Identification and immunolocalization of superoxide dismutase isoenzymes of olive pollen

Gametophytic tissues of plants are an area largely neglected in the broad literature on free radical processes in plants. In order to study the mechanisms of protection against oxidative stress in pollen, the presence of the key antioxidative enzyme superoxide dismutase (SOD; EC 1.15.1.1) was investigated. Crude extracts of olive tree ( Olea europaea L.) pollen were subjected to native PAGE in 10% polyacrylamide gels. The SOD activity staining of gels showed the presence of four isoenzymes. All the SODS were completely inhibited by 2 m M KCN and 5 m M H₂O₂, and therefore belong to the family of CuZn‐SODS. Isoelectric focusing (pH 3.5‐7) of crude extracts and further detection of SOD activity allowed determination of isoelectric points for the four isoforms, namely 4.60, 4.78, 5.08 and 5.22. The cross‐reactivity of pollen extracts with a polyclonal antibody to cytosolic CuZn‐SOD from spinach leaves was assayed by western blotting. After SDS‐PAGE and immunoblotting, a major polypeptide band of about 16.5 kDa was detected, which is characteristic of the subunit of most CuZn‐SODS. Immunocytochemical studies at TEM level using the same antiserum showed that CuZn‐SOD was localized in the cytoplasm of both vegetative and generative cells, and also in material adhered to the pollen wall. The olive pollen CuZn‐SODS could function in the protection against oxidative stress during pollen development.     

Bioclimatic indices as a tool in pollen forecasting

The use of bioclimatic indices could be a major step forward in the methodology of pollen forecasting. The basis for this proposal is that simple meteorological parameters do not reflect the global status of the atmosphere, but merely some static measurements. However, pollen dispersal is, above all, a dynamic phenomenon, and this fact should be reflected in the variables we used to explain it. Here, we test the two methodologies for routine pollen forecasting by comparing correlation coefficients using the same daily Poaceae airborne pollen data base from León (6 years, from 1994 to 1999) as the dependent variable and either simple daily meteorological variables or compound daily bioclimatic indices as independent variables. Both simple and compound indices reproduced the same profile of evolution of plant eco-physiological requirements, as the length of the study period during the pollen season increased. However, for time frames larger than the main pollen period, bioclimatic indices gave superior coefficients, which seems to indicate that these could be more valuable for pre-season pollen forecasting. The continentality index produced the highest mean coefficient, higher than those generated by any meteorological variable. Furthermore, at least for a Mediterranean climate, site location and evapotranspiration in relation to precipitation seem to be the most promising factors for increasing success when forecasting Poaceae airborne pollen concentration.     

Electrophoretic profiling and immunocytochemical detection of pectins and arabinogalactan proteins in olive pollen during germination and pollen tube growth

Background and Aims

Cell wall pectins and arabinogalactan proteins (AGPs) are important for pollen tube growth. The aim of this work was to study the temporal and spatial dynamics of these compounds in olive pollen during germination.

Methods

Immunoblot profiling analyses combined with confocal and transmission electron microscopy immunocytochemical detection techniques were carried out using four anti-pectin (JIM7, JIM5, LM5 and LM6) and two anti-AGP (JIM13 and JIM14) monoclonal antibodies.

Key Results

Pectin and AGP levels increased during olive pollen in vitro germination. (1 → 4)-β-d-Galactans localized in the cytoplasm of the vegetative cell, the pollen wall and the apertural intine. After the pollen tube emerged, galactans localized in the pollen tube wall, particularly at the tip, and formed a collar-like structure around the germinative aperture. (1 → 5)-α-l-Arabinans were mainly present in the pollen tube cell wall, forming characteristic ring-shaped deposits at regular intervals in the sub-apical zone. As expected, the pollen tube wall was rich in highly esterified pectic compounds at the apex, while the cell wall mainly contained de-esterified pectins in the shank. The wall of the generative cell was specifically labelled with arabinans, highly methyl-esterified homogalacturonans and JIM13 epitopes. In addition, the extracellular material that coated the outer exine layer was rich in arabinans, de-esterified pectins and JIM13 epitopes.

