heat flow during the intracontinental to syntransform basin stage in the region.

Fungal microbiota from rain water and pathogenicity of Fusarium species isolated from atmospheric dust and rainfall dust

In order to determine the presence of Fusarium spp. in atmospheric dust and rainfall dust, samples were collected during September 2007, and July, August, and October 2008. The results reveal the prevalence of airborne Fusarium species coming from the atmosphere of the South East coast of Spain. Five different Fusarium species were isolated from the settling dust: Fusarium oxysporum, F. solani, F. equiseti, F. dimerum, and F. proliferatum. Moreover, rainwater samples were obtained during significant rainfall events in January and February 2009. Using the dilution-plate method, 12 fungal genera were identified from these rainwater samples. Specific analyses of the rainwater revealed the presence of three species of Fusarium: F. oxysporum, F. proliferatum and F. equiseti. A total of 57 isolates of Fusarium spp. obtained from both rainwater and atmospheric rainfall dust sampling were inoculated onto melon (Cucumis melo L.) cv. Piñonet and tomato (Lycopersicon esculentum Mill.) cv. San Pedro. These species were chosen because they are the main herbaceous crops in Almeria province. The results presented in this work indicate strongly that spores or propagules of Fusarium are able to cross the continental barrier carried by winds from the Sahara (Africa) to crop or coastal lands in Europe. Results show differences in the pathogenicity of the isolates tested. Both hosts showed root rot when inoculated with different species of Fusarium, although fresh weight measurements did not bring any information about the pathogenicity. The findings presented above are strong indications that long-distance transmission of Fusarium propagules may occur. Diseases caused by species of Fusarium are common in these areas. They were in the past, and are still today, a problem for greenhouses crops in Almería, and many species have been listed as pathogens on agricultural crops in this region. Saharan air masses dominate the Mediterranean regions. The evidence of long distance dispersal of Fusarium spp. by atmospheric dust and rainwater together with their proved pathogenicity must be taken into account in epidemiological studies.

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.

Provision of ecological infrastructures to increase pollinators and other beneficial organisms in rainfed crops in Central Spain

In sustainable intensive agriculture, the biodiversity of monoculture fields can be increased by managing the field margins to provide ecological infrastructures that serve as refuges and resources for beneficial organisms (pollinators and natural enemies). In the present work we summarize two years of field trials following the goal to increase biodiversity of beneficial fauna in a barley field in Central Spain by sowing different herbaceous mixtures in the field margins. The presence of arthropods visiting flowers on plots sown with different types of seed mixtures and unsown natural flora (control plot) was compared by visual sampling every week between April and June. The results showed that a combination of herbaceous big-size seeds was the most successful mixture emerging under our experimental conditions and achieved a higher number of visits of beneficial arthropods than the unsown natural vegetation.

Análisis de las diferencias en el crecimiento vegetativo, en los componentes del rendimiento y en la composición de las uvas y vinos del cv. cabernet sauvignon (vitis vinifera L.), producidos por el déficit hídrico durante pre-envero y post-envero

