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PubMed1998

Seasonal increase in specific IgE in serum induced by natural Japanese cedar pollen exposure in asymptomatic and symptomatic sensitized individuals.

The aim of this study was to determine whether there is a difference in the specific IgE response to pollen exposure between asymptomatic and symptomatic subjects who were already sensitized to Japanese cedar pollen. Sixty-four subjects with detectable serum levels of cedar pollen-specific IgE, which were apparent even before the pollen season in 1997, were enrolled in the study. Thirty-five had typical symptoms of seasonal allergic rhinitis during the pollen season in 1997 (symptomatic group) and the remainder had no seasonal aggravation during the pollen season in 1997 (asymptomatic group). Serum samples were collected twice from each subject, before and during the pollen season in 1997, to determine specific IgE by the lumiward immunoassay system. In both groups, the serum levels of specific IgE during the pollen season were significantly higher than those before the pollen season, and the rates of seasonal increase in specific IgE did not differ significantly between the groups. In conclusion, the specific IgE response during the pollen season is not a hallmark of clinical allergy and does not discriminate between symptomatic and asymptomatic individuals sensitive to the pollen.

Allergens↗
PubMed2005

Characterization of pollen tube development in Pinus strobus (Eastern white pine) through proteomic analysis of differentially expressed proteins.

The differentially expressed proteins in pollen tubes indicate their specific roles in this stage of male gametophyte development. To isolate these proteins, 2-DE was done using ungerminated pollen and 2-day-old pollen tubes of Pinus strobus. Results show that 645 and 647 protein spots were clearly resolved from pollen grains and pollen tubes, respectively. Thirty-eight protein spots were expressed only in pollen tubes, while 19 increased in intensity. MALDI-TOF MS was used to generate tryptic peptide masses that were submitted to Mascot for identification. Of the differentially expressed proteins, 12% matched with hypothetical proteins, 33% did not hit any protein, and for the 55%, a putative function was assigned based on similarity of sequences with previously characterized proteins. Therefore, pollen tube development can be characterized by the cellular activities that involve metabolism, stress/defense response, gene regulation, signal transduction, and cell wall formation. This study expands our understanding of the changes in protein expression associated with pollen tube development and provides insights into the molecular programs that separate the development of the pollen tubes from pollen grains. This is the first report that describes a global analysis of differentially expressed proteins from the pollen tube of any seed plant.

Cell Wall↗
PubMed2002

Pollen competition and seed-siring success in Picea abies.

The aim of the present work was to study pollen-tube competition in Picea abies. Controlled crossings were performed with pollen mixtures including pairs of pollen lots with fast and slowly elongating pollen-tubes. Paternity analysis using isozyme markers was performed on the progenies in order to study whether the in vitro pollen-germination vigour corresponds to the proportion of seeds sired by the pollen donor. Paternal success was found to be unequal, 15 out of 23 crossings producing progeny that differed significantly from the hypothetical ratio of 1:1. The paternal contribution in the majority of the crossings was as expected: the pollen parent with more-vigorous in vitro germination sired more seeds than the less-vigorous pollen. In the case of two pollen mixtures, however, the seed-siring success summed over the maternal trees was the opposite to the expected value. Despite these aberrations, the results support the hypothesis that pollen-tube competition is one of the factors contributing to male fitness in P. abies.However, when all the other factors affecting pollination and seed set under natural conditions are taken into account, it is clear that the seed-siring success of a particular paternal genotype cannot be predicted reliably by measuring only the in vitro pollen vigour.

Journal Article↗
PubMed2002

Genotype-environment interactions in pollen competitive ability in an anemophilous tree, Betula pendula Roth.

