Pablo J. Hidalgo

Supercritical fluid extraction of natural antioxidants from rosemary : Comparison with liquid solvent sonication

Supercritical fluid extraction (SFE) and liquid solvent sonication, in combination with two different sample treatments, were compared for the extraction of natural antioxidants from rosemary leaves. Dried, ground, and sieved rosemary leaves (20 mg) were subjected to SFE with CO(2) at 355 bar at 100 °C (CO(2) density 0.72 g/mL) for 20 min at a liquid flow rate of 4 mL/min. The analytes were concentrated on an ODS trap and subsequently eluted with acetone. Antioxidants in the SF and liquid solvent extract were analyzed by HPLC. Compounds of known antioxidant activity such as carnosol, carnosic acid, and methyl carnosate were identified by mass spectrometry of the HPLC fractions collected. Freezing and grinding the samples in liquid nitrogen resulted in decreased carnosic acid recoveries. Supercritical CO(2) extraction provided the highest recovery of carnosic acid from rosemary leaves (35.7 mg/g), the lowest relative standard deviation (4.4%), and the cleanest extract [Formula: see text] no cleanup prior to HPLC was required. Among the liquid solvents studies, only acetone provided comparable results (73% recovery relative to SC-CO(2) extraction); however, it required decoloration with active carbon prior to HPLC analysis.

Pollen Production in the Poaceae Family.

Total pollen production per inflorescence was studied in the most important species of the Poaceae family in the city of Córdoba in order to further our knowledge of the partial contribution of each species of this family to the total amount of pollen released into the atmosphere. The contribution of grasses in a given area was estimated by counting the number of inflorescences in an area of one square meter. Four different representative areas of the city were selected. The number of pollen grains per anther and flowers per inflorescence was also estimated in order to obtain total pollen production per inflorescence. Pollen production per inflorescence ranged from 14,500 to more than 22,000,000 pollen grains, the amount being clearly higher in the perennial species. Pollen production per square meter was higher in the mountains near the city and lower in areas of abandoned crops. Only a few species are responsible for the majority of pollen produced. A phenological study is necessary in order to determine the temporal distribution of this pollen production and subsequent shedding.

Pollen production of the genus Cupressus

The study of potential pollen emission in the genus Cupressus was carried out in the city of Cordoba (Spain). Pollen production of 227 trees of the represented species of Cupressus in the area (Cupressus sempervirens, C. macrocarpa and C. arizonica) was studied by counting the number of pollen grains per flower, the number of flowers per branch, and the number of branches per m2 of surface area. Total surface area was estimated by considering the shape of the tree as a cone and estimating the average height and radius of each species. Results indicated that there was little variation in the number of pollen grains produced by a flower within each species. Equally, there were few differences in the number of grains per flower among the species, although C. arizonica had slightly fewer than the other species because it possessed fewer stamens per flower. However, there were clear differences in the number of flowers per branch, with C. macrocarpa having the highest floral density, considerably lower in C. a...

Pollen biology in four Mediterranean Quercus species

Potential pollen production, viability and germination were studied in the most important species of Quercus in the mountains of Córdoba to determine the contribution of each species to the total amount of airborne pollen. The results were compared over two consecutive years with different rainfall patterns. The viability of pollen grains was determined at anther opening, and during the pollination period, in order to determine potential pollination capacity. Results indicated that there were differences in the number of pollen grains produced by stamen in the four species. Equally, there were differences in the number of flowers among the species, being Q. suber the species with higher number of catkins groups and flowers per individual tree. Total pollen production per tree can be summarized in the following proportion: 1: 3: 3: 6 (Q. coccifera, Q. ilex ssp. ballota, Q. faginea and Q. suber). Potential pollen viability was estimated using the Fluorochromatic Reaction (FCR) and a germination assay. The results have shown that Quercus potential pollen viability is high and declines slowly with time. Q. coccifera was the species with the highest percentage of germination, with Q. suber being the lowest.

Development of a semi-automatic system for pollen recognition

A semi-automatic system for pollen recognitionis studied for the european project ASTHMA. The goal of such a system is to provideaccurate pollen concentration measurements. This information can be used as well by thepalynologists, the clinicians or a forecastsystem to predict pollen dispersion. At first,our emphasis has been put on Cupressaceae, Olea, Poaceae and Urticaceae pollen types. The system is composed of two modules: pollengrain extraction and pollen grain recognition. In the first module, the pollen grains areobserved in light microscopy and are extractedautomatically from a pollen slide coloured withfuchsin and digitized in 3D. In the secondmodule, the pollen grain is analyzed forrecognition. To accomplish the recognition, itis necessary to work on 3D images and to usedetailed palynological knowledge. Thisknowledge describes the pollen types accordingto their main visible characteristerics and tothose which are important for recognition. Somepollen structures are identified like the porewith annulus in Poaceae, the reticulum in Oleaand similar pollen types or the cytoplasm inCupressaceae. The preliminary results show therecognition of some pollen types, likeUrticaceae or Poaceae or some groups of pollentypes, like reticulate group.

An automated system for surveying and forecasting Olea pollen dispersion

An automated system for Olea pollensurveying and forecasting is being developedfor the province of Cordoba, Spain, within theframework of the European ASTHMA project.Required input data have been split into threecategories: static data, calibration parametersand dynamic data. A 200 × 200 km potential pollenproduction map, centred on the city of Cordoba,was drawn up using digital high-resolutionsatellite data and reflecting average plantdensity, as derived from the observed spatialseparation of trees in 28 different locations.Based on archives of pollen data from 1982, thestart of the pollen season was determined byapplying both heat accumulation methods andneural network analysis including furthermeteorological parameters. The forecast andsurvey system is based on a set of twoelementary atmospheric physics modules andbiological models: emission and dispersionmodules. The system can run in different modes,allowing forecasting of the floweringstart-date and, from that date, charting theemission and dispersion of pollenconcentrations in the area. Also, wind andtemperature fields are regularly computed andupdated using deterministic numerical models tochart the spatial and temporal concentration ofOlea pollen.

