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PubMed2002

Pollen tube development and competitive ability are impaired by disruption of a Shaker K(+) channel in Arabidopsis.

Sexual reproduction in plants requires elongation of the pollen tube through the transmitting tissues toward the ovary. Tube growth rate is a major determinant of pollen competitive ability. We report that a K(+) channel of the Shaker family in Arabidopsis, SPIK, plays an important role in pollen tube development. SPIK was found to be specifically expressed in pollen. When SPIK was heterologously expressed in COS cells, its product formed hyperpolarization-activated K(+) channels. Disruption (T-DNA insertion) of the SPIK coding sequence strongly affected inwardly rectifying K(+)-channel activity in the pollen-grain plasma membrane. Measurements of membrane potential in growing pollen tubes yielded data compatible with a contribution of SPIK to K(+) influx. In vitro pollen germination assays were performed, revealing that the disruption results in impaired pollen tube growth. Analysis of the transmission rate of the disrupted allele in the progeny of heterozygous plants revealed a decrease in pollen competitive ability, the probability of fertilization by mutant pollen being approximately 1.6 times lower than that by wild-type pollen. The whole set of data supports the hypothesis that functional expression of SPIK plays a role in K(+) uptake in the growing pollen tube, and thereby in tube development and pollen competitive ability.

Animals↗
PubMed2004

Ca2+-permeable channels in the plasma membrane of Arabidopsis pollen are regulated by actin microfilaments.

Cytosolic free Ca2+ and actin microfilaments play crucial roles in regulation of pollen germination and tube growth. The focus of this study is to test the hypothesis that Ca2+ channels, as well as channel-mediated Ca2+ influxes across the plasma membrane (PM) of pollen and pollen tubes, are regulated by actin microfilaments and that cytoplasmic Ca2+ in pollen and pollen tubes is consequently regulated. In vitro Arabidopsis (Arabidopsis thaliana) pollen germination and tube growth were significantly inhibited by Ca2+ channel blockers La3+ or Gd3+ and F-actin depolymerization regents. The inhibitory effect of cytochalasin D (CD) or cytochalasin B (CB) on pollen germination and tube growth was enhanced by increasing external Ca2+. Ca2+ fluorescence imaging showed that addition of actin depolymerization reagents significantly increased cytoplasmic Ca2+ levels in pollen protoplasts and pollen tubes, and that cytoplasmic Ca2+ increase induced by CD or CB was abolished by addition of Ca2+ channel blockers. By using patch-clamp techniques, we identified the hyperpolarization-activated inward Ca2+ currents across the PM of Arabidopsis pollen protoplasts. The activity of Ca2+-permeable channels was stimulated by CB or CD, but not by phalloidin. However, preincubation of the pollen protoplasts with phalloidin abolished the effects of CD or CB on the channel activity. The presented results demonstrate that the Ca2+-permeable channels exist in Arabidopsis pollen and pollen tube PMs, and that dynamic actin microfilaments regulate Ca2+ channel activity and may consequently regulate cytoplasmic Ca2+.

Actin Cytoskeleton↗
PubMed2005

VANGUARD1 encodes a pectin methylesterase that enhances pollen tube growth in the Arabidopsis style and transmitting tract.

In flowering plants, penetration of the pollen tube through stigma, style, and transmitting tract is essential for delivery of sperm nuclei to the egg cells embedded deeply within female tissues. Despite its importance in plant reproduction, little is known about the underlying molecular mechanisms that regulate the navigation of the pollen tube through the stigma, style, and transmitting tract. Here, we report the identification and characterization of an Arabidopsis thaliana gene, VANGUARD1 (VGD1) that encodes a pectin methylesterase (PME)-homologous protein of 595 amino acids and is required for enhancing the growth of pollen tubes in the style and transmitting tract tissues. VGD1 was expressed specifically in pollen grain and the pollen tube. The VGD1 protein was distributed throughout the pollen grain and pollen tube, including the plasma membrane and cell wall. Functional interruption of VGD1 reduced PME activity in the pollen to 82% of the wild type and greatly retarded the growth of the pollen tube in the style and transmitting tract, resulting in a significant reduction of male fertility. In addition, the vgd1 pollen tubes were unstable and burst more frequently when germinated and grown on in vitro culture medium, compared with wild-type pollen tubes. Our study suggests that the VGD1 product is required for growth of the pollen tube, possibly via modifying the cell wall and enhancing the interaction of the pollen tube with the female style and transmitting tract tissues.