This study describes genotype-environment interactions in pollen competitive ability expressed as pollen-tube growth rate and seed-siring success in Betula pendula Roth. A factorial crossing design was applied using the same maternal and paternal clones in two different environmental conditions, in a B. pendula seed orchard established in a greenhouse and at an outdoor clone collection. Both single donor and two-donor pollinations were employed. Female inflorescences were collected after a fixed time of germination, pollen-tube lengths were measured for each cross, and paternity of the seeds sired by two-donor pollen mixtures was analyzed using isozyme markers. The pollination site had a significant influence on pollen-tube growth rate and seed-siring success. Significant interactions between pollination site and pollen donor indicated genotype-environment interactions in pollen-tube growth rate and seed-siring success. A highly significant positive correlation between pollen-tube growth rate and seed-siring success was found in the greenhouse but not at the outdoor clone collection. These results suggest that the pollen-tube growth rate can be a predictor of seed-siring success in controlled greenhouse conditions, where differences among maternal plants are mainly of genetic origin, but not in more heterogeneous outdoor conditions. In natural birch stands, environmental maternal effects probably diminish the significance of pollen competition for sexual selection in Betula pendula. At seed orchards, the effects of environmental conditions on pollen competitive ability can have important consequences for the genetic composition of the seed crop.

Journal Article↗
PubMed2000

Modern pollen rain from the Biligirirangan-Melagiri hills of southern Eastern Ghats, India.

A total of 39 soil surface samples collected between 11 degrees 30'N 76 degrees 45'E and 12 degrees 45'N 78 degrees 15'E from the mainly deciduous forests in the Biligirirangan-Melagiri hills of the southern Eastern Ghats were analysed for their pollen content. The samples are distributed among four different deciduous and evergreen vegetation types between 210 and 1700m altitudes and fall within three distinct rainfall regimes. The aims of this paper are to provide new data on the modern pollen rain from the Southern Eastern Ghats, a region characterized by a unique and complex climate and vegetation, and to interpret these data using multivariate statistics and the diagram of pollen percentages. We could distinguish first between the deciduous and the evergreen forests and then also between different types of deciduous forest. The distinction between the evergreen and deciduous forests was based on a humidity gradient and that among the deciduous forests on a physiognomic gradient identified through correspondence analysis. The above analysis also allowed us to identify a set of 14 pollen taxa markers and 11 associated pollen taxa that help differentiate the evergreen from deciduous forests. Similarly, a set of 12 pollen taxa markers and six associated pollen taxa was demarcated to help distinguish woodland formations from scrub and thicket formations, among the deciduous vegetation. We could also differentiate amongst the four distinct vegetation types sampled, on the basis of distinct associations of both tree and herb pollen taxa according to their relative abundance in the pollen diagram as well as on the proportion of total arboreal pollen. The ground cover of grasses and other herbaceous plants in the deciduous forests is effectively demonstrated by percentages of non-arboreal pollen varying between 40 and 70%. The 1000m altitude limit reflecting a gradient of humidity and the physiognomic gradient among deciduous forests seem to be important in this region.

Journal Article↗
PubMed2000

Evaluation of Holocene pollen records from the Romanian Plain.

This study is a critical review of pollen analyses carried out on Holocene sequences from 15 sites in and near the Romanian Plain. Three sites come from natural sediments, 10 sites are from anthropogenic deposits and two are from both anthropogenic and natural settings. The general reconstruction is of a steppe-forest-steppe vegetation through the Holocene. The nature of the deposits, however, casts doubts on this reconstruction. Deposits of archaeological sites generally yield pollen spectra that are influenced by human activities and thus unsuitable for vegetation reconstructions. Loess deposits are also unfavorable for pollen preservation because of high pH and porosity. Consequently, pollen spectra from loess deposits are strongly biased by selective pollen destruction. Research and experiments carried out by several authors suggest that spectra dominated by Asteraceae, Poaceae, Chenopodiaceae or Pinus pollen in soils and loess are a result of selective pollen destruction, especially if low pollen concentrations, progressive pollen deterioration or high frequencies of deteriorated or unidentifiable pollen are evidenced. The fact that pollen records from the Romanian Plain come from loess, alkaline peat or archaeological sites reduces their reliability for reconstructions of vegetation. The vegetation history of similar regions in Hungary, Bulgaria and Turkey suggests that early Holocene steppe vegetation was gradually replaced by forest or forest-steppe vegetation in the late Holocene. Records from lake sediments are required to find out whether the Holocene vegetation history of the Romanian Plain was similar.

Journal Article↗
PubMed2003

Actin-depolymerizing factor mediates Rac/Rop GTPase-regulated pollen tube growth.

