Influence of arginine,ornithine, DFMO and polyamines on division of the generative nucleus in cultured pollen tubes of <Emphasis Type="Italic">Aechmea fasciata</Emphasis> (Bromeliaceae)

During in vitro pollen tube growth of Aechmea fasciata the second pollen mitosis (PM II) that produces two sperm cells was influenced by exogenous amino acids. Arginine (Arg) as single amino acid was the limiting factor for the second mitosis of the generative nucleus and thus the formation of sperm cells in cultured pollen tubes of A. fasciata. The involvement of Arg was probably related to protein synthesis. The need for Arg was not related to polyamine (PA) biosynthesis, since PA added to the germination medium were unfavourable for sperm cell production. Both ornithine (Orn) and difluoromethylornithine (DFMO) inhibited the second mitosis in cultured pollen tubes of A. fasciata. The addition of Arg during the first 2 h of pollen germination was necessary to establish the division of the generative nucleus 6 h later.     

<Emphasis Type="BoldItalic">Betula</Emphasis> spp. pollen in the atmosphere of Novi Sad (2000–2002)

Pollen of Betula spp. is one of the main European aeroallergens. The aim of this study was to determine characteristics and occurrence of the Betula pollen in Novi Sad atmosphere, based on 3-year observations (2000–2002), and to compare pollen season start dates calculated by different methods. Pollen samples have been collected by Hirst volumetric method with a 7-day Burkard spore trap. Four methods (Sum 75, 2.5%, 30 and 1 pg/m ³) have been used for determination of the start dates of the Betula pollen season and the results have been compared. The total annual pollen sum increased during the observed period. In 2000, 2001 and 2002, the highest daily pollen concentrations were 97, 137 and 1034 pg/m ³, respectively. The earliest Betula pollen season start has been calculated by the 1 pg/m ³ method.     

Mating System in a Low-density Natural Population of the Dioecious Wind-pollinated Patagonian Cypress

The mating system of a species is an essential factor that determines the genotypic frequencies of its populations in each generation. Thus, knowledge in this regard is important in the application of practical measures in conservation and forest tree breeding. Austrocedrus chilensis (D.DON) PIC. SER. ET BIZZARRI is a member of the Cupressaceae native to the Andean–Patagonian Forest of Argentina and Chile with regional economic importance. It is a dioecious tree with pollen and seeds distributed by wind. Isozyme markers were utilized to study the mating system of this species through the survey of individual effective pollen clouds in a natural low-density population. If random mating occurs, effective pollen clouds that pollinate single seed trees are expected to be genetically homogeneous. The classical HardyWeinberg test of panmixia was also performed. Although it failed to reject random mating, the analysis of the pollen clouds showed that this mating pattern does not occur. Evidence of locally distributed heterogeneous pollen clouds is presented. The two methods are critically compared.     

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.     

Immature maize spikelets develop and produce pollen in culture

Immature maize spikelets have been successfully grown in vitro. Culture conditions were refined to maximize development of normal pollen grains. Kinetin was not required for normal development, in contrast to the absolute requirement for this plant growth regulator for in vitro tassel development. Development occured in all stages sampled, from premeiosis to postvacuolation, and there was no lag in progression through the various stages of development as compared to greenhouse-grown material. Cultured spikelets produced pollen that appeared morphologically normal, accumulated starch and had the normal two sperm nuclei and single vegetative nucleus.     

