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Origins and genetic nonvariability of the proteins which diffuse from maize pollen.

The major function of pollen is to deliver the sperm nuclei to the embryo sac. It does this by germinating and producing a pollen tube and thus provides a relatively simple developmental system for study. Mutants for many pollen functions are accessible, as it is a haploid cell. Mature pollen was fractionated into diffusible proteins, soluble proteins, and proteins insolubly associated with membrane or wall; these protein fractions have been quantified and cataloged by native and SDS polyacrylamide gel electrophoresis. Diffusible proteins are localized in the pollen grain wall whereas soluble proteins are cytoplasmic. The roles of haploid and diploid genomes in specifying these proteins is discussed. Pollen from maximally divergent maize lines was examined for quantitative and qualitative variation in the diffusible proteins. A surprising conservation was found for these proteins indicating some functional role which is, at present, unknown. Initial experiments on the incorporation of 35S-methionine into germinating pollen indicate that major representatives of the diffusible proteins are made within the pollen grain itself. They are presumably included in the pollen wall during development and diffuse out through the pore region. Studies with pollen mRNA and experiments on incorporation of 35S-methionine into developing anthers are underway and will identify the origin of these proteins. A knowledge of the basic developmental biology of maize pollen is a prerequisite to its judicious use as a monitor of environmental mutagens.

Diffusion↗

[Usefulness and practical use of an automatic counter for scattering pollen in spring].

BACKGROUND: In this decade some automatic pollen counters have been developed and showed them useful in some reports. However, the results of automatic counting have not been established. METHOD: We have compared scattering cedar and cypress pollen counts with Durham pollen samplers and a newly developed automatic pollen counter (KH-3000) in the spring from 2000 to 2004 in Tokyo. RESULTS: The relationship between the data of the automatic counter and that of a Durham sampler method is so good that this automatic counter seems useful. The best relationship between two methods was seen from late February to early April in the season of high amounts of pollens scatters. The ratio of pollen counts measured with automatic counter and Durham sampler was about 3.5 on average in the period of enough cedar and cypress pollen dispersion. The avoidance of pollens seems possible with real-time pollen information because reliability of automatic counting is highly evaluated when enough pollens scatter. CONCLUSION: In future, the standardization of automatic counting should be necessary for the accurate evaluation of scattering pollens.

Air Pollutants↗

The STUD gene is required for male-specific cytokinesis after telophase II of meiosis in Arabidopsis thaliana.

During male meiosis in wild-type Arabidopsis the pollen mother cell (PMC) undergoes two meiotic nuclear divisions in the absence of cell division. Only after telophase II is a wall formed which partitions the PMC into four microspores. Each microspore undergoes two subsequent mitotic divisions to produce one vegetative cell and two sperm cells in the mature pollen grain. In this paper we describe the isolation and the phenotypic characterization of mutations in the STUD (STD) gene, which is specifically required for male-specific cytokinesis after telophase II of meiosis. Although the male meiotic nuclear divisions are normal in std mutant plants, no walls are formed resulting in a tetranucleate microspore. Despite the absence of cell division in the PMC, postmeiotic development in the coenocytic microspore proceeds relatively normally, resulting in the formation of large pollen grains which contain four vegetative nuclei and up to eight sperm cells. Interestingly, these enlarged pollen grains which contain multiple vegetative nuclei and extra sperm cells behave as single male gametophytes, producing only single pollen tubes and resulting in partial male fertility in std mutant plants. Characterization of the process of pollen development and pollen function in std mutants thus reveals two different types of developmental regulation. Each of the four nuclei found in a std microspore following meiosis is capable of independently undergoing the complete mitotic cell division (including cytokinesis) which the single nucleus of a wild-type microspore would normally undertake. The ability of the four meiotic products to independently continue through mitosis does not depend on their division into separate cells, but is controlled by some subcellular component found within the coenocytic microspore. By contrast, the mature std pollen grain functions as a unit and produces only a single pollen tube despite the presence of multiple nuclei within the vegetative cell, suggesting that this process is controlled at the cellular level independently of the extra subcellular components.

