Taphonomic features of Arctic pollen

The composition of the primary pollen changes during pollen transfer and redistribution over the Earths surface and fossilization. These processes are particularly pronounced in the tundra zone. Patterns of pollen spectra formation in the subaerial and subaqueous deposits of the tundra zone were considered and the differences in the content of palynomorphs with exine rupture were revealed. The changes in the effect of regional and local taphonomic factors on Arctic tundra slopes were traced. Pollen conservation in the deposits of the tundra zone, snowfields, and wedge ice was considered.Translated from Izvestiya Akademii Nauk, Seriya Biologicheskaya, No. 2, 2005, pp. 240–252.Original Russian Text Copyright © 2005 by Vasilchuk.     

Airborne pollen dispersal and survival on mount sutton (Canada)

Summary In order to find whether or not a pattern exist in both pollen concentration and viability along altitudinal transects, samples were collected volumetrically (VPPS) each 25 m, from 500 to 825 m, on Mount Sutton (45°04N; 72°32W) (970 m). Both minimum concentration and minimum viability were found at 725 m. Airborne pollen viability was species dependent, while airborne concentrations were not specific. Sampling height influence was also investigated volumetrically (BPS), by comparing paired samples at ground and 10 m levels. Again, pollen concentration pattern was found quite stable, while viability was found to be more height influenced, particularly at 725 m. The 725 m hinge altitude is located just above the June Mean Cloud Base altitude (657 m).     

Modern pollen and stomate deposition in lake surface sediments from across the treeline on the Kola Peninsula, Russia

We sampled and analyzed surface sediments from 31 lakes along a latitudinal transect crossing the coniferous treeline on the Kola Peninsula, Russia. The major vegetation zones along the transect were tundra, birch-forest tundra, pine-forest tundra, and forest. The results indicate that the major vegetation types in our study area have distinct pollen spectra. Sum-of-squares cluster analysis and principal components analysis (PCA) groupings of pollen sites correspond to the major vegetation zones. PCA ordination of taxa indicates that the first axis separates taxa typical of the forest zone (Pinus, Picea) from taxa typical of tundra and forest-tundra zones (Polypodiaceae, Ericaceae, and Betula). The current position of the coniferous treeline, defined in our region by Pinus sylvestris, occurs roughly where Pinus pollen values reach 35% or greater. Arboreal pollen (AP)/non-arboreal pollen (NAP) ratios were calculated for each site and plotted against geographic distance along the transect. AP/NAP ratios of 7 or greater are found within pine-forest tundra and forest vegetation zones. Pinus stomates (dispersed stomatal guard cells) are absent from sites north of the coniferous treeline and all but two samples from the forested sites contain stomates. Stomate concentrations among the samples are highly variable and range from 10 to 458 per ml and positively correlate with the changing Pinus pollen values.     

The partial contribution of specific airborne pollen to pollen induced allergy

The air that we inhale contains simultaneously a multiple array of allergenic pollen. It is well known that such allergens cause allergic reactions in some 15 of the population of the Western World. However little is known about the quantitative aspect of this phenomenon. What is the lowest concentration of pollen that might trigger allergic responses? As people are exposed to heterogeneous and variable environments, clarification of the partial contribution of each of the major airborne pollen allergens and determination of its role in invoking allergy are of prime importance. Objectives: (1) Assessment of a possible correlation between the concentration of airborne pollen and incidence of allergy. (2) Estimation of the lowest average concentrations for various species of airborne pollen that elicit allergic symptoms when exceeded. (3) Determination of the extent of the variations in manifestation of allergy symptoms that can be explained by fluctuations in the concentration of individual species of airborne pollen. Methods: The study was conducted during 14months with a rural population in Israel. The participants completed a detailed questionnaire and were skin prick tested with the common airborne allergens. The appearance of clinical symptoms, i.e. nasal, bronchial, ocular or dermal, were reported daily by the patients. Concentrations of the airborne pollen and spores were monitored in the center of activity of the residents during one day every week, using three Rotorod pollen traps. The pollen grains were identified by light microscopy. Results: The pollen spectrum was divided into time-blocks presenting the main pollination periods of the investigated species. The correlation between the concentration of airborne pollen of the relevant species and the clinical symptoms of the patients was determined for each time block. The correlation differed for different clinical symptoms and for different pollen allergens. Highest correlation with airborne pollen counts was found for patients with nasal and bronchial symptoms. The onset of the clinical symptoms by sensitive patients started, in each of the relevant groups, once the weekly average concentration of the airborne pollen crossed a threshold level. Under the limitations of the present study, this level was estimated to be 2–4 pollen m^–3 air for olive, 3–5 pollen m^–3 air for grasses, 4–5 pollen m^–3 air for Artemisia, 10–20 pollen m^–3 air for pecan and 50–60 pollen m^–3 air for cypress. Conclusions: Fluctuations in specific airborne pollen grains explained up to 2/3 of the variation in clinical allergy responses. Those were: 69 of the variation for cypress (March–April), 66 for the grasses (March–April), 49 for the pecan (May–June) and 62 for Artemisia (Autumn).     

