A first assessment of genetic variation in Welwitschia mirabilis Hook

Welwitschia mirabilis is a monotypic member of the family Welwitchiaceae which, along with Ephedra and Gnetum species, comprises the gymnospermous order Gnetales. While the monophyly of this order is now widely accepted, the relationship of the Gnetales to other seed plants is still contentious. Despite the unique phylogenetic position of W. mirabilis and its extraordinary physiological and anatomical adaptations, little is known about the plant's phylogeny or its current distribution in isolated locations throughout the Namib Desert. As a preliminary step in the design of an more extensive phylogeographic study, we analyzed 37 random amplified polymorphic DNA (RAPD) loci from 59 plants distributed among five sites separated by distances of 6-440 km. Cluster analysis and analysis of molecular variance (AMOVA) revealed significant levels of variation within and between populations and little evidence of inbreeding. Genetic differences between populations reflect the geographic distances separating them. Three of the populations formed discernable genetic clusters, suggesting that little gene flow occurs between populations separated by > or = 18 km. In contrast, gene flow is occurring between two populations separated by only 6 km, supporting previous observations that pollen dispersal is primarily local and that seeds are not readily wind-born over the large distances separating most W. mirabilis populations. As a working hypothesis, we propose that W. mirabilis had a continuous distribution across its current range as much as 105 million years ago, and that as a consequence of subsequent drying trends and physical disturbance, populations became progressively isolated, accounting for their current distribution.     

CO2 gas exchange and transpiration of Welwitschia mirabilis Hook. fil. in the central Namib desert

Summary The diurnal course of CO₂ gas exchange, ¹⁴CO₂ incorporation, malate and citrate content, and traspiration of Welwitschia mirabilis were measured in one of its natural habitats, the Welwitschia-Vlakte in the central Namib desert (Namibia), in order to decide which CO₂ fixation pathway is used by this gymnosperm.The CO₂ gas exchange of Welwitschia is that of a C₃ plant under arid conditions. Younger leaf parts show a two-peaked pattern of photosynthetic CO₂ uptake whereas in older parts the morning peak is followed by net CO₂ release during the rest of the day. The maximum rates of net photosynthesis decrease from 3.4 mol m^-2 s^-1 in 1-year-old parts to 1 mol m^-2 s^-1 in 7-year-old parts. No net CO₂ uptake was detected during the night. The diurnal CO₂ balance indicates that the old leaf parts live at the expense of the younger ones. Irrigation of Welwitschia plants resulted in an increased CO₂ uptake throughout the light period with maximum rate of 4.1 mol m^-2 s^-1. ¹⁴CO₂ was only incorporated during the day.The water loss of Welwitschia by transpiration is considerable, reaching a peak value of 1.9 mmol m^-2 s^-1 around noon. Leaf conductance corresponds with the twopeaked pattern of CO₂ uptake.Although there is no sign of a crassulacean acid metabolism in Welwitschia the leaf contains rather high amounts of malate (up to 200 mol g^-1 dry matter) and citrate (up to 250 mol g^-1 dry matter), which depend on leaf age but do not show any significant day-night oscillation.In spite of all this the ¹³C values are in the range of-17.77 to-19.64. Possible reasons for such a high ¹³C content in a C₃ plant are discussed.Dedicated to Prof. H. Walter, the pioneer of ecophysiological studies in the Namib desert     

Photocontrol of seed germination in Impatiens wallerana Hook. f.

General characteristics of light sensitivity of Impatients wallerana seeds were investigated. Germination was absolutely dependent on light, irrespective of temperature. High percentages of germination were obtained by exposure to long periods of illumination or, alternatively, to several repeated short irradiations with red light. In this case, responsiveness to light was not altered by increasing either the initial incubation period in darkness or the dark intervals between short exposures. Effects of red light were reversed by far-red light, thus demonstrating the involvement of phytochrome. Evidence was presented for an interactive effect, of unknown physiological nature between red and far-red light on the germination of the seeds.Abbreviations P_r phytochrome, red light absorbing form - Pfr phytochrome far-red absorbing form     

Environmental control of crassulacean acid metabolism in Welwitschia mirabilis Hook. Fil. in its range of natural distribution in the Namib desert

