A comprehensive aerobiological study of the airborne pollen in the Irish environment

Respiratory allergies triggered by pollen allergens represent a significant health concern to the Irish public. Up to now, Ireland has largely refrained from participating in long-term aerobiological studies. Recently, pollen monitoring has commenced in several sampling locations around Ireland. The first results of the pollen monitoring campaigns for Dublin (urban) and Carlow (rural) concerning the period 2017–2019 and 2018–2019, respectively, are presented herein. Additional unpublished pollen data from 1978–1980 and, 2010–2011 were also incorporated in creating the first pollen calendar for Dublin. During the monitoring period over 60 pollen types were identified with an average Annual Pollen Integral (APIn) of 32,217 Pollen × day/m³ for Dublin and 78,411 Pollen × day/m³ for Carlow. The most prevalent pollen types in Dublin were: Poaceae (32%), Urticaceae (29%), Cupressaceae/Taxaceae (11%), Betula (10%), Quercus (4%), Pinus (3%), Fraxinus (2%), Alnus (2%) and Platanus (1%). The predominant pollen types in Carlow were identified as Poaceae (70%), Urticaceae (12%), Betula (10%), Quercus (2%), Fraxinus (1%) and Pinus (1%). These prevalent pollen types increased in annual pollen concentration in both locations from 2018 to 2019 except for Fraxinus. Although higher pollen concentrations were observed for the Carlow (rural) site a greater variety of pollen types were identified for the Dublin (urban) site. The general annual trend in the pollen season began with the release of tree pollen in early spring, followed by the release of grass and herbaceous pollen which dominated the summer months with the annual pollen season coming to an end in October. This behaviour was illustrated for 21 different pollen types in the Dublin pollen calendar. The correlation between ambient pollen concentration and meteorological parameters was also examined and differed greatly depending on the location and study year. A striking feature was a substantial fraction of the recorded pollen sampled in Dublin did not correlate with the prevailing wind directions. However, using non-parametric wind regression, specific source regions could be determined such as Alnus originating from the Southeast, Betula originating from the East and Poaceae originating from the Southwest.

     

Growth and development of conifer pollen tubes

Conifer pollen tubes are an important but underused experimental system in plant biology. They represent a major evolutionary step in male gametophyte development as an intermediate form between the haustorial pollen tubes of cycads and Ginkgo and the structurally reduced and faster growing pollen tubes of flowering plants. Conifer pollen grains are available in large quantities, most can be stored for several years, and they grow very well in culture. The study of pollen tube growth and development furthers our understanding of conifer reproduction and contributes towards our ability to improve on their productivity. This review covers taxonomy and morphology to cell, developmental, and molecular biology. It explores recent advances in research on conifer pollen and pollen tubes in vivo, focusing on pollen wall structure, male gametophyte development within the pollen wall, pollination mechanisms, pollen tube growth and development, and programmed cell death. It also explores recent research in vitro, including the cellular mechanisms underlying pollen tube elongation, in vitro fertilization, genetic transformation and gene expression, and pine pollen tube proteomics. With the ongoing sequencing of the Pinus taeda genome in several labs, we expect the use of conifer pollen tubes as an experimental system to increase in the next decade.     

REPRODUCTIVE COSTS OF SELF-POLLINATION IN IPOMOPSIS AGGREGATA (POLEMONIACEAE): ARE OVULES USURPED?

In common with many cosexual angiosperms, the hummingbird-pollinated montane herb Ipomopsis aggregata (Polemoniaceae) is likely to experience self-pollination. Does this incur a fecundity cost even in such a highly self-sterile (presumably self-incompatible) species? Histological studies showed that self pollen germinates, and its tubes penetrate ovules almost as often as those of outcross pollen. Ovules penetrated by self tubes are especially likely to show milky callose occlusion 24–48 hr after pollination, and several observations suggest an association between occlusion and degeneration. Compared to flowers receiving only outcross pollen, seed set was reduced by 42% on average when self pollen was applied along with outcross, either by hand (in emasculated flowers) or by natural autodeposition (in unemasculated flowers). Reductions were statistically indistinguishable whether self pollen was applied 9 hr before outcross pollen, or at the same time. Unemasculated flowers accumulated substantial self pollen loads, and this autodeposition persisted when flowers were probed to mimic hummingbird visitation. Geitonogamy also is substantial, judging from field estimates of pollen transfer. Thus natural self pollen deposition may be sufficient to “usurp” ovules that otherwise could mature. In this light, late-acting self-rejection in I. aggregata seems decidedly less efficient than an early-acting system that would block pollen germination or tube growth.     

