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.     

High Humidity and Heat Stress Causes Dissociation of Endoplasmic Reticulum in Tobacco Pollen

High humidity (95 % RH) and temperature (38/45 °C) stress for 4h applied to pollen grains of Nicotiana tabacum did not affect pollen viability, assessed on the basis of the fluorochromatic reaction test, but affected in vitro germination; pollen grains treated at 38 °C showed marked delay in germination, while those treated at 45 °C failed to germinate in vitro. The major ultrastructural effect of the stress was on RER. Stacks of RER, characteristically present in fresh pollen, were largely dissociated in the stressed pollen. The extent of dissociation of RER was greater in pollen samples stressed at 45 °C than at 38 °C. The generative cell did not show any obvious change in the stressed pollen. RER was restored in pollen grains which showed germination following culture; but not in those which failed to germinate. Apart from affecting other RER-related functions the dissociation of RER is likely to result in the destruction of long-lived mRNA and thus affect the ability of pollen grains to initiate protein synthesis needed for germination.     

In vitro germination and pollen tube growth of maize (Zea mays L.) pollen

Summary Pollen grains containing either theWx,wx,Su ₁,Su ₁,Sh ₂orsh ₂alleles were stored for 0, 1, 2, 3, 4 or 5 days at 2 °C. After each storage period, a portion of each genotype was cultured on a 15% sucrose, 0.6% bacto-agar, 0.03% calcium nitrate and 0.01% boric acid medium, while another portion was placed on receptive silks, the number of kernels produced being a measure of fertilization ability. Regardless of the allele present in the pollen grain, 1 day of storage greatly increased the germination percentage and significantly increased pollen tube length. After 4 days of storage, there was noin vitro germination but some fertilization ability was found. The experiment was designed so that comparisons free from genetic background effects could be made between alleles at each locus. Significant differences at each storage period and a differential response to storage were obtained at some loci for germination percentage, ruptured percentage, pollen tube length and fertilization ability. A relationship between dominance of the allele and response to storage was detected only for fertilization ability. Since alleles at these loci affect the biochemical composition of pollen grains containing them, the results suggest that differences inin vitro germination characteristics and fertilization ability may be associated with biochemical composition.Journal Series Paper No. 3950, Florida Agricultural Experiment Station.     

Pollen morphology and germination inNarcissus

Pollen grains of several species and varieties ofNarcissus were examined with a light microscope and a scanning electron microscope. Normal pollen grains were kidney- or spindle-shaped with a germination furrow and a reticulate structure similar to that of the pollen grains ofAmaryllis. Pollen grains germinated within 2 to 3 hr. Percentage of germination was dependent upon temperature and treatment. Pollen tubes grew in length up to 1,000 μm and branched occasionally or behaved in strange fashion. Fresh pollen grains germinated more in distilled water at lower temperature than in sucrose-aqueous medium. Both in the presence and in the absence of stigmatic exudate calcium increased the percentage of germination. Gibberellic acid, abscisic acid and coumarin inhibited the pollen germination. Plasmoptysis occurred in all species and in all media tested except in a medium containing coumarin without stigmatic exudates. Plasmoptysis did not seem to be induced by hypotonic medium alone. Pollen of high germination capacity showed a high percentage of plasmoptysis. Based on the results obtained, evolution and sterility of theNarcissus plant was discussed.     

紫花含笑（♀）、灰岩含笑（♂）及其杂种F1代花粉萌发试验研究

对紫花含笑(Michelia crassipes)、灰岩含笑(M.calcicola)及其杂种F1代花粉生活力进行了研究,为基于紫花含笑和灰岩含笑杂种F1代的含笑属观赏植物新品种培育与种质创新提供科学数据及研究资料.研究发现,亲本(紫花含笑和灰岩含笑)新鲜花粉萌发率均可达90％以上,杂种F1代花粉萌发率从38％到79％不等,平均为57.7％,低于双亲.亲本及其杂种F1代花粉萌发的最适温度为25℃,温度过高花粉管的伸长受到抑制,并导致花粉管顶端破裂.亲本及多数杂种F1代的新鲜花粉在100 g/L和150 g/L的蔗糖浓度下萌发率都较高;经-20℃贮藏后的花粉对蔗糖浓度的敏感性要高于新鲜花粉.杂种F1代及其亲本的花粉在离体培养中均会出现双萌发管现象.番红染料对液体培养基中的花粉有致死和染色作用,有利于统计杂种F1代及其亲本的花粉萌发率.     

