The effect of lead contamination of soils and air on its accumulation in pollen

Summary Concentrations of heavy metals in the honey, derived fromAster tripolium was the starting-point for this investigation. Lead content of pollen inAster tripolium andTaraxacum officinale is mainly due to airborne lead. Accumulation of other heavy metals such as zinc and copper in pollen occur mainly via a translocation process from roots to flowers.     

Zinc deficiency and pollen fertility in maize (Zea mays)

Zinc deficiency decreased pollen viability in maize (Zea mays L. cv. G2) grown in sand culture. On restoring normal zinc supply to zinc-deficient plants before the pollen mother cell stage of anther development, the vegetative yield of plants and pollen fertility could be recovered to a large extent, but the recovery treatment was not effective when given after the release of microspores from the tetrads. If zinc deficiency was induced prior to microsporogenesis it did not significantly affect vegetative yield and ovule fertility, but decreased the fertility of pollen grains, even of those which visibly appeared normal. If the deficiency was induced after the release of microspores from the tetrads, not only vegetative yield and ovule fertility but pollen fertility also remained unaffected.     

The parallel expression of metal tolerance in pollen and sporophytes of Silene dioica (L.) Clairv., S. alba (mill.) krause and Mimulus guttatus DC

Summary The purpose of this paper was to determine if heavy metal tolerance was expressed in pollen and if its expression was correlated with the tolerance of the pollen source. Clones of Silene dioica, tolerant to zinc, closely related but nontolerant S. alba and clones of Mimulus guttatus tolerant and sensitive to copper were grown in the greenhouse in either standard potting soil or nutrient culture. Pollen was collected shortly after dehiscence, hydrated, and tested over a broad range of metal concentrations. The tolerance of the pollen source was determined by comparing root growth in solutions with and without heavy metals. In both Silene species and M. guttatus, the tolerance of the parental clone was expressed in its pollen. Pollen from tolerant individuals was able to germinate and grow at concentrations of metals which markedly inhibited pollen from nontolerants.     

Zinc in pollen and its incorporation into seeds

Summary A study on the distribution of radioactive zinc (⁶⁵Zn) in flowers of Vicia faba L. and Nicotiana tabacum L. showed the highest activity, expressed as cpm/mg dry weight, in pollen Also, using a histochemical method, it was observed that a portion of the naturally occurring zinc present in pollen grains was transferred into their growing tubes during germination. Calculations in based on the specific activity in ⁶⁵Zn-labelled pollen grains and in seeds from flowers pollinated with this pollen showed that about 68% of the radioisotope content of the pollen was incorporated into the seeds. The significance of these results and the possible role of zinc in fertilization is discussed.     

Suitability of pollen sources for the development and reproduction of Coleomegilla maculata (Coleoptera: Coccinellidae) under simulated drought conditions

Laboratory experiments compared the nutritive value of various pollen sources for the development of Coleomegilla maculata DeGeer under conditions of continuous water availability and simulated drought. When water was continuously available, larval survival was not different from 100% on diets of frozen eggs of Ephestia kuehniella Zeller, corn pollen, sorghum pollen, or pulverized bee pollen, whereas survival of larvae was significantly reduced on the latter three diets in the simulated drought treatment. Pollen of cultivated sunflower, Helianthus annus L., proved fatal to both larvae and adults; its surface structure caused clumping and accumulation on the insect cuticle that led to death from exhaustion/desiccation in petri dishes. The Ephestia egg diet yielded shorter developmental times and heavier adult weights than any pollen diet in both treatments. The drought treatment increased developmental time on all diets with a significant treatment–diet interaction. Drought reduced the adult weight of females on the sorghum pollen diet, and that of both sexes on the bee pollen diet, again with a significant treatment–diet interaction. Initial water content was highest in corn pollen (36.8%), followed by Ephestia eggs (29.2%), sorghum pollen (25.3%), sunflower pollen (8.7%), and bee pollen (4.6%), but did not appear correlated with C. maculata larval survival on pollen sources under drought conditions. Reproductive adult females that received corn or sorghum pollen as a supplement to Ephestia eggs did not differ in fecundity or fertility from those fed only Ephestia eggs.     