Viability and sunlight sensitivity of oak pollen and its implications for pollen-mediated gene flow

Pollen-mediated gene flow and the male reproductive success of wind-pollinated trees depend on the initial viability of the pollen and the changes that occur in its viability during transport in the atmosphere. The viability of Quercus robur pollen was determined before and during exposure to sunlight by in vitro germination and the fluorescein diacetate reaction (FCR) in 2002 and 2003, respectively. These experiments allowed us to calculate initial pollen viability and pollen sensitivity to sunlight. The germination test revealed a lower initial pollen viability (25–65%) than the FCR (53–92%). Following 9.5 h of irradiation the viability was reduced to 75–100% as determined by the in vitro germination test or to 40–70% as determined by the FCR. The actual values of initial pollen viability and pollen sensitivity to sunlight were used to define a range of values for modelling pollen dispersal using the mesoscale meteorological model METRAS. The deposition patterns of viable pollen varied by as much as a factor of 14 by changing the viability parameters in the range of the observed values. This suggests significant differences in male reproductive success. Variations in initial pollen viability have stronger effects on the gene-flow pattern than do variations in pollen sensitivity to sunlight. In particular, pollen distribution throughout the local environment is shaped by the initial pollen viability, while pollen sensitivity to sunlight mainly influences long-distance pollen dispersal.This revised version was published online in February 2005 with corrections to an incorrect sentence in the Results section. Under Pollen Sensitivity it should read: decreased faster when assessed by the germination test than by the FCR.     

Representation of tundra vegetation by pollen in lake sediments of northern Alaska

Aim To understand better the representation of arctic tundra vegetation by pollen data, we analysed pollen assemblages and pollen accumulation rates (PARs) in the surface sediments of lakes. Location Modern sediment samples were collected from seventy‐eight lakes located in the Arctic Foothills and Arctic Coastal Plain regions of northern Alaska. Methods For seventy of the lakes, we analysed pollen and spores in the upper 2 cm of the sediment and calculated the relative abundance of each taxon (pollen percentages). For eleven of the lakes, we used ²¹⁰Pb analysis to determine sediment accumulation rates, and analysed pollen in the upper 10–15 cm of the sediment to estimate modern PARs. Using a detailed land‐cover map of northern Alaska, we assigned each study site to one of five tundra types: moist dwarf‐shrub tussock‐graminoid tundra (DST), moist graminoid prostrate‐shrub tundra (PST) (coastal and inland types), low‐shrub tundra (LST) and wet graminoid tundra (WGT). Results Mapped pollen percentages and multivariate comparison of the pollen data using discriminant analysis show that pollen assemblages vary along the main north–south vegetational and climatic gradients. On the Arctic Coastal Plain where climate is cold and dry, graminoid‐dominated PST and WGT sites were characterized by high percentages of Cyperaceae and Poaceae pollen. In the Arctic Foothills where climate is warmer and wetter, shrub‐dominated DST, PST and LST were characterized by high percentages of Alnus and Betula pollen. Small‐scale variations in tundra vegetation related to edaphic variability are also represented by the pollen data. Discriminant analysis demonstrated that DST sites could be distinguished from foothills PST sites based on their higher percentages of Ericales and Rubus chamaemorus pollen, and coastal PST sites could be distinguished from WGT sites based on their higher percentages of Artemisia. PARs appear to reflect variations in overall vegetation cover, although the small number of samples limits our understanding of these patterns. For coastal sites, PARs were higher for PST than WGT, whereas in the Arctic Foothills, PARs were highest in LST, intermediate in DST, and lowest in PST. Main conclusion Modern pollen data from northern Alaska reflect patterns of tundra vegetation related to both regional‐scale climatic gradients and landscape‐scale edaphic heterogeneity.     

