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Kinase partner protein interacts with the LePRK1 and LePRK2 receptor kinases and plays a role in polarized pollen tube growth.

The pollen-specific receptor kinases LePRK1 and LePRK2 have localization and expression profiles that strongly suggest they play roles in pollen germination and tube growth. To identify downstream components of LePRK signaling, we used their cytoplasmic domains (CDs) as baits in yeast two-hybrid screens of a tomato pollen cDNA library. A pollen-specific protein we named kinase partner protein (KPP) interacted with the CDs of both LePRK1 and LePRK2 in yeast and in an in vitro pull-down assay, and with LePRK2 in a co-immunoprecipitation assay. KPP is a peripheral membrane protein and is phosphorylated in pollen. Pollen tubes over-expressing KPP developed balloon-like tips with abnormal cytoplasmic streaming and F-actin arrangements and plants over-expressing KPP exhibited impaired transmission of the transgene through the male. KPP-like genes are found only in plants; the 14 family members in Arabidopsis thaliana exhibit diverse expression patterns and potentially play roles in signaling pathways in other tissues.

Flowers↗

A simplified method for measuring basophil histamine release and blocking antibodies in hay fever patients. Basophil histamine content and cell preservation.

A simplified method for measuring basophil histamine release in grass pollen hay fever patients has been developed. Leukocytes were challenged in vitro with extracts of Phleum pratense (timothy) and the release of histamine was determined indirectly as the residual histamine in the cell sediment. Several steps to purify histamine thus became superfluous and histamine was directly conjugated with o-phthaldialdehyde to form a fluorophore. The simplified method showed a basophil histamine content which was in accordance with results obtained by more specific methods. No difference in basophil histamine content was found between normal and allergic persons. For the histamine liberation assay blood could be adequately preserved for transport for 48 h at room temperature by adding cell culture medium. Basophil histamine release technique allows evaluation of cell sensitivity for determination of the degree of allergy as well as the level of blocking antibodies.

Adult↗

Early changes in airway smooth muscle hyperresponsiveness.

To study asthmatic airway smooth muscle we developed a canine model of ragweed pollen sensitized, airway hyperresponsiveness because of the difficulties in obtaining human tissue. Tracheal and bronchial smooth muscles from sensitized dogs were shown to possess greater ability to shorten and higher maximum shortening velocity (Vo), both of which contribute to the excessive narrowing of airways typical of human asthma. However, maximum force production remained normal, demonstrating the dissociation between the behaviour of shortening and force. Because we found no evidence of inflammation, hypertrophy, or hyperplasia in the sensitized airway smooth muscles, we felt this is a model of early disease and should provide insight into early and perhaps primary pathogenetic mechanisms. Vo is known to be determined by actomyosin ATPase, which in smooth muscle is activated via phosphorylation of the 20-kDa myosin light chain (MLC20) by myosin light chain kinase (MLCK). Therefore, ATPase activity, MLC20 phosphorylation, and MLCK were investigated. Sensitized tracheal and bronchial smooth muscles showed significantly higher ATPase activity, and a higher level of MLC20 phosphorylation, resulting from increased MLCK activity, a consequence of the measured increase in total quantity of MLCK rather than in specific activity. Since MLCK is activated by binding with Ca(2+)-calmodulin complex, intracellular Ca2+ concentration and calmodulin activity were also assessed, but no difference was found between sensitized and control animals. Our study suggests that increased MLCK quantity may be the cause of airway hyperresponsiveness found in sensitized animals, and future investigation should be focused on depicting the reason for the elevated MLCK.

Animals↗

Self-incompatibility system of Oenothera organensis for the detection of genetic effects at low radiation doses.

The self-incompatibility system of Oenothera organensis is used to analyze the frequency of S locus mutations induced by low doses (2.5-20 rad) of fast neutrons and x-rays. Th principles and methods of the screening system for detection of low dose effects have been presented. The results show that low doses induced significantly higher frequencies of seeds and seedlings than those produced spontaneously. The analyses of part of the progenies derived from the control and treated series suggested that they were due to revertible mutations. A modified in vitro method of culturing pollinated cut styles has been developed for rapid screening of compatible pollen tubes.

Culture Techniques↗

An approach for the conquest of the sugi pollinosis using plant growth regulation.