三倍体丹参杂交种的花粉形态研究

三倍体丹参是以二倍体丹参为父本、人工染色体加倍的四倍体白花丹参为母本杂交选育的杂交种。为深入了解三倍体丹参花粉的特性,以及为三倍体种质利用提供孢粉学依据,该文以二倍体丹参为对照,研究了三倍体丹参杂交种花粉的形态变异规律。利用光学显微镜和扫描电镜对二倍体和三倍体丹参的花粉萌发沟、外壁纹饰、花粉粒形状等特征进行了显微和超微形态观察,综合进行了花粉形态差异比较,并对花粉大小和形状数据进行了差异显著性分析和正态检验。结果表明:(1)二倍体丹参为6沟花粉,三倍体花粉萌发沟有6沟和8沟两种类型,沟内疣状颗粒分布不匀,出现畸形萌发沟。(2)二倍体和三倍体花粉外壁均为网状雕纹。二倍体花粉网眼内具多个多边形穿孔,穿孔大; 6沟和8沟两种类型的三倍体花粉网眼无穿孔或仅有几个小穿孔,6沟和8沟花粉的外壁雕纹相同。(3)三倍体花粉的极轴长(P)和赤道宽(E)均值显著小于二倍体花粉,花粉大小呈偏正态分布,P*E的差异系数大于二倍体花粉,且有极值存在。三倍体和二倍体丹参的萌发沟和雕纹存在差异,而花粉形状差异不显著。综上结果表明三倍体丹参花粉在倍性效应和杂合性的双重影响下发生了形态变异,且有多种形态变化。     

Morphological and functional characterization of BcMF13 in the antisense-silenced plants of Brassica campestris ssp. chinensis var. parachinensis

The gene Brassica campestris male fertility 13 (BcMF13, GenBank accession number EF158459) was isolated as a reproductive organ-specific gene from Chinese cabbage (Brassica campestris L. ssp. chinensis Makino, syn. B. rapa ssp. chinensis). It is exclusively expressed in stage four and five flower buds of fertile lines and is most strongly expressed in stamens. Here, we report a functional characterization of this BcMF13 gene in the antisense-silenced plants. The inflorescence of the BcMF13 mutant was compacted with anthers curved outside. The fertility of this mutant was greatly reduced with less than 5 seeds per silique. Under scanning electron microscopy, the mutant demonstrated numerous shriveled pollen grains with deep invaginations. The frequency of normal pollen grains was just 45.34%. The pollen mother cell, the tetrad, and the mature pollen of the BcMF13 mutant were abnormal resulting in the poor pollen vitality. Germination test in vivo suggested BcMF13 delayed the pollen tubes’ extension in the style. All these indicated BcMF13 had a vital role in pollen development of Chinese cabbage.     

Interspecific pollen transfer as a mechanism of competition: Consequences of foreign pollen contamination for seed set in the alpine wildflower,Polemonium viscosum

Summary Earlier studies showed that under natural conditions foreign pollen receipt is correlated with loss of seed set in populations of P. viscosum. Here, we report on experimental pollinations using foreign pollen from two co-flowering species that share pollinators with P. viscosum. Experiments were designed to ascertain whether improper pollen transfer causes reduced seed set under controlled conditions and, if so, to determine the stage at which interference occurs. In flowers pre-treated by pollination with either Mertensia or Castilleja pollen, adherence of subsequently deposited conspecific pollen grains was unaffected. However, conspecific grains had significantly lower germination and ovule fertilization success in flowers receiving foreign pollen than in controls. These results suggest that pollinator infidelity in high alpine plant communities can reduce female reproductive success of P. viscosum by inhibiting the performance of conspecific pollen grains. Negative effects of Mertensia and Castilleja pollen were comparable despite large interspecific differences in pollen grain morphology. Accordingly, susceptibility to foreign pollen interference appears to be an intrinsic property of P. viscosum that is not dependent on the unique structural properties of pollen produced by competitive species.     

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.     

Pollen dimorphism in threeTeucrium species (Lamiaceae)

In the Iberian Peninsula and Balearic Islands pollen dimorphism was found inTeucrium fruticans L.,T. pseudochamaepitys L., andT. rotundifolium Schreber. Generally, this dimorphism shows two sizes of pollen grains, the smaller more or less collapsed. The percentage of pollen viability is calculated. Differences in size between viable and nonviable pollen grains are similar, being about 40% inT. fruticans andT. pseudochamaepitys, and about 26% inT. rotundifolium. With regard to pollen viability, the percentage of male sterility is higher inT. fruticans, in which from male sterile (ms) plants, 100% nonviable pollen was obtained from every flower observed. InT. pseudochamaepitys andT. rotundifolium with rare exceptions, the percentage of nonviable pollen does not seem to be significant.     

