Methods in phenological mapping

Summary Phenological maps of flowering provide useful information about both spatial and temporal patterns of pollen emission, and their use could bring a substantial improvement of aerobiological forecasts. This paper presents a method for preparing flowering maps by computer on the basis of phenological data and topography. Data drawn from topographic maps are processed with the aid of an empirical model, based on the relationships between phenology and environment, for obtaining a phenological delay matrix. From this matrix it is possible to derive automatically various kinds of maps (chronological, synoptical and differential). An application is described, relating to the blooming patterns of a set of wild plants in a mountainous area of Northern Italy.     

Some details about ragweed airborne pollen in Hungary

Ambrosia elatior – the only species living inHungary – appeared at the beginning of this centuryon the southern border of the country, along theDanube. Since then, unfortunately, it has become themost common weed in Hungary, and the quantity of itspollen is many times higher than the European average(Jäger, 1998; Juhász, 1998). Based on theresults of our ten-year (1989–1998) monitoring inHungary, there is no doubt than more than 60% of latesummer-early autumn pollen originates from ragweed,causing the most serious pollinosis in Hungary. Thequantity and ratio of ragweed pollen is much higherand the length of Ambrosia pollen season islonger in the southern than in the northern part ofHungary.     

Monitoring of airborne pollen and pollen calendar of Cosenza,Southern Italy

Summary From May 1986 to May 1989 surveys of airborne biological particles have been performed in the atmosphere of Cosenza, Italy with the aim of monitoring the presence of airborne pollen. The survey station is situated at Arcavacata of Rende, a hilly area 474 meters above sea level, 7 km north-west of the main town. The sampler (VPPS 2000) is located about 20 meters above ground level. The monitoring, performed in accordance with the criteria suggested by the Italian Association of Aerobiology (A.I.A.), enabled the identification of 26 different pollen types. The pollination graphics show: 1) a late winter period in which pollen from trees appears; 2) a spring period with a prevalence of Gramineae, Fagaceae, Oleaceae andParietaria pollen; 3) a late spring-summer period during which Fagaceae,Pinus and Compositae pollen grains are present; 4) a late summer-autumn and winter period characterized by a drastic reduction of airborne pollen. A study of the calendar reveals, furthermore, that: 1) Gramineae pollen is, amongst those of allergological importance, the most representative; 2)Parietaria is not perennial and is present in modest concentrations; 3)Olea reaches a high peak in June, but not as high as in other southern regions; 3) pollen from arboreal plants is prevalent compared to that of herbaceous plants.     

Seasonal fluctuations of the airborne pollen spectrum in Murcia (SE Spain)

During six consecutive years (1993–1998), aBurkard volumetric pollen trap was continuouslyoperated to sample pollen from the air of thecity of Murcia. The aim of the study was toelucidate the spectra of airborne pollen andthe variations during the year, and toelaborate a pollen calendar. This time spanincludes the end of the period with severedrought from 1990–1995, which particularly affected the south-eastern region of Spain.The total sum of daily average pollenconcentrations amounted to 148,645 pollen grainsbelonging to 93 different taxa. A daily averageof 74 pollen grains/m³ and 11 taxa wererecorded, with maxima of 1157 and 27respectively. The total pollen amountregistered in a year correlated with yearlyrainfall, but there was no relation with meanannual temperature. As for annual fluctuations,there seemed to be no influence by totalrainfall or temperature. Spring and winter werethe seasons with the highest pollen counts andpollen diversity.From the 93 identified taxa, 36 are included inthe pollen calendar. Noteworthy findings are:(i) the presence of Thymelaeaceae,Robinia, Betula, Castanea,Zygophyllum, Caryophyllaceae andCannabis, (ii) a long pollen season ofChenopodiaceae/Amaranthaceae, Urticaceae,Poaceae, Arecaceae and Plantago, (iii)the occurrence of summer, autumn and winterflowering of Artemisia, (iv) the lateappearance of Corylus pollen, and (v) theminor presence of Casuarina pollen duringthe mid winter and late spring.     