Summary Within the area of its natural distribution in South West Africa, Welwitschia mirabilis has a less negative ¹³C value than C₃ plants and a more negative ¹³C value than C₄ species. This indicates that Welwitschia m. assimilates CO₂ partially via CAM when growing in its natural habitat. The difference between the ¹³C values of Welwitschia m. and of the C₃ species is significant in the savanna, whereas it is only small and statistically not significant in the grassland zone. The proportion of CO₂ fixed via CAM is largest in the coastal desert zone. There was no correlation between the ¹³C values and the Cl^- or ash content of the tissue. Thus, CAM in Welwitschia m. seems not to be induced by salt stress. There is no change in the ¹³C values along the persistent Welwitschia m. leaf. The present data indicate that on a broad geographical scale in the area of distribution temperature regime, and water stress as a modifying factor, determine CAM in Welwitschia m. The ecological implications are discussed by comparing the behaviour of Welwitschia m. with other CAM, C₃ and C₄ species of the accompanying flora.     

In vitro pollen germination and pollen tube growth differences among Quercus robur L. clones in response to meteorological conditions

The impact of meteorological conditions on in vitro pollen germination and pollen tube growth during the initial phases of the development of male flowers in the Pedunculate Oak, Quercus robur, is studied. Phenological observations of male flowers and pollen sampling were performed on the field trial established with grafted Pedunculate Oak clones. During the investigation, weather conditions (absolute minimum and maximum daily air temperature, minimum absolute relative humidity of air and amount of precipitation) were recorded by an automatic meteorological station installed at the field trial. Influence of meteorological conditions on pollen germination and pollen tube growth was studied in the following stages of male flower: (I) during the last ten days of flower bud dormancy, (II) during swelling of the buds, (III) during bud burst and beginning of male catkins elongation, (IV) during the final stage of male flower catkins elongation. High temperatures and low relative air humidity during the bud burst and beginning of the male catkins elongation reduced pollen germination and pollen tube growth. Weather conditions did not significantly affect pollen germination and pollen tube growth during the swelling of flower buds, or in the final stage of male catkins elongation.     

Pollen size evolution: correlation between pollen volume and pistil length in Asteraceae

Based on the assumptions that pollen tube length is predetermined by provisions in the pollen and that it is a function of pistil length, I hypothesise that species with longer pistils will have larger pollen grains than species with shorter pistils, and that pistil length and pollen size will be positively correlated in a linear manner. To test this hypothesis, the relationship between pollen grain volume and pistil length was compared in 43 Asteraceae species from Argentina. A positive linear correlation was found between pollen volume and pistil length. This correlation remained significant even after potential effects of phylogenetic relatedness were removed. The maintenance of this correlation suggests that in Asteraceae the association between pistil length and pollen volume may reflect a functional rather than a phyletic relationship. In addition, the pistil length: pollen volume ratio (PPR) was analysed in relation to the phylogenetic position of the species. High values of PPR would imply a reduction of the male gametophyte in relation to the minimal volume that a pollen grain must have to grow and fertilise an ovule. Thus, the general pattern of pollen volume reduction in relation to pistil length previously found among many angiosperm families will be also present within a family, i.e., PPR values of derived Asteraceae would be higher than those of basal species. Results indicated that reduction of pollen volume in derived Asteraceae was three times greater than the concomitant shortening of pistil length. Consequently, PPR increased with the phylogenetic position of the taxa. This work supports the correlation between pistil and pollen characters previously found for other plant families and confirms the influence of post-pollination processes on pollen size evolution. Received: 4 November 1999 / Revision accepted: 14 February 2000     

Pollen morphology of the genera Polygonum s. str. and Polygonella (Polygoneae: Polygonaceae)