Analysis of cedar pollen time series: no evidence of low-dimensional chaotic behavior

Much of the current interest in pollen time series analysis is motivated by the possibility that pollen series arise from low-dimensional chaotic systems. If this is the case, short-range prediction using nonlinear modeling is justified and would produce high-quality forecasts that could be useful in providing pollen alerts to allergy sufferers. To date, contradictory reports about the characterization of the dynamics of pollen series can be found in the literature. Pollen series have been alternatively described as featuring and not featuring deterministic chaotic behavior. We showed that the choice of test for detection of deterministic chaos in pollen series is difficult because pollen series exhibit power spectra. This is a characteristic that is also produced by colored noise series, which mimic deterministic chaos in most tests. We proposed to apply the Ikeguchi–Aihara test to properly detect the presence of deterministic chaos in pollen series. We examined the dynamics of cedar (Cryptomeria japonica) hourly pollen series by means of the Ikeguchi–Aihara test and concluded that these pollen series cannot be described as low-dimensional deterministic chaos. Therefore, the application of low-dimensional chaotic deterministic models to the prediction of short-range pollen concentration will not result in high-accuracy pollen forecasts even though these models may provide useful forecasts for certain applications. We believe that our conclusion can be generalized to pollen series from other wind-pollinated plant species, as wind speed, the forcing parameter of the pollen emission and transport, is best described as a nondeterministic series that originates in the high dimensionality of the atmosphere.     

Entrapment of long-distance transported pollen grains by various moss species in coastal Victoria Land,Antarctica

Summary In northern Victoria Land (continental Antarctica, between 72° and 76°S, 162° and 169°E), 18 moss samples have been collected and analysed for the presence of pollen. In turfs and cushions of 8 different moss species, at least 27 pollen taxa could be identified. The pinus-type pollen and those of grasses were very common. More than 60% of the total grains were damaged or could not be identified. There is evidence that the Antarctic continent could act as a sink for wind-transported pollen from sub-Antarctic islands or from plants (native or cultivated) in South America, Australia, New Zealand and South Africa. However, the pollen concentration in air ( 1 pollen grain/100 m³) and its entrapment rate on moss (about 0.12 grain/cm²/year) result in a very low pollen density in these plants.     

An efficient method for flow cytometric analysis of pollen and detection of 2n nuclei in Brassica napus pollen

A simple and reliable method was developed for isolating pollen nuclei from Brassica napus and Triticum aestivum for DNA analysis using flow cytometry. The nuclei were released from pollen by ultrasonic treatment. The isolated nuclei following filtration through nylon mesh and a purification procedure were suitable for flow cytometric analysis as well as for isolating genomic DNA. Ultrasonic treatment time was optimized for B. napus pollen at different developmental stages. The method is effective and suitable for the preparation of many samples. We analyzed the nuclear DNA levels in pollen of B. napus at three major developmental stages as well as in mature wheat pollen. Only a single 1C peak representing the haploid DNA level was detected in the nuclei isolated from Brassica uninucleate microspores as well as in mature Triticum pollen. Interestingly, diploid nuclei were detected in both binucleate and mature pollen of B. napus. The possible origins of the diploid nuclei are discussed.Abbreviations DAPI 4,6-Diamidino-2-phenylindole - NIB Nuclear isolation buffer     

Modern pollen studies in the territory of Sagalassos (Southwest Turkey) and their use in the interpretation of a Late Holocene pollen diagram