鱼腥草花粉母细胞减数分裂特征及其育性研究

为深入了解鱼腥草花粉母细胞的减数分裂特征与花粉育性的关系,该研究采用卡宝品红染色法对2个鱼腥草居群花粉母细胞的减数分裂过程进行观察,并采用氯化三苯基四氮唑(TTC)染色法、I2-KI染色法、B-K培养基培养法及荧光显微镜观察法来检测鱼腥草花粉的活力及萌发率。结果发现:(1)鱼腥草减数分裂的进程与花序大小、花药颜色、花药长度均有密切的关系。(2)2个居群的鱼腥草中花粉母细胞减数分裂过程正常占88.2%,有11.8%的花粉母细胞减数分裂异常。(3)减数分裂异常表现在减数分裂过程中出现微核、落后染色体、染色体桥、不均等分离、多分体等现象,并发现在二分体阶段及单核花粉发育过程中存在细胞融合。(4)2个居群的鱼腥草花粉活力均不超过1.5%,花粉几乎不萌发。研究认为,鱼腥草花粉育性低的主要原因是单核花粉的发育过程异常,而非鱼腥草花粉母细胞减数分裂异常所致。     

Characterization of a pollen-expressed gene encoding a putative pectin esterase ofPetunia inflata

From a pollen tube cDNA library ofPetunia inflata, we isolated cDNA clones encoding a protein, PPE1, which exhibits sequence similarity with plant, bacterial, and fungal pectin esterases. Genomic clones containing thePPE1 gene were isolated using cDNA for PPE1 as a probe, and comparison of the cDNA and genomic sequences revealed the presence of a single intron in thePPE1 gene. During pollen development,PPE1 mRNA was first detected in anthers containing uninucleate microspores; it reached the highest level in mature pollen and persisted at a high level inin vitro germinated pollen tubes. The observed expression pattern of thePPE1 gene suggests that its product may play a role in pollen germination and/or tube growth.     

May the definition of pollen season influence aerobiological results?

This paper reviews the terms and major criteria used to define and limit the pollen season. Pollen data from Cordoba (Spain), Ourense (Spain) and Bologna (Italy) were used to ascertain the extent to which aerobiological results and pollen curves are modified by the criteria selected. Results were analysed using Spearmanȁ9s correlation test. Phenological observations were also used to determine synchronization between pollen curves and plant phenology. The criteria for limiting the shortest and longest pollen season periods, as well as the earliest and latest start and end dates, varied according to the city and the taxon under study; in many cases, results for a given taxon also depended on the year. The smallest differences were obtained for Platanus and the greatest for Poaceae.     

Aeropalynological study of <Emphasis Type="Italic">Vitis vinifera</Emphasis> in the Braga region (1999–2003)

This study shows the results of Vitis vinifera pollen season in the atmosphere of the Braga region (Portugal) for the last 5 years (1999–2003). The fluctuations in the airborne pollen concentration, the synchronic rhythms in the annual pollen trends and their relationship to the main meteorological parameters were investigated. The vineyard pollen season extends from late May to early July, presenting always a bimodal pattern. During the studied period several pollen types such as Poaceae, Castanea, Olea, Urticaceae, Quercus, Plantago, Rumex, Asteraceae, Pinaceae, Ericaceae and Eucalyptus were also observed. Airborne pollen concentration is highly influenced by mean temperature, rain, number of days with rain and wind direction.     

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.     

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.     

Notes on the floral morphology in Vivianiaceae (Geraniales)

The functional floral morphology of the three genera of Vivianiaceae (= Ledocarpaceae, Geraniales), Rhynchotheca, Viviania and Balbisia, is compared. Likely pollination mechanisms are inferred from morphology and field observations. The flowers of Viviania are nectariferous and apparently zoophilous with nectar as the (primary) pollinator reward. Balbisia has pollen flowers without nectaries, its showy corolla indicates that it is also zoophilous with pollen as sole pollinator reward; bees were observed as flower visitors. One taxon (B. gracilis) may be anemophilous. Rhynchotheca has flowers without petals, with large, pendulous anthers and lacks nectaries. It shows synchronous mass flowering in its natural populations and is evidently anemophilous. A comparison with other Geraniales shows that nectar flowers with small anthers are likely the ancestral condition in Vivianiaceae. This suggests that the pollen flowers with larger anthers of Balbisia and Rhynchotheca may represent an apomorphic condition. The documentation of pollen flowers and anemophily in Vivianiaceae expands the range of known floral and pollination syndromes in Geraniales.     