The pollen of 30 taxa (27 species, one subspecies and two varieties) in two genera, viz Polygonum s. str. and Polygonella was investigated with LM and SEM, and some selected taxa with TEM. In all genera investigated the pollen is prolate to spheroidal, and the aperture is mostly tricolporate, rarely panto-hexacolporate (especially Polygonum section Polygonum). The exine sculpturing pattern is the most variable feature. Three types of exine can be recognized. Type 1 (Avicularia-Type, sensu Hedberg) - All species of section Polygonum and section Tephis share the smooth tectate exine with spinules, sometimes the surface is more or less rough (Polygonum afromontanum in section Tephis). Type 2 (Pseudomollia-Type, sensu Hong) - Pollen of Polygonum molliaeforme (section Pseudomollia) has the exine, which is verrucose on both poles and nearby the mesocolpium, and mostly psilate around the ectoaperture. Type 3 (Duravia-Type, sensu Hedberg) - Pollen grains of Polygonum section Duravia and Polygonella have the exine which is semitectate-reticulate at the mesocolpium and the poles, and rugulate/reticulate or sometimes foveolate with microspinules around the ectoapertures. The pollen grains in four taxa (viz Polygonum section Pseudomollia, P. section Duravia and genus Polygonella) have a well-marked dimorphism of the ektexine, which is considered to be a synapomorphic condition. The differences of pollen grain between the genus Polygonella and Polygonum section Duravia are almost non existent and clearly interrelated. It is therefore postulated that the similarity in pollen of both taxa is not the result of convergency, but is interpreted as a homology. It is noteworthy that the pollen of Polygonum molliaeforme (section Pseudomollia) appears as intermediate between the Avicularia-type and the Duravia-type, and is well supported the value of separated section for its own. Additionally, in TEM, some exine ultrastructures (e.g. columellae, foot layer, endexine) appear to be valuable characters for comparison between/among taxa. The systematic potentialities of the pollen data of the studied taxa at various systematic levels are also discussed.     

Pollen in wetlands: using simulations of pollen dispersal and deposition to better interpret the pollen signal

Unlike most other ecosystems, many wetlands preserve an archive of their own history in their accumulating sediments. Reading this archive can be complex and challenging. This paper explores some of the challenges faced when trying to interpret the pollen record preserved in sediment archives from wetlands. It uses simulations of pollen dispersal and deposition in wetland scenarios to demonstrate that relatively simple models can replicate many of the features of the wetland pollen signal, and explores the application of these models to developing better understanding and interpretation of the palaeo-record.     

Leaf architecture of the genus Didymaea Hook. f. (Rubiaceae)

We examined 532 specimens representing eight species of Didymaea (Rubiaceae) to describe their leaf architecture. Characters observed in the herbarium specimens and cleared leaves were leaf base, apex, and margin; trichome type and distribution; vein orders from primary to higher order veins, veinlets, and ultimate marginal vein. Six morphometric variables were analyzed using variance and principal component analysis to identify which characters are useful to distinguish species. Most of the species present brochidodromous venation pattern, except D. hispidula, which has eucamptodromous venation pattern; Didymaea floribunda is unique in having cylindrical and bicellular trichomes. Principal component analysis revealed that 78% of the total variation is mainly related to leaf maximum width. Variance analysis showed significant differences between D. microflosculosa and the other species. Architectural features were useful for differentiating all the species, and an identification key for the species of Didymaea based only on leaf characters is presented.     

Pollen development of Rheum nobile Hook.f. & Thomson (Polygonaceae), with reference to its sterility induced by bract removal

The Himalayan alpine herb, Rheum nobile , terminates in a stout conical compound raceme concealed by large translucent bracts. It bears many fruits even under hostile conditions such as low temperature or persistent cloudy weather. To clarify the role of the bracts, the structure and the development of the pollen grains were examined after removing the bracts to expose the flowers to the open air for 9 days. Half of the individuals with bracts removed showed 0 to 1 % of pollen stainability and the pollen grains were variable in shape and size. It was also observed that the bracts of Rheum nobile increased the temperature of inflorescence by about 10deg;C above ambient daytime temperatures. These results suggest that one of the causes for the inhibition of pollen development was low temperature. The remainder, however, indicated high stainability of 70–100%. It is suggested that the extreme difference of pollen stainability between two groups of Rheum nobile exposed to the surroundings may be related to the stage of microsporogenesis. Bracts of Rheum nobile might play an important role in normal reproduction under low temperature at high altitudes.     