Amino Acid Sequence↗
PubMed1994

Airborne birch pollen antigens in different particle sizes.

Two particle samplers for ambient air, situated together: a static size-selective bio-aerosol sampler (SSBAS) and a Burkard pollen and spore trap were compared in sampling intact birch pollen grains through one flowering period of Betula (a total of 44 days). The SSBAS trapped pollen grains three times more efficiently than the Burkard trap, but the variations in pollen counts were significantly correlated. In contrast, birch pollen antigenic activity and the pollen count in the Burkard samples were not closely correlated. The antigenic concentration was occasionally high both before and after the pollination period. There was a high birch pollen antigenic activity in particle size classes where intact pollen grains were absent, even on days when the pollen count was very low. Correspondingly, on days with high birch pollen counts in the air, pollen antigenic activity was on several occasions low, indicating that pollen grains were empty of antigenic material. The small particle size classes are especially important to allergic patients because they are able to penetrate immediately into the alveoli and provoke asthmatic reactions. Therefore, aerobiological information systems based on pollen and spore counts should be supplemented with information concerning antigenic activities in the air.

Air Pollutants↗
PubMedDate not supplied

Reduction in allergenicity of grass pollen by genetic engineering.

BACKGROUND: Hay fever and allergic asthma triggered by grass pollen allergens affect approximately 20% of the population in cool temperate climates. Ryegrass is the dominant source of allergens due to its prodigious airborne pollen production. Lol p 5 or group 5 is among the most important and widespread grass pollen allergen because it reacts with IgE antibodies of more than 90% of grass pollen-allergic patients, contains most of the grass pollen-specific IgE epitopes and elicits strong biological responses. Significant efforts have been made in developing diagnostic and therapeutic reagents for designing new and more effective immunotherapeutic strategies for treatment of allergic diseases. An alternative approach to this problem could be to reduce the amount of allergen content in the source plant. METHODS: High velocity microprojectile bombardment was used to genetically engineer ryegrass. Antisense construct targeted to one of major allergen, Lol p 5, was introduced. The expression of antisense RNA was regulated by a pollen-specific promoter. Pollen was analysed for IgE reactivity. RESULTS: Analysis of proteins with allergen-specific monoclonal and polyclonal antibodies did not detect Lol p 5 in the transgenic pollen. The transgenic pollen showed remarkably reduced allergenicity as reflected by low IgE binding capacity of pollen extract as compared to control pollen. The transgenic ryegrass plants in which Lol p 5 gene expression is perturbed showed normal fertile pollen development. CONCLUSIONS: Our studies showed that it is possible to selectively 'switch off' allergen production in pollen of ryegrass demonstrating feasibility of genetic engineering of plants for reduced allergenicity.

Allergens↗
PubMedDate not supplied

Secretion of proinflammatory eicosanoid-like substances precedes allergen release from pollen grains in the initiation of allergic sensitization.