The influence of relative humidity on the swelling of pollen grains in vitro

The volume of hydrated pollen grains of Petunia hybrida L. during swelling in germination medium increased three times. The volume of desiccated pollen grains increased only two times after transfer to the same medium. This difference in swelling ability is attributed to different rigidities of the pollen grain wall,ccaused by the different hydration states. The relationship between pollen grain swelling and germination metabolism with regard to relative humidity is discussed.Abbreviation RH relative humidity     

Early in vitro flowering and seed production in culture in <Emphasis Type="Italic">Dendrobium</Emphasis> Chao Praya Smile (Orchidaceae)

Plantlets of Dendrobium Chao Praya Smile maintained in vitro were induced to flower, which produced viable seeds within about 11 months. A two-layer (Gelrite-solidified layer topped with a layer of liquid medium of the same volume and composition) culture system containing benzyladenine (BA) at 11.1 μM induced the highest percent of flowering (45%) in plantlets within 6 months from germination. The percentage of inflorescence induction was increased to 72% by pre-selecting morphologically normal seedlings prior to two-layer culture. Plantlets in culture produced both complete (developmentally normal but smaller than flowers of field grown plants) and incomplete flowers. Pollen and female reproductive organs of in vitro-developed complete flowers were morphologically and anatomically similar to flowers of field grown plants. In addition, 65% of the pollen grains derived from in vitro-developed flower were tetrad suggesting that regular meiosis occurred during microsporogenesis. The percentage of germination of pollen grains derived from in vitro-developed flowers and flowers of field grown plants, incubated on modified Knops’ medium for 8 days, were 18.2 and 52.8%, respectively. Despite a lower percentage of germination of the pollen grains derived from in vitro-developed flowers, flowers induced in culture could be self-pollinated and developed seedpods with viable seeds. Nearly 90% of these seeds developed into protocorms on germination in vitro. These seedlings were grown in culture and induced to flower in vitro again using the same procedure.     

Pollen-pistil interaction in Brassica oleracea

Quantitative studies of the adhesion of pollen grains to the stigma in Brassica oleracea revealed that self-pollen is initially less firmly bound than cross-pollen. The pollen grain tryphine, believed to be important in the adhesion process, has been shown to differ in mobility following self- and cross-pollination when observed using fluorescent probes. The hydration of the pollen grains has been investigated in vitro by measuring the changes in shape, volume and fresh weight of the imbibing grains. Whilst little change in volume could be detected there was a considerable increase in fresh weight together with a change of shape. The significance of these events, which occur prior to pollen germination, is discussed in relation to their effect upon subsequent germination and expression of self-incompatibility.Abbreviations RH relative humidity - SI self-incompatibility - ConA concanavalin A - I-ANS I-anilino-napthyl-sulfonic acid     

Pollen biology in four Mediterranean Quercus species

Potential pollen production, viability and germination were studied in the most important species of Quercus in the mountains of Córdoba to determine the contribution of each species to the total amount of airborne pollen. The results were compared over two consecutive years with different rainfall patterns. The viability of pollen grains was determined at anther opening, and during the pollination period, in order to determine potential pollination capacity. Results indicated that there were differences in the number of pollen grains produced by stamen in the four species. Equally, there were differences in the number of flowers among the species, being Q. suber the species with higher number of catkins groups and flowers per individual tree. Total pollen production per tree can be summarized in the following proportion: 1: 3: 3: 6 (Q. coccifera, Q. ilex ssp. ballota, Q. faginea and Q. suber). Potential pollen viability was estimated using the Fluorochromatic Reaction (FCR) and a germination assay. The results have shown that Quercus potential pollen viability is high and declines slowly with time. Q. coccifera was the species with the highest percentage of germination, with Q. suber being the lowest.     