Pollen tube elongation is a rapid tip growth process that is driven by a dynamic actin cytoskeleton. A ubiquitous family of actin binding proteins, actin-depolymerizing factors (ADFs)/cofilins, bind to actin filaments, induce severing, enhance depolymerization from their slow-growing end, and are important for maintaining actin dynamics in vivo. ADFs/cofilins are regulated by multiple mechanisms, among which Rho small GTPase-activated phosphorylation at a terminal region Ser residue plays an important role in regulating their actin binding and depolymerizing activity, affecting actin reorganization. We have shown previously that a tobacco pollen-specific ADF, NtADF1, is important for maintaining normal pollen tube actin cytoskeleton organization and growth. Here, we show that tobacco pollen grains accumulate phosphorylated and nonphosphorylated forms of ADFs, suggesting that phosphorylation could be a regulatory mechanism for their activity. In plants, Rho-related Rac/Rop GTPases have been shown to be important regulators for pollen tube growth. Overexpression of Rac/Rop GTPases converts polar growth into isotropic growth, resulting in pollen tubes with ballooned tips and a disrupted actin cytoskeleton. Using the Rac/Rop GTPase-induced defective pollen tube phenotype as a functional assay, we show that overexpression of NtADF1 suppresses the ability of NtRac1, a tobacco Rac/Rop GTPase, to convert pollen tube tip growth to isotropic growth. This finding suggests that NtADF1 acts in a common pathway with NtRac1 to regulate pollen tube growth. A mutant form of NtADF1 with a nonphosphorylatable Ala substitution at its Ser-6 position [NtADF1(S6A)] shows increased activity, whereas the mutant NtADF1(S6D), which has a phospho-mimicking Asp substitution at the same position, shows reduced ability to counteract the effect of NtRac1. These observations suggest that phosphorylation at Ser-6 of NtADF1 could be important for its integration into the NtRac1 signaling pathway. Moreover, overexpression of NtRac1 diminishes the actin binding activity of green fluorescent protein (GFP)-NtADF1 but has little effect on the association of GFP-NtADF1(S6A) with actin cables in pollen tubes. Together, these observations suggest that NtRac1-activated activity regulates the actin binding and depolymerizing activity of NtADF1, probably via phosphorylation at Ser-6. This notion is further supported by the observation that overexpressing a constitutively active NtRac1 in transformed pollen grains significantly increases the ratio of phosphorylated to nonphosphorylated ADFs. Together, the observations reported here strongly support the idea that NtRac1 modulates NtADF1 activity through phosphorylation at Ser-6 to regulate actin dynamics.

Actin Depolymerizing Factors↗
PubMed2006

Proteome analysis of maize pollen for allergy-relevant components.

Over the last few decades, the cultivation of maize (Zea mays) has strongly increased in Central Europe. We therefore decided to study the allergen composition and the allergenic potency of its pollen in comparison with pollen from timothy grass (Phleum pratense), a typical representative of the native grasses. We found that 65% of the sera reactive to timothy pollen also bound to maize pollen proteins. By using 2-DE immunoblotting, followed by incubation with mAbs directed against known allergens or protein sequencing, those IgE-reactive components were further classified. Although novel, maize-specific pollen allergens could not be found, the presence of crossreacting allergens belonging to groups 1 and 13 (Zea m 1 and 13), both having high IgE prevalence, as well as the presence of the less important group 3 and 12 allergens was found. The structural variability of Zea m 1 and Zea m 13 was determined by sequencing clones isolated from a maize pollen cDNA library. This revealed sequence identities of 72 and 70%, respectively, to the corresponding Phl p 1 and Phl p 13 allergens of timothy grass pollen. IgE-crossreactivity was further studied using immunoblot inhibition tests. Here, timothy pollen extract completely blocked IgE binding to maize, whereas maize pollen extract blocked IgE reactivity to only some timothy pollen allergens.

Allergens↗
PubMed1995

Expression of heat shock factor and heat shock protein 70 genes during maize pollen development.