Arabidopsis↗

Cloning and expression pattern of Hor v 9, the group 9 pollen isoallergen from barley.

In this study we report the cloning, sequence, and characterization of Hor v 9 allergen cDNAs from barley (Hordeum vulgare) pollen. Structural homologues of Kentucky bluegrass (Poa pratensis) group 9 pollen allergens were identified in a cDNA library of barley pollen expressed mRNAs. The Hor v 9 cDNA clone (hvp9742) contained an open reading frame encoding 313 amino acids which included a putative 27-residue signal peptide and one asparagine sequon for glycosylation. The mRNA corresponding to clone hvp9742 was produced abundantly in pollen during the late stages of anther development. The protein encoded by clone hvp974 was synthesized as a fusion protein in the E. coli expression vector pMAL. Immunoblots using antibodies to this recombinant allergen, rHor v 9, showed that Hor v 9 protein accumulated during pollen development and was produced maximally at pollen maturity. Using these antibodies, we also provide evidence that Hor v 9 protein localized to the extracellular matrix of mature pollen. Southern blots suggested that Hor v 9 allergens exist as multiple isoforms in barley. Sequence comparisons showed that the Hor v 9 cDNA clones were also homologous to group 5 allergens of Timothy grass (Phleum pratense) pollen and canary grass (Phalaris aquatica) pollen, and the group 9 allergen of ryegrass (Lolium perenne) pollen.

Allergens↗

A rice-based edible vaccine expressing multiple T cell epitopes induces oral tolerance for inhibition of Th2-mediated IgE responses.

Peptide immunotherapy using multiple predominant allergen-specific T cell epitopes is a safe and promising strategy for the control of type I allergy. In this study, we developed transgenic rice plants expressing mouse dominant T cell epitope peptides of Cry j I and Cry j II allergens of Japanese cedar pollen as a fusion protein with the soybean seed storage protein glycinin. Under the control of the rice seed storage protein glutelin GluB-1 promoter, the fusion protein was specifically expressed and accumulated in seeds at a level of 0.5% of the total seed protein. Oral feeding to mice of transgenic rice seeds expressing the T cell epitope peptides of Cry j I and Cry j II before systemic challenge with total protein of cedar pollen inhibited the development of allergen-specific serum IgE and IgG antibody and CD4(+) T cell proliferative responses. The levels of allergen-specific CD4(+) T cell-derived allergy-associated T helper 2 cytokine production of IL-4, IL-5, and IL-13 and histamine release in serum were significantly decreased. Moreover, the development of pollen-induced clinical symptoms was inhibited in our experimental sneezing mouse model. These results indicate the potential of transgenic rice seeds in production and mucosal delivery of allergen-specific T cell epitope peptides for the induction of oral tolerance to pollen allergens.

Allergens↗

The importance of the pecan tree pollen in allergic manifestations.

BACKGROUND: Pecan tree pollen is considered to be highly allergenic. However, no specific scientific data about its role in causing allergic diseases are available. OBJECTIVE: To study the role of pecan tree pollen in the development of allergy. METHODS: The presence of pecan tree pollen was determined by weekly and monthly counting of airborne grains. The incidence of pecan tree pollen atopy and clinical manifestations were studied in 395 participants, aged 4-70 years, who comprised 78.2% of the whole eligible population of a rural community. The participants were skin tested for different extracts of allergens, completed detailed questionnaires, and their medical files were evaluated. RESULTS: During May, pecan tree pollen grains comprised 70% of the total airborne grains. A positive skin-prick test (SPT) to pecan was shown by 46 (11.6%) participants, constituting 25.4% of the atopic population. Of those who were found atopic to one or more allergens 50.3% had symptoms, whereas the parallel figure for those atopic to pecan pollen was 76.1% (P < 0.005); 58.7% of the pecan atopic participants had hay fever, 43.5% had asthma, and 31.5% had both hay fever and asthma. Among pecan atopic participants the incidence of hay fever increased with age (P = 0.05), while the incidence of bronchial asthma, as a sole manifestation of allergy, decreased in the > 17-year-old age group (P < 0.01). Of the pecan atopics 65.2% had clinical symptoms coinciding only with the pecan pollen season and an additional 10.9% had perennial symptoms. CONCLUSION: Pecan tree releases highly allergenic pollen grains, which are correlated to the incidence of hay fever in the exposed population. The contribution of pecan tree pollen to the symptoms was highly significant after discounting olive and cypress trees that also pollinate in the spring. In children, the pecan tree constitutes a possible etiologic agent for the development of asthma.

