In vitro pollination in poplar of section Populus

Two methods of in vitro pollination of aspen were tested and compared, first with moist pollen by transferring swollen pollen grains from a wet agar surface to the stigma and, second, using dry pollen grains. In vitro pollination with dry pollen grains appeared to be more efficient and this method was applied in three subsequent years. Additionally, pollination was performed with selected pollen grains in order to obtain triploid plants. Diploid pollen was induced by heat-treatment of pollen mother cells, but the enrichment of unreduced pollen from natural pollen mixtures was also successful. The selection of diploid pollen, which is larger in diameter than haploid pollen, was performed by sieving using micro sieves. Five triploid plants from different poplar combinations were obtained from 1,227 zygotic embryos cultured after in vitro pollination of 2,676 ovaries with dry pollen over 3?years. Embryo rescue was used to facilitate the development of immature zygotic embryos. The results of this study demonstrate the possibility to effect pollination using selected single pollen grains with the help of in vitro techniques.     

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–pollen interactions in Betula pendula in vitro

Direct pollen interactions, as well as interactions mediated by a recipient, can have a remarkable influence on pollen fertilization ability. Under conditions of pollen competition it could be advantageous if pollen grains interfered with the germination of other pollen. The aim of this study was to find out if there are direct negative or positive pollen–pollen interactions between pollen grains from genetically slightly different donors. The in vitro germinability of the pollen from several Betula pendula Roth clones was investigated. The pollen interactions between the clones were examined pairwise by using equal pollen mixtures. In three of the eight cases the germination percentage of the pollen mixture was significantly higher than the average germination percentage of the separate clones that formed the mixture, which indicates some type of interaction between the pollen populations. We found only positive interactions between the pollen of clones. This study also documented density-dependent germination of pollen grains in vitro (=pollen population effect). Adding an aqueous pollen extract to the incubation medium increased the germination percentages of poorly germinating pollen and small pollen populations. Germination-stimulating effects were found to exist both with fresh and dead pollen. Such direct pollen–pollen interactions could be explained by specific water-soluble substances diffusing from pollen grains.     

POLLEN-POLLEN AND POLLEN-STYLE INTERACTIONS DURING POLLEN TUBE GROWTH IN ERYTHRONIUM GRANDIFLORUM (LILIACEAE)

Using pairs of pollen donors, I analyzed the growth of pollen tubes growing in different channels of the same style in Erythronium grandiflorum. After 24 hr the length of pollen tubes of randomly selected pollen donors was affected by the presence of other donors. The mean pollen tube lengths of donors did not differ when taken across all of the donor pairings, but in individual pairs, pollen from one donor was often significantly longer than pollen from the other donor when tested across several recipients. Pollen tube lengths were also consistently longer for pollen paired with pollen from the same donor than when paired with pollen from other donors, apparently because of mutual stimulation between the pollen populations from the same plant. In a second experiment, the amount of pollen tube attrition after five days of growth of pollen from a donor growing near (within 3 m) the recipient depended upon the source of other pollen growing in the same style. Local pollen experienced more attrition paired with self pollen than when paired with outcross pollen. Pollen from different outcross distances also modified the attrition of local pollen, but local pollen usually outcompeted pollen from greater distances. Since the growth of local pollen was modified depending upon the source of other pollen growing in the same style, it is probable that recipient styles are selectively inhibiting pollen tubes to produce the patterns of pollen tube interaction observed. The results from these two experiments indicate that the amount of attrition for pollen can be dependent on the composition of the pollen pool. Both direct pollen tube interactions and mediation by the stylar tissue appear to affect the growth rate and attrition of pollen in Erythronium.     

Microgametophyte population sizes and plant reproductive output in the insect-pollinated Prunella grandiflora (Lamiaceae)

Intraspecific variation in pollen deposition and number of pollen tubes per style is rarely quantified, but is essential for assessing the occurrence of pollen limitation and pollen competition and their evolutionary implications. Moreover, pollen deposition, pollen tube growth, and the fate of fertilized ovules are rarely distinguished in field studies. Here we present such a study in eight natural populations of Prunella grandiflora. We quantified microgametophyte population sizes and inferred pollen limitation when the number of fertilizable ovules exceeded pollen tubes, and assessed seed set and fate after open pollination. Two and three populations had on average significantly fewer pollen grains and pollen tubes per flower, respectively, than the fixed number of fertilizable ovules per fruit, while one population experienced significant pollen competition. Style length was positively correlated with the number of pollen tubes. While pollen availability was very variable, seed abortion was significantly less frequent in denser populations, and in one population the proportion of well-developed seeds was significantly, positively correlated with the number of pollen tubes in the style. Less pollen deposition, lower numbers of pollen tubes reaching the base of the style, lower pollen quality and therefore increased abortion of fertilized ovules can all reduce seed set in natural P. grandiflora stands. Substantial intraspecific variability implies that microgametophyte competition also occurs in this species. Finally, style morphology may affect pollen receipt.     