The sugi (Cryptomeria japonica) pollinosis becomes representative allergic disease in early spring in Japan. However, effective treatment for the sugi pollinosis and countermeasure against pollen of C. japonica at its source have not been developed in a practical sense. In this paper, the research aiming to prevent dispersion of pollen of the C. japonica is introduced on application and practical application to the field from the laboratory using the growth regulation of the plant. We found that formation of male flower bud in C. japonica could be suppressed by TNE, since the 3 beta-hydroxylase is inhibited by the action of Trinexysapacethyl, TNE.

Allergens↗

Potato MADS-box gene POTM1-1 transcripts are temporally and spatially distributed in floral organs and vegetative meristems.

The POTM1-1 gene is abundantly expressed in both vegetative and reproductive organs of potato. We performed in situ hybridization and RNA blotting analysis to investigate the patterns of POTM1-1 gene expression in the flower development and the early tuber development. In the early flowers, POTM1-1 transcripts were accumulated abundantly in the developing reproductive organs, including the placentae of carpels and the pollen sacs of stamens. In contrast, the pattern of POTM1-1 distribution during late flower development was different from that of the early flower development. The POTM1-1 transcripts were abundant in the sepals and petals of late flowers, but were minimally expressed in the stamens and carpel. In the shoot apical meristem of the vegetative organs, transcripts were distributed throughout meristem domes, young leaves, and developing vascular cambium. In the early tuberization, the transcripts were widely distributed in the swollen tips of the stolons. Taken together, the results suggest that POTM1-1 gene expression is temporally and spatially regulated in active growing tissues of both vegetative and floral organs with specific distribution patterns dependent upon the developmental stages of the tissue.

Flowers↗

Pollen movement in declining populations of California Valley oak, Quercus lobata: where have all the fathers gone?

The fragmented populations and reduced population densities that result from human disturbance are issues of growing importance in evolutionary and conservation biology. A key issue is whether remnant individuals become reproductively isolated. California Valley oak (Quercus lobata) is a widely distributed, endemic species in California, increasingly jeopardized by anthropogenic changes in biota and land use. We studied pollen movement in a savannah population of Valley oak at Sedgwick Reserve, Santa Barbara County, to estimate effective number of pollen donors (Nep) and average distance of effective pollen movement (delta). Using twogener, our recently developed hybrid model of paternity and genetic structure treatments that analyses maternal and progeny multilocus genotypes, we found that current Nep = 3.68 individuals. Based on an average adult density of d= 1.19 stems/ha, we assumed a bivariate normal distribution to model current average pollen dispersal distance (delta) and estimated delta= 64.8 m. We then deployed our parameter estimates in spatially explicit models of the Sedgwick population to evaluate the extent to which Nep may have changed, as a consequence of progressive stand thinning between 1944 and 1999. Assuming that pollen dispersal distance has not changed, we estimate Nep was 4.57 individuals in 1944, when stand density was 1.48. Both estimates indicate fewer effective fathers than one might expect for wind-pollinated species and fewer than observed elsewhere. The results presented here provide a basis for further refinements on modelling pollen movement. If the trends continue, then ongoing demographic attrition could further reduce neighbourhood size in Valley oak resulting in increased risk of reproductive failure and genetic isolation.

California↗

Use of plots to define pollen-vegetation relationships in densely forested ecosystems of Tropical Africa.

Modern soil samples from South Congo were analyzed for pollen content and compared to forest inventories to define modern pollen-vegetation relationships. A correspondence analysis (CA) was applied independently to botanical and pollen data and a hierarchical cluster analysis to pollen data only. Subsequently, a CA using a presence-absence approach has been made to directly compare the two types of data. Results show that the pollen rain and floristic composition of the sampled sites are not directly linked to altitudinal or precipitation gradients, but clear evidence of variation in relation to hygromorphy and soil type is detected. The forests occurring in swampy environments are well differentiated from the forests developed on well-drained soils by pollen and floristic data. Among forests on well-drained soils, a good distinction can be made between those growing on sandy soils and those growing on ferralitic soils. The comparison between pollen spectra and vegetation shows site-to-site variations in pollen assemblages in relation to the floristic heterogeneity of forests, and it appears that few taxa show a good correlation between plant cover and pollen abundance.

Journal Article↗

[Double-blind randomized trial of the effect of Bidro on hay fever in children].