It is commonly believed that allergic sensitization starts when an allergen contacts the surface of an antigen-presenting cell in mucosal or skin epithelia. Most studies dealing with this aspect use allergen extracts as stimulus. Under natural exposure conditions, however, the bioavailability of allergen depends on allergen liberation from internal binding sites within the allergen carrier, e.g. pollen grains. In comparing total protein and major allergen release from timothy grass (Phleum pratense L.) pollen freshly collected on rural meadows or near high-traffic roads, there was a striking difference between the pollen, with higher allergen release rates from rural meadow pollen grains. Thus, allergen release does not explain the higher prevalence rates of atopic sensitization and disease observed in many epidemiological studies in children exposed to automobile exhaust. Therefore, other possible effectors from pollen grains were investigated. Pollen grains incubated in protein- free buffer were found to secrete significant amounts of eicosanoid-like substances, namely leukotriene (LT) B(4)-like and prostaglandin E(2)-like substances, in a pH-, time- and temperature-dependent fashion. The highest values of eicosanoid secretion were found in birch, grass and mugwort pollen, while pine (Pinus sylvestris L.) pollen showed only marginal eicosanoid-like secretion. Additionally, the release of these substances was significantly higher from pollen which had been collected near roads with heavy traffic, indicating a stronger proinflammatory activity of these pollen grains. In order to investigate the effects of air pollutants, native pollen grains were exposed in a dose- and time-dependent fashion in a fluidized bed reactor to traffic-related pollutants, e.g. volatile organic compounds (toluene, m-xylene), leading again to a significant increase in the secretion of LTB(4)-like immunoreactivity, in contrast to exposure with sulfur dioxide. This finding opens a new dimension of understanding of the early events in allergic sensitization, indicating that proinflammatory effects of the allergen carrier, e.g. the pollen grain itself, can lead to activation of the mucosal membrane. These findings might help to also explain the higher prevalence rates of pollen allergy in areas with high automobile exhaust emissions. Furthermore, the allergenic 'potency' of various allergens has to be redefined at the allergen carrier level with regard to different stages of allergen and mediator release prior to the contact with the host's immune system.

Air Pollution↗
PubMed2001

Multiple pollen sensitization: a molecular approach to the diagnosis.

BACKGROUND: Sensitization to multiple pollen species is a frequent diagnostic event. Several allergenic molecules with a high level of homology have been identified in divergent pollen families and named panallergens. METHODS: We sought to define the criteria to evaluate the prevalence of the multiple pollen sensitization, to identify specific markers of this condition, and to correlate them with the underlying allergic disease. Patients presenting an allergic respiratory disease underwent skin testing with 23 pollens. Patients fulfilling predefined selection criteria were grouped and classified as having multiple pollen sensitization. Patients in each subgroup were tested for IgE to rBet v 2, rJun o 2, rBet v 1, rPhl p 5 and bromelain. Demographical, allergological and clinical data were recorded in the subgroup of patients with multiple pollen sensitization. RESULTS: Seventeen percent of the pollen-sensitized patients formed the multiple pollen-sensitized subgroup. These subjects were positive for most of the pollen species tested regardless of known exposure to them. None of the subjects sensitized to less than six pollen species were positive to panallergens, whereas 55% of the sera of the multiple pollen-sensitized group were positive to rBet v 2, and 15% to rJun o 2. IgE to rBet v 1 and rPhl p 5 were found positive in all the subgroups. Age, gender, bronchial asthma, oral allergy syndrome, skin test reactivity and previous specific immunotherapy differed significantly when these two subsets were considered. CONCLUSIONS: Allergy diagnosis based on allergenic molecules is crucial in the patient with multiple pollen sensitization. This condition appears to be determined by the sensitization to defined allergenic components (panallergens) rather than by pollen of multiple species as such. Detection of IgE to nonpanallergenic molecules allows to identify more relevant allergenic sources. Clinical aspects of the underlying allergic disease (e.g. asthma and oral allergy syndrome) seem to be differently related to IgE reactivity to panallergens.

Adolescent↗
PubMed1993

Properties of tree and grass pollen allergens: reinvestigation of the linkage between solubility and allergenicity.