How does secondary pollen presentation affect the fitness of Polygala vayredae (Polygalaceae)?

Secondary pollen presentation is the relocation and presentation of pollen in floral structures (termed pollen presenters) other than the anthers. These pollen presenters are often found close to the stigma and have been hypothesized to increase the accuracy of pollen transfer, although no experimental studies have been done. We examined the function of the pollen presenter and its efficiency in pollen dispersal, female fitness, and the degree of interference created by self-pollen in the shrublet Polygala vayredae, an insect-pollinated species with secondary pollen presentation. Herkogamy, a mechanism generally involved in the reduction of self-interference, was also evaluated. Significant pollen was lost (49% of total pollen) during the secondary relocation in the pollen presenter. However, pollen was exported from the pollen presenter, and subsequent pollen losses were similar to those in species with primary pollen presentation. Despite the presence of a self-incompatibility system, the numbers of developed pollen tubes as well as fruit and seed production were significantly reduced by the self-pollen interference created at the stigmatic papillae level. The extent of herkogamy correlated positively with female fitness. The secondary pollen presentation mechanism may in fact be an accurate system for pollen transport, but it may also have its costs. Further comparative studies involving species with primary and secondary pollen presentation are needed to fully understand the advantages and disadvantages of secondary pollen presentation.     

Cytoskeleton in Pollen and Pollen Tubes of Ginkgo biloba L.

The distribution of F-actin and microtubules was investigated in pollen and pollen tubes of Ginkgo biloba L. using a confocal laser scanning microscope after fluorescence and immunofluorescence labeling. A dense F-actin network was found in hydrated Ginkgo pollen. When Ginkgo pollen was germinating,F-actin mesh was found under the plasma membrane from which the pollen tube would emerge. After pollen germination, F-actin bundles were distributed axially in long pollen tubes of G. biloba. Thick F-actin bundles and network were found in the tip of the Ginkgo pollen tube, which is opposite to the results reported for the pollen tubes of some angiosperms and conifers. In addition, a few circular F-actin bundles were found in Ginkgo pollen tubes. Using immunofluorescence labeling, a dense microtubule network was found in hydrated Ginkgo pollen under confocal microscope. In the Ginkgo pollen tube, the microtubules were distributed along the longitudinal axis and extended to the tip. These results suggest that the cytoskeleton may have an essential role in the germination of Ginkgo pollen and tube growth.     

Airborne ragweed (Ambrosia artemisiifolia L.) pollen concentrations in Croatia, 2002–2004

The aim of this study was to determine the onset, length and end of the ragweed pollen season, taking into account diurnal, day-to-day, monthly and annual pollen variations, the effect of some meteorological parameters on atmospheric pollen concentrations and possible differences in the airborne pollen season and concentration due to sampling site. Airborne pollen was collected at three sites in central Croatia (Zagreb, Samobor and Ivanić Grad) during three pollen seasons (2002–2004). Seven-day Hirst-type volumetric pollen traps were used for pollen sampling. Ragweed pollen was the third most abundant pollen type to occur in the atmosphere of central Croatia. Total Ambrosia pollen concentration was the highest in the 2003 pollen season and the lowest in 2004 at all sampling sites. Maximum emissions were restricted to August and September. Intradiurnal periodicity showed a peak from 1000 to 1200 hours. The concentration of ragweed pollen during the pollen season was greatly influenced by temperature and precipitation: on rainy days accompanied by temperature decline, the air pollen concentration decreased abruptly. The results of this study are aimed at helping to alleviate the symptoms of allergic reactions in individuals with ragweed pollen hypersensitivity, thus improving their quality of life.     