Analysis of Stomatal and Nonstomatal Components in the Environmental Control of CO(2) Exchange in Leaves of Welwitschia mirabilis

In well-watered plants of Welwitschia mirabilis, grown in the glass-house under high irradiance conditions, net CO₂ assimilation was almost exclusively observed during the daytime. The plants exhibited a very low potential for Crassulacean acid metabolism, which usually resulted in reduced rates of net CO₂ loss for several hours during the night. In leaves exposed to the diurnal changes in temperature and humidity typical of the natural habitats, CO₂ assimilation rates in the light were markedly depressed under conditions resembling those occurring during midday, when leaf temperatures and the leaf-air vapor pressure differences were high (36°C and 50 millibars bar⁻¹, respectively). Studies on the relationship between CO₂ assimilation rate and intercellular CO₂ partial pressure at various temperatures and humidities showed that this decrease in CO₂ assimilation was largely due to stomatal closure. The increase in the limitation of photosynthesis by CO₂ diffusion, which is associated with the strong decline in stomatal conductance in Welwitschia exposed to midday conditions, may significantly contribute to the higher ¹³C content of Welwitschia compared to the majority of C₃ species.     

Effects of oxybarbiturates on the germination behaviour of Impatiens bolstii pollen.

Swelling of and cytoplasmic streaming within the grain, germination pore expansion, and pollen tube emergence, in that order, constitute the normal germination pattern of Impatiens holstii pollen grains. This normal sequence of events becomes manifest when the pollen grains are placed on water-agar, barbituric acid-agar, and on barbital-agar media whereas this sequence of events is interfered with when the pollen grains are incubated on amytal-agar or on seconal-agar media. Evidence is presented which demonstrates that the normal sequence of events which make up the normal germination pattern of Impatiens holstii pollen grains can be separated from each other by varying the concentrations of the oxybarbiturates amytal and seconal used. Evidence is also presented which indicates that the actions of barbituric acid and the oxybarbiturates barbital, amytal, and seconal on Impatiens holstii pollen germination behaviour correlate well with certain aspects of the pharmacological literature concerning the actions of these organic compounds on the central nervous system (CNS).     

Pollen development in orchids 4. Cytoskeleton and ultrastructure of the unequal pollen mitosis inPhalaenopsis

Summary The unequal first mitosis in pollen ofPhalaenopsis results in a small generative cell cut off at the distal surface of the microspore and a large vegetative cell. No preprophase band of microtubules is present, but polarization of the microspore prior to this critical division is well marked. A generative pole microtubule system (GPMS) marks the path of nuclear migration to the distal surface, and the organelles become unequally distributed. Mitochondria, plastids and dictyosomes are concentrated around the vegetative pole in the center of the microspore and are almost totally excluded from the generative pole. The prophase spindle is multipolar with a dominant convergence center at the GPMS site. The metaphase spindle is disc-shaped with numerous minipoles terminating in broad polar regions. In anaphase, the spindle becomes cone-shaped as the spindle elongates and the vegetative pole narrows. These changes in spindle architecture are reflected in the initial shaping of the telophase chromosome groups. F-actin is coaligned with microtubules in the spindle and is also seen as a network in the cytoplasm. An outstanding feature of orchid pollen mitosis is the abundance of endoplasmic reticulum (ER) associated with the spindle. ER extends along the kinetochore fibers, and the numerous foci of spindle fibers at the broad poles terminate in a complex of ER.Abbreviations CLSM confocal laser scanning microscope/microscopy - DMSO dimethyl sulfoxide - ER endoplasmic reticulum - FITC fluorescein isothiocyanate - GPMS generative pole microtubule system - MBS m-maleimidobenzoic acidN-hydroxysuccinimide ester - PPB preprophase band of microtubules - RhPh rhodamine palloidin - TEM transmission electron microscope/microscopy     

Pollen morphology and ultrastructure of the Bennettitales: In situ pollen of Cycadeoidea