Modern pollen precipitation studies were carried out on a regional scale in the territory of the classical city of Sagalassos (Southwest Turkey). 57 moss cushions and 7 soil samples from different vegetation types were analysed palynologically. The resulting pollen spectra were grouped into 14 vegetation groups, which were examined using discriminant analysis and principal components analysis. These numerical procedures are applied to display the structure in the data set in a comprehensive way. Some of the modern spectra groups seem to be well-defined, whereas others are difficult to distinguish from each other. After the relationships between the modern pollen spectra had been analysed, the fossil data from a core from the marsh of Gravgaz were added to the data set so that any similarities between fossil and modern spectra could be seen. A joint principal components analysis of the modern and fossil pollen spectra revealed that the pollen spectra from the earliest (ca. 2600-2500 BP; Cal. 830-590 BC till 790-520 BC) and the latest (starting at ca. 1300 BP; Cal. 660-780 AD) pollen assemblage zones have modern counterparts. On the other hand, the pollen spectra from the pollen assemblage zone dominated by Artemisia (ca. 2500-2300 BP; Cal. 790-520 BC till 410-210 BC) and from the pollen assemblage zone that indicated the practice of arboriculture (ca. 2300-1300 BP; Cal. 410-210 BC till 660-780 AD) have no close modern analogues, although individual tabulated pollen percentages revealed some similarities with some modern spectra groups.     

A comparison of sedimentary DNA and pollen from lake sediments in recording vegetation composition at the Siberian treeline

Reliable information on past and present vegetation is important to project future changes, especially for rapidly transitioning areas such as the boreal treeline. To study past vegetation, pollen analysis is common, while current vegetation is usually assessed by field surveys. Application of detailed sedimentary DNA (sedDNA) records has the potential to enhance our understanding of vegetation changes, but studies systematically investigating the power of this proxy are rare to date. This study compares sedDNA metabarcoding and pollen records from surface sediments of 31 lakes along a north–south gradient of increasing forest cover in northern Siberia (Taymyr peninsula) with data from field surveys in the surroundings of the lakes. sedDNA metabarcoding recorded 114 plant taxa, about half of them to species level, while pollen analyses identified 43 taxa, both exceeding the 31 taxa found by vegetation field surveys. Increasing Larix percentages from north to south were consistently recorded by all three methods and principal component analyses based on percentage data of vegetation surveys and DNA sequences separated tundra from forested sites. Comparisons of the ordinations using procrustes and protest analyses show a significant fit among all compared pairs of records. Despite similarities of sedDNA and pollen records, certain idiosyncrasies, such as high percentages of Alnus and Betula in all pollen and high percentages of Salix in all sedDNA spectra, are observable. Our results from the tundra to single‐tree tundra transition zone show that sedDNA analyses perform better than pollen in recording site‐specific richness (i.e., presence/absence of taxa in the vicinity of the lake) and perform as well as pollen in tracing vegetation composition.     

Modern pollen rain in mangroves from San Andres Island, Colombian Caribbean

The precise characterisation of present-day mangrove ecosystems from modern pollen rain facilitates the accurate use of fossil pollen data for late Quaternary sea level and environmental reconstructions. Here, we investigate whether the analysis of pollen rain data corroborates existing floristic and structural characterisation of different mangrove types at the Caribbean island of San Andrés, Colombia. At 82 plots along 20 transects of four distinct mangrove types, samples were obtained of (i) surface sediments for pollen analysis, and (ii) a range of environmental parameters (including inundation levels, salinity and pH). This information was compared to previously sampled mangrove composition and tree basal area. In surface sediment samples 82 pollen taxa were found, from which 19 were present in the vegetation plots. However, because pollen may be transported by wind and/or watercourses, the overall floristic composition of the different forest types may not necessarily be reflected by the pollen spectra. Local vegetation (i.e. mangroves and beach) represented > 90% of the pollen spectra, while the regional one (i.e. hinterland forests) represented < 5% of it. Unlike the four mangrove types that were previously described in the vegetation, the analysis of pollen samples suggested only three distinct types of forest.The groups were characterised based on (i) the dominance of at least one of the true mangrove species from pollen data ordination and the presence of associated species, and (ii) their relationship with environmental parameters. Rhizophora was present in all plot samples, but did not contribute to forest type separation. In fact, just three true mangrove species proved reliable indicators of (i) high salinity and fringe mangroves (i.e. Avicennia), (ii) high pH levels and landward mangroves (i.e. Conocarpus), and (iii) natural or anthropogenic caused disturbance of forest stands (Laguncularia and associated Acrostichum fern). Hence our study confirms that mangrove pollen spectra can be accurately used to describe different mangrove environments for fossil based palaeoecological reconstructions.     