Conclusions

Pectins and AGPs are newly synthesized in the pollen tube during pollen germination. The synthesis and secretion of these compounds are temporally and spatially regulated. Galactans might provide mechanical stability to the pollen tube, reinforcing those regions that are particularly sensitive to tension stress (the pollen tube–pollen grain joint site) and mechanical damage (the tip). Arabinans and AGPs might be important in recognition and adhesion phenomena of the pollen tube and the stylar transmitting cells, as well as the egg and sperm cells.     

Predicting the intensity of the birch pollen season

We present a model for the prediction of the magnitude ofBetula flowering and pollen dispersal which may be used in the management of birch pollinosis and in the planning of clinical trials. The pollen sum during the flowering season is regressed on the temperature sum from May 1st to July 20th during the initiation year, the pollen sum of the initiation year, and the temperature sum during the main pollen season in the flowering year. We suggest that the fluctuating flowering pattern inBetula alba-species is primarily determined by the availability of assimilation products during inflorescence initiation and development during the spring one year before anthesis. When inflorescences, which are initiated during the previous year, elongate in the beginning of anthesis, they act as strong sinks to stored carbohydrates, and thus compete with developing leaves and shoots. The result is an initially reduced photosynthetic capacity in years with intense flowering, and a limited potential for the initiation of new inflorescences for the following year. The ambient temperature during catkin initiation affects assimilation efficiency and is a determinant of about equal importance to flowering intensity as is the magnitude of the flowering in the initiation year. The amount of pollen dispersed is also dependent on the weather during anthesis, which is not possible to predict until about one month in advance. The two other independent variables are available during the previous summer, making it possible to give a sufficiently valid prediction to allergologists about the magnitude of the next birch pollen season, according to its botanical determinants. We suggest that the varying reproductive output inBetula alba should not be described as true masting. A more parsimonious explanation to the flowering pattern is that an individual continually maximizes reproductive effort, according to what is possible, but that reproduction is often constrained by the environment.     

Predicting the intensity of the birch pollen season

We present a model for the prediction of the magnitude ofBetula flowering and pollen dispersal which may be used in the management of birch pollinosis and in the planning of clinical trials. The pollen sum during the flowering season is regressed on the temperature sum from May 1st to July 20th during the initiation year, the pollen sum of the initiation year, and the temperature sum during the main pollen season in the flowering year. We suggest that the fluctuating flowering pattern inBetula alba-species is primarily determined by the availability of assimilation products during inflorescence initiation and development during the spring one year before anthesis. When inflorescences, which are initiated during the previous year, elongate in the beginning of anthesis, they act as strong sinks to stored carbohydrates, and thus compete with developing leaves and shoots. The result is an initially reduced photosynthetic capacity in years with intense flowering, and a limited potential for the initiation of new inflorescences for the following year. The ambient temperature during catkin initiation affects assimilation efficiency and is a determinant of about equal importance to flowering intensity as is the magnitude of the flowering in the initiation year. The amount of pollen dispersed is also dependent on the weather during anthesis, which is not possible to predict until about one month in advance. The two other independent variables are available during the previous summer, making it possible to give a sufficiently valid prediction to allergologists about the magnitude of the next birch pollen season, according to its botanical determinants. We suggest that the varying reproductive output inBetula alba should not be described as true masting. A more parsimonious explanation to the flowering pattern is that an individual continually maximizes reproductive effort, according to what is possible, but that reproduction is often constrained by the environment.     

Cell wall acyl-lipids, proteins and polysaccharides in mature and germinated olive pollen

Proteins, acyl-lipids and polysaccharides from cell walls of mature and germinated olive pollen were studied. In general, hemicelluloses are the most abundant polysaccharides, arabinose in mature and glucose in germinated pollen being the main components of these macromolecules. Protein content and its amino acid composition are very similar in walls from mature and germinated pollen, these compounds showing a weak acid character. Free-fatty acids are the most abundant lipid molecules in mature and germinated pollen walls and a decrease in acyl-lipids, especially in polar lipids, as well as a higher unsaturation of their fatty acid components are observed after germination.     