Pollen precedence and stigma closure: a mechanism of competition for pollination between Delphinium nelsonii and Ipomopsis aggregata

Summary Previous experiments showed that the sympatric herbs Delphinium nelsonii and Ipomopsis aggregata compete for hummingbird pollination and that deleterious effects of the former species on seed set of the latter involve interspecific pollen transfer. However, seed set was not reduced when pollen of both species was applied simultaneously to I. aggregata stigmas. Hence a competitive effect may require arrival of foreign pollen before conspecific pollen. To explore this possibility we subjected I. aggregata flowers to a competition treatment in which they received D. nelsonii pollen 6 h before I. aggregata pollen, or to a control in which they received only the conspecific pollen. Foreign pollen precedence decreased mean seed set by almost 50%, which is consistent with effects observed in previous experiments. Reduced seed set can be explained by the fact that foreign pollen often caused stigma lobes to close together within 1.5–6 h, reducing subsequent receptivity. Stigma closure was also elicited by conspecific pollen, but not by mechanical stimulation, and was influenced by size of the pollen load and identity of the plant being pollinated.     

Precision of the daily pollen count. Identifying sources of variation using variance component models

Summary During the pollen season, a daily account of airborne pollen is reported by radio and newpapers in Denmark as «Today's pollen count». The Aerobiological Group under the Danish Asthma and Allergy association is responsible for the daily identification and counting of pollen. In 1984 the Group set up a trial to investigate the reproducibility of the pollen count.Three trained pollen counters independently examined the same specimens from two Burkard pollen traps. The traps were located on the roof of the Meterological Institute in Copenhagen 15 meters above ground level in the northern outskirts of central Copenhagen. This is the usual sampling site. Specimens from each trap from 20 days during the grass pollen season were selected. Four species groups were identified in sufficient numbers for statistical analysis. These were, in order of occurrence, spruce/pine (Picea/Pinus), grass (Poaceae), nettle (Urtica) and mugwort (Artemisia).Identification of sources of variation and assessment of their relative importance were carried out using variance components models. A detailed account of the method is given.Qualitatively equivalent conclusions were reached for each group: i) The expected seasonal variation was identified; ii) The counters contributed significantly to the variation of pollen counts; iii) The traps did not contribute to the variance, but a day-trap interaction was identified. This interaction was interpreted. as a problem of instrumental variation over time of traps of variation in meteorological conditions; iv) The total variance was larger than originally expected. The relative uncertainty was greater than 50 per cent.The variance of the daily pollen count cannot be reduced much by reducing counter variation or day-trap interaction. The variation among pollen counters is small, and the day-trap interaction is difficult to fully comprehend. The basic measurement error can be reduced by examining a larger area of each specimen. Although this implies a greater cost, it is probably the most effective method of reducing the uncertainty of «Today's pollen count».     

Aerobiological study of Fagaceae pollen in the middle-west of Spain

The concentration of airborne Fagaceae pollen in Salamanca and the correlations with some meteorological parameters have been examined. Castanea and Quercus pollen grains were collected from 1998 to 2004 using a Burkard spore trap. No pollen grains of Fagus were found. The main pollen season took place in April and May for Quercus and in June and July for Castanea. Yearly variations on these dates could be related to the influence of meteorological factors such as rainfall, temperature, or dominant winds. The highest values appeared in the year 2004 for both taxa. The Fagaceae airborne content was mainly due to Quercus pollen, Castanea having a scarce pollen content in the city of Salamanca. The highest counts of Fagaceae pollen grains were found from mid May to early June due to the pollen behavior of oaks. The cumulative counts varied over the years, with a mean value of 2,384 pollen grains, a highest total of 6,036 in 2004 and a lowest total of 954 in 2001. No cyclic variations were observed. Daily pollen concentrations presented positive correlation with temperature, negative with relative humidity and slightly negative with rainfall using Spearman's correlation coefficients, only in the case of Castanea, because the particular hourly distribution of rainfall during the spring might affect Quercus airborne pollen.     

Comparison of different techniques for gene transfer into mature and immature tobacco pollen

The particle gun, cocultivation withAgrobacterium tumefaciens, and imbibition in DNA solutions were compared as methods to transfer DNA into mature and immature pollen ofNicotiana tabacum. Bombardment of mature pollen with the β-glucuronidase gene cloned behind the pollen-specific PA2 promoter of the chalcone isomerase gene ofPetunia hybrida resulted in the expression of the β-glucuronidase gene in 0.025% of the pollen grains. Bombardment of younger stages followed byin vitro maturation also resulted in the formation of mature pollen that expressed β-glucuronidase, although at a lower frequency. Cocultivation of pollen duringin vitro maturation orin vitro germination withAgrobacterium tumefaciens did not yeild β-glucuronidase-expressing pollen. In these cases, an intron-containing β-glucuronidase gene was used which effectively prevented β-glucuronidase expression in the bacteria. Imbibition of mature, dry pollen in various DNA solutions of the same constructs also did not lead to the formation of β-glucuronidase expressing pollen.     

