Developmental changes in DNA methylation of the two tobacco pollen nuclei during maturation

Changes in DNA methylation during tobacco pollen development have been studied by confocal fluorescence microscopy using a monoclonal anti-5-methylcytosine (anti-m5C) antibody and a polyclonal anti-histone H1 (anti-histone) antibody as an internal standard. The specificity of the anti-m5C antibody was demonstrated by a titration series against both single-stranded DNA and double-stranded DNA substrates in either the methylated or unmethylated forms. The antibody was found to show similar kinetics against both double- and single-stranded DNA, and the fluorescence was proportional to the amount of DNA used. No signal was observed with unmethylated substrates. The extent of methylation of the two pollen nuclei remained approximately constant after the mitotic division that gave rise to the vegetative and generative nuclei. However, during the subsequent development of the pollen, the staining of the generative nucleus decreased until it reached a normalized value of \documentclass[12pt]{minimal} \usepackage{wasysym} \usepackage{amsmath} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} \begin{equation*}\frac{1}{5}\end{equation*}\end{document} of that of the vegetative nucleus. The use of a confocal microscope makes these data independent of possible focusing artefacts. The anti-histone antibody was used as a control to show that, while the antibody staining directed against 5-methylcytosine changed dramatically during pollen maturation, the histone signal did not. We observed the existence of structural dimorphism amongst tobacco pollen grains, the majority having three pollen apertures and the rest with four. However, the methylation changes observed occurred to the same extent in both subclasses.

Directional Guidance of Nicotiana alata Pollen Tubes in Vitro and on the Stigma1

Pollen tubes navigate the route from stigma to ovule with great accuracy, but the cues that guide them along this route are not known. We reproduced the environment on the stigma of Nicotiana alata by immersing pollen in stigma exudate or oil close to an interface with an aqueous medium. The growth of pollen in this culture system mimicked growth on stigmas: pollen grains hydrated and germinated, and pollen tubes grew toward the aqueous medium. The rate-limiting step in pollen germination was the movement of water through the surrounding exudate or oil. By elimination of other potential guidance cues, we conclude that the directional supply of water probably determined the axis of polarity of pollen tubes and resulted in growth toward the interface. We propose that a gradient of water in exudate is a guidance cue for pollen tubes on the stigma and that the composition of the exudate must be such that it is permeable enough for pollen hydration to occur but not so permeable that the supply of water becomes nondirectional. Pollen tube penetration of the stigma may be the most frequently occurring hydrotropic response of higher plants.

The late Pliocene pollen record of ODP Site 108-658 off Cape Blanc, Atlantic Ocean

A 200 m long marine pollen record from ODP Site 658 (21°N, 19°W) reveals cyclic fluctuations in vegetation and continental climate in northwestern Africa from 3.7 to 1.7 Ma. These cycles parallel oxygen isotope stages. Prior to 3.5 Ma, the distribution of tropical forests and mangrove swamps reached Cape Blanc, 5°N of the present distribution. Between 3.5 and 2.6 Ma, forests occurred at this latitude during irregular intervals and nearly disappeared afterwards. Likewise, a Saharan paleoriver flowed continuously until isotope Stage 134 (3.35 Ma). When river discharge ceased, wind transport of pollen grains prevailed over fluvial transport. Pollen indicators of trade winds gradually increased between 3.3 and 2.5 Ma. A strong aridification of the climate of northwestern Africa occurred during isotope Stage 130 (3.26 Ma). Afterwards, humid conditions reestablised followed by another aridification around 2.7 Ma. Repetitive latitudinal shifts of vegetation zones ranging from wooded savanna to desert flora dominated for the first time between between 2.6 and 2.4 Ma as a response to the glacial stages 104, 100 and 98. Although climatic conditions, recorded in the Pliocene, were not as dry as those of the middle and Late Pleistocene, latitudinal vegetation shifts near the end of the Pliocene resembled those of the interglacial-glacial cycles of the Brunhes chron.

Early Cretaceous spores and pollen of ODP Leg 103 holes

About 80 species of spores and pollen grains were recorded during detailed palynological investigations of selected Lower Cretaceous sections from Holes 638B and 638C and the bottom of Hole 641C. Most of them are long-ranging taxa with worldwide distribution. However, on the Iberian margin and in the southern European basins, Trilobosporites canadensis, Trilobosporites bernissartensis, Parvisaccites amplus, Foveosporites subtriangularis, and Ephedripites multicostatus seem to be index species of the Valanginian to late Aptian interval. Clavatipollenites was not recovered in the Barremian marginal marine sediments.