Adolescent↗

Genome-wide identification of genes expressed in Arabidopsis pistils specifically along the path of pollen tube growth.

Plant reproductive development is dependent on successful pollen-pistil interactions. In crucifers, the pollen tube must breach the stigma surface and burrow through the extracellular matrix of the stigma epidermal cells and transmitting tract cells before reaching its ovule targets. The high degree of specificity in pollen-pistil interactions and the precision of directional pollen tube growth suggest that signals are continually being exchanged between pollen/pollen tubes and cells of the pistil that line their path. However, with few exceptions, little is known about the genes that control these interactions. The specialized functions of stigma epidermal cells and transmitting tract cells are likely to depend on the activity of genes expressed specifically in these cells. In order to identify these genes, we used the Arabidopsis (Arabidopsis thaliana) ATH1 microarray to compare the whole-genome transcriptional profiles of stigmas and ovaries isolated from wild-type Arabidopsis and from transgenic plants in which cells of the stigma epidermis and transmitting tract were specifically ablated by expression of a cellular toxin. Among the 23,000 genes represented on the array, we identified 115 and 34 genes predicted to be expressed specifically in the stigma epidermis and transmitting tract, respectively. Both gene sets were significantly enriched in predicted secreted proteins, including potential signaling components and proteins that might contribute to reinforcing, modifying, or remodeling the structure of the extracellular matrix during pollination. The possible role of these genes in compatible and incompatible pollen-pistil interactions is discussed.

Arabidopsis↗

The link between allergic rhinitis and allergic asthma: a prospective population-based study. The Copenhagen Allergy Study.

BACKGROUND: It has been hypothesized that allergic rhinitis and allergic asthma are manifestations of the same disease entity. We aimed to investigate the relationship between allergic rhinitis and allergic asthma. METHODS: Participants in a population-based study of 15-69-year-olds in 1990 were invited to a follow-up in 1998. A total of 734 subjects were examined on two occasions eight years apart. Allergic rhinitis to pollen was defined as a history of nasal symptoms on exposure to pollens and IgE specific to pollen. Allergic asthma to pollen was defined as a history of lower airway symptoms on exposure to pollens and IgE specific to pollen. Similarly, diagnoses of allergic rhinitis and allergic asthma to animals or mite were defined. RESULTS: At follow-up, all subjects with allergic asthma to pollen (n = 52) had in addition allergic rhinitis to pollen. In the longitudinal analysis, there were a total of 28 new (incident) cases of allergic asthma to pollen. They all had allergic rhinitis to pollen at baseline, or had developed allergic rhinitis to pollen at follow-up. Accordingly, allergic rhinitis to animals and mite were ubiquitous in subjects with allergic asthma to animals and mite, respectively. CONCLUSIONS: The results support the hypothesis that allergic rhinitis and allergic asthma are manifestations of the same disease entity.

Adolescent↗

Pollen embryogenesis to induce, detect, and analyze mutants.

The development of fully differentiated plants from individual pollen grains through a series of developmental phases that resemble embryogenesis beginning with the zygote was demonstrated during the mid-1960's. This technology opened the door to the use of haploid plants (sporophytes with the gametic number of chromosomes) for plant breeding and genetic studies, biochemical and metabolic studies, and the selection of mutations. Although pollen embryogenesis has been demonstrated successfully in numerous plant genera, the procedure cannot as yet be used routinely to generate large populations of plants for experiments. Practical results from use of the technology in genetic toxicology research to detect mutations have failed to fully realize the theoretical potential; further developments of the technology could overcome the limitations. Pollen embryogenesis could be used to develop plants from mutant pollen grains to verify that genetic changes are involved. Through either spontaneous or induced chromosome doubling, these plants can be made homozygous and used to analyze genetically the mutants involved. The success of this approach will depend on the mutant frequency relative to the fraction of pollen grains that undergo embryogenesis; these two factors will dictate population size needed for success. Research effort is needed to further develop pollen embryogenesis for use in the detection of genotoxins under both laboratory and in situ conditions.