A study on pollen grains ofCrocus cartwrightianus (Iridaceae)

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

Interaction between floral color change and gender transition in the protandrous weed Saponaria officinalis

Natural selection has directed the evolution of floral traits so that pollinator visits are manipulated to maximize the fitness of individual plants by directing which other individual sires its seeds. In some plants, flowers change color over time and may have the ability to direct pollinators to rewarding flowers. In addition, by varying when pollen is available and when stigmas are receptive, protandrous plants can show variation in selfing rates. In this study, the association between color change and gender transition in flowers of Saponaria officinalis was examined. Anthocyanins were extracted from flowers of each gender stage to measure color using spectrophotometry. Female‐phase flowers were found to have significantly higher anthocyanin concentration than male‐phase flowers in both natural populations and experimental plots. This color change corresponded to a decrease in male sexual function, which was measured by the percentage of pollen grains stained as viable by lactophenol aniline blue and germinated on Brewbaker–Kwack media. Color change was phenotypically plastic. Plants grown in full sun had a more extensive color change than those grown in shaded experimental plots, and this effect was reversed the following year when the shading was removed. Pollinator observations documented both diurnal and nocturnal insect visitation. Fruit and seed set were equivalent on inflorescences bagged during daylight versus night, indicating that both diurnal and nocturnal insects are effective pollinators. If pollinators discriminate based on color, this could potentially reduce within‐plant floral visits and also geitonogamy. This study is the first to document flower color change and moth pollination in Saponaria officinalis.     

Kanamycin resistance of germinating pollen of transgenic plants

The influence of kanamycin on the percentage of pollen germination and on tube growth of pollen from non-transformed and transformed plants of various species containing a chimaeric kanamycin resistance gene (NPTII) was investigated. Pollen grains isolated from kanamycin resistant plants expressed resistance when germinating in vitro, whereas kanamycin impaired tube growth of pollen from non-transformed plants. Pollen grains from transgenic plants were less sensitive and produced significantly longer tubes. mRNAs of the chimaeric gene are probably presynthesized concurrently with the other mRNAs during microsporogenesis, and kanamycin resistance is expressed by mRNA translation during pollen tube elongation. Received: 24 August 1999 / Revision accepted: 20 October 1999     

L'effetto Di Popolazione Nelle Colture Del Polline: Alcuni Aspetti Della Sua Natura

Abstract

Pollen population effect in artificial culture: some aspect of its nature. — The ability of pollen to grow on several rather different artificial media has not yet allowed investigators to determine the best conditions for each species. However a common requirement for all kinds of pollen is a high density of pollen grains on the culture medium, that stimulates the pollen germination and the pollen tube growth. Many investigators dealing with the causes of that phenomenon failed to reach a convincing conclusion on its causes. Our experiment was set to test the ability of an unknown pollen growth factor (PGF) to diffuse from growing pollen into the medium. Consequently after removal of the pollen culture a second pollen population grown on the same medium should be able to use the PGF eventually dissolved in it. Widely scattered populations of Malus pollen were grown onto membrane filters in contact with such «preconditioned» media and tested for their ability to germinate. Because the percentage of germination were lower than expected, wide-pored membranes (8.0 μ instead of 0.8 μ) were tested, thus achieving a rate of germination as high as that of the crowded populations of pollen. Hence it was argued that the PGF is water soluble, but is absorded on the membrane before reaching the contact of the pollen. However if the pores surface is small enough to be saturated by part of the PGF the free remainder is still enough to stimulate the small population of pollen. To test the assumption (BREWBAKER and KWACK, 1963) that simply Ca++ supported by other cations is the PGF, Ca++, Mg++ and K+ have been supplemented to the Standard medium as sulfates at a 10—⁴ M concentration, combined in a factorial design. The results are reported in table 2 and can be briefly summarized as follow. The most efficient ion appears to be K+, followed by Ca++ and Mg++. Each one is able to increase significantly the rate of germination with respect to the following ion, but two negative interactions (Ca × K and Mg × K, P: 0.01) were found to be significant. Because the only significant interactions were with K+, and because the mixtures containing K+ significantly differ but from treatment with Ca++ and Mg++, and not from K+, it could be tentatively stated that the negative interaction is at expenses of the K+ stimulatory effect (see fig. 1). The joint evaluation of both experiments strongly suggests the idea that the PGF is an organic rather than an inorganic molecule, with high affinity with the membrane on which it can be adsorbed. Most probably this organic growth factor needs an inorganic ion to display its stimulatory effect, but this cannot be identified with the PGF. In fact none of the tested inorganic molecules is adsorbed on the narrow pored membrane and they display full stimulatory activity through it. The second experiment seems to suggest the idea that such inorganic ion is K+.     