Miocene pollen of ODP Leg 127 sites

Pollen floras were obtained from Miocene sediments recovered at four sites drilled during Ocean Drilling Program Leg 127. The local pollen floras of each site were correlated to the standard pollen zones of northeast Japan by using the concept of the essential members for each pollen zone. At Site 797, the complete floral range was obtained for recognition of the NP2 zone and the pollen components of the NP1 zone were also clarified continuously. The ages of the boundaries between pollen zones NP4/NP3, NP3/NP2, and NP2/NP1 are estimated to be about 7 Ma, 13 Ma, and 17-18.5 Ma, respectively. Even in the same pollen zone, the ratios of major pollen taxa vary with the location. This variation is expressed on maps representing two different times during the Miocene.

Atmospheric Poaceae pollen frequencies and associations with meteorological parameters in Brisbane, Australia: a 5-year record, 1994-1999

Grass pollen is an important risk factor for allergic rhinitis and asthma in Australia and is the most prevalent pollen component of the aerospora of Brisbane, accounting for 71.6% of the annual airborne pollen load. A 5-year (June 1994-May 1999) monitoring program shows the grass pollen season to occur during the summer and autumn months (December-April), however the timing of onset and intensity of the season vary from year to year. During the pollen season, Poaceae counts exceeding 30 grains m(-3) were recorded on 244 days and coincided with maximum temperatures of 28.1 +/- 2.0degreesC. In this study, statistical associations between atmospheric grass pollen loads and several weather parameters, including maximum temperature, minimum temperature and precipitation, were investigated. Spearman's correlation analysis demonstrated that daily grass pollen counts were positively associated (P < 0.0001) with maximum and minimum temperature during each sampling year. Precipitation, although considered a less important daily factor (P < 0.05), was observed to remove pollen grains from the atmosphere during significant periods of rainfall. This study provides the first insight into the influence of meteorological variables, in particular temperature, on atmospheric Poaceae pollen counts in Brisbane. An awareness of these associations is critical for the prevention and management of allergy and asthma for atopic individuals within this region.

The effects of nitrogen application and assimilate availability on engorged pollen production and spikelet sterility in rice

Increased rates of nitrogen fertilizer application lead to increased spikelet sterility. A field experiment was conducted to investigate the effects on engorged pollen production and spikelet sterility, of nitrogen and assimilate availability during microspore development, in two rice cultivars (Doongara and Amaroo) grown under two different water depths. Despite the temperature not being low enough during microspore development to cause spikelet sterility, the number of engorged pollen grains was lower in cv. Doongara than in cv. Amaroo. Nitrogen application decreased the number of engorged pollen grains per anther through increased spikelet density. Nitrogen application increased spikelet sterility as a result of increased panicle density showing pronounced indirect effect of N on spikelet sterility. Engorged pollen number was also closely related (r = -0.636*) to the nitrogen content of the leaf blade, indicating a direct negative effect of plant N status on engorged pollen production. The results suggest that the intrinsic pollen producing ability is the key element in the difference in cold tolerance between the two cultivars, particularly under high N rates. Opening the canopy for increased solar radiation interception by the treated plants increased the level of engorged pollen, indicating the importance of immediate assimilate availability for engorged pollen production. Shading reduced crop growth rate, but did not effect engorged pollen production. There was no effect of variation in assimilates production on spikelet sterility.

Pollen flux off Cape Blanc

Fluxes of airborne freshwater diatoms (FD), phytoliths (PH), and pollen grains (PO) collected with sediment traps off Cape Blanc, northwest Africa, from 1988 till 1991 are presented. Both continental rainfall variations and wind mean strength and direction play a key role in the temporal fluctuations of the fluxes of eolian traces in the pelagic realm. Drier conditions in Northern Africa in 1987 could have preceded the high lithogenic input and moderate FD flux in 1988. The PH peak in summer 1988 was probably caused by increased wind velocity. Wetter rainy seasons of 1988/89 might have promoted a significant pollen production in summer 1989, and FD in late 1989 and early 1990, as well as contributed to the reduction of the lithogenic flux in 1989/90. Decreased fluxes of FD, PH and PO, and higher contribution of the 6-11 µm lithogenic fraction in 1991 would mainly reflect minor intensity and decreased amount of continental trade winds. Air-mass backward trajectories confirm that the Saharan Air Layer is predominantly involved in the spring/summer transport. Trade winds play a decisive role in the fall/winter months, but also contribute to the transport during late spring/summer. Origin of wind trajectories does not support a direct relationship between transporting wind-layers and material source areas in Northern Africa. High winter fluxes of eolian tracers and high amount of trade winds with continental origin in summer warn against a simplistic interpretation of the seasonal eolian signal preserved in the sediments off Cape Blanc, and the wind layer involved in its transport.