Haploidy↗

The cytological mechanism of low fertility in the naked seed rice.

The low fertility of naked seed rice (NSR) was investigated by the following observations: somatic chromosome constitute, behavior of pollen mother cells (PMCs), the germination of mature pollen grains, the development of male and female gametes and the structure of the anther opening. The results indicated that somatic chromosomal number was 2n = 24, behavior of PMCs were normal and most of pollen grains could regularly develop further to mature male gametophytes in NSR. And dehiscence chamber and thickened endothecium cell (TEC) in numerous anthers of the NSR were developed abnormally after dicaryotic phase, result in few anthers complete opening and most partly opening or failure to opening, therefore much fewer of pollen grains attach on the stigma as compared with normal variety. Furthermore most of embryo sacs possessed abnormal structure and were sterile. All of above illustrated that the failure of the anther opening and the abortion of female gametophyte were main factors controlling the low seed-setting rate of the NSR.

China↗

Development of ovulate cones from initiation of reproductive buds to fertilization in Cephalotaxus wilsoniana Hay.

Cephalotaxus wilsoniana Hay, is endemic to Taiwan. This study was performed morphologically and anatomically to investigate reproduction in this species for the purpose of conservation. The duration from reproductive bud formation to fertilization in C. wilsoniana lasts about one year and five months. Buds are initiated in late January and differentiate into one vegetative bud and 3 female cones in late February. A female cone is constructed with 4 pairs of decussate opposite bracts. A small ridge-like secondary axis sits on the axil of each bract. Two ovules are borne on both sides of each secondary axis. A lysogenous pollen chamber begins to be formed from the degenerative tissues on the top end of the nucellus in early March. In late March the megasporogenous tissue is differentiated in the core center of the nucellus, and the micropyle closes gradually after pollination. By late July, pollen tubes have developed in the pollen chamber, and the megaspore mother cell appears. Then the functional megaspore becomes active in mid-October. The 8 free nucleate macrogametophyte appears in late December. From January to late March of the following year, the elliptical cyst-like female gametophyte keeps growing through continuous divisions of its free nuclei. The cyst layer of protoplast thickens in early April. In mid-April, cell walls begin to form among free nuclei. The archegonia are initiated in late April. Pollen tubes extend their tips to the macrogametophyte in early May, and each tube with 2 spermatozoids reaches a mature archegonium with an egg needed to perform fertilization in late May. Generally, only 1-(3) ovules in each cone can become mature.

Plants, Medicinal↗

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

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

Climate↗

The major olive pollen allergen (Ole e I) shows both gametophytic and sporophytic expression during anther development, and its synthesis and storage takes place in the RER.

The distribution of Ole e I (the major olive pollen allergen) and its transcripts was investigated in the anther from premeiotic stages until the dehiscent pollen stage. Crude protein extracts were analyzed by immunoblotting and probed with a monoclonal antibody to Ole e I. The protein, with three variants, was found to accumulate from the early microspore stage onwards. In addition to the previously reported localization of the protein, Ole e I has been immunolocalized for the first time within the pollen wall and in the tapetum. Reverse transcription-polymerase chain reaction analysis using specific oligonucleotides and RNA extracted from whole anthers revealed that the Ole e I gene is expressed from the late tetrad stage onwards. No expression was found in control tissues such as petals, roots or leaves. Light microscopy in situ hybridization on developing flower buds and dehiscent pollen confirmed the transcripts to be present in both the microspores and the sporophytic tissue (tapetum). Labeling was found primarily in the tapetum, reaching the highest concentration in the cytoplasm of the developing and mature pollen, once tapetum started to degenerate. In situ hybridization at the transmission electron microscope level showed the transcripts to accumulate on ribosomes of the rough endoplasmic reticulum. These studies, together with others carried out previously by us, indicated that both synthesis and storage of Ole e I take place in the endoplasmic reticulum, coincidentally with the conspicuous changes suffered by this membrane system during pollen development. This process is most likely controlled at the transcriptional level. The localization of the protein in the pollen ectexine bring new insights into the function of the allergen, which are discussed.

