Stable isotope ratios of carbon and nitrogen in pollen grains in order to characterize plant functional groups and photosynthetic pathway types

Measurements of delta(13)C, delta(15)N and C : N ratios on modern pollen grains from temperate plants, including whole grains as well as extracted sporopollenin, were analysed in order to characterize physiological plant types at the pollen level and to determine the variation of these parameters in modern pollen grains of the same climatic area. Measurements are presented for 95 batches of whole modern pollen from 58 temperate species and on the stable fraction of modern pollen grains, chemically extracted sporopollenin, for two modern species. Fourier transform infrared (FTIR) and cross-polarization and magic-angle spinning (CP/MAS) sporopollenin spectra were conducted in parallel. C(3) and C(4) plants can be separated by delta(13)C measurements based on pollen. Probabilistic assignments to plant functional groups (herbaceous, deciduous woody, evergreen woody) of C(3) plants by the means of a discriminant analysis can be made for C : N ratios and for delta(13)C. The results are related to other studies on sporopollenin in order to use this method in future work on fossil samples. Stable isotope measurements on pollen allow improved pollen diagrams, including forms that cannot be differentiated at species level, increasing the accuracy and resolution of plant physiological type distribution in quaternary and older fossil sediments.     

A study on pollen grains ofCrocus cartwrightianus (Iridaceae)

Crocus cartwrightianus andC. cartwrightianus cv.albus pollen have been studied from structural and ultrastructural points of view and the germination assayed in vivo and in vitro.C. cartwrightianus pollen is regularly shaped and sized, has a low percentage of anomalous grains and has a high germination rate in vitro, whileC. cartwrightianus cv.albus is less regularly shaped with some variation in size and has a high percentage of anomalous grains and a low germination percentage in vitro. Ultrastructural observations have revealed, in the pollen of both the taxa, the presence of a thin elongated vegetative nucleus and a generative cell surrounded by a thin membrane. However,C. cartwrightianus pollen shows a thicker intine, andC. cartwrightianus cv.albus shows numerous pollen germination anomalies which are in common withC. sativus.     

The use of sieving during standard pollen pre-treatment of samples of fossil deposits to enhance the concentration of large pollen grains

Small pollen grains and fine particles of the organic matrix in samples of a coarse detritus mud and a blanket peal deposit were removed by sieving through 30μ polyester mesh after digestion with KOH and acetolysis during standard pollen pre-treatment. The technique enhanced the number of large pollen grains m the sample. There was also an increase in the range of taxa which produce huge grains. The method provided a means of estimating the ratio of any scarce large pollen grain to the total grain sum in samples prepared by standard methods of pre-treatment.     

An aerobiological study of pollen grains and fungal spores of Barcelona (Spain)

Summary A study of concentration of airborne pollen grains and fungal spores has been carried out in Barcelona (Spain) during 1989–90. The volumetric method of filtration, previously described for airborne pollen analysis (Suarez-Cervera and Seoane-Camba, 1983) has been used. In this case, the filters have also been cultivated in Czapecdox-agar, Sabouraud-agar and Sabouraud-agar with streptomycin for the identification of the fungal colonies. Analysis of the number of fungal spores growing on the filter shows that the maxima of colonies of spores developed in culture per m³ of air filtered, correspond to September–December. Pollen and spore concentrations start from November–December, reach a maximum in March–April and decline progressively until September–October. Therefore, in the city of Barcelona, the greatest concentration occurs in spring and the lowest in autumn.     

New details on the morphology of fossil onagraceous pollen grains

Fossil onagraceous pollen grains from two Upper Miocene localities in E. Austria were investigated by LM and EM. Exine structure and sculpture as well as viscin threads suggest affinities with the extant genusCircea.     

Modern surface pollen assemblages from the Middle and High Atlas,Morocco: insights into pollen representation and transport

Thirty-three modern surface samples were collected in the environmentally and climatologically contrasting regions of the Middle and High Atlas Mountains, Morocco. Samples representing forest and steppe montane environments (1935–2760 m above sea level) are clustered around study sites at Lake Tislit (High Atlas, semi-arid oro-Mediterranean bioclime) and Lake Sidi Ali and Michliffen (Middle Atlas, sub-humid montane Mediterranean bioclime). Good discrimination between regional pollen spectra is evident, with Middle Atlas samples reflecting higher arboreal cover (Cedrus and evergreen Quercus) and High Atlas samples with high abundances of non-arboreal taxa, including Artemisia and Fabaceae. These four taxa (Cedrus, evergreen Quercus, Artemisia and Fabaceae) are furthermore shown to be reliable indicators of local source vegetation within a 100 m² quadrat, taking into account threshold abundances of 7%, 20%, 4% and 10%, respectively. Deciduous Quercus, Olea and Phillyrea show long-distance pollen dispersal across both regions, contributing to non-trivial arboreal pollen (AP) values of up to 35% (typically 20–30%) in the High Atlas spectra. In the Middle Atlas, AP values of 40 to 50% occur in open sampling locations and > 60% under forest canopy cover. These insights should be taken into account when interpreting ancient pollen spectra from regional lakes and bogs for palaeoenvironmental reconstruction.     

