Origins of prehistoric flints: The neocortex memory revealed by scanning electron microscopy

Flint has been widely used during prehistoric times and the question of determining the origin of specific pieces is recurrent in archaeological discussions. We have run a scanning electron microscope examination of the surfaces of both natural flints from geological deposits and flaked artefacts excavated from prehistoric sites in the Massif Central (France). These samples exhibit mineralogical textures resulting from genomorphic and phenomorphic processes, dispersed alteration in the form of erosion as well as the dissolution and migration of more or less mobile elements. Rocks used to manufacture artefacts at Sainte-Anne 1 (Haute-Loire, France) and Payre (Ardèche, France) show various kinds of alteration related to the range of phases of transport and deposition to which they were subjected, beginning with their first exposure in a geological context and following through to the post-depositional phase, (i.e., following abandonment as artefacts in an archaeological site). Later phenomena do not however completely obliterate stigma imprinted prior to their prehistoric collection, because the main late-phase change that occurs on flat surfaces prior to their collection is preserved in the hollows of remnant natural surfaces on archaeological tools. With the scanning electron microscope, five distinct morphologies representative of different environments have been characterised by establishing associations between the different forms and by plotting their regional distribution.     

Historical biogeography of Eusattus muricatus (Coleoptera: Tenebrionidae) within the Great Basin, western North America

Genetics of Eusattus muricatus LeConte (Coleoptera: Tenebrionidae) populations within the Lahontan Basin and the Lahontan Trough of the Great Basin, western North America were assayed using protein electrophoresis. Two similar population phenograms were developed using neighbour-joining and maximum likelihood algorithms. An area cladogram was developed using geological data. Maximum likelihood phenogram and area cladogram were compared to test for congruence between geology and phylogeny. Pairwise regressions of estimated gene flow measures ( Nm ) yielded a significant relationship for populations within the Lahontan Basin but not for the Lahontan Trough. Protein molecular clock calculations showed that populations within the Lahontan Basin were primarily differentiating according to the dune/subbasin isolation events predicted by the area cladogram.     

Ultrastructure of Proechinophthirus zumpti (Anoplura,Echinophthiriidae) by scanning electron microscopy

The ultrastructure of Proechinophthirus zumpti Werneck, 1955, mainly the external chorionic features of the egg, is described through electronic microscopy techniques. This species was first cited in Argentina, infesting Arctocephalus australis (Zimmermann, 1873). The morphological adaptations of adults and nymphs are described in both species of Proechinophthirus parasitic on Otariidae: P. fluctus (Ferris, 1916) and P. zumpti.     

Sex determination and the evolution of dioecy from monoecy in Sagittaria latifolia (Alismataceae)

The role of mutations of small versus large effect in adaptive evolution is of considerable interest to evolutionary biologists. The major evolutionary pathways for the origin of dioecy in plants (the gynodioecy and monoecy-paradioecy pathways) are often distinguished by the number of mutations involved and the magnitude of their effects. Here, we investigate the genetic and environmental determinants of sex in Sagittaria latifolia, a species with both monoecious and dioecious populations, and evaluate evidence for the evolution of dioecy via gynodioecy or monoecy-paradioecy. We crossed plants of the two sexual systems to generate F1, F2 and backcross progeny, and grew clones from dioecious populations in low-and high-fertilizer conditions to examine sex inconstancy in females and males. Several lines of evidence implicate two-locus control of the sex phenotypes. In dioecious populations sex is determined by Mendelian segregation of alleles, with males heterozygous at both the male- and female-sterility loci. In monoecious populations, plants are homozygous for alleles dominant to male sterility in females and recessive to female sterility in males. Experimental manipulation of resources revealed sex inconstancy in males but not females. These results are consistent with predictions for the evolution of dioecy via gynodioecy, rather than the expected monoecy-paradioecy pathway, given the ancestral monoecious condition.     

Gender variation of sequential inflorescences in a monoecious plant Sagittaria trifolia (Alismataceae)

In protogynous plants, female flowers of early blooming plants are at a reproductive disadvantage because they cannot set fruit due to the lack of available pollen. To study this phenomenon, gender expression of the monoecious herb Sagittaria trifolia was investigated over the entire flowering season in two field and two cultivated populations in Hubei and Hunan Provinces, China. In racemes of S. trifolia, flowers open sequentially from bottom to top, with female flowers opening first followed by male flowers. This creates a temporal separation of sexes in the species. Under field conditions small plants are often male, with production of both male and female flowers increasing with plant size. Femaleness increased among sequential inflorescences since female flower production increased whereas male flower production did not. Seed production was greater in large inflorescences because they contain more female flowers, and the number of ovules increased in female flowers at basal positions within the raceme. A consistent pattern of high seed set was observed in flowers from both field and cultivated populations. About 1 % of unfertilized ovules resulted from no pollination and 2 % of the seeds produced were only partly developed due to resource limitation. In the first inflorescence of the six experimental populations, 6.7-40.0 % of individuals produced only male flowers, and female flowers of 1.9-6.5 % individuals were aborted. The occurrence of male flowers in early blooming inflorescences could be an adaptive strategy to conserve resources and enhance pollination of female flowers in protogynous S. trifolia.     