Pollen morphology and its effect on pollenl collection by honey bees,Apis Mellifera L. (Hymenoptera: Apidae), with special Reference to Upland Cotton,Gossypium Hirsutum L. (Malvaceae)

Honey bees, Apis mellifera, forage readily on flowers of upland cotton, Gossypium hirsutum, to harvest nectar. The abundant pollen gets caught in the haircoat of the bees, but cotton pollen is nevertheless rarely collected. Honey bee pollen collection effectiveness was therefore investigated in a flight room using cotton and five other spheroidal pollen taxa presented in sequence. Honey bees visited all pollen dishes, but okra pollen (Abelmoschus esculentus) was never packed successfully by the bees landing in the pollen dish. Cotton pollen was collected by 16% of the landing foragers, pumpkin pollen (Cucurbita pepo) by 71%, and pollen of corn (Zea mays), pigweed (Amaranthus palmeri), and sunflower (Helianthus annuus) were readily collected by nearly all foragers. The amount of time spent in the pollen dish was always short (1 to 9 seconds) and homogeneous among all pollen taxa, indicating that none of them was strongly repellent to the bees. The reduced effectiveness with which honey bees collected cotton pollen was demonstrated by the longer amount of time needed for pollen grooming and packing between two consecutive landings in the pollen dish and the small size of cotton pollen pellets (averages of 0.42 mg and 8.23 mg per pellet for cotton and corn pollen, respectively). This reduced efficiency in cotton pollen collection was associated primarily with the length of the spines on cotton pollen which physically interfered with the pollen aggregating process used by honey bees.     

First volumetric airborne pollen sampling in Montevideo City,Uruguay

A volumetric aeropalynological sampling was carried out for the first time in MontevideoUruguay, from October 2000 to September 2001, using a Rotorod sampler Model 40. During the year 76 pollen types were identified. Airborne pollen was recorded over the year but a maximum pollen period was observed from August to April. For the rest of the months, pollen concentration was below 1% of the total annual pollen (TP). The pollen spectrum was characterized by the dominance of herbaceous pollen (NAP), which represented 68% of the TP and dominates the spectrum from November to March. Poaceae was the most frequent and abundant pollen type accounting for 45% of TP. The pollen spectrum reflected the floristic diversity of the city and most of the sources of airborne pollen are present in local and regional flora. Fourteen pollen types reach more than 1% of the TP and most of them are cited as allergenic pollen in other regions. These results may prove important for future medical research.     

用激光扫描共聚焦显微镜观察雪松花粉和花粉管

为更直观地观察和显示花粉和花粉管中细胞结构及其细胞核的状态与行为。雪松花粉和花粉管经卡诺液固定,分别以埃氏苏木精、曙红、Hoechst 33243单染和曙红-Hoechst 33342双染后,用冬青油整体透明,在激光扫描共聚焦显微镜下观察。4种染色法观察效果不同;以曙红-Hoechst 33342双染的样品观察效果最佳,在紫外光激发下清晰地显示出细胞核,在488 nm激光激发下不仅能清晰看到花粉和花粉管壁结构,且能分辨管细胞、柄细胞及体细胞的结构特点和空间位置关系。建立了一种快速简便的适于在激光扫描共聚焦显微镜下观察花粉和花粉管中成员细胞结构及其细胞核的状态、行为的制片技术;激光扫描共聚焦显微镜具有独特的共轭成像装置、连续光学扫描、图像三维重组和多通道检测等功能,极好地展示了雪松花粉和花粉管的结构特点,相比于传统的光学显微镜和荧光显微镜,其观察到的图像更清晰、更直观、更具立体感。     

In vitro pollen competitive ability in Viola tricolor: temperature and pollen donor effects

In this study on Viola tricolor pollen, the competitive ability of 16 pollen donors originating from a wild population was analysed in a set of greenhouse and germination temperatures. The aim was to examine the consistency in donor pollen performance across temperatures and to see whether variation in performance was random or due to individual differences in the plastic response to temperature. Pollen tube growth rate in vitro was investigated in two greenhouse temperatures (on the day pollen was collected) and in four germination temperatures. In addition, pollen tube growth rate was assessed in vivo (in one temperature) to examine the relationship between in vivo and in vitro growth. A temperature difference of 5 K - corresponding to natural variation in time and space detected in the field - affected pollen tube growth rate. For both temperature components, significant pollen donor by temperature interactions were found and rank order of pollen donors changed across treatments. Although pollen competitive ability in violets was strongly influenced by both temperature components, the occurrence of pollen donor by temperature interactions indicates that donor siring ability varies with temperature. This, in turn, may suggest a means to maintain pollen competitive ability despite selection for this trait.     

