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Pollen flow in the wildservice tree, Sorbus torminalis (L.) Crantz. II. Pollen dispersal and heterogeneity in mating success inferred from parent-offspring analysis.

Knowing the extent of gene movements from parents to offspring is essential to understand the potential of a species to adapt rapidly to a changing environment, and to design appropriate conservation strategies. In this study, we develop a nonlinear statistical model to jointly estimate the pollen dispersal kernel and the heterogeneity in fecundity among phenotypically or environmentally defined groups of males. This model uses genotype data from a sample of fruiting plants, a sample of seeds harvested on each of these plants, and all males within a circumscribed area. We apply this model to a scattered, entomophilous woody species, Sorbus torminalis (L.) Crantz, within a natural population covering more than 470 ha. We estimate a high heterogeneity in male fecundity among ecological groups, both due to phenotype (size of the trees and flowering intensity) and landscape factors (stand density within the neighbourhood). We also show that fat-tailed kernels are the most appropriate to depict the important abilities of long-distance pollen dispersal for this species. Finally, our results reveal that the spatial position of a male with respect to females affects as much its mating success as ecological determinants of male fecundity. Our study thus stresses the interest to account for the dispersal kernel when estimating heterogeneity in male fecundity, and reciprocally.

Conservation of Natural Resources↗

The Arabidopsis actin-related protein 2 (AtARP2) promoter directs expression in xylem precursor cells and pollen.

To investigate the role of the actin cytoskeleton in plant growth and development, we have cloned and determined the DNA sequence of a gene encoding an actin-related protein (arp) from Arabidopsis thaliana (AtARP2) and studied its expression patterns. A. thaliana appears to have only one AtARP2 gene which contains 14 introns, an unusually large number when compared to 4 or 5 introns in the actin genes isolated so far. The predicted protein shows high homology to and shares typical peptide insertions with arp2 proteins from other organisms. The AtARP2 transcript is present in all plant tissues, at a very low level, and is down-regulated by light. Promoter-GUS expression studies showed that the AtARP2 promoter directs activity predominantly in a very small number of cells immediately adjacent to the xylem in all organs. In addition, strong expression was observed in pollen grains. We discuss the potential role of an arp2/3 complex in plant development.

Actin-Related Protein 2↗

Quantitation of the major allergen of several Parietaria pollens by an anti-Par 1 monoclonal antibody-based ELISA. Analysis of crossreactivity among purified Par j 1, Par o 1 and Par m 1 allergens.

BACKGROUND: Plants of the genus Parietaria, Urticaceae family, represent a major cause of pollinosis in the Mediterranean area. Different Parietaria species crossreact to a great extent, but studies on the crossreactivity among the major allergens of these pollens have not been carried out so far. OBJECTIVE: To develop an immunochemical method to quantify the major Parietaria judaica allergen, Par j 1, as well as to verify the presence of Par j 1-like proteins in different Urticaceae pollens. These proteins would be purified in order to study the cross-reactivity among them. METHODS: Immunoaffinity chromatography with a monoclonal antibody, solid-phase enzyme-linked immunoassays and SDS-PAGE. RESULTS: A monoclonal antibody-based ELISA for the quantification of Par j 1 has been developed. The assay has a sensitivity of 0.2 ng/mL and shows a high correlation with the allergenic activity of P. judaica extracts determined by radioallergosorbent assay (RAST) inhibition. By means of this assay, proteins homologous to Par j 1 were detected in P. officinalis and P. mauritanica. These proteins (Par o 1 and Par m 1, respectively) were purified by affinity chromatography using the same monoclonal antibody employed in the ELISA. Crossed-inhibition experiments demonstrated that Par j 1, Par o 1, and Par m 1, competed for the binding of specific IgE from a P. judaica-sensitive patients serum pool. CONCLUSION: The results here described suggest that shared allergenic epitopes are present in the three main allergens investigated, which may simplify the diagnosis and therapy for Parietaria allergy.

Allergens↗

Environmental factors as a cause for the increase in allergic disease.