Induction of embryogenic pollen grains in situ and subsequent in vitro pollen embryogenesis in Nicotiana tabacum by treatments of the pollen donor plants with feminizing agents

Tobacco plants ( Nicotiana tabacum L., var. Badischer Burley) were treated with chemicals (sprays and soil drenches) known to affect sex expression in other species. Their effect was tested on sex balance, pollen sterility, embryogenic pollen grain (P-grain) formation in situ, and on pollen plant formation in anther and pollen cultures after anther preculture. Napthalene acetic acid (NAA) increased the length of pistils and stamens and shifted sex balance towards femaleness when the plants were raised in long or short days at 24 or 15°C. In parallel, pollen sterility, P-grain frequency in situ and pollen plant production from anther and pollen cultures were increased by NAA. Alar 85 redueed the length of pistils and stamens and shifted sex balance towards femaleness when the plants were raised in long days at 24°C, but shifted it towards maleness in short days and/or at 15°C. In parallel, pollen sterility, P-grain frequency in situ, and pollen plant production in vitro were increased when plants in long days at 24°C were treated with Alar 85, but decreased when plants in short days and/or at 15°C were treated. Ethrel, Cycocel, and GA₃ applied in a similar manner, were ineffective. Water sprays and nitrogen starvation shifted sex balance towards femaleness in long days at 15°C and increased pollen sterility, P-grain frequency in situ and pollen plant production in vitro. At 24°C, water sprays and nitrogen starvation had no effect.     

外源性糖对四种木本观赏植物花粉离体培养的影响

分别以单、双、多糖作用于紫荆、紫丁香、桃、连翘四种木本观赏植物花粉的离体培养,检测其对花粉萌发、花粉管生长及花粉管胼胝体形成的影响,结果表明糖的类型和浓度对花粉萌发有较大影响,同时不同植物的花粉对糖的敏感性也存在差异;花粉管长度明显受蔗糖浓度的影响,尤其是连翘的花粉管长度,与蔗糖浓度成反比;在四种植物的花粉管生长中后期,均观察到胼胝体的形成,胼胝体形成的数量与蔗糖浓度及花粉管长度存在一定的联系;在花粉管中胼胝体的形成没有特定位点。     

Inhibition of the activity of aminoacyl-tRNA synthetases in pollen grains of maize

Extracts from pollen grains of maize ( Zea mays L.) show a low activity of aminoacyltRNA synthetases (EC 6. 1. 1). They also contain a specific factor inhibiting the activity of these enzymes. The molecular mass of this factor, which may be a short peptide, is about 3000 Da as determined by column chromatography on Sephadex G-25 Fine. The Michaelis constant (K_m), determined for the amino acid in the presence of this factor, suggests its allosteric influence on the affinity of the enzyme. Short-term incubation of the factor with pronase R resulted in conversion of the inhibiting action into a stimulating one. Kinetics of aminoacylation reactions confirm inhibitory and stimulative influences of the effector on the enzyme activity. High performance liquid chromatography shows that inhibition of the activity of aminoacyl-tRNA synthetases is affected by a group of compounds of similar molecular masses.     

Electron tomography of structures in the wall of hazel pollen grains

The three-dimensional structure of channels and bacula cavities in the wall of hazel pollen grains was investigated by automated electron tomography in order to explore their role in the release of allergen proteins from the pollen grains. 3D reconstructions of 100–150 nm thick resin-embedded sections, stabilized by thin platinum–carbon coating, revealed that the channels aimed directly towards the surface of the grain and that the bacula cavities were randomly sized and merged into larger ensembles. The number and the dimensions of the ensembles were quantitatively determined by neighboring voxel analysis on thresholded reconstructed volumes. To simulate the allergen release, allergen proteins were approximated by a hard sphere model of a diameter corresponding to the largest dimension of the known 3D structure of the major birch allergen, Bet v 1, whose amino acid sequence is highly similar to the amino acid sequence of the major hazel allergen, Cor a 1. The analysis of positions where the hard sphere fits into the resolved channels and bacula cavity structures revealed that unbound allergens could freely traverse through the channels and that the bacula cavities support the path of the allergens towards the surface of the grain.     