Microsporogenesis in male sterile Rosmarinus officinalis L. (Lamiaceae), an ultrastructural study

Within an extensive reproductive biology program on Rosmarinus officinalis a study of microsporogenesis in male sterile plants has been carried out. Two events, related to the two types of male sterile flowers found in this species (MS; Male sterile and INT; Intermediate male sterile flowers), are described. The first event is characterized by the early appearance of necrotic areas in the anther tissues during the final differentiation phases of the MS flowers, before meiosis takes place. Initially, these necrotic areas are small, later on they enlarge affecting the sporogenous tissue and tapetum. All the anthers in which necrosis has begun finally become aborted and lack pollen grains. The second event is manifested in the anthers of the INT flowers. Previous to the release of the microspores from the callosic envelope, vacuolisation of the tapetal cells takes place. The tapetum does not properly carry out its secretory function. It accumulates some substances in vacuoles and starch granules in plastids....

A tree species range in the face of climate change: cork oak as a study case for the Mediterranean biome

Species distribution models are feasible methods for projecting theoretical responses of living organisms’ occurrence under several future climate change scenarios. The major interest is focused on trees, which regulate the equilibrium within ecosystems and guarantee the survival of many life forms on the Earth. The repercussions of climatic drivers are expected to pose the strongest threats for the Mediterranean biome, an acknowledged hotspot of biodiversity. Here, we focused on cork oak (Quercus suber L.), a keystone species of many landscapes, sustaining a rich biodiversity, ecological processes and economic incomes. Results of 8 combined ecological modelling techniques and two Global Circulation Models highlight a broad contraction of the species potential range over the twenty-first century, both under intermediate and high emissions scenarios. Coupled northward and upward shifts are predicted, mostly pertaining Iberia and North Africa. The potential areas detected at Levantine will likely undergo disappearance. To exacerbate the impacts of climate change, the future of the ecosystems linked to cork oak remains uncertain, because of the expected implications on the phenotypic plasticity or evolutionary responses. A synergy among niche-based, physiological and eco-genetic investigations is strongly needed in the field of applied research, to improve the assessment of conservation and reforestation actions.

Trends in evergreen oak suitability from assembled species distribution models: assessing climate change in south-western Europe

The expected changes to the environmental conditions have concerned the scientific community over the last few decades. A rise in the mean temperature and a variation in rainfall patterns could modify the current distribution of plant species. In this study, we analysed four evergreen oaks (Quercus ilex subsp. ilex, Q. ilex subsp. ballota, Q. suber and Q. coccifera) by means of species distribution models. Three algorithms were used: maximum entropy, logistic regression and environmental distance. Taxa occurrences were taken, chiefly from the National Forest Inventories, and climate data was retrieved from the WorldClim 1.4 project. The present period and four future scenarios were studied. The latter were carried out by averaging thirteen global circulation models (GCMs). Area under the curve was used for validating the models. Maps indicating the suitability and cosuitability among the evergreen oaks were developed. The potential distribution of evergreen oaks in the present period was found to be wider than the actual distribution. Simulations indicate that climate change would increase the cosuitability of western temperate areas for Mediterranean oaks. The use of different algorithms and GCMs, as well as the high validation values obtained, make the study robust. Oaks are an important source of income, especially Q. ilex subsp. ballota and Q. suber. Our findings contribute to our understanding of the dynamics of oaks, and can be considered for management programmes aimed at conserving this natural heritage.

Predicting suitability of forest dynamics to future climatic conditions: the likely dominance of Holm oak [Quercus ilex subsp. ballota (Desf.) Samp.] and Aleppo pine (Pinus halepensis Mill.)

Key messageComposite logistic regression models simulating the potential effect of global climate change on forests dynamics in the southern Iberian Peninsula identify Holm oak [Quercus ilexsubsp.ballota(Desf.) Samp.] and Aleppo pine (Pinus halepensisMill.) as the chief beneficiaries of the anticipated environmental shifts, whereas other oak species and conifers suffer a decline.ContextThe ten most important tree species (five oaks and five conifers) in Southern Spain were selected for the study. The study area, corresponding to the region of Andalusia, is located in an interesting position between Central European and North African climates. The territory also exhibits the most extreme patterns of rainfall in the Iberian Peninsula.AimsThis study aims to model the potential distribution of the ten species in response to climate change, in several time periods, including the present and two future twenty-first century dates.MethodsThe potential distributions within the different scenarios were simulated using logistic regression techniques based on a set of 19 climate variables from the WorldClim 1.4 project. The scenarios were drawn from the RCP 2.6 and 6.0 in the CCSM4 Global Circulation Model. The resolution of the output maps was 30 arc-seconds.ResultsThe simulation predicted increased distribution areas for Q. ilex and P. halepensis under the four future scenarios as compared to present. The eight remaining taxa suffered a severe retraction in potential distribution.ConclusionGlobal climate change is likely to have a significant impact on forest dynamics in southern Spain. Only two species would benefit to the detriment of the others. Logistic Regression is identified as a robust method for carrying out management and conservation programmes.