CONTRIBUTION OF CRYPTIC INCOMPATIBILITY TO THE MATING SYSTEM OF EICHHORNIA PANICULA TA (PONTEDERIACEAE)

Tristylous populations of the annual aquatic Eichhornia paniculata have high levels of outcrossing and intermorph mating despite being fully self- and intramorph compatible. Experimental studies of pollen germination, ???pollen-tube growth, and pollinations with mixtures of genetically marked pollen were used to determine whether postpollination processes contribute to the observed mating patterns. Differences in pollen germination were small and did not contribute to differences in pollen siring ability. The fraction of pollen tubes first entering the ovary, however, was greater for legitimate outcross pollen than for either of the other two pollen types (self or outcross illegitimate pollen) in all recipient morphs. Moreover, legitimate pollen had higher siring success when in competition with illegitimate pollen types (self or outcross) in each recipient style morph. The ranking of pollen performance for different pollen-style combinations was the same for both the pollen-tube growth and marker-gene experiments indicating that differences in pollen-tube growth rate are the principal cause of differences in pollen siring ability. Cryptic incompatibility in E. paniculata may represent a weak heteromorphic incompatibility system because the observed patterns of pollen-tube growth parallel pollen-tube growth and seed-set patterns that occur in related species with strong trimorphic incompatibility. The ability to produce mostly outcrossed progeny when pollinators are abundant, but to reliably produce seed under a variety of environmental and demographic conditions may be favored in E. paniculata because of its colonizing life history and occurrence in ephemeral habitats. Cryptic incompatibility may be more likely to occur in species subject to wide fluctuations in population size and levels of pollinator service.     

Airborne Grass Pollen Antigens in a Grassland as Studied by Immunoblotting with Anti-Lol p I Antibody

Airborne grass pollen in a grassland was measured using a Burkard air sampler and immunoblotting. The pollen antigens collected on the tape of the air sampler were transferred onto a nitrocellulose membrane, and visualized as purple spots by immunoenzymatic staining with anti-Lol p I antibody. These spots were quantified automatically using an image processing system. The airborne grass pollen antigens in the grassland where four species of grass were growing were prevalent mainly from evening to early morning during the first half of the pollen season, but in the daytime during the latter half of the season. The antigen dispersion time was confined to a few hours of the day. The flowering time of several grass species which were hydroponically cultured in the laboratory were also studied. Each species had a different flowering time. We believe that the time when an antigen is present during the day is related to the different flowering times of the various grass species growing in the grassland.     

Endogenous polyamine content during in vivo maturation and in vitro culture of maize pollen

Changes in polyamine content during in vivo maturation and in vitro culture of maize (Zea mays L.) pollen were studied. The endogenous content of free, conjugated and bound polyamines was analyzed during 30 days of pollen evolution, in both developmental pathways (microsporogenesis and androgenesis). The induction of androgenesis from cold-pretreated uninucleate pollen results, in most of cases, in a lower total polyamine content than that of the in vivo uninucleate pollen. These differences indicate that polyamine metabolism is altered during the induction of androgenesis, and this could be a consequence of increased polyamine assimilation. In general, pollen stages that involve cell division (tetrades, pre-anthesis pollen and four-day cultured pollen) are characterized by a predominance of free Spd. The increase of Spd and Spm in 15-day cultured pollen, when the first embryoids are formed, outline the possible implication of these polyamines in embryogenetic processes. Furthermore, these findings may contribute to the improvement of maize androgenesis yield, especially in recalcitrant genotypes, by the exogenous application of polyamines or polyamine-inhibitors to the culture medium.Abbreviations PAs polyamines - Put putrescine - Spd spermidine - Spm spermine - S free polyamine fraction - SH conjugated polyamine fraction - PH bound polyamine fraction     