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.     

国产鸡血藤属和崖豆藤属(豆科)的花粉形态及其系统学意义

为探讨国产豆科(Leguminosae)鸡血藤属(Callerya Endl.)和崖豆藤属(Millettia Wight et Arn.)的属间和属下分类,在扫描电镜下观察了鸡血藤属11种5变种和崖豆藤属6种2变种的花粉形态。结果表明,鸡血藤属和崖豆藤属植物的花粉都是3孔沟或3沟的单粒,辐射对称,赤道面观主要为类球形或近扁球形,极面观主要为圆形或圆状三角形。鸡血藤属植物的花粉大小为中等,而崖豆藤属的为小型。两属植物花粉的沟膜呈现或多或少的隆起或凹陷,附有细微的或粗糙的颗粒,花粉外壁纹饰可分为6种类型。花粉形态不支持Schot合并所有灰毛鸡血藤复合体中除了亮叶鸡血藤[C.nitida(Benth.)R.Geesink]与喙果鸡血藤[C.cochinchinensis(F.P.Metcalf)Z.WeiPedley]为灰毛鸡血藤[C.cinerea(Benth.)Schot]的观点,花粉大小支持将鸡血藤属从广义崖豆藤属中分离。     

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。     

Pollen-ovule patterns in tribeTrifolieae (Leguminosae)

Relative pollen and ovule production in the genera of the legume tribeTrifolieae is explored particularly as to howMedicago with its explosive pollination mechanism compares with its allies.Medicago produces much larger, although much fewer pollen per ovule than the other five genera in the tribe; this is interpreted as a consequence of its highly specialized, irreversible pollination mechanism, which allows only one effective exchange of pollen with pollinators.Melilotus andTrifolium produce a comparatively large quantity of pollen; this can be related to their floral characteristics requiring abundant pollen to achieve fertilization. InMedicago andTrigonella, annuals convert a higher proportion of pollen grains and ovules into seeds than perennials.     

Overexpression of <Emphasis Type="Italic">Zm401</Emphasis>, an mRNA-like RNA,has distinct effects on pollen development in maize

We previously identified a 0.7 Kb cDNA fragment of Zm401, a novel pollen-specific gene in maize (Zea mays). However, little information is known about the function of Zm401 in pollen development. The full-length of Zm401 cDNA was amplified by 5′ RACE and 3′ RACE and both sequence analysis and in vitro translation of Zm401 showed that it belonged to an mRNA-like non-coding gene. To analyze its possible biological roles in pollen development, the Zm401 cDNA was overexpressed in transgenic maize under the control of a pollen specific promoter Zm13 or a CaMV 35S promoter. RT-PCR and RNA gel blot analysis indicated that the expression level of Zm401 in leaves and anthers of transgenic plants was much higher than that of non-transformants. Compared with the non-transformed maize, transgenic maize showed distinct phenotypes, such as abnormal tassels and degenerate anthers. The histological observation showed that the development of pollen grains and anthers in transgenic plants were abnormal. These abnormalities include delayed degradation of tapetum, asynchronous fusion of pollen sacs, and aborted pollen grain development. Furthermore, the pollen viability in six transgenic plants ranged from 1.24% to 6.63%. The reduced pollen viability cosegregated with the transgene in a selfed progeny. These results suggest that Zm401 is involved in the regulation of pollen development. This article demonstrated Zm401, as a non-coding RNA, plays an essential role in pollen development. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

An efficient method for flow cytometric analysis of pollen and detection of 2n nuclei in Brassica napus pollen

A simple and reliable method was developed for isolating pollen nuclei from Brassica napus and Triticum aestivum for DNA analysis using flow cytometry. The nuclei were released from pollen by ultrasonic treatment. The isolated nuclei following filtration through nylon mesh and a purification procedure were suitable for flow cytometric analysis as well as for isolating genomic DNA. Ultrasonic treatment time was optimized for B. napus pollen at different developmental stages. The method is effective and suitable for the preparation of many samples. We analyzed the nuclear DNA levels in pollen of B. napus at three major developmental stages as well as in mature wheat pollen. Only a single 1C peak representing the haploid DNA level was detected in the nuclei isolated from Brassica uninucleate microspores as well as in mature Triticum pollen. Interestingly, diploid nuclei were detected in both binucleate and mature pollen of B. napus. The possible origins of the diploid nuclei are discussed.Abbreviations DAPI 4,6-Diamidino-2-phenylindole - NIB Nuclear isolation buffer