LEARNING OBJECTIVES: To be able to understand the interaction among genetic factors, environmental exposure to allergens, and nonspecific adjuvant factors contributing to the increase in atopic diseases in developed countries. DATA SOURCES: Peer-reviewed literature identified by searching medical databases. STUDY SELECTION: Careful review of epidemiologic cross-sectional, sequential, and longitudinal population studies and, when appropriate, intervention studies. The criteria used to accept a study reporting environmental factors influencing the prevalence of allergic diseases were adopted from the report published by the US Department of Health and Education in 1964 (Hill AB, Principles of Medical Statistics, 9th Ed. New York: Oxford University Press, 1971, p. 323) RESULTS: There is ample evidence that specific environmental factors may cause sensitization and development of allergic symptoms and disease in susceptible individuals. It is unclear when and how long a sufficient exposure will result in clinical symptoms related to the immunoglobulin E-sensitizing agents. CONCLUSIONS: Environmental factors play an important role for the development and manifestation of allergic conditions in genetically predisposed subjects. It is well documented that increased exposure to indoor allergens and selected outdoor allergens (eg, grass pollen and molds) and smoking are important risk factors for development of asthma and allergic sensitization. The importance of other environmental factors is less clear and which environmental factors that cause the increase in prevalence of allergic disease is still unknown.

Allergens↗

Microsporogenesis in the endangered species Cupressus dupreziana A. Camus: evidence for meiotic defects yielding unreduced and abortive pollen.

To understand the reproductive biology of Cupressus dupreziana A. Camus (Cupressaceae), a highly endangered Mediterranean conifer, the processes of microsporogenesis and pollen differentiation were investigated cytologically. Pre-meiotic development proved to be similar to the coniferous pattern: the microsporangia differentiated sporogenous tissue in which microsporocytes separated and underwent meiosis. As the meiotic steps proceeded, unexpected irregularities were observed concerning chromosomal and nuclear behaviour. This mainly included: abnormal chromosome segregation and cytokinesis, and nuclear fusion of the meiotic products. The result was the formation, in the same microsporangium, of heterogeneous microspore populations arranged in monads, dyads, triads, tetrads, and polyads, and cytoplasts giving rise to pollen grains of different sizes. This indicates that in C. dupreziana both abortive and unreduced pollen grains are generated. The significance of the finding is discussed in relation to reproductive biology and vulnerability to extinction.

Cell Differentiation↗

Rop GTPase: a master switch of cell polarity development in plants.

Cell polarity is fundamentally important to plant growth and development, yet the mechanism governing its development is understood poorly. Several studies have revealed a role for Rop GTPases in pollen polar tip growth. Rop is also localized to the future site of root hair development and the tip of root hairs, and expression of constitutively active Rop mutants impacts on the morphogenesis of tip-growing root hairs as well as on non-tip-growing cells. These findings highlight the importance of Rop as a common switch in cell polarity control in plants.

Cell Polarity↗

The effect of cycloheximide on dictyosome activity in Tradescantia pollen tubes determined using cytochalasin D.

Dictyosome activity in Tradescantia pollen tubes has been determined using a recently developed method based on the assumption that the rate of vesicle accumulation around the dictyosomes, after treatment with cytochalasin D, is equivalent to the actual rate of vesicle production. In tubes germinated in the presence of 1.0 micrograms/ml cycloheximide, reduced dictyosome activity could be detected as early as 10 min after sowing, although tube extension was not halted until later. After 30 min vesicle production had completely ceased. These observations are discussed in relation to previous reports on the effect of cycloheximide on pollen tube growth, and in relation to the synthesis and transfer of membrane proteins to secretory vesicles and the plasma membrane. It is concluded that the ability of pollen to germinate and produce short tubes in the presence of cycloheximide, does not necessarily indicate that protein synthesis is not a requirement for early pollen tube growth, as protein shortages would not be expected to become apparent over time periods less than the dictyosome turnover time and the secretory vesicle residence time.

Cycloheximide↗

Identification of pronp1, a tobacco profilin gene activated in tip-growing cells.