Aerobiology, allergenicity and biochemistry of three pollen types in Berhampore town of West Bengal, India

An aerobiological survey was performed in Berhampore town of West Bengal, India, to know the frequency of three common airborne pollen, namely Acacia auriculiformis, Eucalyptus citriodora and Madhuca indica using an ASTIR one day volumetric sampler. Acacia pollen showed its peak concentration in September, followed by Madhuca in April, while Eucalyptus showed its two peaks between September–October and January–April. Meteorological factors like temperature, RH, rainfall played an important role in release and dispersal of pollen. Skin prick tests with the antigenic extracts of the three pollen types, showed their allergenic potentialities. The highest markedly positive reactions were exhibited by Eucalyptus (34.04%), followed by Madhuca (22.93%) and Acacia (21.87%). 30–60% (NH₄)₂ SO₄ cut fraction (Fraction II) of each pollen type showed maximum positivity in skin prick test. Biochemical analysis showed that Acacia pollen was richer in protein and carbohydrate, than the other two types. The total protein component of the above types were studied by SDS-PAGE showing different protein bands with a range of molecular weight 29–110 KD. In isolated fraction II (allergenically most potent) of Eucalyptus and Madhuca different protein band of 43–200 KD were obtained, while a single protein band of 57 KD was obtained for Acacia. The IgE specific allergenic reactivity was confirmed by Dot-blotting technique. This revised version was published online in June 2006 with corrections to the Cover Date.     

Pollen viability and transgene expression following storage in honey

Transgenic plants of tobacco andArabidopsis that produce genetically marked pollen, expressing the reporter geneuidA (gusA), were generated to determine whether pollen proteins can be expressed and stable in honey, a potential route by which foreign proteins might enter the wider environment. Hydrated tobacco pollen was found to lose viability rapidly in honey, while pollen in the natural dehydrated form remained viable for at least several days and in some cases several weeks, as determined by FDA staining activity and germinability. Dehydrated pollen was found to be capable of transient foreign gene expression, following microprojectile bombardment, after incubation in honey for at least 120 h. PCR amplification of transgene sequences in pollen of transgenic plants revealed that pollen DNA can remain relatively intact after 7 weeks in honey. GUS enzyme activity analysis and SDS-PAGE of pollen proteins revealed that foreign and native pollen proteins are stable in pollen incubated in honey for at least 6 weeks. We conclude that pollen may represent an ecologically important vector for transgenic protein products.     

Molecular and physiological characterisation of a 14-3-3 protein from lily pollen grains regulating the activity of the plasma membrane H+ ATPase during pollen grain germination and tube growth

A 14-3-3 protein has been cloned and sequenced from a cDNA library constructed from mRNAs of mature pollen grains of Lilium longiflorum Thunb. Monoclonal antibodies (MUP 5 or MUP 15) highly specific against 14-3-3 proteins recognised a 30-kDa protein in the cytoplasmic fraction of many various lily tissues (leaves, bulbs, stems, anther filaments, pollen grains, stigmas) and in other plants (Arabidopsis seedlings, barley recombinant 14-3-3). In addition, 14-3-3 proteins were detected in a microsomal fraction isolated from pollen grains and tubes, and the amount of membrane-bound 14-3-3 proteins as well as the amount of the plasma membrane (PM) H⁺ ATPase increased during germination of pollen grains and tube growth. No change was observed in the cytoplasmic fraction. A further increase in the amount of 14-3-3 proteins in the microsomal fraction was observed when pollen grains were incubated in germination medium containing 1 μM fusicoccin (FC) whereas the number of 14-3-3s in the cytoplasmic fraction decreased. Fusicoccin also protected membrane-bound 14-3-3 proteins from dissociation after washing with the chaotropic salt KI. Furthermore, FC stimulated the PM H⁺ ATPase activity, the germination frequency and the growth rate of pollen tubes, thus indicating that a modulation of the PM H⁺ ATPase activity by interaction with 14-3-3 proteins may regulate germination and tube growth of lily pollen. Received: 20 June 2000 / Accepted: 2 October 2000     