La presente Tesis Doctoral se realizó con el fin de estimar conjuntamente la respuesta agronómica y fisiológica de la vid (Vitis vinifera L.), así como los efectos sobre la evolución de la maduración, composición y la calidad de la uva y del vino, bajo la aplicación de diferentes déficit hídricos en pre-envero y post-envero, dentro de un marco de referencia de cambio climático. La variación climática que prevén los estudios sobre el cambio climático, resulta un factor decisivo en la eficiencia del uso del agua en la vid. En zonas cálidas, las estrategias de cultivo del viñedo frente al cambio climático deben de ir dirigidas a atenuar sus efectos sobre el crecimiento y el desarrollo de la vid, haciéndose imprescindible el estudio pormenorizado del déficit hídrico como factor decisivo en la obtención de las uvas adecuadas, ya que son la clave indispensable para el éxito en la elaboración del vino, y de forma muy especial en los vinos enfocados a un sector de alta calidad. El ensayo se llevó a cabo en un viñedo comercial de Bodegas Licinia, en la Comunidad de Madrid, durante los años 2010 y 2011. La variedad estudiada fue Cabernet sauvignon / 41 B, plantada a un marco de plantación de 3 m x 1 m, con un guiado vertical de la vegetación. El dispositivo experimental fue totalmente al azar, y se establecieron 4 tratamientos experimentales con 4 grados de disponibilidad hídrica, déficit moderado continuo (T0,45-0,6), déficit severo continuo (T0-0,3), déficit severo después de envero (T0,45-0,3) y déficit severo antes de envero (T0-0,6). En cada tratamiento se distribuyeron 3 repeticiones. El año 2010 fue el más lluvioso de los años de ensayo, con 478 mm de precipitaciones anuales, lo que supuso 146 mm más que en el año 2011. Su distribución a lo largo del ciclo fue más homogénea en el año 2010, mientras que en 2011 las precipitaciones contabilizadas en el período de maduración de la uva fueron nulas. La temperatura media subió 0,9ºC en 2011, respecto a 2010 y en cuanto a la integral térmica eficaz, en 2011 se acumularon, desde el 1 de abril hasta el final de ciclo, 217 grados•día más que en 2010. El déficit hídrico en pre-envero, modificó notablemente el crecimiento vegetativo y la producción de cosecha de la parcela de ensayo, no así la fertilidad de las yemas. El tratamiento con mayor disponibilidad hídrica (T0,45-0,6) obtuvo el mayor peso de baya, y los tratamientos con menor déficit hídrico en pre-envero (T0,45-0,6 y T0,45-0,3) registraron los mayores rendimientos de cosecha, mientras que las menores tasas de cuajado correspondieron al tratamiento con un déficit severo continuo (T0-0,3). La parcela de ensayo se caracterizó por un exceso de vigor y un alto crecimiento vegetativo. El pH del mosto se vio afectado por el déficit hídrico, disminuyendo su valor en el tratamiento de déficit hídrico severo antes de envero (T0-0,6). Organolépticamente, no se percibieron diferencias significativas en los vinos elaborados en función del déficit hídrico, y respecto a su composición físico-química, solo existieron diferencias en la concentración de ácido L-Málico, con mayores concentraciones en los tratamientos sin déficit hídrico en pre-envero, T0,45-0,6 y T0,45-0,3. El déficit hídrico modificó notablemente el color del vino, aumentando los valores de las coordenadas CIELAB a* y b*, la luminosidad (L*), croma (C*) y tonalidad (H*), para los tratamientos con un déficit severo en pre-envero (T0-0,3 y T0-0,6) y disminuyendo estas en el tratamiento con mayor disponibilidad hídrica (T0,45-0,6). Del mismo modo, mediante el análisis de color por métodos tradicionales, IPT e IC de los vinos, aumentó en los tratamientos con mayor déficit hídrico en pre-envero (T0-0,3 y T0-0,6), respecto a los tratamientos de mayor disponibilidad (T0,45-0,6 y T0,45-0,3). La concentración de taninos de la baya en vendimia, no se vio afectada por el déficit hídrico, aunque sí estuvo relacionada positivamente con el tamaño de las bayas. Organolépticamente, los hollejos del año 2011 resultaron con menor frescura, acidez, afrutado, sensación herbácea e intensidad tánica, aunque con mayor astringencia respecto a 2010. Las pepitas fueron más astringentes y aromáticas pero menos crujientes, sin llegar a los niveles de madurez del año 2010. El catador relacionó los taninos con la calidad del vino, asociándolos con un mayor cuerpo, acidez, intensidad, equilibrio gustativo, amargor y menor astringencia en la fase gustativa. La concentración de taninos en los vinos