Applying modern analogs to understand the pollen content of coprolites

Most scientists working with coprolites from archaeological contexts assume that human fecal specimens reflect the mixing of the pollen ingested during the period in which the contribution to the coprolite, 19 to 37.5 h, was ingested, that the amount of pollen in a fecal sample directly reflects the amount of pollen originally ingested during that interval, and that differences between the amounts of pollen in different fecal specimens reflect differences in the quantities of pollen ingested at different times. These assumptions were tested and found wanting in an experiment in which two persons sequentially ate separate quantities of 15 pollen types in meals over a four-day interval. The pollen was retrieved and analyzed from feces produced during those four days and five days of subsequent fecal production. Pollen ingested first appeared in relatively small amounts, usually the day after it was ingested. Its concentration per gram of sample then increased rapidly and remained high over a one to three day interval relative to the amounts in previous and subsequent fecal specimens deposited. When pollen concentrations declined some pollen was retained in the gastrointestinal system and much lower concentrations per gram of sample of each type continued to appear in fecal samples for several days. These relatively low pollen concentrations appeared in fecal samples approximately twice as often as did higher concentrations. Our results indicate that comparatively high pollen concentrations can be used to determine that a given pollen type was ingested, but comparisons between pollen concentrations of the same pollen type in different fecal specimens or between different pollen types in the same fecal specimen, cannot be used to determine whether different amounts of pollen were ingested, or what was the relative amount of each ingested. Because pollen concentrations per gram of sample varied widely with time since ingestion, percentages of given pollen types did not occur in predictable patterns and could actually increase as the concentration of the pollen type decreases. Hence, percentages should not be used in coprolite pollen analysis. The experimental results also suggest that variations in the pollen content of different portions of a coprolite are meaningful only in terms of the overall pattern of a sequential group of coprolites.     

Pollen-pistil size correlation and pollen size-number trade-off in species of Argentinian Nyctaginaceae with different pollen reserves

Data on pollen and pistil traits from 14 Argentinean Nyctaginaceae species with starch or lipids as pollen reserves are presented. We expect differences in the traits between these two groups of species, but the same pattern within each group for (a) the relationship between pollen size and pistil length assuming that pollen tube length is predetermined by provisions in the pollen independently of the pollen reserve type, and for (b) a trade-off between size and number because available resources for male function are not unlimited. In particular we expect that (a) species with longer pistils will have larger pollen grains, (b) pollen grain size and the number of pollen grains per flower will be negatively correlated. Significant differences in the mean pollen number per flower and mean pollen size were observed between species with different pollen reserve type but not for pistil length. Then, correlation analyses were performed for species with starchy pollen or with pollen with lipids separately. Pollen size - pistil length correlation was positive and significant for species with starchy pollen but not for species with pollen with lipids. On the other hand, pollen size - number correlation was not significant for both starchy and oil-rich species. Results suggest that pollen reserve type would be a relevant factor that constraint pollen size in species of Nyctaginaceae, and that this pollen trait should be considered when studying pollen-pistil relationships, mainly between species of those families with mixed pollen reserves.     

野慈姑花粉限制评估的影响因素

以野慈姑为材料,研究补充授粉的处理水平(单花、花序)和花粉质量(自花花粉、异花花粉)对花粉限制评估的影响.结果表明: 补充授粉的处理水平会造成花粉限制的程度不同(P=0.0021),单花水平的授粉会导致花朵之间的资源重新分配,从而影响结实;补充授粉的花粉质量会对结实造成影响,补充异花花粉比自花花粉能产生更多的种子(P=0.0497),且萌芽率更高(P=0.0428),表明该体系的花粉限制不是数量的限制,而是优质花粉的限制.不同的试验水平和花粉质量可能对花粉限制的评估造成影响,相关研究应尽量对整个花序或者植株进行补充授粉,材料允许时还应考虑自异花粉的对比试验,有利于更客观地认识和检测花粉限制.     

Differentiation of embryogenic pollen in cold-treated buds ofNicotiana tabacum var. Badischer burley and nutritional requirements of the isolated pollen to form embryos

Summary Embryogenic pollen were selectively isolated from buds after cold treatment at 10 °C for 10 days; it was immaterial whether the buds were taken from short day and low temperature (SD and LT; 8 hours light, 18 °C) or long day and high temperature (LD and HT; 16 hours light, 24 °C) plants. However, in buds from SD and LT plants the differentiation of embryogenic pollen could be detected as early as 7 days after the cold treatment, and pollen from these plants formed embryos at higher frequency (up to 4% of cultured pollen) than those from LD and HT plants (up to 1% only).The embryogenic pollen, in isolated buds, differentiated by way of pollen dimorphism. During cold treatment a fraction of pollen remained small, retained clear cytoplasm and was capable of embryogenesis in comparison to gametophytic pollen which enlarged and acquired granular cytoplasm. In our experiments cold treatment was a key factor in the induction of pollen dimorphism. This aspect of cold treatment in pollen embryogenesis is reported for the first time and was possible on the basis of selection of embryogenic pollen by density gradient centrifugation. The ratio of embryogenic pollen was about one fifth of the total population.The nutritional requirements of isolated pollen for embryogenesis were rather simple. These pollen formed embryos which readily developed into plantlets on a mineral medium supplemented with sucrose provided the pH was 6.8.     