Diurnal patterns of pollen emission in Ambrosia, Phleum, Zea, and Ricinus

Hourly measurements of pollen emission were made from cultivated plots of Ambrosia, Phleum, Zea, and Ricinus over the course of several pollination seasons as part of a study of pollen dispersion from known sources. A characteristic diurnal emission pattern was found for each genus. Ambrosia pollen emission normally begins an hour or two after sunrise, peaks a few hours later, and decreases through the afternoon. Phleum starts during the night, peaks about 2 hr after sunrise, and declines slowly through the day. Zea emits pollen fairly uniformly during the period from 2 hr after sunrise to about sunset, while Ricinus pollen was collected from several hours after sunrise to late afternoon with a peak in mid-morning. Daily patterns often vary from the seasonal mean in response to changing meteorological conditions.     

A new technique for monitoring pollen flow in orchids

Summary The orchid Prasophyllum fimbria is pollinated by nectar-feeding native bees and wasps. The pollinia are patially separated from the viscidium by a stipe so that pollinia can be labelled with coloured histochemical stains without interfering with pollinarium removal. Pollen flow was monitored by following the movement of the coloured pollen in several populations of P. fimbria in Western Australia. Statistical analysis confirmed that pollen labelling did not interfere with pollinarium removal or subsequent pollination of the labelled flower. Fifty eight labelled pollinaria were removed by vectors from 16 test spikes, with a total of 125 flowers on 47 spikes receiving labelled pollen. An average of 2 flowers received pollen for every pollinium removed but up to 6 flowers received pollen from a single collinium. No significant differences between mean vector flights and pollen flow distances were detected. On average, geitonogamous transfers only accounted for 22% of all pollinations. This is a simple and inexpensive technique for the direct labelling of pollen with minimal disruption to the pollination system and may have applications in other plant families.     

THE CONTRIBUTION OF MATING SYSTEM VARIATION TO REPRODUCTIVE ISOLATION IN TWO CLOSELY RELATED CENTAURIUM SPECIES (GENTIANACEAE) WITH A GENERALIZED FLOWER MORPHOLOGY

In closely related plant species that display strong similarities in phenology and pollinator communities, differences in breeding system and associated shifts in floral traits may have important effects on the magnitude and direction of heterospecific pollen flow and hybridization. Here, we quantified the strength of several pre‐ and postzygotic barriers acting between the facultatively outcrossing Centaurium erythraea and the predominantly selfing C. littorale via a suite of experiments, and estimated the frequency of hybridization in the field using molecular markers. The reproductive barriers primarily responsible for preventing hybridization were essentially prezygotic and these acted asymmetrically. Due to differences in floral display, pollen production, and pollen transfer rates, heterospecific pollen flow occurred predominantly from C. erythraea to C. littorale. In C. littorale, on the other hand, close anther–stigma positioning and resulting higher capacity for autonomous selfing functioned as an efficient barrier to counterbalance the higher risk for hybrid mating. In both species the action of all reproductive barriers resulted in a small opportunity for hybrid establishment, which was confirmed by the occurrence of only ～1% putative hybrids in the field. Our findings confirm that differences in breeding system affect heterospecific pollen transfer patterns and that autonomous selfing may efficiently prevent hybridization.     

Molecular cloning,functional expression and characterization of two serine/threonine-specific protein kinases from<Emphasis Type="Italic"> Nicotiana tabacum</Emphasis> pollen