INTRODUCTION: The natural product royal jelly (trade name Bidro) is widely used for the treatment and prevention of allergic symptoms. Several case reports suggest marked clinical benefits. Our aim was to assess whether treatment with royal jelly modifies the development of allergic hay fever symptoms in children with pollen allergy. MATERIALS AND METHODS: Eighty children aged 5-16 years with hay fever symptoms due to birch, grass and/or mugwort participated in a placebo-controlled, double-blind, randomized, controlled trial. Treatment with royal jelly or placebo was administered three to six months before and throughout the pollen season. RESULTS: The primary outcome was the occurrence of symptoms of rhinitis and conjunctivitis in the pollen season. A secondary outcome was symptom severity, as measured by symptom score, by visual analog scale (VAS) and by the need for additional hay fever treatment. Sixty-four children completed the study; 34 of them had been treated with royal jelly and 30 with placebo. All of the patients in both groups developed hay fever symptoms during in the pollen season. The severity of hay fever symptoms was similar in the two groups (VAS = 1.6 in the placebo group and 1.4 in the royal jelly group; the mean number of combined daily nasal symptoms was 2.83 in the placebo group and 2.82 in the royal jelly group). The need for additional hay fever treatment was similar in the two groups. DISCUSSION: We conclude that royal jelly has no effect on the occurrence of hay fever in the pollen season, nor does it modify the severity of hay fever symptoms.

Child↗

Attractive and repulsive interactions between female and male gametophytes in Arabidopsis pollen tube guidance.

Sexual reproduction in plants, unlike that of animals, requires the action of multicellular haploid gametophytes. The male gametophyte (pollen tube) is guided to a female gametophyte through diploid sporophytic cells in the pistil. While interactions between the pollen tube and diploid cells have been described, little is known about the intercellular recognition systems between the pollen tube and the female gametophyte. In particular, the mechanisms that enable only one pollen tube to interact with each female gametophyte, thereby preventing polysperm, are not understood. We isolated female gametophyte mutants named magatama (maa) from Arabidopsis thaliana by screening for siliques containing half the normal number of mature seeds. In maa1 and maa3 mutants, in which the development of the female gametophyte was delayed, pollen tube guidance was affected. Pollen tubes were directed to mutant female gametophytes, but they lost their way just before entering the micropyle and elongated in random directions. Moreover, the mutant female gametophytes attracted two pollen tubes at a high frequency. To explain the interaction between gametophytes, we propose a monogamy model in which a female gametophyte emits two attractants and prevents polyspermy. This prevention process by the female gametophyte could increase a plant's inclusive fitness by facilitating the fertilization of sibling female gametophytes. In addition, repulsion between pollen tubes might help prevent polyspermy. The reproductive isolations observed in interspecific crosses in Brassicaceae are also consistent with the monogamy model.

Arabidopsis↗

Isolation and characterization of a polymorphic stigma-specific class III peroxidase gene from Senecio squalidus L. (Asteraceae).

A novel stigma-specific class III peroxidase gene, SSP (Stigma-Specific Peroxidase), has been isolated from the self-incompatible daisy Senecio squalidus L. (Asteraceae). Expression of SSP in flower buds is developmentally regulated, with maximal levels of expression coinciding with anthesis, when stigmas are most receptive to pollen and when self-incompatibility is fully developed. In situ hybridization revealed SSP expression to be localized exclusively to the specialized secretory epidermal cells (papillae) of the stigma, which receive and discriminate pollen. SSP is therefore the first tissue-specific and cell-specific peroxidase gene identified in a plant. SSP belongs to a distinct clade of class III plant peroxidases that possess two introns, instead of the more normal situation of three conserved introns. The deduced amino acid sequence of SSP revealed a 27 amino acid signal peptide, suggesting that the SSP protein is secreted to the cell wall of the stigmatic papillae. In-gel peroxidase activity assays showed that SSP has relatively low peroxidase activity compared to other, as yet uncharacterized, peroxidases present in stigmatic extracts. Six SSP alleles have been cloned from different lines of S. squalidus carrying a range of self-incompatibility (S)-alleles but there was no consistent association between the presence of a particular SSP allele and S-genotype indicating that SSP is not the female determinant of SSI in S. squalidus. Nevertheless, the precise expression of SSP in stigmatic papillae suggests that it may have a more general function in pollen-stigma interactions, or alternatively in protection of stigmas from pathogen attack. Extensive database screens have identified homologues of SSP in other plant species, but available expression data for these genes indicates that none are flower-specific, suggesting that SSP represents a new functional type of class III peroxidase specific to the stigma. We discuss the possible function(s) of S. squalidus SSP in pollen-stigma interactions and in protection of stigmas from pathogen attack.