Improving early-season estimates of olive production using airborne pollen multi-sampling sites

The aim of this study was to improve the accuracy of aeropalynological models to forecast yields in areas with heterogeneous characteristics by applying principal component analysis to integrate the airborne pollen sampled from more than one trap. The sampling was performed during the past seven years (1998–2004) in the main northeast olive regions of Portugal. Annual crop production was forecasted on the basis of airborne pollen concentration measured at flowering, comparing the performance of three different independent variables: total airborne pollen concentration sampled in each trap and a derived variable that was determined by principal component analysis of the total airborne pollen concentration sampled. The best predictive results were obtained using a logarithmic relationship with airborne pollen concentration principal component scores describing about 97% of olive fruit production variability over the last seven years. The use of this technique improved the ability of pollen to explain the production interannual variations by about 13%. The comparison between actual reported and the adjusted production showed an average spread deviation of 5%.     

Correlation between large-scale atmospheric fields and the olive pollen season in Central Italy

Olives are one of the largest crops in the Mediterranean and in central and southern Italy. This work investigates the correlation of the Olea europaea L. pollen season in Perugia, the capital city of the region of Umbria in central Italy, with atmospheric parameters. The aim of the study is twofold. First, we study the correlation between the pollen season and the surface air temperature of the spring and late spring in Perugia. Second, the correlation between the pollen season and large-scale atmospheric patterns is investigated. The average surface temperature in the spring and late spring has a clear impact on the pollen season in Perugia. Years with higher average temperatures have an earlier onset of the pollen season. In particular, a 1°C higher (lower) average surface temperature corresponds to an earlier (later) start of the pollen season of about 1 week. The correlation between the pollen season and large-scale atmospheric patterns of sea level pressure and 500-hPa geopotential height shows that the cyclonic activity in the Mediterranean is unequivocally tied to the pollen season in Perugia. A larger than average cyclonic activity in the Mediterranean Basin corresponds to a later than average pollen season. Larger than average cyclonic activity in Northern Europe and Siberia corresponds to an earlier than average pollen season. A possible explanation of this correlation, that needs further investigation to be proven, is given. These results can have a practical application by using the seasonal forecast of atmospheric general circulation models.     

Spatiotemporal models for predicting high pollen concentration level of Corylus,Alnus, and Betula

Corylus, Alnus, and Betula trees are among the most important sources of allergic pollen in the temperate zone of the Northern Hemisphere and have a large impact on the quality of life and productivity of allergy sufferers. Therefore, it is important to predict high pollen concentrations, both in time and space. The aim of this study was to create and evaluate spatiotemporal models for predicting high Corylus, Alnus, and Betula pollen concentration levels, based on gridded meteorological data. Aerobiological monitoring was carried out in 11 cities in Poland and gathered, depending on the site, between 2 and 16 years of measurements. According to the first allergy symptoms during exposure, a high pollen count level was established for each taxon. An optimizing probability threshold technique was used for mitigation of the problem of imbalance in the pollen concentration levels. For each taxon, the model was built using a random forest method. The study revealed the possibility of moderately reliable prediction of Corylus and highly reliable prediction of Alnus and Betula high pollen concentration levels, using preprocessed gridded meteorological data. Cumulative growing degree days and potential evaporation proved to be two of the most important predictor variables in the models. The final models predicted not only for single locations but also for continuous areas. Furthermore, the proposed modeling framework could be used to predict high pollen concentrations of Corylus, Alnus, Betula, and other taxa, and in other countries.     

Altitudinal fluctuations in the olive pollen emission: an approximation from the olive groves of the south-east Iberian Peninsula

The possible existence of altitudinal fluctuations in the seasonal behaviour of the olive pollen emission was studied. Three pollen volumetric samplers distributed in olive groves all over the altitudinal cliseries of the province of Jaén (south-east Spain) were used. Pollen emission data were recorded during a 3-year period (2007–2009). This research has revealed the effect of altitude on consecutive olive pollen season in the province of Jaén. The first pollen grains were detected in the olive growing areas located within the area of the Guadalquivir River, where are found the lowest levels of altitude into the province. A notable delay in the pollination season of the olive groves located at higher altitudes was observed. Geographical fluctuations on both daily pollen concentrations and number of critical days were also detected. Accumulated variables of temperature and precipitation since the start of the pre-flowering period have been shown to be two of the main factors affecting olive pollen levels. The fluctuations observed in the olive pollen season may similarly occur in the case of other allergenic plant species such as cypress (Cupressaceae), plane tree (Platanaceae) or grasses (Poaceae). Furthermore, and for the clinical consequences of the findings presented in this study, we believe that it would be advisable to install a micro-aerobiological network permanently in the province of Jaén.     