We have analysed the expression of heat shock protein 70 (HSP70) and heat shock factor (HSF) gene during maize pollen development, HSFs being the transcriptional activators of hsp genes. In order to eliminate the sporophytic tissues of anthers, we have isolated homogeneous cell populations corresponding to five stages of maize pollen development from microspores to mature pollen. We show that in the absence of heat stress, hsp70 genes are highly expressed late-bicellular pollen as compared to other stages. HSP70 transcripts are significantly accumulated in response to a heat shock at the late microspore stage but to a much lower extent than in vegetative tissues. The latest stages of pollen development, i.e. mid-tricellular and mature pollen, do not exhibit heat-induced accumulation of HSP70 transcripts. Therefore, we analysed the expression of hsf genes throughout pollen development. We demonstrate that at least three hsf genes are expressed in maize and that transcripts corresponding to one hsf gene, whose expression is independent of temperature in somatic as well as in microgametophytic tissues, are present at similar levels throughout pollen development. In addition, we show that the expression of the two other hsf genes is heat-inducible in maize vegetative tissues and is not significantly increased after heat shock at any stage of pollen development. These results indicate that the loss of hsp gene expression at late stages of pollen development is not due to a modification of hsf gene expression at the mRNA level and that hsf gene expression is differentially regulated in vegetative and microgametophytic tissues.

Amino Acid Sequence↗
PubMed2006

Pollen foraging: learning a complex motor skill by bumblebees (Bombus terrestris).

To investigate how bumblebees (Bombus terrestris) learn the complex motor skills involved in pollen foraging, we observed naïve workers foraging on arrays of nectarless poppy flowers (Papaver rhoeas) in a greenhouse. Foraging skills were quantified by measuring the pollen load collected during each foraging bout and relating this to the number of flowers visited and bout duration on two consecutive days. The pollen standing crop (PSC) in each flower decreased drastically from 0530 to 0900 hours. Therefore, we related foraging performance to the changing levels of pollen available (per flower) and found that collection rate increased over the course of four consecutive foraging bouts (comprising between 277 and 354 individual flower visits), suggesting that learning to forage for pollen represents a substantial time investment for individual foragers. The pollen collection rate and size of pollen loads collected at the start of day 2 were markedly lower than at the end of day 1, suggesting that components of pollen foraging behaviour could be subject to imperfect overnight retention. Our results suggest that learning the necessary motor skills to collect pollen effectively from morphologically simple flowers takes three times as many visits as learning how to handle the most morphologically complex flowers to extract nectar, potentially explaining why bees are more specialised in their choice of pollen flowers.

Animals↗
PubMed2005

In vitro germination and transient GFP expression of American chestnut (Castanea dentata) pollen.

The development of the male reproductive structures of American chestnut (Castanea dentata) is described to advance our understanding of its reproductive behavior. This information has been vital in the development of a strategy to collect pollen grains from male catkins suitable for in vitro germination and transformation experiments. Cutting male catkins into small segments and rolling them over a culture plate resulted in evenly dispersed and large amounts of pollen with minimal unwanted accessory floral parts. To optimize pollen viability, the effect of various storage conditions on in vitro germination was examined. Our results showed that initial storage at 4 degrees C for 2 weeks significantly increased percent germination as compared to freshly collected pollen and those stored directly at -20 degrees C or -80 degrees C. This also means that for long-term storage of American chestnut pollen, the catkins should first be kept at 4 degrees C for a couple of weeks and then at -80 degrees C. The use of pollen grains with high viability is necessary for the transformation of American chestnut pollen. To optimize pollen transformation via particle bombardment, the effects of target distance, target pressure, and pollen developmental stage were examined. Statistical analysis showed that bombardment of ungerminated pollen at 1,100 psi resulted in the highest percent transient GFP expression (4.1%).

Fagaceae↗
PubMed2004

Transport of airborne pollen into the city of Thessaloniki: the effects of wind direction, speed and persistence.

We examined the effect of the wind vector analyzed into its three components (direction, speed and persistence), on the circulation of pollen from different plant taxa prominent in the Thessaloniki area for a 4-year period (1996-1999). These plant taxa were Ambrosia spp., Artemisia spp., Chenopodiaceae, Corylus spp., Cupressaceae, Olea europaea, Pinaceae, Platanus spp., Poaceae, Populus spp., Quercus spp., and Urticaceae. Airborne pollen of Cupressaceae, Urticaceae, Quercus spp. and O. europaea make up approximately 70% of the total average annual pollen counts. The set of data that we worked with represented days without precipitation and time intervals during which winds blew from the same direction for at least 4 consecutive hours. We did this in order to study the effect of the different wind components independently of precipitation, and to avoid secondary effects produced by pollen resuspension phenomena. Factorial regression analysis among the summed bi-hourly pollen counts for each taxon and the values of wind speed and persistence per wind direction gave significant results in 22 cases (combinations of plant taxa and wind directions). The pollen concentrations of all taxa correlated significantly with at least one of the three wind components. In seven out of the 22 taxon-wind direction combinations, the pollen counts correlated positively with wind persistence, whereas this was the case for only two of the taxon-wind speed combinations. In seven cases, pollen counts correlated with the interaction effect of wind speed and persistence. This shows the importance of wind persistence in pollen transport, particularly when weak winds prevail for a considerable part of the year, as is the case for Thessaloniki. Medium/long-distance pollen transport was evidenced for Olea (NW, SW directions), Corylus (NW, SW), Poaceae (SW) and Populus (NW).