Characters in the book lungs of Scorpiones (Chelicerata,Arachnida) revealed by scanning electron microscopy

The fine structure of the book lungs in 29 species representing ten monophyletic taxa of the Scorpiones (Arachnida) was investigated using scanning electron microscopy (SEM). Scorpion lungs are not homogeneous across the group. Here we describe and score three sets of phylogenetically informative characters: (1) the surface ornament of the lung lamellae, (2) the distal margins of the lamellae and (3) the fine structure of the spiracle margin. Provisional results suggest that reticulation on the surface of the lung lamellae is characteristic of the Buthidae. By contrast, non-buthid scorpions maintain the air space between adjacent lamellae using projecting trabeculae. Typically they are simple struts, but the trabeculae are distally branched in all investigated Scorpionidae, plus at least one species belonging to the Liochelidae. Simple thorns on the lamellar margins probably represent the plesiomorphic condition, while more complex, branched, arcuate morphologies appear to be homoplastic, occurring sporadically in numerous scorpion sub-groups. The tightly packed, hexagonal pillars around the posterior margin of the spiracle support a close relationship between Scorpionidae and Liochelidae, to the exclusion of the Urodacidae.     

Protoperidinium (Dinophyceae) from Greece as seen by scanning electron microscopy

The marine dinoflagellates Protoperidinium ventricum (Abé) Balech, P. diaholus (Cleve) Balech, P. tristylum (Stein) Balech and P. divergens (Ehrenberg) Balech, collected from neritic plankton of the gulfs of Saronikos and Korinthiakos, Greece, have been examined by SEM. Morphological characteristics of their thecae are described and evaluated. Results are compared with patterns established by earlier SEM studies on related taxa.     

Sex in advertising: dioecy alters the net benefits of attractiveness in Sagittaria latifolia (Alismataceae)

The flowers and inflorescences of animal-pollinated dioecious plants are generally small and inconspicuous in comparison with outcrossing cosexual species. The net benefits of an attractive floral display may be different for dioecious compared to cosexual populations because dioecious species experience a more severe reduction in pollen delivery when pollinators forage longer on fewer individuals. Here, we develop a model that predicts the decrease in pollen delivery in dioecious relative to cosexual populations from female-female, female-male and male-male visit sequences as the number of individuals visited varies. To evaluate the predictions of our model we conducted a common garden experiment with dioecious and monoecious (cosexual) arrays of the insect-pollinated herb Sagittaria latifolia. We find that, although increasing the advertisements of floral rewards (i.e. increasing floral display) attracts more pollinators to individuals, the probability that these pollinators subsequently deliver pollen to neighbouring plants depends on sexual system. Because the number of individual plants visited per foraging trip did not increase significantly with floral display, the relative pollination success of dioecious versus monoecious populations decreases with increased floral display. We propose that this could explain why dioecy is strongly correlated with reduced floral display among angiosperm species.     

The effects of density on size-dependent gender plasticity in the monoecious species Sagittaria potamogetifolia (Alismataceae)

Aim

To test the fitness-gain curve model proposes that cosexual plants adjust their sex ratios and resource allocation depending on their size. In this study, the monoecious species Sagittaria potamogetifolia was used as a model to determine the effects of plant size and density on gender modification and reproductive allocation.

Aerenchyma develops by cell lysis in roots AND CELL SEPARATION IN LEAF PETIOLES IN Sagittaria lancifolia (Alismataceae)

Aerenchyma gas spaces are important for plants that survive flooding because these spaces provide an internal pathway for oxygen transport to the root zone. The objective of this study was to characterize the development of aerenchyma gas spaces in Sagittaria lancifolia L., a dominant species in freshwater wetlands adjacent to the Gulf of Mexico. Tissue at different developmental stages was collected from hydroponically grown plants, embedded in plastic, and sections were observed with a light microscope. In S. lancifolia roots, lysigeny (cell lysis) produced gas spaces that increased in volume from the root meristem to the most mature root tissue. Shoot aerenchyma occurred in the large petioles of S. lancifolia and through the blade midrib, but not in the laminar portion of the blade. In contrast to the roots, gas spaces in the petiole were formed by schizogeny (cell separation during development). Shoot initials produced cells that formed interlocking cylinders in the cortex and diaphragm cells that bridged the central portion of the cylinders. Division and expansion of both these cell types increased the diameter of the cylinders and created schizogenous gaps between diaphragm layers that produced large gas spaces in mature tissue. Therefore, aerenchyma development occurs by two different processes in S. lancifolia.     