Pollen grain morphology ofCoris (Primulaceae)

Pollen grain morphology, sculpturing, and wall ultrastructure are investigated in two species ofCoris (Primulaceae),C. monspeliensis L. andC. hispanica Lange. The study includes both acetolysed and unacetolysed pollen. No evidence of any major palynological difference is recorded between these two species, apart from a somewhat larger pollen inC. monspeliensis. However,Coris can be distinguished from the remaining members of thePrimulaceae by the conjunction of relatively large pollen grains, prominent margo, and particular tectal pattern causing a reticulate surface with minute luminal perforations decreasing towards the colpi. From both these distinctive features, and others typically primulaceous, some evolutionary considerations are inferred. Finally, the higher proportion of irregular grains inC. hispanica is interpreted in light of environmental stress.     

杉木花粉发育过程中形态结构的变化研究

该研究以扬州地区成年杉木为材料,采用半薄切片法以及扫描电镜和透射电镜观察法,对杉木花粉发育过程进行了观察。结果表明:杉木的造孢细胞形成于10月,持续至翌年的1月底至2月初。造孢细胞位于小孢子囊最里面,呈多边形,数量多且彼此排列紧密;细胞壁很薄,细胞核较大,内含丰富的细胞器和脂类物质。造孢细胞发育成熟后彼此分离形成小孢子母细胞。小孢子母细胞的减数分裂开始于2月下旬,经两次减数分裂,先后形成二分染色体和四分染色体。小孢子从四分体释放后,体积迅速增大,发育形成花粉,这个过程中,伴随着明显的营养物质变化。杉木成熟花粉壁由薄壁区和厚壁区两部分组成,薄壁区中央有一个乳头状突起,突起上有一萌发孔。成熟花粉外壁着生了许多瘤状颗粒。该研究结果为杉木生殖发育、有性繁殖以及系统演化研究提供了重要依据。     

Division of the generative nucleus in cultured pollen tubes of the Bromeliaceae

Pollen germination, division of the generative nucleus and position of the generative nucleus in the pollen tube during in vitro germination were examined for six bromeliad cultivars. The influence of mixed amino acids (casein hydrolysate) and individual amino acids (Arg, Asn, Asp, Glu, Gly, Met, Phe, Orn, Tyr) were tested. Aechmea fasciata and A. chantinii pollen tubes showed more generative nuclear division in cultured pollen tubes than the other four cultivars tested. Casein hydrolysate did not stimulate generative nuclear division. In general arginine (1 mM) improved division of the Aechmea generative nucleus and to a lesser extent this of Vriesea `Christiane', Guzmania lingulata and Tillandsia cyanea. A concentration of 2 mM arginine reduced pollen tube growth of Aechmea. The vegetative nucleus was ahead of the generative nucleus in approximately 50% of the pollen tubes of all cultivars studied. In about 25% of the pollen tubes, the generative nucleus was ahead and in ±25% pollen tubes the vegetative and generative nuclei were joined together. The distance between the two generative nuclei and the distance from the generative nuclei to the pollen tube tip differed significantly for Aechmea fasciata and A. chantinii. The influence of different amino acids for Aechmea fasciata and A. chantinii varied with respect to pollen germination and generative nuclear division. Arg and Met improved nuclear division of both Aechmea cultivars. Pollen germination and sperm cell production were not linked. This information is important to ameliorate in vitro pollination methods used to overcome fertilization barriers in Bromeliaceae and other higher plants.     