se vio favorecida con el déficit hídrico en pre-envero y post-envero. Los tratamientos con mayor déficit hídrico en pre-envero, T0-0,6 y T0-0,3, obtuvieron las menores concentraciones de potasio en mostos y vinos. Las relaciones entre la concentración de potasio, ácido L-Málico y el porcentaje de color rojo puro (dA(%)) resultaron altamente significativas, de modo que las mayores tasas de potasio en el vino se asociaron a los valores más bajos de color rojo y a los mayores de ácido L-Málico. ABSTRACT The present Doctoral Thesis has been done in order to estimate the grapevine (Vitis vinifera L.) agronomic and physiologic performance or response as well as the impact in the grape and wine maturity, composition and quality evolution, with different water deficits. The variation in climate that the global warming studies for seen is a key factor for the grapevine water use efficiency. In warm areas the farming vineyards strategy to face the climatic change, should be focused on diminish the effects on the grapevine growth and development, so that the water deficit detailed analysis becomes decisive to obtain the appropriate grapes, that are the main subject for a successful wine production and especially for top quality wines. The trial was carried out in a commercial vineyard in Chinchón (Madrid), Licinia winery, during the 2010 and 2011 seasons. The grape variety studied was Cabernet Sauvignon grafted onto 41B with a vine spacing 3m x 1m trained as VSP. Experimental design consisted on 4 irrigation treatments with 3 replications totally randomized. Irrigation treatments were: moderate regulated deficit (T0,45-0,6), severe continuous deficit (T0-0,3), severe post-veraison deficit (T0,45-0,3) and severe pre veraison deficit (T0-0,6). The 2010 was rainier year than the 2011; Total annual rain in 2010 was 478 mm, which resulted in 146 mm more than in 2011. The distribution along the vine cycle was more homogeneous in the 2010, whereas precipitations in 2011 along the grape maturity period were nonexistent. The average temperature in 2011 was 0,9ºC higher than that of the 2010 and regarding to the thermal integral, in the 2011 from 1st April to the end of the growing cycle, was 217 degrees•day higher than that in 2010. Water deficit significantly modified the vegetative growth and yield but, it did not modified bud fertility. The treatment with the highest water availability (T0,45-0,6) got the highest berry size, the lowest berry set rates were found in the severe continuous deficit treatment (T0-0,3). The plot studied in this trial was characterized by both excessive vigour and vegetative growth. Water deficit modified the pH must by, reducing it in the severe water deficit during pre-veraison (T0-0,6). There were not differences in wine tasting between the water deficits treatments. Regarding to the physical-chemical composition, it only existed differences in the L-malic acid concentration, resulting higher concentrations in the water deficit pre-veraison treatments: T0,45-0,6 y T0,45-0,3. Water deficit significantly modified wine colour by, increasing the CIELAB coordinates a* and b*, the brightness (L*), croma (C*) and tonality (H*), in the lower water availability pre-veraison treatments (T0-0,3 y T0-0,6), and reducing them in the in the moderate continuous water deficit ones (T0,45-0,6). By means of traditional wine colour parameters analyses, red colour percentage, TPI, they became higher in the lower water availability pre-veraison treatments (T0-0,3 y T0-0,6), than in those with higher availability (T0,45-0,6 y T0,45-0,3). At harvest, berry tannins concentrations was not affected by the water deficit although it did in a positive way, in the berry size. Berry tasting in 2011, resulted in a lower freshness, acidity, fruity, herbaceous flavour and tannic intensity, but with higher astringency respect to the 2010 season. Seeds, in 2011, were more astringent and aromatic as in the 2010, but less crunchy, without getting to the point of maturity. The taster linked the tannins to wine quality, associating them with a higher bodiest wine, acidity, intensity, taste balance, bitterness and with a lower astringency in the tasting stage. Treatments with a higher water deficit up to veraison T0-0,6 y T0-0,3 got less musts and wines potassium concentration. The relation between L-malic acid and the full red color percentage (dA(%)), were highly related, resulting the higher potassium content the lower wine quality.