Pollen from 50 lake surface samples in Brandenburg, Germany, in 2009

Past changes in plant and landscape diversity can be evaluated through pollen analysis, however, pollen based diversity indexes are potentially biased by differential pollen production and deposition. Studies examining the relationship between pollen and landscape diversity are therefore needed. The aim of this study is to evaluate how different pollen based indexes capture aspects of landscape diversity. Pollen counts were obtained from surface samples of 50 small to medium sized lakes in Brandenburg (Northeast Germany) and compiled into two sets, with one containing all pollen counts from terrestrial plants and the second restricted to wind-pollinated taxa. Both sets were adjusted for the pollen production/dispersal bias using the REVEALS model. A high resolution biotope map was used to extract the density of total biotopes and different biotopes per area as parameters describing landscape diversity. In addition tree species diversity was obtained from forest inventory data. The Shannon index and the number of taxa in a sample of 10 pollen grains are highly correlated and provide a useful measure of pollen type diversity which corresponds best to landscape diversity within one km of the lake and the proportion of non-forested area within seven km. Adjustments of the pollen production/dispersal bias only slightly improve the relationships between pollen diversity and landscape diversity for the restricted dataset as well as for the forest inventory data and corresponding pollen types. Using rarefaction analysis, we propose the following convention: pollen type diversity is represented by the number of types in a small sample (low count e.g. 10), pollen type richness is the number of types in a large sample (high count e.g. 500) and pollen sample evenness is characterized by the ratio of the two. Synthesis. Pollen type diversity is a robust index that captures vegetation structure and landscape diversity. It is ideally suited for between site comparisons as it does not require high pollen counts. In concert with pollen type richness and evenness, it helps evaluating the effect of climate change and human land use on vegetation structure on long timescales.

ELUCIDATING ENIGMATIC FLORAL ISSUES IN COPAIFERA LANGSDORFFII DESF. (LEGUMINOSAE, CAESALPINIOIDEAE)

This study focuses on the floral development of Copaifera langsdorffii to elucidate uncertain features in its floral morphology, such as the tetramerous calyx, lack of petals, blackened anthers and their supposed sterility, as well as polyembryony. Buds and flowers were dissected and prepared for examination under scanning electron and light microscopes. The floral apex initiates two bracteoles, five sepals, five petals, five outer stamens, five inner stamens, and one carpel. Order is helical for sepals, reversed unidirectional for the petals, and unidirectional for two whorls of stamens. The tetramerous calyx results from the union of two adaxial sepal primordia, which forms one large sepal and three other smaller sepals. Although the flower lacks petals, the petal primordia are initiated but do not elongate like the other floral organs, remaining as petal rudiments. Ten stamens are formed in two distinct whorls. Formation within each whorl is almost simultaneous, and the inner whorl is formed shortly after the outer. During organ elongation, the inner stamen primordia bases are reoriented outward, resulting in a single whorl of stamens. The darkened anthers have viable pollen grains. Thus, there is no relation between sterility and the dark coloration of the anthers. No signs of extranumerary embryos are observed; therefore, polyembryony is not confirmed. Although studies on floral development of Detarieae have been reported, few Neotropical genera of the tribe (such as Copaifera) have been studied.