In plant cells, several cellular processes depend on rapid reorganization of a dynamic network of actin cytoskeletal elements in response to internal and environmental stimuli. Profilins, ubiqitous eukaryotic actin monomer-binding proteins with highly conserved three-dimensional structures, regulate the actin cytoskeleton and are considered to link the microfilament system with signal transduction pathways. Plant profilins have been grouped into two distinct classes, gametophytic (pollen-specific) and sporophytic. Here we report the isolation of a profilin gene that seems to be activated during tip growth of specialized cells of gametophytic as well as sporophytic origin. Identification of a genomic DNA clone containing a tobacco profilin gene, pronp1, and analysis of the pronp1 promoter-uidA fusion gene in transgenic Nicotiana tabacum plants revealed a prominent expression of pronp1 in mature pollen and elongating pollen tubes and significant activity in root hairs of developing seedlings. This expression pattern was distinct from that of any other profilin gene isolated so far. Pronp1 thus represents a unique profilin gene that is activated at the transcriptional level in two kinds of tip-growing cells, pollen tubes and root hairs, both of which require rapid organization of the actin cytoskeleton. The isolation of such a gene has fundamental importance for our understanding of modulation of the actin cytoskeleton at the molecular level.

Contractile Proteins↗

Correlation between date of birth and pollen sensitivity.

Attempts by previous investigators have failed to show significant correlation between seasonal pollens and the birth dates of individuals as evidence for the development of tolerance in immunologically immature neonates. Using a large clinic population with adequate controls, significant correlation between ragweed, grass or tree pollen sensitivity and the dates of birth was not obtained.

Alternaria↗

RNAi-mediated silencing of OsGEN-L (OsGEN-like), a new member of the RAD2/XPG nuclease family, causes male sterility by defect of microspore development in rice.

We have cloned a new member of the RAD2/XPG nuclease family, OsGEN-L (OsGEN-like), from rice (Oryza sativa L.). OsGEN-L possesses two domains, the N- and I-regions, that are conserved in the RAD2/XPG nuclease family. Database searches and phylogenetic analyses revealed that OsGEN-L belongs to class 4 of the RAD2/XPG nuclease family, and OsGEN-L homologs were found in animals and higher plants. To elucidate the function of OsGEN-L, we generated rice OsGEN-L-RNAi transgenic plants in which OsGEN-L expression was silenced. Most of the OsGEN-L-RNAi plants displayed low fertility, and some of them were male-sterile. OsGEN-L-RNAi plants lacked mature pollen, resulting from a defect in early microspore development. A OsGEN-L-green fluorescent protein (GFP) fusion protein was localized in the nucleus, and the OsGEN-L promoter was specifically active in the anthers. Furthermore, a recombinant OsGEN-L protein possessed flap endonuclease activity and both single-stranded and double-stranded DNA-binding activities. Our results suggest that OsGEN-L plays an essential role in DNA metabolism required for early microspore development in rice.

Cell Nucleus↗

Hazards of imidacloprid seed coating to Bombus terrestris (Hymenoptera: Apidae) when applied to sunflower.

Seed coating treatments of sunflower by the systemic insecticide imidacloprid was suspected of affecting honey bees and bumblebees. The hypothesis raised was whether imidacloprid could migrate into nectar and pollen, then modify flower attractiveness, homing behavior, and colony development. Our greenhouse and field experiments with Bombus terrestris L. were aimed at the following: the behavior of workers foraging on treated and control plants blooming in a greenhouse, the homing rate of colonies placed for 9 d in a treated field compared with colonies in a control field, and the development of these 20 colonies under laboratory conditions when removed from the fields. In the greenhouse, workers visited blooming heads of treated and control plants at the same rate and the mean duration of their visits was similar. In field colonies, analysis of pollen in hairs and pellets of workers showed that in both fields 98% of nectar foragers visited exclusively sunflowers, whereas only 25% of pollen gatherers collected sunflower pollen. After 9 d, in the control and treated field, 23 and 33% of the marked foragers, respectively, did not return to hives. In both fields, workers significantly drifted from the center to the sides of colony rows. During the 26-d period under field and laboratory conditions, the population increase rate of the 20 colonies was 3.3 and 3.0 workers/d in hives of the control and treated field, respectively. This difference was not significant. New queens were produced in eight colonies in either field. The mean number of new queens per hive was 17 and 24 in the control and treated field, respectively. Their mating rate was the same. It was concluded that applying imidacloprid at the registered dose, as a seed coating of sunflowers cultivated in greenhouse or in field, did not significantly affect the foraging and homing behavior of B. terestris and its colony development.

Animals↗

Experimental allergic conjunctivitis in guinea pigs induced by Japanese cedar pollen.