The micromorphology and ultrastructure of in situ pollen from Cycadeoidea dacotensis are described from permineralized specimens collected from the Lower Cretaceous of North America. Pollen grains are ovoid and relatively small, averaging 25 μm in length and 12 μm in width. Grains are monosulcate with the exine typically invaginated in apertural regions. Exine ornamentation ranges from punctate to psilate. The exine averages 0.73 μm in thickness and is composed of a light-staining sexine and a dark-staining nexine. The sexine consists of a thin, homogeneous tectum, typically with a well-defined inner boundary, and a thicker granular infratectum. The infratectal granules are relatively uniform in size, however, variation occurs in the arrangement of granules. In some grains, the sexine appears homogeneous because there is little lacunal space between the individual granules. The granular infratectum is in direct contact with the underlying nexine. The nexine is uniform in thickness in both apertural and nonapertural regions, and it lacks lamellae throughout. Pollen morphology and ultrastructure are compared with those of the bennettitalean genus Leguminanthus and the dispersed genus Monosulcites. In addition, the fine structure of Cycadeoidea pollen is compared to that of the gymnosperm groups with which the Bennettitales are regarded to be most closely related, including Gnetales, Pentoxylales, and Eucommiidites-type pollen-producing plants.     

Molecular analyses of the Chinese herb Leigongteng (Tripterygium wilfordii Hook.f.)

Tripterygium wilfordii Hook.f., known as Leigongteng (Thunder God Vine) in traditional Chinese medicine, has attracted much attention for its applications in relieving autoimmune disorders such as rheumatoid arthritis and systemic lupus erythematosus, and for treating cancer. Molecular analyses of the ITS and 5S rDNA sequences indicate that T. hypoglaucum and T. doianum are not distinct from T. wilfordii, while T. regelii should be recognized as a separate species. The results also demonstrate potential value of rDNA sequence data in forensic detection of adulterants derived from Celastrusangulatus in commercial samples of Leigongteng.     

Pollen dispersal and long distance transport: the case of thermophilic pollen in subarctic Canada

At Kuujjuarapik, on the eastern coast of Hudson Bay (Canada), thermophilic tree pollens, all of long distance origin, are found in low concentrations, but throughout the pollen season. A Cour impaction pollen trap has allowed us to obtain statistically significant concentrations. The analysis of the origins of synchronous air masses by back-trajectories shows that the weekly variations of thermophilic pollens can be linked to air masses coming either from northern regions (Alaska or Beaufort Sea) for minima, and southern regions (Gulf of Mexico) for maxima.     

罗布红麻和罗布白麻生态地理特性探讨

根据文献资料和我们多年的研究,发现罗布红麻和罗布白麻的各地居群具有多样的生态地理特性。通过在陕西关中地区的引种栽培试验,证实了各地罗布红麻和罗布白麻居群生态地理特性的多样性。这对于其引种栽培和开发利用都有极为重要指导意义。     

In vitro Arabidopsis pollen germination and characterization of the inward potassium currents in Arabidopsis pollen grain protoplasts

The focus of this study is to investigate the regulatory role of K(+) influx in Arabidopsis pollen germination and pollen tube growth. Using agar-containing media, in vitro methods for Arabidopsis pollen germination have been successfully established for the first time. The pollen germination percentage was nearly 75% and the average pollen tube length reached 135 microm after a 6 h incubation. A decrease in external K(+) concentration from 1 mM to 35 microM resulted in 30% inhibition of pollen germination and 40% inhibition of pollen tube growth. An increase in external K(+) concentration from 1 mM to 30 mM stimulated pollen tube growth but inhibited pollen germination. To study how K(+) influx is associated with pollen germination and tube growth, regulation of the inward K(+) channels in the pollen plasma membrane was investigated by conducting patch-clamp whole-cell recording with pollen protoplasts. K(+) currents were first identified in Arabidopsis pollen protoplasts. The inward K(+) currents were insensitive to changes in cytoplasmic Ca(2+) but were inhibited by a high concentration of external Ca(2+). A decrease of external Ca(2+) concentration from 10 mM (control) to 1 mM had no significant effect on the inward K(+) currents, while an increase of external Ca(2+) concentration from 10 mM to 50 mM inhibited the inward K(+) currents by 46%. Changes in external pH significantly affected the magnitude, conductance, voltage-independent maximal conductance, and activation kinetics of the inward K(+) currents. The physiological importance of potassium influx mediated by the inward K(+)-channels during Arabidopsis pollen germination and tube growth is discussed.