A study of Gramineae and Urticaceae pollen in the Derby area

Summary Gramineae and Urticaceae seasonal variations and diurnal rhythms for Derby in 1986, 1987 and 1988 are shown and discussed.The time of highest hourly concentration of Gramineae for Derby in June was 1900 hours. Maximum hourly counts for June of Urticaceae pollen occurred between 1400 and 1600 hours.Urticaceae has a second period of high concentration in August. Gramineae and Urticaceae pollens from 1969–1990 show no significant long term trends.     

Trends in the frequency of different pollen types in Vienna from 1976 to 1989

Summary For allergological purposes it is of interest whether the number of allergenic pollen will change in a significant order. Linear regression and correlation of different pollen types show that the annual totals of most of the allergenic pollen remain constant. None of the pollen grains show distinct and significant disappearance, but four types show significant increase of annual totals: Varia, Sambucus, Cupressaceae and Rubicaea. Pollen of Cupressaceae and Castanea fairly will become of more allergological interest within the next years, if the upward-trend should prove to be continous. No significant change was found in the number of the investigated spores.     

A study of Quercus pollen in the Derby area, UK

The major allergenic pollen prevalent in the Derby air in May is Quercus pollen which has been monitored volumetrically from 1970–1997. Quercus pollen levels in Derby are increasing, showing an established long term trend, with 1995 being an exceptionally high year. There is now an earlier start date and a longer seasonal duration. The mean Quercus diurnal periodicity for 1991–1997 shows a peak at 15.00 hours.A detailed study of the 1990–1997 seasons established that a maximum temperature of 20 °C or above, at the usual time of flowering, occasions the start of the Quercus pollen season. Average May temperature and drought in the previous June and July are important factors in determining Quercus pollen totals. Predictions for the forthcoming seasons were produced which compared favourably with the actual pollen totals.     

Distribution of loosely-bound calcium in the vegetative and generative cells of the pollen grains in Chlorophytum elatum

We used chlorotetracycline fluorescence, alizarin staining and potassium pyroantimonate methods, as well as X-ray microanalysis, to demonstrate the differential localization of Ca2+ during pollen maturation. Level of loosely-bound Ca2+ ions was higher in the generative cell than in the vegetative cell of the mature pollen grain which is one of the symptoms of metabolic differentiation of the two sister cells. This revised version was published online in July 2006 with corrections to the Cover Date.     

A correlation study between airborne pollen and cases of pollinosis in humans

This study aims to establish themathematical relationship between allergenicpollen in the air and clinical cases ofpollinosis (hay fever) in humans, and toevaluate the immediate effect of wind and rainon such cases. The pollen was collected inFigueres, Catalonia, Spain (42°30 Nand 3° E), through a filtration method,in the spring of 1998. Skin tests revealed thatthe antigenic taxa are Olea europaea,Plantago sp., Parietaria officinalisand Gramineae. A linear correlation –with the number of days with presence of pollenas the abscissa and the number of days withcoincidence of pollinosis as the ordinate – wasperformed. The linear correlation coefficientwas 0.9285, at the limit of a directcorrelation by two degrees of freedom.Furthermore, a negative relationship wasobserved between the number of days with rainand number of days with pollinosis, and apositive relationship between the number ofdays with fixed wind speed and number of dayswith pollinosis.     

A histochemical study of pollen digestion in the alimentary canal of honeybees (Apis mellifera L.)

Histochemical staining of embedded sections of the alimentary canal of honeybees indicated that carboxylated polysaccharides could be efficiently digested and absorbed, except when sequestered within unswollen and unbroken pollen grains that appeared to be impervious to enzymes of the gut. Hemicelluloses and pectic acids of the pollen wall structure underwent partial digestion, but pollen wall cellulose and sporopollenin were not digested. A separation of pollen walls from colloidal semisolids of the pollenkitt and disgorged protoplasm was apparent. The layers of slurry created by this separation remained into the rectum, and consisted primarily of saturated organic compounds which do not stain with weakly basophilic or acidophilic dyes.     

Cellular changes during the acquisition of embryogenic potential in isolated pollen grains ofNicotiana tabacum

Summary We used electron microscopical techniques to study ultrastructural changes during the acquisition of embryogenic competence in immature pollen grains ofNicotiana tabacum, isolated at the early- or mid-bicellular stage and cultured in vitro under starvation conditions. Cytoplasmic and nuclear changes during the starvation treatment are reported. Dedifferentiation of plastids, dilation of the wall of the generative cell, the appearance of a large vacuole, loss of nuclear pores in the vegetative nucleus, changes in chromatin and nucleolar structure, and a decrease in the size of the nucleolus were observed. We suggest that these events are the first step in the switch from generative to vegetative generation during pollen embryogenesis.