The existence of different kinds of kinases in pollen and pollen tubes suggests that kinase-mediated signaling pathways are likely involved in regulating pollen germination and pollen tube growth during the life cycle of higher plants. We have used RT-PCR and RACE to isolate full-length cDNAs for two pollen-expressed kinases, named NtPK1 and NtPK2, of Nicotiana tabacum. NtPK1 and NtPK2 encode proteins of 365 and 369 amino acids with calculated molecular masses of 39.2 kDa and 39.5 kDa, respectively, and both proteins possess the 12 sub-domains that are conserved among protein kinases. The nucleotide and deduced amino acid sequences of NtPK1 and NtPK2 share 88% and 91% identity, respectively, with the C-terminal region being the most conserved. RT-PCR analysis revealed that NtPK1 was specifically expressed in pollen and pollen tubes, and that NtPK2 was also expressed in pistil and petal. Immunoblot analysis using anti-NtPK1 and anti-NtPK2 antibodies confirmed that both NtPK1 and NtPK2 were produced in pollen and pollen tubes, and that NtPK2 was also produced in developing male gametophytes and other floral tissues. Biochemical fractionation experiments showed that, in all the tissues examined, NtPK1 and NtPK2 were present in the cytosolic fraction and not in the microsomal fraction. NtPK1 and NtPK2 were found to autophosphorylate on threonine and, for NtPK2, on serine as well. All the results taken together suggest that NtPK1 and NtPK2 are novel receptor-like cytosolic serine/threonine kinases, and could mediate signaling pathways required for pollen germination and/or pollen tube growth.The nucleotide sequence data of NtPK1 and NtPK2 reported in this paper will appear in the EMBL/GenBank/DDBJ nucleotide sequence databases under the accession numbers AJ608156 and AJ608157, respectively     

BUMBLE BEE POLLINATION AND FLORAL MORPHOLOGY: FACTORS INFLUENCING POLLEN DISPERSAL IN THE ALPINE SKY PILOT,POLEMONIUM VISCOSUM (POLEMONIACEAE)

Pollen dispersal success in entomophilous plants is influenced by the amount of pollen produced per flower, the fraction of pollen that is exported to other flowers during a pollinator visit, visitation frequency, and the complementarity between pollen donor and recipients. For bumble bee-pollinated Polemonium viscosum the first three determinants of male function are correlated with morphometric floral traits. Pollen production is positively related to corolla and style length, whereas pollen removal per visit by bumble bee pollinators is a positive function of corolla flare. Larger-flowered plants receive more bumble bee visits than small-flowered individuals. We found no evidence of tradeoffs between pollen export efficiency and per visit accumulation of outcross pollen; each was influenced by unique aspects of flower morphology. Individual queen bumble bees of the principal pollinator species, Bombus kirbyellus, were similar in male, female, and absolute measures of pollination effectiveness. An estimated 2.9% of the pollen that bumble bees removed from flowers during a foraging bout was, on average, deposited on stigmas of compatible recipients. Significant plant-to-plant differences in pollen production, pollen export per visit, and outcross pollen receipt were found for co-occurring individuals of P. viscosum indicating that variation in these fitness related traits can be seen by pollinator-mediated selection.     

Immunolocalization of the cell wall components inPinus densiflora pollen

Summary In order to compare cell wall formation in gymnosperm pollen with that in angiosperm pollen, the distribution of cell wall constituents in the pollen grain and pollen tube ofPinus densiflora was studied immunocytochemically with monoclonal antibodies JIM 5 (against non- or poorly esterified pectin), JIM 7 (against highly esterified pectin), JIM 13 (against arabinogalactan proteins, AGPs), and LM 2 (against AGPs containing glucuronic acid). In the pollen grain wall, only the outer layer of the intine was labeled with JIM 5 and weakly with JIM 7. The tube wall was scarcely labeled with JIM 5 and very weakly labeled with JIM 7. In contrast, the whole of both the intine and the tube wall was strongly labeled with JIM 13 and LM 2, and the generative-cell wall was also labeled only with LM 2. The hemicellulose B fraction, which is the main polysaccharide fraction from the pollen tube wall, reacted strongly with JIM 13 and especially LM 2, but not with antipectin antibodies. These results demonstrate that the wall constituents and their localization inP. densiflora pollen are considerably different from those reported in angiosperm pollen and suggest that the main components of the cell wall ofP. densiflora pollen are arabinogalactan and AGPs containing glucuronic acid.Abbreviations AGPs arabinogalactan proteins - ELISA enzymelinked immunosorbent assay - MAbs monoclonal antibodies     

Enzymatic, isoelectric, and molecular-weight characterization of water-soluble maize-pollen proteins