Allergens↗

Hypereosinophilia, neurologic, and gastrointestinal symptoms after bee-pollen ingestion.

A patient developed hypereosinophilia (13,440 cells per cubic millimeter) 6 weeks after beginning the ingestion of bee pollen. Symptoms included generalized malaise, headache, nausea, abdominal pain diarrhea, generalized pruritus, and decreased memory. Evaluation revealed no other known cause for the patient's hypereosinophilia, which resolved after bee-pollen ingestion was stopped. The product contained a mixture of entomophilous and anemophilous pollens to which the patient was skin test positive. An open challenge with the bee pollen later reproduced the presenting symptoms with a concomitant rise of the eosinophil count from 207 to 890 cells per cubic millimeter. The patient has since remained well avoiding bee pollen. This study strongly suggests that hypereosinophilia with attendant pathophysiologic disturbances may be an adverse reaction to bee-pollen ingestion in atopic individuals.

Adult↗

[Peculiarities of embryonal development in vitro of Brassica napus pollen grains].

Literature data concerning the problem of the embryogenesis in microspore cultures of Brassica were reviewed. The influence of stress factors on the microspore cultures was examined. The some cytological and morphological peculiarity microspore entering in the embryogenesis developmental pathway were described.

Brassica napus↗

[Development of allergic reactivity to Artemesia pollen during combined sensitization to pollen and microbes].

Some regularities of formation of hypersensitivity of the immediate type to the pollen of Artemisia absinthium were studied under conditions of combined hypersensitivity to pollen and Brucella abortus 19-BA vaccine strain; the latter was administered 3, 12, and 28 days after the pollen. The degree of specific allergic reconstruction to the pollen was studied by passive skin anaphylaxis after Ovary, indirect degranulation of mast cells of healthy rats, and by general anaphylaxis in response to intravenous injection of the Artemisia absinthium pollen water-salt extract. Early formation of allergy to the pollen was observed in the groups of animals with combined hypersensitivity to the pollen and brucellae. The degree of allergic reactivity to the pollen allergen was more expressed in the groups with combined allergy than in those with pure pollen hypersensitivity at all the stages of this experiment.

Anaphylaxis↗

Experimental analysis of protogyny in Aquilegia canadensis (Ranunculaceae).

Dichogamy is very common in flowering plants and is widely thought to reduce pollen-pistil interference, especially self-pollination. Yet, the functional significance of dichogamy has rarely been studied. We investigated the nature and functioning of dichogamy in eastern Ontario populations of Aquilegia canadensis, a highly selfing columbine previously described as protogynous. We then manipulated flowers to determine whether increased protogyny would reduce self-fertilization. Contrary to previous reports, A. canadensis is not dichogamous. Controlled pollinations in a greenhouse showed that pollen tubes generally begin to develop after anther dehiscence. Although stigmas can collect pollen early in floral development, naturally pollinated flowers collected from four populations had few pollen grains on stigmas and almost no pollen tubes in styles until after anther dehiscence. Limited pollen deposition before anther dehiscence was also associated with low nectar availability and limited sepal expansion. Because inbreeding depression is strong in this species, selection may favor increased protogyny if it reduces selfing. We tested this hypothesis by comparing the level of selfing in flowers rendered protogynous by the removal of the first 19 (of 39) anthers to develop, with nonprotogynous control flowers. Contrary to expectations, protogyny did not reduce selfing. Our results emphasize the importance of detailed field observations and manipulative experiments in understanding the nature and functional significance of dichogamy.

Journal Article↗