An autoradiographic study of RNA synthesis during maturation and germination of pollen grains ofHyoscyamus niger

Summary The pattern of RNA synthesis during maturation and germination of pollen grains ofHyoscyamus niger was studied using³H-uridine autoradiography. Incorporation of label during pollen maturation was periodic with peak RNA synthesis occurring in the uninucleate, nonvacuolate pollen grains and in the vegetative cell of the bicellular pollen grains. During the early stages of germination, isotope incorporation occurred predominantly in the nucleus of the vegetative cell with little or no incorporation in the generative cell. With the appearance of the pollen tube, incorporation of³H-uridine in the vegetative cell nucleus decreased and completely disappeared at later stages of germination. No incorporation of isotope was observed in the sperms formed in the pollen tube by the division of the generative cell. From a comparison of the results of this study with those of previous works on RNA synthesis during pollen embryogenesis in cultured anthers ofH. niger, it is concluded that in contrast to embryogenic development, there is no requirement for sustained RNA synthesis by the generative cell nucleus for normal gametophytic development.     

Airborne pollen and spores on the Arctic island of Jan Mayen

A survey of airspora collected on Jan Mayen, an isolated North Atlantic island (71°N, 8°30′W), using a Burkard seven-day volumetric trap from 24th April to 31th August, 1988, revealed only very small concentrations. A total of 10 different pollen types were recorded, constituting a seasonal sum of 29 pollen grains. The local pollen season was confined to July, with Oxyria digna and Salix as the most numerous pollen types recorded. Exotic pollen grains, namely Betula, Pinus and Castanea type, were recorded in three periods during June and July. Studies of back trajectories indicate North America and/or Iceland and Greenland as possible source areas for the Betula pollen. There were more diatoms than pollen in the local airspora. Fungal spores mainly occurred in late July and August. Cladosporium constituted less than 5% of the total seasonal sum of fungal spores, while basidiospores contributed nearly 12%. The highest diurnal average of Cladosporium was 27 spores m^?3 air. The seasonal maximum of unidentified fungal spores reached a diurnal average of 639 spores m^?1 air on 27th August.     

POLLEN LONGEVITY AND STIGMA PRE-EMPTION IN CLARKIA

The ability of pollen to effect seed set decreases with age when stored at room temperature or on the plant, but not when stored at 5 C. Seed set declines rapidly with pollen stored 10 days or more at room temperature. Seed set declines rapidly with pollen stored 4 days or more on the plant. The degree of pollen germination on the stigma and the number of pollen tubes in the style correlate positively with seed set. Pollen germination on the stigma and pollen tube growth in the style are strongly correlated with seed set for pollen stored at room temperature, but not, or only weakly, correlated for pollen stored at 5 C. In the latter case, seed sets are lower than expected, indicating a possible viability reduction associated with a post-pollination function. Inviable pollen grains temporarily interfere with normal pollen germination on the stigma. Essentially no seeds are produced when in viable pollen, immediately followed by viable pollen, is placed on a fresh stigma. However 27 to 65 seeds are produced when in viable pollen, followed in 1 to 6 days by viable pollen, is placed on fresh stigmas. New stigma surface is exposed at the center as expanding stigma surfaces carry the interfering inviable pollen toward the edge of the stigma.     