In this study we reinvestigated the kinetics of allergen release from birch pollen (Betula verrucosa) and timothy grass pollen (Phleum pratense) using different protein extraction procedures, immunoblotting with specific antibodies and immune electron microscopy. Pollen allergens such as the major birch pollen allergen, Bet v I, the major timothy grass pollen allergens, Phl p I and Phl p V, group-II/III allergens from timothy grass and profilins were released rapidly and in large amounts from hydrated pollen. Within a few minutes pollen allergens could be detected in aqueous supernatants prepared from birch and grass pollen with serum IgE or specific antibodies. In parallel the allergen content in the pollen pellet fractions decreased. A nonallergenic protein such as heat shock protein 70 can be extracted in sufficient amounts only with harsh extraction procedures. Immune electron microscopy of dry and rehydrated birch pollens showed that after short hydration, the major birch pollen allergen, Bet v I, migrated into the exine and to the surface of intact pollen grains, whereas profilin, against which a lower percentage of patients is sensitized, was retained in the pollen grain. Comparing the amino acid composition and hydrophilicity of the tested allergens with a nonallergenic protein such as heat shock protein 70, no significant difference was noted. In agreement with earlier observations we conclude that the allergenic properties of proteins are rather linked to the amount and speed of solubility from airborne particles than to intrinsic properties.

Allergens↗
PubMedDate not supplied

Clinico-immunologic evaluation of allergy to Himalayan tree pollen in atopic subjects in India--a new record.

Exposure to local pollen allergens has a direct bearing on the prevalence of allergic symptoms among the inhabiting atopic population. The populations in the Himalayas and around it are exposed to a variety of pollen grains from trees growing in the region, but the pollen-population interaction has not been clinically investigated. Himalayan tree pollen from five different taxa, i.e. Alnus nitida (AN), Betula utilis (BU), Cedrus deodara (CD), Mallotus phillipensis (MP) and Quercus incana (QI) were evaluated for their allergenicity in the Indian population by in vivo (skin prick test) and in vitro (ELISA) clinico-immunological methods. The presence of specific IgE against these tree pollen in the sera of skin test positive patients was taken as evidence for sensitization to these pollen. The average skin positivity in atopic populations recorded at different allergy centers in India varied from 2.2% against AN, to 4.7% against MP pollen. Significantly raised specific IgE against these pollen were observed in the sera of hypersensitive patients. The sensitization pattern to Himalayan tree pollen in these atopic populations varied. It was concluded that skin prick test positivity and raised IgE antibodies specific to AN, BU, CD, MP and QI established Himalayan tree pollen as important sensitizers in the atopic populations of India. A high incidence of skin sensitivity was observed to pollen antigens of Cedrus deodara, Mallotus phillipensis and Quercus incana in patients of Chandigarh residing in the hills and foothills of the Himalayas while Alnus nitida, Betula utilis and Cedrus deodara were important sensitizers in Delhi patients. The skin sensitization pattern against these pollen was in accordance with the level of exposure to these pollen of the subjects residing in that part of the country.

Adolescent↗
PubMed2005

[Prediction of atmospheric Japanese cedar pollen counts in Oita University Faculty of Medicine Complex].

BACKGROUND: We investigated the method of predicting atmospheric Japanese cedar pollen counts and the first day of pollen release at Oita University Faculty of Medicine Complex in this study. METHOD: We set up a Durham sampler on the roof (30 m from the ground) of Oita University Faculty of Medicine complex and investigated atmospheric pollen counts day by day from January 1 to April 30 from 1990 through 2004. RESULTS: The total pollen counts per year correlated very well with averaged temperature, averaged relative humidity, and the sunshine duration during the previous July. The sunshine duration yielded the highest correlation coefficient. Multiple regression analysis showed the sunshine duration during the previous July and total pollen count of the previous year to be independent predictors of the current year's total pollen counts. Multiple correlation coefficients was 0.9518 (p<0.001). We considered the Hita area to be the source of the pollen, according to meteorological conditions. Prediction of total pollen counts was more accurate if based on factors within the previous 10 years than on factors within the previous 5 years. There was no correlation between the first day of pollen release and averaged temperature in January. Our data also showed that the first day of pollen release could not be predicted by the cumulative thermal constant method and threshold temperature for the development of male Japanese cedar flowers. Multiple regression analysis showed the minimum temperature from November 1 to the first day of pollen release, the date of minimum temperature, and the averaged temperature for the January to be independent predictors of the number of days from January 1 to the first day of pollen release. CONCLUSION: We concluded that it is important to establish a method of predicting atmospheric Japanese cedar pollen counts that is based on the characteristics of each area.