Green fluorescent protein (GFP) as a marker during pollen development

The transient expression of three mutant forms of green fluorescent protein (GFP) genes, GFP4, GFP5ER, and GFP4S65C, under several constitutive and pollenspecific promoters throughout pollen development in Nicotianatabacum, thaliana and Antirrhinummajus is described. Immature pollen of tobacco, Arabidopsis and snapdragon, isolated at different developmental stages, were bombarded with plasmids containing the GFP and cultured in vitro for several days until maturity. The expression of GFP was monitored every day during in vitro maturation, germination and pollination, as well as after in situ pollination. The expression pattern of each construct was compared in parallel experiments to that of ßglucuronidase (GUS) constructs expressed by the same promoters. The results show that the expression level of all three GFP mutant forms was dependent on the strength of the promoter used. The strongest promoter was the DC3 promoter, and no notable differences in the intensity and brightness of all three versions of GFP were observed. GFPexpressing pollen from tobacco and snapdragon developed in vitro for several days until maturity and germinated in vitro as well as on the surface of stigmata, strongly suggesting that all three GFPs are not toxic for the development of functional pollen. Furthermore, stably transformed tobacco plants expressing GFP under the control of the strong pollenexpressed DC3 and LAT52 promoters were not impaired in reproductive function, confirming that GFP can be used as a nondestructive marker for plant reproductive biology and development.     

Quantitative analysis of the distribution of organelles in tobacco pollen tubes: implications for exocytosis and endocytosis

Summary The present study provides the first quantitative analysis on the distribution of organelles in pollen tubes ofNicotiana tabacum L. Organelles were studied on living pollen tubes by means of fluorescence confocal laser scanning microscopy and on cryo-fixed, freeze-substituted and serially sectioned material by electron microscopy. In the tip a 300 nm to 400 nm thick wall was secreted that proximately gradually separated into a wall with an opaque inner side and a more translucent, layered outer side. Tubular endoplasmic reticulum was particularly abundant in the tip of the tube, surrounding the region where secretory vesicles (SV) accumulated. Mitochondria were randomly distributed throughout the cytoplasm, no accumulations were present. Dictyosomes, however, showed an increased abundance at 25–30 m behind the tip. The accumulation of coated pits (CP) in a zone 6–15 m behind the tip identifies this zone as the major site of endocytosis: 50% of all CP occur in this zone. Quantification of exo- and endocytosis showed that only part of the membrane material of the SV can be retrieved after exocytosis. The typical zonation in endocytotic activity may serve to maintain a difference in membrane protein composition between the tip and the tube.     

Spatial variation of airborne pollen over south-east France: characterization and implications for monitoring networks management

The French airborne pollen monitoring network (RNSA) is currently regrouping 70 Hirst-type pollen traps covering the whole French territory. The aim of this paper is to introduce a simple statistical methodology that can be used to characterize pollen spatial variation. This pilot study is restricted to a limited portion of the RNSA network (18 monitoring stations), eight taxa of allergenic relevance, and a 3-year period (2003–2005). The first step of the approach consisted in quantifying the trap-to-trap pollen similarities on the basis of an original index, called mean Pollinic Distance (mPD), that relies on the comparison of pollen concentration time series. Regression analyses were next conducted with different spatial variables. Distance, latitude and altitude differences were identified as significant predictors of pollen variations, as measured by mPD. In order to further characterize pollen spatial properties, cluster analysis was performed with mPD as the distance estimate. Interestingly, the clusters of sites identified on the basis of the similarity of their pollen profiles, correspond to distinct geographic areas that might be interpreted as homogeneous air masses. The results have major implications for monitoring networks management since they provide an objective basis (1) for choosing the relevant scale to elaborate and supply pollen-related information, and (2) for optimizing networks configuration.