Alleles↗

Assessing the effects of Bt Maize on the predatory mite Neoseiulus cucumeris.

The investigation of Neoseiulus cucumeris in the context of the ecological risk assessment of insect resistant transgenic plants is of particular interest as this omnivorous predatory mite species is commercially available and considered important for biological control. In a multitrophic feeding experiment we assessed the impact of Bt maize on the performance of N. cucumeris when offered spider mites (Tetranychus urticae) reared on Bt (Bt11, Syngenta) or non-Bt maize (near isogenic line) and Bt or non-Bt maize pollen as a food source. Various parameters including mortality, development time, oviposition rate were measured. Spider mites were used as a prey for N. cucumeris, since these herbivores are known to contain similar levels of Cry1Ab toxin, when reared on Bt maize, as those found in the transgenic leaf material. In contrast, toxin levels in pollen of this transgenic cultivar are very low. No differences in any of the parameters were found when N. cucumeris was fed with spider mites reared on Bt and non-Bt maize. Pollen was shown to be a less suitable food source for this predator as compared to spider mites. Moreover, subtle effects on female N. cucumeris (9% longer development time and 17% reduced fecundity) were measured when fed with pollen originating from Bt maize as compared to non-Bt maize pollen. Our findings indicate that the predatory mite N. cucumeris is not sensitive to the Cry1Ab toxin as no effects could be detected when offered Bt-containing spider mites, and that the effects found when fed with Bt maize pollen can be assigned to differences in nutritional quality of Bt and non-Bt maize pollen. The significance of these findings is discussed with regard to the ecological relevance for risk assessment of transgenic plants.

Animals↗

[Prediction of the day on which sugi (Cryptomeria japonica D. Don) pollen scattering will begin--from view point of plant physiology].

We investigated the cumulative thermal constant and threshold temperature for the development of sugi (Cryptomeria japonica D. Don) male flowers to predict the day on which pollen scattering would begin. The threshold temperature for the development of sugi male flowers was 0.17 degrees C and the cumulative thermal constant was 184 degrees C/day. Using these results, we determined how long it would take for sugi to be awaken from dormancy and we investigated the meteorological date. Reactivation from dormancy requires 3 days, where the minimum temperature is less than 3 degrees C with a mean temperature of less than 6 degrees C; the following day must have a maximum temperature of more than 10 degrees C, and these conditions must be maintained for at least 2 days. From these results, it is not difficult to predict the day on which sugi pollen scattering will begin.

Forecasting↗

The promoter of the wheat puroindoline-a gene (PinA) exhibits a more complex pattern of activity than that of the PinB gene and is induced by wounding and pathogen attack in rice.

Puroindolines form the molecular basis of wheat grain hardness. However, little is known about puroindoline gene regulation. We previously reported that the Triticum aestivum puroindoline-b gene (PinB) promoter directs beta-glucuronidase gene (uidA) seed-specific expression in transgenic rice. In this study, we isolated a puroindoline-a gene (PinA), analyzed PinA promoter activity by 5' deletions and compared PinA and PinB promoters in transgenic rice. Seeds of PinA-1214 and PinB-1063 transgenic plants strongly expressed uidA in endosperm, in the aleurone layer and in epidermis cells in a developmentally regulated manner. The GUS activity was also observed in PinA-1214 embryos. Whereas the PinB promoter is seed specific, the PinA promoter also directed, but to a lower level, uidA expression in roots of seedlings and in the vascular tissues of palea and pollen grains of dehiscent anthers during flower development. In addition, the PinA promoter was induced by wounding and by Magnaporthe grisea. By deletion analysis, we showed that the "390-bp" PinA promoter drives the same expression pattern as the "1214-bp" promoter. Moreover, the "214-bp" PinA promoter drives uidA expression solely in pollen grains of dehiscent anthers. The presence of putative cis-regulatory elements that may be related to PinA expression is discussed from an evolutionary point of view. By electrophoretic mobility shift assay, we showed that putative cis-elements (WUN-box, TCA motifs and as-1-like binding sites) whose presence in the PinA promoter may be related to wounding and/or the pathogen response form complexes with nuclear extracts isolated from wounded wheat leaves.