Cryptomeria↗
PubMed2005

[Analysis of pine pollen by using FTIR, SEM and energy-dispersive X-ray analysis].

Infrared spectroscopy (IR), scanning electron microscope (SEM) and energy-dispersive X-ray analysis (EDX) were used to analyze nutrients in four pine pollen powder samples. The IR fingerprints showed that each of the samples, pinus massoniana, pinus yunnanensis, pinus tabulaeformis, and pinus densiflora, respectively had its own characteristic infrared spectrum. Based on the difference of the relative intensity of those characteristic absorption peaks, the IR fingerprints can be used for the identification of the four kinds of pine pollen samples. The broken pollen of pinus was more easily to release nutritional components for the distinct difference IR fingerprints of natural and broken masson pine pollen samples. As a result of SEM, four kinds of pollen grains were oblong or subspheroidal in distal face and proximal face. The exine sculpture of the four kinds of samples were granulous and almost the same, but there was some difference of the size of pollen grains. The main morphologic change of the broken pollen was that the air bags were separated from pollen particles, and part of the main body of pollen particles was broken. The energy-dispersive X-ray analysis results showed that eleven elements, including Mg, Se, Si, Sr, P, S, Cl, K, Ca, Mn, and Fe, existed and the highest content in pollen of pinus was K element. The contents of trace elements were different in different kinds of pollen of pinus. The element intensity in broken masson pine pollen was distinctlyhigher than that innatural masson pine pollen.

Microscopy, Electron, Scanning↗
PubMed1996

[The pollen count of Cryptomeria japonica and nasal symptom of Cryptomeria japonica pollinosis patients in recent three years in Mie Prefecture].

Airborne scattering of Cryptomeria japonica pollen depends on climate. Total pollen count and scattering pattern varies from one year to another. We counted Cryptomeria japonica pollen in February and March from 1993 to 1995 at several points in Mie Prefecture. One-third to two-thirds of all the patients who visited our clinic for the first time in the pollen period were occupied by allergic patients. The pollen count in 1995 amounted to 17943/cm2 at Tsu and 99512/cm2 in the whole prefecture, which is the largest in the past 12 years. The number of days in which patients severely suffered was correlated to the pollen count for the past three years. It was shown that nasal symptoms developed after certain amount of pollen had scattered, but the severity of symptoms thereafter largely depended on the total pollen count. At Tsu in 1995, we scarcely had pollen scattering in February but had tremendous amount of pollen count in March. Percentage of development of nasal symptoms among thirty patients was well correlated to the logarithm of accumulated pollen count. When accumulated pollen count reached to 100/cm2, half the patients developed symptoms. Most of the patients developed symptoms when it reached to 1000/cm2. Since there is a great interindivisual difference in when and how much patients have nasal symptoms, the choice of therapy should be based not only on the pollen count but also on the symptom and condition of each patient.

Adult↗
PubMed1998

Effects of pollen quality and genotype on the dance of foraging honey bees.

Animals assess the quality and quantity of food and choose among different foods based on these assessments. We explored whether there was genetic variation for assessment of pollen quality by foraging honey bees, Apis mellifera. Honey bees derived from two genotypic strains foraged for pollen of varying quality from a petri dish placed inside an outdoor flight cage. The strains were the result of a colony-level, two-way selection on amount of stored pollen. We used the forager's round dance to quantify the assessments of pollen quality by individually marked worker bees. The dance rate (number of 180 degrees turns per minute) and the probability of dancing were each greater when bees foraged for pure pollen compared with a lower-quality mixture of pollen and alpha-cellulose (1:1 by volume). Bees from the high-pollen genotypic strain had a higher dance rate than those from the low-pollen strain, suggesting different assessments. Bees from the low-pollen strain, however, had a higher probability of dancing than did bees from the high-pollen strain. Dance duration was not affected by a bee's strain or by the quality of pollen. We conclude that the dance rate may be used to quantify a forager's subjective evaluation of pollen quality and that this evaluation has a genetic component. Our results also suggest that the dance may function at the colony level to recruit bees to more profitable pollen sources. Copyright 1998 The Association for the Study of Animal Behaviour.