Environmental Monitoring↗
PubMed2006

A 30-day-ahead forecast model for grass pollen in north London, United Kingdom.

A 30-day-ahead forecast method has been developed for grass pollen in north London. The total period of the grass pollen season is covered by eight multiple regression models, each covering a 10-day period running consecutively from 21 May to 8 August. This means that three models were used for each 30-day forecast. The forecast models were produced using grass pollen and environmental data from 1961 to 1999 and tested on data from 2000 and 2002. Model accuracy was judged in two ways: the number of times the forecast model was able to successfully predict the severity (relative to the 1961-1999 dataset as a whole) of grass pollen counts in each of the eight forecast periods on a scale of 1 to 4; the number of times the forecast model was able to predict whether grass pollen counts were higher or lower than the mean. The models achieved 62.5% accuracy in both assessment years when predicting the relative severity of grass pollen counts on a scale of 1 to 4, which equates to six of the eight 10-day periods being forecast correctly. The models attained 87.5% and 100% accuracy in 2000 and 2002, respectively, when predicting whether grass pollen counts would be higher or lower than the mean. Attempting to predict pollen counts during distinct 10-day periods throughout the grass pollen season is a novel approach. The models also employed original methodology in the use of winter averages of the North Atlantic Oscillation to forecast 10-day means of allergenic pollen counts.

Climate↗
PubMed2006

Examining Ambrosia pollen episodes at Poznań (Poland) using back-trajectory analysis.

The pollen grains of Ambrosia spp. are considered to be important aeroallergens in parts of southern and central Europe. Back-trajectories have been analysed with the aim of finding the likely sources of Ambrosia pollen grains that arrived at Poznań (Poland). Temporal variations in Ambrosia pollen at Poznań from 1995-2005 were examined in order to identify Ambrosia pollen episodes suitable for further investigation using back-trajectory analysis. The trajectories were calculated using the transport model within the Lagrangian air pollution model, ACDEP (Atmospheric Chemistry and Deposition). Analysis identified two separate populations in Ambrosia pollen episodes, those that peaked in the early morning between 4 a.m. and 8 a.m., and those that peaked in the afternoon between 2 p.m. and 6 p.m.. Six Ambrosia pollen episodes between 2001 and 2005 were examined using back-trajectory analysis. The results showed that Ambrosia pollen episodes that peaked in the early morning usually arrived at Poznań from a southerly direction after passing over southern Poland, the Czech Republic, Slovakia and Hungary, whereas air masses that brought Ambrosia pollen to Poznań during the afternoon arrived from a more easterly direction and predominantly stayed within the borders of Poland. Back-trajectory analysis has shown that there is a possibility that long-range transport brings Ambrosia pollen to Poznań from southern Poland, the Czech Republic, Slovakia and Hungary. There is also a likelihood that Ambrosia is present in Poland, as shown by the arrival of pollen during the afternoon that originated primarily from within the country.

Air↗
PubMed2000

Meteorological factors affecting daily urticaceae pollen counts in southwest Spain.

The influence of meteorological factors on daily Urticaceae pollen counts were studied in Córdoba (southwest Spain) in 1996 and 1997. The daily Urticaceae pollen concentrations were obtained by using a Hirst-type volumetric sampler, and meteorological data were obtained from the Córdoba airport, located near the sampling site. The highest correlation between pollen concentration and meteorological parameters was obtained during non-rainy seasons. Temperature was found to be the most important meteorological parameter influencing pollen counts in spring, as temperature is the main reason for the increase of pollen concentration in the atmosphere. In autumn, humidity was another important parameter influencing pollen counts. Rain, however, did not appear to be significant. The influence of the pollen concentration of the 2 previous days and the pollen concentration of the previous day has been studied. During periods with low precipitation, the pollen concentration of the previous day was a useful predictor of Urticaceae pollen concentrations for the following day.