Identification of aqueous pollen extracts using surface enhanced Raman scattering (SERS) and pattern recognition methods

Aqueous pollen extracts of varying taxonomic relations were analyzed with surface enhanced Raman scattering (SERS) by using gold nanoparticles in aqueous suspensions as SERS substrate. This enables a selective vibrational characterization of the pollen water soluble fraction (mostly cellular components) devoid of the spectral contributions from the insoluble sporopollenin outer layer. The spectra of the pollen extracts are species‐specific, and the chemical fingerprints can be exploited to achieve a classification that can distinguish between different species of the same genus. In the simple experimental procedure, several thousands of spectra per species are generated. Using an artificial neural network (ANN), it is demonstrated that analysis of the intrinsic biochemical information of the pollen cells in the SERS data enables the identification of pollen from different plant species at high accuracy. The ANN extracts the taxonomically‐relevant information from the data in spite of high intra‐species spectral variation caused by signal fluctuations and preparation specifics. The results show that SERS can be used for the reliable characterization and identification of pollen samples. They have implications for improved investigation of pollen physiology and for allergy warning.

     

Phosphorylation of pollen proteins in relation to self-incompatibility in rye (Secale cereale L.)

The gametophytic two-locus self-incompatibility (SI) system in rye was investigated in view of a possible involvement of protein phosphorylation and Ca²⁺ as constituents of a signal transduction mechanism. Phosphorylation kinetics in pollen grains was found to be significantly different after in vitro treatment of pollen with either cross or self stigma proteins, with a pronounced phosphorylation activity in self-treated pollen grains. Loss of SI in self-compatible (SC) mutants was associated with a significantly decreased basic phosphorylation activity in untreated pollen grains as compared to SI genotypes. Separation of phosphorylated pollen proteins by SDS-PAGE reveals four major proteins in the MW range of 43–82 kDa which were differently phosphorylated in SI vs SC genotypes as well as in cross vs self-treated pollen grains. Application of different protein kinase inhibitors and the Ca²⁺ antagonists verapamil and La³⁺ to isolated stigmas resulted in an inhibition of the SI response in in vitro self-pollination. The role of protein kinases and Ca²⁺ as constituents of a putative SI-specific signal transduction mechanism is discussed.     

Macronutrient input from pollen in two regenerating pine stands in southeast Korea

This study examined macronutrient input from pollen in two naturally regenerating pine stands in southeast Korea. Durham gravity pollen collectors were used to measure pine pollen deposition and the macronutrients in the collected pine pollen were analyzed. In 1998, pine pollen deposition began just before 18 April and lasted for approximately 2weeks. Total pine pollen deposition differed between the two sampling sites; 27.5kgha^–1 was collected from the mature stand and 17.7kgha^–1 was collected from the young stand. The values for nutrient deposition from pine pollen are 549gha^–1 N, 78gha^–1 P, 240gha^–1 K, 45gha^–1 S and 22gha^–1 Mg at the mature stand and 353gha^–1 N, 51gha^–1 P, 151gha^–1 K, 27gha^–1 S and 14gha^–1 Mg at the young stand, suggesting that nutrients from pine pollen contribute to forest nutrient cycling. The pine pollen deposition values obtained from our study (17.7–27.5kg^–1ha^–1year^–1) are approximately 1/115–180-fold that of pine litterfall in Korea. If we take pollen nutrients into account, the contribution rate of pollen to the annual nutrient input is very high in our study (N 1/30, P 1/5, K 1/9 that of litterfall). Macronutrient deposition from pine pollen is concentrated temporally in spring. Although the annual contribution of nutrient mass by pollen is small compared to that of litterfall, the rapid turnover rate of pollen nutrients combined with episodic deposition suggests that pollen may play a disproportionate role in temperate pine forest nutrient cycling.     

Concentration of airborne pollen of Vitisvinifera L. and yield forecast: a case study at S.Michele all'Adige,Trento, Italy

The possibility of creating a forecast model for grapeproduction using airborne pollen concentrations ofVitis vinifera L. was investigated. The studyarea is located in the province of Trento (46°11,11°08), North Italy. Because of its prestigious wineproduction, the area is quite important on a nationalscale.Airborne pollen concentration data (P/m³) werecollected for a five year period (1993–1997) with aHirst type sampler (VPPS 2000 – Lanzoni) following thestandard methodology proposed by the ItalianAeroallergen Network. For the same five-year period,grape production data (kg/ha) from vineyards adjacentto the pollen sampler were used along with provincialdata. Regression lines were constructed using the Lnof the annual sum of daily pollen concentrations (Ip)as the independent variable and grape productionvalues as the dependent variable. Correlations betweenpollen data (pollen index, beginning and duration ofmain pollen season) and meteorological data(temperature and rainfall) are shown.     

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.