Journal Article↗
PubMed2000

Pollen input to, and incorporation in, two crater lakes in tropical northeast Australia.

Pollen input to the water surfaces of maar crater lakes Barrine and Eacham (1km(2) and 0.5km(2) respectively, ca. 720m a.s.l.), surrounded by rainforest in northeastern Australia, were measured in floating Tauber traps for a variety of periods spanning the years 1978 to 1985. The mean weekly total pollen catch is 75% of that estimated for the surrounding region of varied vegetation. Regardless of their sources, the influxes of pollen taxa are unevenly distributed over the lakes' surfaces. There is similar variation through time, with some indication that this might be smoothed over periods of 5years or more. There are no inflow streams and pollen input down surrounding slopes is negligible. Mean total pollen influx to seston traps close to the lakes' bottoms (1979 to 1988) is <5% of that to the Tauber traps, a discrepancy attributed to trap design. In contrast, mean annual influx measured by the Tauber traps is similar to that estimated from dated sediment samples younger than 1966 AD at Lake Barrine. The proportions of different pollen taxa are less affected by trap position or period of exposure than are their influxes; their mean percentages for Tauber traps, seston traps and sediment samples are all rather similar.The similarity between the mean contribution of exclusively rainforest pollen to the lakes' surfaces (56%) and to Oldfield traps in rainforest interiors (51%) emphasizes the importance of the rainforests, which clothe the crater walls, as sources of pollen to the lakes. The impact of pollen from other vegetation in the region, although present, is much smaller than at interfaces between rainforest and other vegetation types. All this pollen is mixed within and above the crater and delivered to the lakes' surfaces by gravity and frequent rainfall. Limnological processes redistribute it in the water body before it is incorporated into the deep-lake sediments.The value and limitations of current pollen transport and accumulation theory are noted in relation to sites with morphologies similar to these two crater lakes and to the irregular flowering and floristic inhomogeneity of tropical rainforest. The potential for the use of modern pollen input data in the interpretation of pollen analyses from the sediments of such sites is explored.

Journal Article↗
PubMed2000

Spatial variation of modern pollen in Oregon and southern Washington, USA.

Surface sediments from 95 lakes provide information on the spatial variation of modern pollen spectra in Oregon and southern Washington. Percentages for 13 pollen types were compared within and between vegetation zones to characterize regional patterns of pollen spectra. The percentage data were also compared with climate variables to determine relationships between pollen percentages and regional climate gradients. The composition of modern pollen spectra corresponds well with the distribution of the pollen producers. Most pollen assemblages were generally dominated by Pinus, but those west of the Cascade Range were dominated by Alnus. Low percentages of Pseudotsuga/Larix, Tsuga mertensiana, Abies, and Picea pollen coincided with local occurrence of the trees. The distributions of the pollen data were arranged along gradients of temperature and effective moisture. West of the Cascade Range, Alnus, Tsuga heterophylla, Pseudotsuga/Larix, and Cupressaceae pollen were abundant and correlate well with moderate temperature and high effective moisture. In the shrub-steppe and woodlands east of the Cascade Range, where effective moisture is low, Artemisia, Cupressaceae, and Pinus pollen were dominant. At high elevations, Pinus, T. mertensiana, Abies, and Picea were common pollen types in areas with short growing seasons and high effective moisture. Pollen percentages collected from lake surface sediments, moss polsters, and soils were compared within a number of vegetation types to assess their similarity. The three types of sample yielded similar results for forested areas, but lake sediment samples from upper- and lower-treeline sites captured a more regional picture of the vegetation.

Journal Article↗
PubMed2004

Investigation of short-range cedar pollen forecasting.