Possibility of real-time measurement of an airborne Cryptomeria japonica pollen allergen based on the principle of surface plasmon resonance

In order to establish a real-timemeasurement system for the concentration ofairborne pollen allergens, we examined Cry j 1,one of the major allergens of Cryptomeriajaponica pollen, as an example to establishthe system. The feasible system consisted of:collection of airborne pollen allergens usingthe Virtual Impactor or Cyclone sampler,extraction of Cry j 1 using 10mM HEPES buffercontaining 0.125M NH₄HCO₃ andfinally, real-time measurement of Cry j 1 usingthe BIACORE 3000 system. The sensitivity of thesystem was 5 ng/ml, and 0.1 ml sample volumeand at least 500 pg of Cry j 1 were requiredfor each measurement. Quantification of Cry j 1in the air can be determined 30 min aftercollection, i.e. 15 min for extraction, 10 minfor separation from particulate matters and 5min for the measurement.     

Antibacterial activity and phytochemical evidence for the plant origin of Turkish propolis from different regions

Honeybees collect propolis from practically any abundant plant source in the neighborhood of the hive, be it populus, eucalyptus, pine, sugarcane, cashew nut or orange trees. We have described that the origin plants of Turkish propolis are Populus sp., Eucalyptus sp. and Castanea sativa. In our previous study, propolis samples from Middle Anatolia displayed the typical pattern of “poplar” propolis: they contained pinobanksin, caffeic and ferulic acids and their esters. The propolis samples examined in this study were shown not to contain polar phenolics. The main components of Eucalyptus propolis were aromatic acids, mainly cinnamic acid and its esters, that are usually found in Eucalyptus species resins. The second distinct sample originated from West Anatolia. Although it contained low amounts of phenolic substances and aromatic acids, its main components were sugars and glycosides. The study revealed that there was no significant difference between propolis samples in antibacterial activity, however the yeasts were shown to be more sensitive to eucalyptus-propolis. Gram negative bacteria were susceptible to none of the samples tested.     

Annual, daily and intradiurnal variation of Celtis pollen in the city of La Plata, Argentina

The behaviour of Celtis airborne pollenwas studied for a period of three years(1998–2000) in the city of La Plata. The pollengrains were captured with a Lanzoni trap andthe maximum pollination period was observed tooccur during the end of winter and spring inthe three years. The annual values of pollenconcentration varied, and a significantdecrease was observed during 2000. The greatestairborne pollen record was in October with anaverage of 82.3% in relation to its totalconcentration during 1998–2000. Based on theintradiurnal behaviour analysis, it wasobserved that the maximum pollination peakoccurs at 2H when the temperaturereaches its maximum values. Considering thethree sampling years, the meteorologicalvariables that most influenced the processes of pollen emission,dispersion and transportation in the atmospherewere: maximum and minimum temperature andrainfall.     

Natural levels of pollination intensity and effects of pollen loads on offspring quality in females of Thalictrum pubescens (Ranunculaceae)

Females of Thalictrum pubescens produce stamens containing sterile pollen. Earlier studies have shown the presence of stamens does not increase seed set through increased visitation by insects. Insects may, however, increase pollen deposition on stigmas and increase pollen competition. This paper examines: 1) pollen deposition levels in natural populations and 2) whether larger pollen loads lead to higher quality offspring. The majority of stigmas in two populations received less than 15 grains, but a small proportion had large loads. The latter may represent the occasional insect visit. These visits may provide the opportunity for pollen tube competition. In the greenhouse, flowers receiving heavy-pollen loads had higher seed set than flowers receiving light-pollen loads. Seeds from the two treatments were the same mass, had similar germination times, and seedlings had the same mass. In conclusion, it does not appear that pollen tube competition can account for the maintenance of stamens in females of T. pubescens.     

Intrusion of airborne pollen through open windows and doors

The importance of the transport of pollen by air movement into houses was evaluated using six to eight simultaneously collecting rotorod-type samplers, creating either a sampler line from outdoors to inside the room, or a sampler grid inside a room. The number of incoming pollen grains was highly dependent on the outdoor concentration. The highest concentrations inside (1–2 m distance) and outside (1 m) the room were 600 and 3,250 grains/m³, respectively, in the Betula pollen season and 1,980 and 5,080 grains/m³ in the Pinus season. The pollen concentration and the indoor/outdoor (I/O) ratio decreased as the distance from the ventilation opening increased. Inside the room at a distance of 1–2 m 28%, and at a distance of 3–5 m 12%, of the outside concentration was recorded. In the lower part of the opening the mean proportion was 63% and in the upper part of the opening it was 40%. Efficient ventilation with two open windows increased the I/O ratio and enabled the pollen to spread throughout the room. During the Pinus pollen season 3–35% of the outdoor concentration was simultaneously recorded at six locations inside the room with two open windows and only 0.1–3.6% with one open window. At the same point in the room the I/O ratio varied from <1 to 35%, depending on the sampling conditions. Only a minor effect on the I/O ratio was found between small and large ventilation windows and the door, although it was expected that more air and pollen grains would come indoors through a larger opening.     