Maize (Zea mays L. cv. Kansas Drought Synthetic) pollen proteins were fractionated via a continuous tandem cascade ultrafiltration system into three molecular-weight classes. Proteins from the whole extract and each fraction were further separated by polyacrylamide gel electrofocusing in the pH 5 to 9 range and characterized by their enzymatic activity and isoelectric point. Twenty-one protein bands were detected with Fast Green FCF staining. Esterase, peroxidase, leucine aminopeptidase, catalase, and amylase activities were located in 16 bands. Five bands remained unclassified. All the enzymes assayed, except amylase, were polymorphic. The extractions, fractionations and zymograms were reproducible and indicated that several protein bands contained at least two different enzymes with very similar molecular weight class and isoelectric point for each enzyme and protein band     

Effect of genome-plastome interaction on meiosis and pollen development in Oenothera species and hybrids

 Oenothera villaricae Dietrich and Oe. picensis Dietrich, complete translocation heterozygotes, are fully interfertile, giving rise to six discrete classes of true-breeding hybrids from a reciprocal cross. Associated with each parent and hybrid is a characteristic abortive non-staining pollen fraction easily distinguished from fully developed pollen under the light microscope. Pollen abortion has been associated with translocation rings in other angiosperm species, and may characterize such systems. The abortive pollen fraction is significantly different between reciprocal Oenothera hybrids, however (P<0.001), indicative of partial cytoplasmic control. Pollen abortion is most severe in the F₁ hybrid generation, and ameliorates with successive generations of hybrid self-fertilization. Three-way analysis of variance shows significant effects on pollen stainability (a measure of the non-abortive fraction) for nucleus, cytoplasm and selfed hybrid generation, individually or in combinations. This result suggests a combined nucleocytoplasmic basis for the pollen abortion. Correlated with the observation of increased pollen abortion in Oenothera hybrids are meiotic findings of broken chromosome rings (chains, univalents), asymmetric anaphase chromosome distributions and trinucleate tetrads. To test the hypothesis that such anomalous meiotic events play a role in the mechanism of pollen abortion, meiotic disjunction frequency was determined for each parent, F₁ and F₉ selfed hybrid accessions. Three-way analysis of variance shows levels of significance comparable to those noted for pollen stainability (P<0.001) for effects of nucleus, cytoplasm and selfed hybrid generation on disjunction frequency. A high degree of correlation (r ²=0.984) is noted between disjunction frequency and pollen stainability. We conclude that the abortive pollen grains are indeed the products of nondisjunctional meiotic events, which themselves are consequences of hybrid nucleocytoplasmic incompatibility. Received: 28 May 1997 / Revision accepted: 23 July 1997     

Comparative studies of pollen and fluorescent dye transport by bumble bees visiting Erythronium grandiflorum

Summary In the Colorado Rocky Mountains the glacier lily Erythronium grandiflorum exhibits a striking dimorphism in pollen color and is commonly pollinated by the bumble bee Bombus occidentalis. We induced bees to visit sequences of flowers in a flight cage, and compared dispersal of distinctively-colored pollen and fluorescent pigment (dye) that the bee had picked up at a single donor flower. Nonparametric and parametric analyses showed that dispersal properties of pollen and dye differed; consistently less pollen was deposited and it was carried consistently shorter distances than dye. Dye thus does not provide an accurate means of assessing exacty where or how far pollen travels in this plant-pollinator system. On the other hand, both pollen and dye responded similarly to several experimental manipulations of donor and recipient flowers. Hence dye may well be of value for a qualitative investigation of how floral traits influence pollen dispersal.     

Release of proteins from the pollen wall of Linum grandiflorum

Summary The emission of proteins from the pollen wall of Linum grandiflorum stained with Coomassie blue was followed directly in moistened grains as well as in pollen prints. Within the first minute of the grain being moistened exine-borne proteins emerged from both inter-apertural and apertural sites; subsequently, proteins of a different nature were discharged from the apertures only. In a fraction of the grains the release of intine proteins was not preceded by that of exine proteins. Pin and thrum pollen did not differ in terms of mode or site of this protein emission. The presence and emergence of exine proteins from the apertures is explained by the process of infolding of the colpal wall at desiccation and its expansion at rehydration, which causes an initial trapping and subsequent re-exposure of surface materials. This explanation may also account for the occurrence of poral sporophytic proteins in the pollens of many dictoyledons.