Atmospheric conditions during high ragweed pollen concentrations in Zagreb,Croatia

We examined the atmospheric conditions favourable to the occurrence of maximum concentrations of ragweed pollen with an extremely high risk of producing allergy. Over the 2002–2009 period, daily pollen data collected in Zagreb were used to identify two periods of high pollen concentration (> 600 grains/m³) for our analysis: period A (3–4 September 2002) and period B (6–7 September 2003). Synoptic conditions in both periods were very similar: Croatia was under the influence of a lower sector high pressure system moving slowly eastward over Eastern Europe. During the 2002–2009 period, this type of weather pattern (on ~ 70% of days), in conjunction with almost non-gradient surface pressure conditions in the area (on ~ 30% of days) characterised days when the daily pollen concentrations were higher than 400 grains/m³. Numerical experiments using a mesoscale model at fine resolution showed successful multi-day simulations reproducing the local topographic influence on wind flow and in reasonable agreement with available observations. According to the model, the relatively weak synoptic flow (predominantly from the eastern direction) allowed local thermal circulations to develop over Zagreb during both high pollen episodes. Two-hour pollen concentrations and 48-h back-trajectories indicated that regional-range transport of pollen grains from the central Pannonian Plain was the cause of the high pollen concentrations during period A. During period B, the north-westward regional-range transport in Zagreb was supplemented significantly by pronounced horizontal recirculation of pollen grains. This recirculation happened within the diurnal local circulation over the city, causing a late-evening increase in pollen concentration.     

Seed set,pollen morphology and pollen surface composition response to heat stress in field pea

Pea (Pisum sativum L.) is a major legume crop grown in a semi‐arid climate in Western Canada, where heat stress affects pollination, seed set and yield. Seed set and pod growth characteristics, along with in vitro percentage pollen germination, pollen tube growth and pollen surface composition, were measured in two pea cultivars (CDC Golden and CDC Sage) subjected to five maximum temperature regimes ranging from 24 to 36 °C. Heat stress reduced percentage pollen germination, pollen tube length, pod length, seed number per pod, and the seed–ovule ratio. Percentage pollen germination of CDC Sage was greater than CDC Golden at 36 °C. No visible morphological differences in pollen grains or the pollen surface were observed between the heat and control‐treated pea. However, pollen wall (intine) thickness increased due to heat stress. Mid‐infrared attenuated total reflectance (MIR‐ATR) spectra revealed that the chemical composition (lipid, proteins and carbohydrates) of each cultivar's pollen grains responded differently to heat stress. The lipid region of the pollen coat and exine of CDC Sage was more stable compared with CDC Golden at 36 °C. Secondary derivatives of ATR spectra indicated the presence of two lipid types, with different amounts present in pollen grains from each cultivar.     

A new procedure to asses pollen viability

We tested pollen viability of eight species using four vital dyes, a new peroxidase test together with three other established methods (MTT, Baker’s and X-Gal), to determine their potential to differentiate fresh pollen from pollen heated for 2 h and 24 h at 80°C (killed pollen) and compared the results with in vitro germination. We found that two of three dyes previously employed to determine viability also stained killed pollen, while the new peroxidase test and MTT did not. We suggest that the latter two are the best methods to test pollen viability, since they do not normally stain either killed or aborted pollen. Received: 7 May 1999 / Revision accepted: 25 October 1999     

Physiological factors limit fruit set of tomato (Lycopersicon esculentum Mill.) under chronic, mild heat stress

The effects of chronic, mild heat stress on fruit set, fruit production, release of pollen grains, photosynthesis, night respiration and anther dehiscence were examined in tomatoes (Lycopersicon esculentum Mill.) differing in high‐temperature sensitivity. Plants were grown under three temperature regimes: (1) 28/22 or 26/22 °C (optimal temperature); (2) 32/26 °C (high temperature); and (3) 32/26 °C day/night temperatures relieved at 28/22 °C for 10 d before anthesis, then returned to 32/26 °C (relieving treatment). FLA 7156 was the only cultivar with fruit set at 32/26 °C. All five cultivars, however, had fruit set under the relieving treatment (RT). The longer the relief, the higher the percentage of fruit set. Longer periods of relief also increased the number of pollen grains released, and linear regression analysis showed a significant relationship between the number of pollen grains released and the percentage of fruit set. Germination of pollen grains was also lowered in high‐temperature‐grown plants. The number of pollen grains produced, photosynthesis and night respiration did not limit fruit set under chronic, mild heat stress, however. This suggested that cultivar differences in pollen release and germination under heat stress are the most important factors determining their ability to set fruit.