The relationship between the flowering phenophase and airborne pollen of Betula in galicia (N.W. spain)

The aim of this work was to investigate thephenological behaviour of Betula in Galicia, NW Spain, and to examine therelationship between the Betula pollencurves and the flowering phenophase. Threetrees were chosen from the each of ninepopulations of Betula located atdifferent altitudes and phytogeographicpositions. Phenological observations of theflowering periods of Betula were made ineach of them. Environmental factors such asfrequency of mist, latitudinal and topographicposition, proximity of the ocean, degree ofsolar exposure, and altitude result inphenological differences between theinvestigated populations. The correlationbetween the Pollinic Production Index of Betula pollen in Galicia and theaerobiological data of the seven monitoringstations showed that the period in which thehighest concentrations were registered wasalmost synchronous with the flowering times atmost of the phenological stations studied.Other factors such as transport and reflotationshould also be taken into account to provide anadequate interpretation of the aerobiologicaldata of Betula pollen in the atmosphere.     

Identification and linkage mapping of complementary recessive genes causing hybrid breakdown in an intraspecific rice cross

One outcome of hybrid breakdown is poor growth, which we observed as a reduction in the number of panicles per plant and in culm length in an F₂ population derived from a cross between the genetically divergent rice (Oryza sativa L.) cultivars ‘Sasanishiki’ (japonica) and ‘Habataki’ (indica). Quantitative trait locus (QTL) analysis of the two traits and two-way ANOVA of the detected QTLs suggested that the poor growth was due mainly to an epistatic interaction between genes at QTLs located on chromosomes 2 and 11. The poor growth was likely to result when a plant was homozygous for the ‘Habataki’ allele at the QTL on chromosome 2 and homozygous for the ‘Sasanishiki’ allele at the QTL on chromosome 11. The results suggest that the poor growth found in the F₂ population was due to hybrid breakdown of a set of complementary genes. To test this hypothesis and determine the precise chromosomal location of the genes causing the hybrid breakdown, we performed genetic analyses using a chromosome segment substitution line, in which a part of chromosome 2 from ‘Habataki’ was substituted into the genetic background of ‘Sasanishiki’. The segregation patterns of poor growth in plants suggested that both of the genes underlying the hybrid breakdown were recessive. The gene on chromosome 2, designated hybrid breakdown 2 (hbd2), was mapped between simple sequence repeat markers RM3515 and RM3730. The gene on chromosome 11, hbd3, was mapped between RM5824 and RM1341. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

RAPD and RFLP mapping of the bacterial blight resistance gene xa-13 in rice

Bacterial blight (BB) caused by Xanthomonas oryzae pv oryzae (Xoo) is one of the most serious diseases of rice. The recessive gene xa-13 confers resistance to Philippine race 6 of Xoo. To tag xa-13 with molecular markers, RAPD analysis was conducted with the combined use of near-isogenic lines and bulked segregant analysis. From the survey of 260 arbitrary 10-nucleotide primers, one primer (OPAC05) was detected to amplify specifically a 0.9-kb band from the DNA of susceptible plants. The distance between the RAPD marker OPAC05-900 and xa-13 was estimated to be 5.3 cM. The RAPD marker was then mapped on chromosome 8 using a mapping population of doubled haploid lines derived from the cross of IR64/Azucena. The linkage between RFLP markers and the RAPD marker was analyzed using an F₂ population of 135 plants derived from a cross between a near-isogenic line for xa-13, IR66699-5-5-4-2, and IR24. No recombinants were found between RZ28 and CDO116 and their distance from xa-13 was estimated to be 4.8 cM. RG136 was located at 3.7 cM on the other side of xa-13. The mapping of xa-13 with closely linked DNA markers provides the basis for marker-aided selection for rice improvement.Department of Agronomy, South China Agricultural University, Guangzhou, China     

Cytological mechanism of pollen abortion resulting from allelic interaction of F1 pollen sterility locus in rice (Oryza sativa L.)