The effects of pollen load size and donor diversity on pollen performance, selective abortion, and progeny vigor in Mirabilis jalapa

The influence of pollen competitive environment on pollen performance (pollen germination, stigmatic penetration, and pollen tube growth rate), the maturation or abortion of initiated fruit, seed size, and seedling vigor was explored by manipulating the size and diversity of stigmatic pollen loads on Mirabilis jalapa. All aspects of pollen performance significantly increased with the number of pollen grains on a stigma or pollen tubes in a style, but was not influenced by the diversity of pollen donors. Plants tended to mature single-ovulate fruits that came from flowers where pollen load size and diversity were greatest and aborted those where these were lowest. No plants from seeds resulting from pollinations with a single pollen grain survived, but other fitness measures were mostly determined by maternal plant. The data suggest that pollen performance is influenced by pollen competitive environment, and both the genetic diversity of the pollen load and number of competing pollen tubes are important determinants of seed/fruit abortion.     

Contributions of Pollen Morphology to the Phylogeny of Some Ranalean Families

The hypothesis that gene flow northwards by long distance wind dispersal of pollen is a contributing factor in Scots pine (Pinus sylvestris L.) seed crops in northern Scandinavia (Umeå) was examined. Pollen was observed in the air before local pollen shedding began. This pollen has been shown to germinate and is likely to be viable. This early pollen was probably not of local origin. Wind patterns and the geographic pattern of pollen shedding were such that it is likely that the origin of the early pollen was hundreds of km to the south. Since many of the female strobili are receptive to pollination before local pollen shedding occurred, early non-local pollen has an advantage over later local pollen. Thus, the gene flow caused by pollen migration is likely to be large enough to be of evolutionary significance.     

Environmental Effects on Components of Pollen Performance in Faramea occidentalis (L.) A. Rich. (Rubiaceae)1

Studies focusing on gametophytic competition have focused on differences in pollen performance (e.g., pollen germination rate and pollen tube growth rate) among donors in order to examine genetic sources of variation in siring success among them. Donors that produce the fastest growing pollen tubes are expected to fertilize more ovules relative to donors with slow growing pollen tubes under conditions of gametophytic competition. However, the performance of pollen in the field is known to be influenced by environmental conditions in addition to the genotype of donor plants. This field study of Faramea occidentalis was conducted to: (1) determine the effect of environmental conditions during pollination on pollen performance; (2) measure differences among pollen donors in pollen performance; (3) determine if the pollen of different donors responds differently to a variety of environmental conditions surrounding pollinated flowers; and (4) measure differences among pollen recipients in pollen performance. Single-donor crosses were made between four pollen donors and four pollen recipients under a variety of environmental conditions. Pollen performance was then quantified as the growth rate of the fastest pollen tube, the mean pollen tube growth rate, and by a pollen germination index. Pollination environment (the environment surrounding a pollinated flower) and recipient significantly affected all three measures of pollen performance. Pollen donors did not differ overall in pollen performance. However, there was significant among-donor variation in two of the five pollination environment conditions. Future studies of variation in relative siring success may benefit by considering pollination environment in addition to donor and recipient identity.     

A pollen calendar for Chittagong University Campus,Chittagong (Bangladesh)

Summary This work presents a calendar of pollen types found in the atmosphere of Chittagong University Campus, Chittagong, Bangladesh, recorded over the period of March 1988–February 1990. A total of 9,225 pollen/cm² was encountered and classified in 36 pollen types belonging to 26 families. The pollen concentration showed three peak periods, March, October and November and each period was dominated by a specific pollen type. Low frequency of pollen was probably due to heavy rainfall which washed down the pollen. January and February were also marked by lower abundance of pollen and was probably due to the low rate of flowering. This study clearly showed four airborne pollen seasons from October to November, December to February, March to April and June to September. The dominant pollen types were Poaceae, followed by Amaranthaceae, Arecaceae, Asteraceae and Cyperaceae. The pollen calendar was provided to show the range of flowering with the period of occurrence in air of the area studied.