Base Sequence↗

N-Glycan analysis by matrix-assisted laser desorption/ionization mass spectrometry of electrophoretically separated nonmammalian proteins: application to peanut allergen Ara h 1 and olive pollen allergen Ole e 1.

A method has been developed which allows the analysis of glycoproteins separated by SDS-PAGE. The procedure, though applicable to N-glycosylated glycoproteins of any origin, is particularly devised for glycoproteins potentially containing fucose in alpha1,3-linkage to the reducing GlcNAc as may be found in plants and invertebrates, e.g., insects and parasitic helminths. Starting with an established procedure for mass spectrometric peptide mapping, the analysis of N-glycans by matrix-assisted laser desorption/ionization mass spectrometry involved the use of peptide:N-glycosidase A, a triphasic microcolumn for sample cleanup, and a new matrix mixture consisting of 2,5-dihyhydroxybenzoic acid, 1-hydroxyisoquinoline, and arabinosazone. The method was tested on proteins with N-glycans of known structure, i.e., as horseradish peroxidase, zucchini ascorbate oxidase, soybean agglutinin, honeybee venom hyaluronidase, bovine ribonuclease B, and bovine fetuin. An electrophoretic band corresponding to 4 microg of glycoprotein was generally sufficient to allow detection of the major N-glycan species. As an additional benefit, a peptide mass map is generated which serves to identify the analyzed protein. The method was applied to glycoprotein allergens whose glycan structures were unknown. Ara h 1 and Ole e 1, major allergens from peanut and olive pollen, respectively, contained mainly xylosylated N-glycans with the composition Man(3(-4))XylGlcNAc(2) in the case of Ara h 1 and GlcNAc(1-2)Man(3)XylGlcNAc(2) in the case of Ole e 1 where also some GlcNAc(0-2)Man(3)XylFucGlcNAc(2) was found.

Allergens↗

Maternal pollen allergy may be more important than birch pollen exposure during pregnancy for atopic airway disease in the child.

In 1993 extremely high levels of birch pollen were recorded in Stockholm, Sweden. We investigated the effects of this exposure on sensitization and development of atopic airway disease in children. The aim of this study was to assess the influence of maternal birch sensitization and symptoms of pollen allergy, as well as exposure to birch pollen during pregnancy, on sensitization and development of atopic airway disease in children. A total of 387 children with atopic heredity (70% had atopic mothers) and born in Stockholm 1993 or 1994 were investigated at age 4.5-5 yr. The children were examined and skin prick tested with inhalant and food allergens. IgE-antibodies against birch pollen and recombinant birch pollen allergen were analyzed in serum. The same tests were performed on the mothers. Children of mothers with symptoms of pollen allergy more often showed symptoms of rhinoconjunctivitis at age 4.5-5, after both high dose [Odds ratio (OR) 5.3; 95% confidence interval (CI): 2.0-13.7] and low dose (OR 4.0; 95% CI: 1.5-10.9) exposure to birch pollen during pregnancy. Similar tendencies were noted for children of mothers sensitized to birch, where stronger effects were suggested in boys (OR 3.8; 95% CI: 1.3-11.5) than in girls (OR 1.2; 95% CI: 0.2-5.5) in the high-dose exposed group. For asthma symptoms and sensitization to birch in the children the results were less consistent. It may be concluded that, maternal pollen allergy seems to have a stronger influence on the development of rhinoconjunctivitis in children with a family history of atopy than the degree of allergen exposure during pregnancy.

Allergens↗

Pollen grain granuloma: a new clinicopathologic entity.

Chronic ulceration of the ankle developed in a woman who had been stepped on by a cow. Microscopic examination revealed a florid foreign body granulomatous reaction in which pollen grains were found both within giant cells and lying free in fibrous tissue. The grains resembled those of the Asteraceae, or Compositae, family. This is the first report of pollen grains associated with an invasive tissue process. The morphologic features of pollen grains and their differentiation from other foreign bodies are discussed.

Female↗