Pollen abortion is one of the major reasons causing the inter-subspecific F₁ hybrid sterility in rice and is due to allelic interaction of F₁ pollen sterility genes. The microsporogenesis and microgametogenesis of Taichung 65 and its three F₁ hybrids were comparatively studied by using techniques of differential interference contrast microscopy, semi-thin section light microscopy, epifluorescence microscopy and TEM. The results showed that there were differences among the cytological mechanisms of pollen abortion due to allelic interaction at the three F₁ pollen sterility loci. The allelic interaction at S-a locus resulted in microspores unable to extend the protoplasm membrane with the enlargement of the microspore at the middle microspore stage and finally producing empty abortive pollen. The allelic interaction at S-b locus caused asynchronous development of microspores at the middle microspore stage producing stainable abortive pollen. The allelic interaction at S-c locus mainly led to the non-dissolution of the generative cell wall and finally caused the hybrid F₁ mainly producing stainable abortive pollen. Genotypic identification indicated that the abortive pollen were those with S ^j allele.     

Genetic analysis and gene mapping of a rice few-tillering mutant in early backcross populations ( Oryza sativa L.)

A rice mutant,G069, characteristic of few tiller numbers, was found in anther culture progeny from theF ₁ hybrid between anindica-japonica cross, Gui630×02428. The mutant has another two major features: delayed tillering development and yellowing apex and margin on the mature leaves. As a donor parent,G069 was further backcrossed with the recurrent parent,02428, for two turns to develop aBC ₂F₂ population. Genetic analysis in theBC ₂F₂ population showed that the traits of few-tillering and yellowing apex and margin on the mature leaves were controlled by one recessive gene. A pool of equally mixed genomic DNA, from few-tillering individual plants inBC ₂F₂, was constructed to screen polymorphism with simple sequence repeat (SSR) markers in comparison with the02428 genome. One SSR marker and three restriction fragment length polymorphism (RFLP) markers were found possibly linked with the recessive gene. By using these markers, the gene of few-tillering was mapped on chromosome 2 between RFLP marker C424 and S13984 with a genetic distance of 2.4 cM and 0.6 cM, respectively. The gene is designatedft1.     

Comparison between the allergenic airborne pollen in Trieste and at Lozzo di Cadore (Italy) in 1989

Summary A comparative study was carried out between aeroallergenic pollen spectra in Trieste and at Lozzo di Cadore. The two localities were found to be different on a five-day running mean basis as to the 8 pollen taxa monitored by the National Aerobiological Monitoring Network managed by the Italian Association of Aerobiology (AIA). In the zone of Lozzo the pollination curve of the spring flowering taxa showed a lag of about one month due to the colder climate. Lozzo di Cadore showed a poor airborne pollen content,Corylus and Graminaceae being prevailing. Trieste has a higher airborne pollen diversity and longer pollination times because of its higher floristic and vegetational complexity. In Trieste the allergenic pollen spectrum showed great quantities ofOstrya, Quercus, Oleaceae, Graminaceae and of ruderal taxa, widespread over the area, due to man's influence.     

Experimental results about Platanus pollen deposition

A comparison between the aerobiological and floristic data carried out for Platanus are reported. The source was constituted by 60 Platanus trees. When they were pollinating we calculated the total pollen prodution of the source, controlling the number of male inflorescences for each plant, the number of anthers for each inflorescence and the number of pollen for each anther. Aerobiological data were obtained with 16 Durham gravimetric samplers positioned over an area of approximately 25 km² at variable distances and in different directions to capture pollen transported by winds coming from any direction. In the first year of analysis, we carried out two kind of aerobiological monitoring positioning a Hirst volumetric trap near the closest gravimetric sampler. The two methods showed a similar trend in Platanus pollen profiles. In the following years, only Durham gravimetric samplers were used to study pollen dispersion. The data showed that there were high concentrations of pollen on the soil near the source while at distances higher than 800 m pollen concentration decreased dramatically. About 1/4 of all the pollen produced fell in an area within 400 m from the source and at a distance of 2750 m only 9 pollen grains/cm² fell throughout the entire season. This revised version was published online in June 2006 with corrections to the Cover Date.     