Syncyte formation in the microsporangium of <Emphasis Type="Italic">Chrysanthemum</Emphasis> (Asteraceae): a pathway to infraspecific polyploidy

Polyploidy, which is thought to have played an important role in plant evolution and speciation, is prevalent in Chrysanthemum (x = 9). In fact, polyploid series are known in C. zawadskii (2x, 4x, 6x, 8x, and 10x) and C. indicum (2x, 4x, and 6x), but the mechanism by which polyploidization occurs is unknown. Here we show that in diploid individuals of both C. zawadskii and C. indicum, the fusion between two adjacent pollen mother cells (PMCs) occurs at a frequency of 1.1–1.3% early in the first meiotic division. While possessing the chromosomes of both PMCs, the fused cell or syncyte undertakes subsequent meiotic division processes as a single large PMC, producing four 2n pollen grains that are able to germinate. Despite their low frequency, syncyte formation may have played a major role in the production of infraspecific polyploids in Chrysanthemum.     

A comparative biochemical study of conifer pollen allergens

In the order Coniferales, only the family Cupressaceae is regarded as being a significant source of airborne allergens, withJuniperus ashei characterized as the most significat aeroallergen. Pollen of the closely related speciesJ. virginiana has been shown to cross-react withJ. ashei pollen, however,J. virginiana pollen is not considered an important aeroallergen. Although there have been several reports of allergies toPinus pollen, the pollen of this genus is regarded as hypoallergenic. Our previous studies have shown that pollen extracts ofJ. ashei, J. virginiana, J. pinchotii, Cupressus macrocarpa, Pinus echinata andP. taeda all contained several proteins with the same molecular weights including the reported allergen ofJ. ashei. The present study compared the biochemistry ofJ. ashei, J. virginiana andP. echinata pollen. A time course experiment ofJ. ashei, J. virginiana andP. echinata showed thatJ. ashei released a greater quantity of protein within the first minute of moistening. SDS-PAGE analyses showed that the reported allergen ofJ. ashei pollen extracts was released in large quantities within the first minute of extraction. It was also determined that individual pollen grains ofP. echinata contained a greater quantity of protein than the pollen ofJ. ashei andJ. virginiana, but due to the large size of pine pollen there was less protein per gram of pollen. Lipid analysis of these three taxa showed that the pollen ofP. echinata contained more lipid per grain and per gram of pollen. Results indicate that the rapid release of the reported allergen fromJ. ashei pollen contributes to the allergenicity of this species compared to bothJ. virginiana andP. echinata.     

Isolation of Allium pollen protoplasts

Studies on protoplasts isolation were carried out with mature pollen grains of 29 samples of species of Allium aflatunense, A. cepa, A. fistulosum, A. karataviense, A. longicuspis, A. nutans, A. odorum, A. sativum and A. schoenoprasum. Surface sterilized pollen grains drifted from crushed anthers were incubated in an enzyme solution containing 1% (w/v) cellulase Onozuka R-10, 1% (w/v) Macerozyme R-10, 0,5 mol l^-1 sucrose and the basal salts of Nitsch medium. Protoplasts were released within 3 to 120 min, either from the pollen grain, through a slightly disturbed germination pore (narrow aperture), or through a wider aperture, when the exine surrounding the germination pore was disturbed. For the first time, protoplasts were obtained from 13 genotypes of 6 Allium species, at a rate of 1 to 30% of the digested intact pollen grains, depending on the genotype.     

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.     

Agouticarpa, a new neotropical genus of Tribe Gardenieae (Rubiaceae)

Agouticarpa, a new genus of the informalAlibertia group (Rubiaceae), is described and illustrated. It comprises six species, and occurs from Costa Rica to Bolivia. Three species are here described as new:Agouticarpa grandistipula, A. hirsuta, andA. velutina. Additionally, three new combinations are made:A. curvifolia andA. williamsii (previously inGenipa) andA. isernii (previously inAlibertia). Agouticarpa is characterized by being dioecious, having elliptic to abovate, membranaceous stipules, male flowers in a branched dichasial or thyrse-like inflorescence, a poorly developed cup-shaped calyx, pollen grains with 3–7 apertures, and large globose fruits.