Chaperonas moleculares y tolerancia a estrés abiótico en especies arbóreas

Las temperaturas extremas, la sequía y otros estreses abióticos limitan la producción forestal de forma significativa, causando grandes pérdidas económicas en el sector. Los árboles, al ser organismos sésiles, han desarrollado una serie de estrategias para percibir dichos factores, activando respuestas defensivas apropiadas. Entre ellas ocupa un lugar preeminente la síntesis de proteínas con actividad chaperona molecular. Las chaperonas moleculares interaccionan con proteínas desnaturalizadas total o parcialmente, promoviendo su correcto plegamiento y ensamblaje. Las chaperonas moleculares que se sintetizan de forma predominante en plantas, pero no en otros eucariotas, pertenecen a la familia sHSP (small heat-shock proteins). Se trata de una familia inusualmente compleja y heterogénea, cuyos miembros son de pequeño tamaño (16-42 kD) y poseen un dominio “alfa-cristalina” muy conservado. Estas proteínas están implicadas en protección frente a estrés abiótico mediante la estabilización de proteínas y membranas, si bien su mecanismo de acción se conoce de forma incompleta. A pesar del evidente potencial aplicado de las proteínas sHSP, son muy escasos los estudios realizados hasta el momento con un enfoque netamente biotecnológico. Por otra parte, casi todos ellos se han llevado a cabo en especies herbáceas de interés agronómico o en especies modelo, como Arabidopsis thaliana. De ahí que las sHSP de arbóreas hayan sido mucho menos caracterizadas estructural y funcionalmente, y ello a pesar del interés económico y ecológico de los árboles y de su prolongada exposición vital a múltiples factores estresantes. La presente Tesis Doctoral se centra en el estudio de sHSP de varias especies arbóreas de interés económico. El escrutinio exhaustivo de genotecas de cDNA de órganos vegetativos nos ha permitido identificar y caracterizar los componentes mayoritarios de tallo en dos especies productoras de madera noble: nogal y cerezo. También hemos caracterizado la familia completa en chopo, a partir de su secuencia genómica completa. Mediante expresión heteróloga en bacterias, hemos analizado el efecto protector de estas proteínas in vivo frente a distintos tipos de estrés abiótico, relevantes para el sector productivo. Los resultados demuestran que las proteínas sHSP-CI: (i) aumentan la viabilidad celular de E.coli frente a casi todos estos factores, aplicados de forma individual o combinada; (ii) ejercen un rol estabilizador de las membranas celulares frente a condiciones adversas; (iii) sirven para mejorar la producción de otras proteínas recombinantes de interés comercial. El efecto protector de las proteínas sHSP-CI también ha sido analizado in planta, mediante la expresión ectópica de CsHSP17.5-CI en chopos. En condiciones normales de crecimiento no se han observado diferencias fenotípicas entre las líneas transgénicas y los controles, lo que demuestra que se pueden sobre-expresar estas proteínas sin efectos pleiotrópicos deletéreos. En condiciones de estrés térmico, por el contrario, los chopos transgénicos mostraron menos daños y un mejor crecimiento neto. En línea con lo anterior, las actividades biológicas de varias enzimas resultaron más protegidas frente a la inactivación por calor, corroborando la actividad chaperona propuesta para la familia sHSP y su conexión con la tolerancia al estrés abiótico. En lo que respecta a la multiplicación y propagación de chopo in vitro, una forma de cultivo que comporta estrés para las plantas, todas las líneas transgénicas se comportaron mejor que los controles en términos de producción de biomasa (callos) y regeneración de brotes, incluso en ausencia de estrés térmico. También se comportaron mejor durante su cultivo ex vitro. Estos resultados tienen gran potencial aplicado, dada la recalcitrancia de muchas especies vegetales de interés económico a la micropropagación y a la manipulación in vitro en general. Los resultados derivados de esta Tesis, aparte de aportar datos nuevos sobre el efecto protector de las proteínas sHSP citosólicas mayoritarias (clase CI), demuestran por vez primera que la termotolerancia de los árboles puede ser manipulada racionalmente, incrementando los niveles de sHSP mediante técnicas de ingeniería genética. Su interés aplicado es evidente, especialmente en un escenario de calentamiento global. ABSTRACT Abiotic stress produces considerable economic losses in the forest sector, with extreme temperature and drought being amongst the most relevant factors. As sessile organisms, plants have acquired molecular strategies to detect and recognize stressful factors and activate appropriate responses. A wealth of evidence has correlated such responses with the massive induction of proteins belonging to the molecular chaperone family. Molecular chaperones are proteins which interact with incorrectly folded proteins to help them refold to their native state. In contrast to other eukaryotes, the most prominent stress-induced molecular chaperones of plants belong to the sHSP (small Heat Shock Protein) family. sHSPs are a widespread and diverse class of molecular chaperones that range in size from 16 to 42k Da, and whose members have a highly conserved “alpha-crystallin” domain. sHSP proteins play an important role in abiotic stress tolerance, membrane stabilization and developmental processes. Yet, their mechanism of action remains largely unknown. Despite the applied potential of these proteins, only a few studies have addressed so far the biotechnological implications of this protein family. Most studies have focused on herbaceous species of agronomic interest or on model species such as Arabidopsis thaliana. Hence, sHSP are poorly characterized in long-lived woody species, despite their economic and ecological relevance. This Thesis studies sHSPs from several woody species of economic interest. The most prominent components, namely cytosolic class I sHSPs, have been identified and characterized, either by cDNA library screening (walnut, cherry) or by searching the complete genomic sequence (poplar). Through heterologous bacterial expression, we analyzed the in vivo protective effects of selected components against abiotic stress. Our results demonstrate that sHSP-CI proteins: (i) protect E. coli cells against different stressful conditions, alone or combined; (ii) stabilize cell membranes; (iii) improve the production of other recombinant proteins with commercial interest. The effects of CsHSP17.5-CI overexpression have also been studied in hybrid poplar. Interestingly, the accumulation of this protein does not have any appreciable phenotypic effects under normal growth conditions. However, the transgenic poplar lines showed enhanced net growth and reduced injury under heat-stress conditions compared to vector controls. Biochemical analysis of leaf extracts revealed that important enzyme activities were more protected in such lines against heat-induced inactivation than in control lines, lending further support to the chaperone mode of action proposed for the sHSP family. All transgenic lines showed improved in vitro and ex vitro performance (calli biomass, bud induction, shoot regeneration) compared to controls, even in the absence of thermal stress. Besides providing new insights on the protective role of HSP-CI proteins, our results bolster the notion that heat stress tolerance can be readily manipulated in trees through genetic engineering. The applied value of these results is evident, especially under a global warming scenario.