Climate↗
PubMed2006

Ultrastructure and germination of Vitis vinifera cv. Loureiro pollen.

The cultivar Loureiro of Vitis vinifera is one of the most economically important, recommended in almost the totality of the Região Demarcada dos Vinhos Verdes. In vineyards, the grape productivity of this cultivar is normal while in others it is extremely low. The aim of this work was to study the morphology and germination of Vitis vinifera cv. Loureiro pollen with high and low productivity. The pollen grain was examined under light, transmission and scanning electron microscopy. Typically V. vinifera pollen present three furrows but in the cultivar Loureiro we found tricolporated and acolporated (without furrows or pores) pollen grains. Both pollen types present generative and vegetative cells with the usual aspect and a dense cytoplasm rich in organelles. In the acolporated pollen a continuous exine layer and an irregular intine layer were observed. Differences were found in the starch accumulation, since only in tricolporated pollen abundant plastids filled with numerous starch granules were observed. To determine the causes of the low productivity of this cultivar we tested pollen viability by the fluorochromatic reaction and pollen germinability by in vitro assays. We observed that the acolporated pollen grain is viable, but no germination was recorded.

Cell Survival↗
PubMed2006

Promoter activity of a putative pollen monosaccharide transporter in Petunia hybrida and characterisation of a transposon insertion mutant.

For the growth of the male reproductive cells of plants, the pollen, the presence of sufficient sucrose or monosaccharides is of vital importance. From Petunia hybrida a pollen-specific putative monosaccharide transporter designated PMT1 (for petunia monosaccharide transporter) has been identified previously. The present work provides an in-depth analysis and characterisation of PMT1 in the context of pollen development with the GUS reporter gene and an insertion mutant. The promoter of the pollen-specific putative PMT1 gene has been isolated by inverse PCR and sequenced. Analysis of plants transformed with the promoter-GUS fusion confirmed the specificity of this gene, belonging to the late pollen-specific expressed genes. GUS activity was detected even after 24 h of in vitro pollen germination, at the pollen tube tip. To elucidate the importance of PMT1 for gametophyte development and fertilisation, we isolated a mutant plant containing a transposon insertion in the PMT1 gene by the dTph1 transposon-tagging PCR-based assay. The PMT1 mutant contained a dTph1 insertion in position 1474 bp of the transcribing part of the gene, before the last two transmembrane-spanning domains. Analysis of the progeny of the heterozygous mutant after selfing revealed no alterations in pollen viability and fertility. Mature pollen grains of a plant homozygous for the transposon insertion were able to germinate in vitro in a medium containing sucrose, glucose, or fructose, which indicates that PMT1 is not essential for pollen survival. Several explanations for these results are discussed in the present work.

Amino Acid Sequence↗
PubMed2003

Distyly and pollen dimorphism in Damnacanthus (Rubiaceae).

We document distyly in Damnacanthus (Rubiaceae) and investigate pollen morphology in the genus. Within nine taxa in Japan (four species, four varieties and one form), five show distyly. Non-distylous taxa of Damnacanthus always have a long style and short stamens. We examined the pollen morphology of all taxa of Damnacanthus in Japan. The pollen grains of Damnacanthus are 3- to 5-orthocolporate or sometimes loxocolporate. Polymorphic pollen is sometimes observed even within a single flower. Pollen dimorphism is associated with distyly; pollen grains from short-styled (thrum) flowers are significantly larger than those from long-styled (pin) flowers, and the muri are smooth in pin flowers whereas the pollen grains of thrum flowers have minute granules on the top and/or on the sides. In non-distylous taxa, in which the style is longer than stamens, the muri are always provided with minute granules. Interestingly, the pollen grains with granules in the non-distylous taxa contrast with the pollen of the distylous taxa, because granulate pollen grains are found only in thrum flowers. Although Damnacanthus is a stenopalynous genus, the number of colpi is useful for delimiting taxa. Based on palynological characters, we conclude that Damnacanthus should be revised.

Pollen↗