We report a new experimental allergic conjunctivitis with Japanese cedar pollen as antigen in guinea pigs, and the immunological characteristics of this model were also elucidated. Allergic conjunctivitis was developed by immunization in guinea pigs with a mixture containing Japanese cedar pollen and killed Bordetella pertussis. When local application of Japanese cedar pollen suspension 14 d after systemic immunization was performed every 3d, remarkable conjunctivitis was observed from 20 to 35 d. Increase in vascular permeability and decrease in histamine contents of the conjunctiva were also observed after local application of antigen. Passive cutaneous anaphylactic (PCA) reactions revealed that both IgG- and IgE-rich antibodies were produced in this model. Chlorpheniramine, ketotifen and levocabastine were effective in inhibiting cedar pollen-induced conjunctivitis. Although a high concentration was needed, tranilast and amlexanox also showed significant inhibition of conjunctivitis induced by cedar pollen.

Animals↗

Behavioral genomics of honeybee foraging and nest defense.

The honeybee has been the most important insect species for study of social behavior. The recently released draft genomic sequence for the bee will accelerate honeybee behavioral genetics. Although we lack sufficient tools to manipulate this genome easily, quantitative trait loci (QTLs) that influence natural variation in behavior have been identified and tested for their effects on correlated behavioral traits. We review what is known about the genetics and physiology of two behavioral traits in honeybees, foraging specialization (pollen versus nectar), and defensive behavior, and present evidence that map-based cloning of genes is more feasible in the bee than in other metazoans. We also present bioinformatic analyses of candidate genes within QTL confidence intervals (CIs). The high recombination rate of the bee made it possible to narrow the search to regions containing only 17-61 predicted peptides for each QTL, although CIs covered large genetic distances. Knowledge of correlated behavioral traits, comparative bioinformatics, and expression assays facilitated evaluation of candidate genes. An overrepresentation of genes involved in ovarian development and insulin-like signaling components within pollen foraging QTL regions suggests that an ancestral reproductive gene network was co-opted during the evolution of foraging specialization. The major QTL influencing defensive/aggressive behavior contains orthologs of genes involved in central nervous system activity and neurogenesis. Candidates at the other two defensive-behavior QTLs include modulators of sensory signaling (Am5HT(7) serotonin receptor, AmArr4 arrestin, and GABA-B-R1 receptor). These studies are the first step in linking natural variation in honeybee social behavior to the identification of underlying genes.

Animals↗

Silencing of an anther-specific zinc-finger gene, MEZ1, causes aberrant meiosis and pollen abortion in petunia.

MEZ1 (MEiosis-associated Zinc-finger protein 1) was first isolated as an anther-specific cDNA from Petunia hybrida. In the present study, we report its functional characterization, including its spatial and temporal expression profiles and phenotypes in MEZ1-silenced plants. MEZ1 transcripts were specifically localized in pollen mother cells during early stages of anther development, and were later distributed in vegetative tissues in anthers. Silencing of MEZ1 by cosuppression resulted in several anomalies during male meiosis that included inability of chromosomes to condense, loss of meiotic synchrony, and premature and apparently uncontrolled cytokinetic events. Consequently, by the end of meiosis 8-10 cells, instead of the normal 4, with varying DNA contents were formed in the MEZ1-silenced meiocytes. Most of these aborted prematurely, and those that matured had a distinctive morphology. MEZ1-silenced plants were female sterile when pollinated with wild-type pollen but they infrequently produced a few seeds upon self-pollination. Resulting T1 plants had increased ploidy levels and exhibited severe anomalies during male meiosis, rendering them completely sterile. We discuss possible role of MEZ1 in the proper progression of plant meiosis.

Arabidopsis↗

Partial silencing of the NEC1 gene results in early opening of anthers in Petunia hybrida.