Pollen forecasting is of increasing interest as a way to help the general public avoid contact with allergy-inducing pollen. It was recently reported that the dynamics underlying pollen concentration series is very similar to that of low-dimensional deterministic chaos, thus opening up new avenues of development in local pollen forecasting. Our analysis of hourly cedar pollen series for two seasons showed evidence of a small degree of determinism underlying the pollen time-series dynamics. However, we could not confirm that our pollen series was generated by a low-dimensional chaotic system. The nearest-neighbor method using local constant prediction applied to hourly pollen forecasting with a 1-h lead time was effective for small to medium pollen variations, but failed to reproduce large and intermittent pollen bursts. The performance of the nearest-neighbor model was significantly improved by applying a nonlinear filter to the source dataset. Standard time-series techniques such as neural networks did not improve upon these results. The difficulty in fully characterizing and accurately forecasting the pollen series was thought to originate in the nonstationarity of the series and in the large and intermittent pollen bursts that were found to have no apparent time structure. Thus the dynamics of hourly pollen series is probably not strongly tied to a low-dimensional chaotic system.

Journal Article↗
PubMed2000

Pollen competition in a natural population of Cucurbita foetidissima (Cucurbitaceae).

The pollen competition hypothesis predicts that when the number of pollen grains deposited onto stigmas exceeds the number of ovules, selection can operate in the time frame between deposition and fertilization. Moreover, because of the overlap in gene expression between the two phases of the life cycle, selection on microgametophytes may alter the resulting sporophytic generation. The extent to which pollen competition occurs in nature has been unclear, because tests of the predictions of the pollen competition hypothesis have used cultivars and/or artificial growth conditions and hand-pollination techniques. In this study we used a wild species, Cucurbita foetidissima, in its natural habitat (southern New Mexico) to determine the amount and timing of the arrival of pollen onto stigmas, the relationship between pollen deposition and seed number, and the effects of the intensity of pollen competition on progeny vigor. We found that ∼900 pollen grains are necessary for full seed set and that a single visit by a pollinator results in the deposition of 653.0 ± 101.8 pollen grains. About 29% of the flowers receiving a single pollinator visit had 900 or more pollen grains on its stigma. Moreover, within 2 h of anthesis, >4000 pollen grains were deposited onto a typical stigma, indicating that multiple pollinator visits must have occurred. Fruits produced by multiple visits had greater seed numbers (206 vs. 147) than fruits produced by a single visit. Finally, the progeny produced by multiple pollinator visits were more vigorous than those produced by single visits with respect to five measures of vegetative growth (MANCOVA, Wilks' lambda = 0.96, F(6,370) = 2.54, P < 0.02. These data demonstrate that conditions for pollen competition exist in nature and support the prediction that pollen competition enhances offspring vigor.

Journal Article↗
PubMed2004

The CaMV 35S promoter is highly active on floral organs and pollen of transgenic strawberry plants.

We have evaluated the expression of the reporter beta-glucuronidase (GUS) gene driven by the cauliflower mosaic virus 35S (CaMV 35S) promoter in flowers and pollen from 14 independent transgenic strawberry lines. Of the 14 lines evaluated, 13 (92.8%) showed GUS activity--as estimated by the histochemical GUS assay--in some floral organs, with expression being most common in the flower stem, sepals, petals, ovary and stigma. Ten of these thirteen transgenic lines (77%) showed GUS activity in pollen, although the percentages of positive pollen per flower varied greatly among the different lines. A study of the GUS expression during pollen maturation showed that the (CaMV 35S) promoter showed low expression in pollen from flower buds before anthesis but was activated in mature pollen following anther dehiscence. The percentages of pollen grains that showed GUS activity ranged from 2.1% to 46.3%. These percentages were similar or even higher when mature pollen was stored dry at room temperature for 2 weeks. After 5 weeks of storage, the percentages of GUS-positive pollen decreased in two of the six lines analysed but remained at similar values in the other four lines. GUS activity was also measured in protein extracts of mature pollen by means of the fluorometric GUS assay, with the values obtained ranging from 3.8 micromol MU mg protein(-1) h(-1) to 0.26 micromol MU mg protein(-1) h(-1). Contrary to the generally held view that the CaMV 35S promoter is virtually silent in pollen, we conclude that it is highly expressed in transgenic strawberry pollen.

Flowers↗