Two novel reports of semidry stigmatic surface in Asteraceae

Research documents related to the morphology and function of style branches and stigmatic surface of Asteraceae are still rather few, and the literature reports are thus controversial. We report in the present study that the stigmatic surfaces of two non-related species of Asteraceae (Lessingianthus grandiflorus and Lucilia lycopodioides) have features of semidry stigmas. Sporodermis of both species was also analyzed so that we could understand how the stigmatic surface works during pollen deposition and rehydration. Stylar branches and pollen grains (sporodermis) were studied using scanning and transmission electron microscopy (SEM and TEM) and histochemistry techniques. The inner and marginal bands of stylar branches in these species display intermediary features between the dry and wet types of stigma: the cuticle characterizes the dry stigma and cells with secretory activity characterize the wet stigma; these showed differences from what has been described to the Asteraceae family, where stigmatic surface of species from several tribes is considered dry. Pollen grains are medium-size to large with exine ornamentation (echinate and echinolophate) and abundant secretion which latter characterizes pollenkitt. We can assume that two processes might help pollen grain hydration on stigmatic surface in Lessingianthus grandiflorus and Lucilia lycopodioides: (1) the presence of pollenkitt, as observed in the secretory content inside exine cavities and around pollen grains; and (2) the secretory activity of stigmatic surface cells, whose secretion accumulates among intercellular and subcuticular spaces and leads to cuticle disruption during the floral receptive phase. Our results suggest that ultrastructural and histochemical studies should be considered when describing stigmatic surface and that the ""semidry"" feature within Asteraceae should be investigated still more in detail, so that the taxonomic or adaptation value of this trait in the family can be verified. (C) 2010 Elsevier GmbH. All rights reserved.

Stratigraphic and palaeoenvironmental significance of a Pennsylvanian (Upper Carboniferous) palynoflora from the PiauFormation, Parnaba Basin, northeastern Brazil

A well-preserved palynoflora is reported from within a cored interval of a coal-exploratory borehole (1-UN-23-PI of the Geological Survey of Brazil) in the southern part of the Parnaiba Basin, northeastern Brazil. The sample studied is from the lower portion of the Piaui Formation. Its palynoflora is characterized by particular abundance of the trilete cavate/pseudosaccate miospores Spelaeotriletes triangulus Neves and Owens, 1966 and S. arenaceus Neves and Owens, 1966, together with cingulizonate forms mainly attributable to Vallatisporites Hacquebard, 1957 and Cristatisporites R. Potonie and Kremp emend. Butterworth et al., 1964. Radially and bilaterally symmetrical monosaccate pollen grains are also well-represented, chiefly by Plicatipollenites Lele, 1964 and Potonieisporites Bhardwaj, 1954, respectively. Taeniate grains (i.e., monosaccates and bisaccates) are relatively minor constituents of the palynoflora; no marine microplankton were encountered. Several species are described in detail : the trilete apiculate spores Brevitriletes levis (Balme and Hennelly) Bharadwaj and Srivastava, 1969 and Horriditriletes uruguaiensis (Marques-Toigo) Archangelsky and Gamerro, 1979; and the taeniate pollen grains Meristocorpus ostentus sp. nov. and Lahirites segmentatus sp. nov. A Pennsylvanian (Late Carboniferous : late Westphalian) age is adduced for the palynoflora via its correlation with part of the Tapajos Group (specifically, the upper Itaituba Formation) of the Amazonas Basin in northern Brazil. The entirely land-derived palynomorphs, associated with abundant plant debris, corroborate previous suggestions that the lower part of the Piaui Formation accumulated in a nonmarine setting under conditions of aridity.

Permian Palynofloral Assemblages of the Chaco-Paraná Basin (Argentina): Systematics and Stratigraphic Significance

Core samples from an upper Palaeozoic, partly glaciogene borehole section (Ordóñez: YPF Cd O es-1) in the southern Chaco-Paraná Basin (Córdoba Province, northeastern Argentina) have produced variable palynological results. Samples from the lower part of the section (i.e., from the diamictite-bearing upper Ordóñez Formation) proved non-palyniferous. Those from the overlying, essentially post-glacial Victoriano Rodríguez Formation yielded spore-pollen assemblages in varying concentrations and in good to excellent states of preservation, thus providing the material basis for the present account. The palynomorph taxa represented in the assemblages comprise 20 species of spores (distributed among 14 genera) and 25 species of pollen grains (14 genera). The majority of the species are described in systematic detail. One trilete spore species -Convolutispora archangelskyi- is newly proposed. Several other, possibly new species (three of trilete spores, one of monosaccate pollen) are represented insufficiently for other than informal naming. The following new combinations, also of trilete spore species, are instituted: Converrucosisporites confluens (Archangelsky & Gamerro, 1979), C. micronodosus (Balme & Hennelly, 1956), and Anapiculatisporites tereteangulatus (Balme & Hennelly, 1956). Sculptural intergradation (granulate through verrucate) among three species -Granulatisporites austroamericanus Archangelsky & Gamerro, 1979, C. confluens, and C. micronodosus- prompts their informal grouping, proposed herein, as the Converrucosisporites confluens Morphon, which is also recognizable elsewhere in the Gondwanan Permian. The possibility, if not the likelihood, that G. austroamericanus is conspecific with Microbaculispora tentula Tiwari, 1965 is canvassed. The palynologically productive borehole section of the Victoriano Rodríguez Formation studied here is assignable to the middle to upper Cristatisporites Zone and to the succeeding Striatites Zone, thus signifying an Early Permian age for this section and facilitating correlation with strata of the Paraná and Paganzo Basins. From this and prior work, the Ordóñez well sequence embracing the Ordóñez and Victoriano Rodríguez Formations includes, in addition to the latter two zones, the preceding (late Pennsylvanian) Potonieisporites-Lundbladispora Zone which is known from the lower to mid-upper part of the Ordóñez Formation. Thus, the Carboniferous-Permian boundary can be inferred to lie within the upper part of the latter formation