Forecasting olive (Olea europaea) crop production by monitoring airborne pollen

Forecasting harvests of olives destined for the production of olive oil can be based on counts of airborne olive pollen, and meteorological and agronomic observations. This study was carried out during six consecutive years (1990–1995) in the Campiña Alta (an olive-producing region in the province of Córdoba, south-west Spain). Olive pollen totals are the annual sum of the concentrations recorded for the periods that the filters of a Cour trap were exposed. The meteorological data are the values of accumulated rainfall between 1 September and the following 15 April (a date prior to the beginning of olive flowering). The agronomic data are the forecast and actual productions for the province of Córdoba, supplied by the Board of Agriculture of the Andalusian government, and the actual production of the Campiña Alta, supplied at the end of harvest by private olive-growing co-operatives. The data were combined, and four mathematical equations were obtained to forecast the crop 6 months in advance, with varying degrees of reliability. The reliability was very high for an appropriate agricultural area. The most accurate equation isY=?1.90×10⁴+2.35X+53.94 (which forecasts the production of the Campiña Alta), whereY is the olive production (MT),X the olive pollen count,Z the rainfall prior to flowering, anda, b andc are constants. The least accurate equation is that relating olive pollen concentrations with olive production in the province of Córdoba.     

Relationship between pollen emission and fruit production in olive (Olea europaea L.)

Aerobiological data of pollen emission concentrations is used to predict fruit production. The principal aim of this work was to study the relationship between pollen emission patterns, emission homogeneity and fruit production in olive (Olea europaea L.). Data of daily pollen concentrations in the atmosphere during the flowering period, collected over a 20‐year period in Perugia (Central Italy), and the corresponding fruit production data were analysed. Correlation and regression analyses on the partial pollen amounts (subdivisions of the whole flowering period), their statistical variability (expressed as coefficients of variation of the daily pollen concentration), and the production values in the different years demonstrate that pollen emission, during the seven to ten day period immediately preceding the maximum pollen emission day, appears to be most closely related to fruit production. Moreover, the pollen emission homogeneity (minimal variability in daily pollen concentrations) during the “critical” flowering period is very important for fertilization.     

Prediction of the beginning of the olive full pollen season in south-west Spain

Given the clinical and agricultural importance of the olive in SW Spain, we have carried out a study to predict the starting date of its full pollen season. The study covers 6 years of meteorological and palynological observations — the latter using a Cour sampler installed in Huelva (SW Spain). The results obtained show that olive full pollination begins when the plant has accumulated 731C of daily temperature above 5C from the end of its dormant period. The mean duration of this accumulation was 83 days. A positive relationship has been found between mean temperature of the months before the pollen season (February and March) and the date when the season starts (April). From the data available, rainfall registered between 1 September and 31 March (both before pollination), does not affect the starting date of the full pollen season, but can affect total pollen production, particularly in years with prolonged drought.     

Prediction of the beginning of the olive full pollen season in south-west Spain

Given the clinical and agricultural importance of the olive in SW Spain, we have carried out a study to predict the starting date of its full pollen season. The study covers 6 years of meteorological and palynological observations — the latter using a Cour sampler installed in Huelva (SW Spain). The results obtained show that olive full pollination begins when the plant has accumulated 731°C of daily temperature above 5°C from the end of its dormant period. The mean duration of this accumulation was 83 days. A positive relationship has been found between mean temperature of the months before the pollen season (February and March) and the date when the season starts (April). From the data available, rainfall registered between 1 September and 31 March (both before pollination), does not affect the starting date of the full pollen season, but can affect total pollen production, particularly in years with prolonged drought.     