The NEC1 gene, previously isolated from Petunia hybrida, is expressed at high levels in nectaries, and in a very localized fashion in stamens, particularly in the anther stomium cells and the upper part of the filament. To elucidate the function of the NEC1 gene, co-suppression was employed for down-regulation of NEC1 expression, and transposon insertion mutagenesis was used to knock out the NEC1 function. Among the transgenic plants and plants carrying dTph1 inserted in the NEC1 gene, an "early open anther" phenotype was observed. In this mutant phenotype, the anthers already open in young flower buds (1.8 cm) that still contain immature pollen, resulting in poor pollen quality and impaired pollen release. The results obtained indicate that NEC1 might be involved in the development of stomium cells, which are ruptured during the normal process of anther dehiscence to release mature pollen. Southern analysis revealed the presence of a highly homologous NEC1-like gene, named NEC2, in the P. hybrida genome. The presence of NEC2 was confirmed by segregation analysis and sequencing of genomic clones. The implications of these results and possible reasons why no visually obvious phenotype in nectaries could be produced by co-suppression or transposon insertion mutagenesis are discussed.

Alleles↗

Anaphylactic reaction after the ingestion of chamomile tea: a study of cross-reactivity with other composite pollens.

We report a case of an 8-year-old atopic boy in whom ingestion of a chamomile-tea infusion precipitated a severe anaphylactic reaction. The patient suffers from hay fever and bronchial asthma caused by a variety of pollens (grass, olive, and mugwort). This severe reaction was developed after his first ingestion of chamomile tea. Studies revealed the presence of immediate skin test reactivity and a positive passive transfer test to chamomile-tea extract. Moreover, both specific antichamomile-tea extract and anti-Matricaria chamomilla-pollen extract IgE antibodies were detected by an ELISA technique. Cross-reactivity among chamomile-tea extract and the pollens of Matricaria chamomilla, Ambrosia trifida (giant ragweed), and Artemisia vulgaris (mugwort), was demonstrated by an ELISA-inhibition study. These findings suggest a type I IgE-mediated immunologic mechanism as being responsible for the patient's anaphylactic symptoms and also suggest that the patient cross-reacted the pollens of Matricaria chamomilla contained in the chamomile tea because he was previously sensitized to Artemisia pollen.

Anaphylaxis↗

Peptide mapping analysis of group I allergens of grass pollens.

Group I allergens, from eight different clinically important grass pollens of the Pooideae (rye-grass, canary, Kentucky bluegrass, orchard and timothy), Chloridoideae (Bermuda grass) and Panicoideae (Johnson grass, maize) were isolated by sodium dodecyl sulphate polyacrylamide gel electrophoresis and subjected to limited partial proteolysis and cyanogen bromide cleavage. The generated digests were visualised on gels by silver staining. Replicate gels were blotted and screened with Lolp I-specific mAb FMC A1 and IgE binding to an allergic human serum pool, to determine the degree of antigenic and IgE-binding similarities, respectively. The highest antigenic and IgE-binding similarities were between orchard and rye-grass pollens, both in the same tribe Poeae, and among the closely related grass genera. No peptide mapping of major grass pollen allergens has previously been undertaken. This study proves peptide mapping is a powerful method for the preliminary identification and internal amino acid microsequencing of common antigenic and IgE-binding determinants of allergens, providing information that is useful in the development of reagents for the treatment of grass-pollen-associated allergies.

Allergens↗

Structural and functional compartmentalization in pollen tubes.

Eukaryotic cellular functions are achieved by concerted activities in the cytosol and functions compartmentalized in the nucleus and other membrane-bound organelles. Moreover, the cytosol and nucleoplasm are populated with mega molecular ensembles that are specialized for different metabolic and biochemical processes. Pollen tubes are unique plant cells with a dramatic growth polarity. Tube growth is restricted to the tip and is supported by a polarized cytoplasmic organization. The apical region of elongating pollen tubes is a domain occupied exclusively by transport vesicles to support the secretion and endocytic activity needed for the rapid cell expansion at the apex. Larger organelles are predominantly segregated to the cytoplasm distal to the subapical region. Underlying the organelle compartmentalization is an elaborate actin cytoskeleton with distinct structural and dynamics properties at the tip, in the subapical region, and in the cytoplasm subtending it. Cytoplasmic domains with differential ionic conditions and spatially restricted localization of molecules in pollen tubes may also be important for regulating the polar cell growth process. The polarized cellular organization in pollen tubes drives an extremely efficient cell growth process that is responsive to extracellular signals, including directional cues. It may be an amplified framework of the cytoplasmic architecture that supports growth in other plant cell types that involves considerably more subtle and transient differential cell expansion.

Cell Compartmentation↗