Low temperature induced spikelet sterility in rice. I. Nitrogen fertilisation and sensitive reproductive period

Low temperature during panicle development in rice increases spikelet sterility. This effect is exacerbated by high rates of nitrogen (N) application in the field. Spikelet sterility induced by low temperature and N fertilisation was examined in glasshouse experiments to clarify the mechanisms involved. In two glasshouse experiments, 12-h periods of low (18/13degreesC) and high (28/23degreesC) day/night temperatures were imposed over periods of 5-7 days during panicle development, to determine the effects of low temperature and N fertilisation on spikelet sterility. In one experiment, 50% sunlight was imposed together with low temperature to investigate the additive effects of reduced solar radiation and low temperature. The effect of increased tillering due to N fertilisation was examined by a tiller removal treatment in the same experiment. Pollen grain number and spikelet sterility were recorded at heading and harvest, respectively. Although there was no significant effect of low temperature on spikelet sterility in the absence of applied N, low temperature greatly increased spikelet sterility as a result of a reduction in the number of engorged pollen grains per anther in the presence of applied N. Spikelet sterility was strongly correlated with the number of engorged pollen grains per anther. Low temperature during very early ( late stage of spikelet differentiation-pollen mother cell stage) and peak ( second meiotic division stage-early stage of extine formation) microspore development caused a severe reduction in engorged pollen production mainly as a result of reduced total pollen production. Unlike low temperature, the effect of shading was rather small. The increased tillering due to application of high rates of N, increased both spikelet number per plant and spikelet sterility under low temperature conditions. The removal of tillers as they appeared reduced the number of total spikelets per plant and maintained a large number of engorged pollen grains per anther which, in turn, reduced spikelet sterility. The number of engorged pollen grains per anther determined the numbers of intercepted and germinated pollen grains on the stigma. It is concluded that N increased tillering and spikelet number per plant and this, in turn, reduced the number of engorged pollen grains per anther, leading into increased spikelet sterility under low temperature condition.

Low temperature induced spikelet sterility in rice. II. Effects of panicle and root temperatures

Low temperatures impose restrictions on rice (Oryza sativa L.) production at high latitudes. This study is related to low temperature damage that can arise mid-season during the panicle development phase. The objective of this study was to determine whether low temperature experienced by the root, panicle, or foliage is responsible for increased spikelet sterility. In temperature-controlled glasshouse experiments, water depth, and water and air temperatures, were changed independently to investigate the effects of low temperature in the root, panicle, and foliage during microspore development on spikelet sterility. The total number of pollen and number of engorged pollen grains per anther, and the number of intercepted and germinated pollen grains per stigma, were measured. Spikelet sterility was then analysed in relation to the total number of pollen grains per spikelet and the efficiency with which these pollen grains became engorged, were intercepted by the stigma, germinated, and were involved in fertilisation. There was a significant combined effect of average minimum panicle and root temperatures on spikelet sterility that accounted for 86% of the variation in spikelet sterility. Total number of pollen grains per anther was reduced by low panicle temperature, but not by low root temperature. Whereas engorgement efficiency ( the percentage of pollen grains that were engorged) was determined by both root and panicle temperature, germination efficiency (the percentage of germinated pollen grains relative to the number of engorged pollen grains intercepted by the stigma) was determined only by root temperature. Interception efficiency (i.e. percentage of engorged pollen grains intercepted by the stigma), however, was not affected by either root or panicle temperature. Engorgement efficiency was the dominant factor explaining the variation in spikelet sterility. It is concluded that both panicle and root temperature affect spikelet sterility in rice when the plant encounters low temperatures during the microspore development stage.