Frequency and distance of pollen dispersal from transgenic oilseed rape (Brassica napus)

The objective of this study was to evaluate pollen dispersal inBrassica napus (oilseed rape). The selectable marker, used to follow pollen movement, was a dominant transgene (bar) conferring resistance to the herbicide glufosinate-ammonium. Transgenic and non-transgenic plants of the cultivar Westar were planted in a 1.1 ha field trial, with the transgenic plants in a 9 m diameter circle at the centre, surrounded by non-transgenic plants to a distance of at least 47 m in all directions. A 1 m circle of non-transgenic plants was sown in the centre of the transgenic area to allow estimation of the level of pollen dispersal when plants were in close contact. Honeybee hives were placed at the trial site to optimize the opportunity for cross-pollination. During the flowering period, regular observations were made of the number of plants flowering and the number and type of insects present in 60 1 m² areas. These areas were located uniformly around the plot at distances of 1, 3, 6, 12, 24, 36 and 47 m from the edge of the 9 m circle of transgenic plants. Seed samples were harvested from each of the 7 distances so that approximately 20% of the circumference of the plot was sampled at each distance. The centre non-transgenic circle was also sampled. Plants were grown from the seed samples and sprayed with glufosinate to estimate the frequency of pollen dispersal at each distance. In order to screen enough samples to detect low frequency cross-pollination events, seed samples were tested in the greenhouse and on a larger scale in the field. Results were confirmed by testing progeny for glufosinate resistance and by Southern blot analysis. The estimated percentage of pollen dispersal in the non-transgenic centre circle was 4.8%. The frequency was estimated to be 1.5% at a distance of 1 m and 0.4% at 3 m. The frequency decreased sharply to 0.02% at 12 m and was only 0.00033% at 47 m. No obvious directional effects were detected that could be ascribed to wind or insect activity.     

Aerobiology of Castanea pollen in Galicia

The concentration of airborne chestnut pollenhas been investigated at four monitoringstations situated in several cities in Galicia(NW Spain) during 1995–1998. Their pollenseason takes place from mid June to thebeginning of August. The annual total chestnutpollen shows differences between years in eachcity. Likewise there are significantdifferences between cities in each year.The pollen concentrations were closelycorrelated with meteorological parameters. Theyincreased with maximum temperatures and hoursof sunshine and they decreased with rainfalland relative humidity.The diurnal variations of pollen concentrationsshow different patterns in urban and ruralareas. Where the spore trap is surrounded byarboreal masses with chestnut as the dominanttree, the pattern shows two peaks, one in theevening (between seven and mid-night) andanother in the morning (between four and one inthe afternoon).     

Purification and characterization of allergens from Xanthium strumarium pollen

The allergenic components present in whole pollen extract of Xanthium strumarium were isolated by sequential ammonium sulphate precipitation, DEAE Sephadex A50 chromatography and gel filtration. The techniques of RAST inhibition and skin test were utilized to check the allergenicity of fractionated proteins revealing the presence of Xan Ib and Xan VIa as the important allergenic componenets. Xan Ib was found to be devoid of carbohydrate and had a molecular weight of 103 000 daltons. Xan VIa was a glycoprotein of molecular weight 17 000 daltons. The carbohydrate moiety of Xan Vla was found to be associated with allergenicity. The characteristic pattern of whole pollen extract on CIE and TLIEF showed 36 and 21 protein bands, respectively. The use of FPLC in isolation of partially purified allergens from Xanthium is discussed.     

Comparison of some pollen concentrations in Finland and the Estonian SSR

Summary Pollen and spore concentrations were compared in Tartu, Estonia and in three sites in Finland (Turku, Kuopio and Oulu) during May–September 1989. The onset ofQuercus, Pinus, Poaceae,Urtica andRumex flowering started earlier in Tartu than at any site in Finland. The flowering ofJuniperus andArtemisia, on the other hand, began earlier in Turku and Kuopio than in Tartu.Pinus andJuniperus showed a significant correlation (number of pollen grains at the same date) between Tartu and Turku and between Turku and Kuopio. Poaceae andUrtica were correlated between all the sites, as wasRumex except between Tartu and Turku.Artemisia was correlated between Tartu and Turku, Turku and Oulu, and Kuopio and Oulu.Cladosporium correlated between Tartu and Turku. The pollen seasons of Poaceae,Urtica andRumex are prolonged towards the south.