Paternity analysis using microsatellite markers to identify pollen donors in an olive grove

Olive (Olea europaea L.) is a wind-pollinated, allogamous species that is generally not considered to be self-compatible. In addition, cross-incompatibilities exist between cultivars that can result in low fruit set if compatible pollinisers are not planted nearby. In this study, microsatellite markers were used to identify 17 genotypes that were potential pollen donors in a commercial olive orchard. DNA typing with the same primers was also applied to 800 olive embryos collected from five cultivars in the grove over 2 years of study. Pollen donors for the cultivars Barnea, Corregiola, Kalamata, Koroneiki, and Mission were estimated by paternity analysis, based on the parental contribution of alleles in the genotypes of the embryos. The exclusion probability for the marker set was 0.998 and paternity was assigned on the basis of the ‘most likely method’. Different pollen donors were identified for each of the maternal cultivars indicating that cross-compatibilities and incompatibilities varied between the genotypes studied. Cross-pollination was the principal method of fertilization, as selfing was only observed in two of the embryos studied and both of these were from the cultivar Mission. This is the first report where these techniques have been applied to survey the pollination patterns in an olive grove. The results indicate that careful planning in orchard design is required for efficient pollination between olive cultivars.     

The effect of meteorological parameters on diurnal patterns of airborne olive pollen concentration

Aerobiological studies carried out in the atmosphere of Granada using a Hirst-type volumetric spore trap during the period 1993-1996 show that there is not a single diurnal pattern for olive pollen (Olea europaea L.) over the course of the main pollen season. Examination of the behaviour of airborne olive pollen concentration allows the establishment of either regular (54.4% of the studied days) or irregular (45.6% of the time) patterns of diurnal variation. On a given day, the pattern found will depend on a combination of different factors: the origin of the captured pollen (either local or regional), source distribution in relation to the pollen sampler, topography, and different meteorological variables (mean air temperature, sunshine hours, total rainfall, relative humidity, wind speed and direction, and periods of calm). Regional sources were significant contributors to city centre pollen concentrations when moderate (< 10 km/h) winds from the 4th quadrant and warm temperatures (19-26 C) allow swift transport from the W-NW of the province.     

Analysis of the pollen allergen content of twelve olive cultivars grown in Portugal

Olive pollen presents intercultivar variability as regards to its antigenic and allergenic composition. In this study, we report the presence of differences among the SDS-PAGE pollen protein profiles of twelve Portuguese olive cultivars. Though most soluble proteins from these extracts seemed similar, three bands of about 18, 20 and 22 kDa presented sharp differences in intensity among the cultivars analyzed. The dissimilarity of patient’s sera reactivity to these protein extracts and the presence of several allergens already characterized (Ole e 1, Ole e 2, Ole e 5 and Ole e 9) in the extracts were also investigated. Epidemiological data indicated that 53.3 % out of the 428 patients analyzed with reactivity to pollen extracts, presented specific IgE levels to Olea europaea. A representative number of these sera were assayed in immunoblotting experiments. The cultivars ‘Galega’ and ‘Conserva de Elvas’ displayed low reactivity to the sera of atopic patients, whereas the extracts corresponding to the cultivars ‘Cobrançosa’, ‘Ascolana’ and ‘Verdeal de Serpa’ led to higher IgE reactivity. The use of antibodies to the allergens Ole e 1, Ole e 2, Ole e 5 and Ole e 9 in immunoblotting experiments also allowed cultivar discrimination. The cultivar ‘Verdeal de Serpa’ presented the highest Ole e 1, Ole e 5 and Ole e 9 allergen loads but the lowest Ole e 2. ‘Carrasquenha’ was the second cultivar in terms of the higher allergen content. Oppositely, the lowest allergen loads were those of the cultivars ‘Galega’ and ‘Conserva de Elvas’ coincidentally with their low IgE reactivity. These data may help interpret physiological differences in pollen performance for successful olive fertilization and, moreover, to better define future strategies for allergy diagnosis and treatment by specific immunotherapy.