Ornamentation of thecal plates in Protoperidinium (Dinophyceae) as seen by scanning electron microscopy

The thecae of over thirty species of Protoperidinium from aroundthe British Isles have been examined by SEM and the varioustypes of ornamentation described. The simplest type consistsof smooth plates in which the trichocyst pores have an evenor irregularly thickened rim. In one species the pores are situatedon mounds. In another variation the plates are covered withminute pimples. Many species have plates which are thickenedby a surface reticulum and this may be made up of lines of pimples,of irregularly arranged ridges or bars which protrude to formsmall spines. In one species the ridges always surround trichocystpores and in another there are prominent spines which bear noobvious relationship with the pores. It is concluded that thecalornamentation can provide useful taxonomic characters withinthis genus.     

Litter decomposition in grasslands of Central North America (US Great Plains)

One of the major concerns about global warming is the potential for an increase in decomposition and soil respiration rates, increasing CO₂ emissions and creating a positive feedback between global warming and soil respiration. This is particularly important in ecosystems with large belowground biomass, such as grasslands where over 90% of the carbon is allocated belowground. A better understanding of the relative influence of climate and litter quality on litter decomposition is needed to predict these changes accurately in grasslands. The Long‐Term Intersite Decomposition Experiment Team (LIDET) dataset was used to evaluate the influence of climatic variables (temperature, precipitation, actual evapotranspiration, and climate decomposition index), and litter quality (lignin content, carbon : nitrogen, and lignin : nitrogen ratios) on leaf and root decomposition in the US Great Plains. Wooden dowels were used to provide a homogeneous litter quality to evaluate the relative importance of above and belowground environments on decomposition. Contrary to expectations, temperature did not explain variation in root and leaf decomposition, whereas precipitation partially explained variation in root decomposition. Percent lignin was the best predictor of leaf and root decomposition. It also explained most variation in root decomposition in models which combined litter quality and climatic variables. Despite the lack of relationship between temperature and root decomposition, temperature could indirectly affect root decomposition through decreased litter quality and increased water deficits. These results suggest that carbon flux from root decomposition in grasslands would increase, as result of increasing temperature, only if precipitation is not limiting. However, where precipitation is limiting, increased temperature would decrease root decomposition, thus likely increasing carbon storage in grasslands. Under homogeneous litter quality, belowground decomposition was faster than aboveground and was best predicted by mean annual precipitation, which also suggests that the high moisture in soil accelerates decomposition belowground.     

冠果草的性表达状态及其进化含义

通常认为雄花两性花同株是植物性表达两性花进化到雌雄异株的过渡类型之一。慈姑属仅冠果草（Ｓａｇｉｔｔａｒｉａ ｇｕｙａｎｅｎｓｉｓ Ｈ．Ｂ．Ｋ．ｓｕｂｓｐ．ｌａｐｐｕｌａ（Ｄ．Ｄｏｎ）Ｂｏｊｉｎ）的性表达为典型的雄花两性花同株,其他种为雌雄同株。对冠果草野外和实验居群的性配置进行了定量观察。东乡和武夷山居群雄花的比例分别为２．４８％０．９６％。实验表明,栽培条件影响冠果草的性表达：在营养较好的条件下     

Population genetic structure of Sagittaria natans (Alismataceae), an endangered species in China,revealed by nuclear SSR loci analyses

Sagittaria natans (Alismataceae) is an endangered aquatic herb in China. The nuclear microsatellite (SSR) loci variation of the 11 extant populations of S. natans was investigated in the present study. A relative high level of genetic diversity was found in this species. The Wilcoxon’s signed rank tests did not indicate a recent bottleneck in any of the populations. The Analysis of molecular variance (AMOVA) showed a low level of population differentiation (FST = 0.284) among the populations of S. natans. This result was supported by STRUCTURE analysis (K = 1), the NJ analyses and Mantel test (r = ?0.0221, p > 0.5). The genetic structure could be contributed to an earlier period of more pronounced gene flow when the species had a more continuous distribution. The results of this study could be used as the basis for conservation guidelines for the management of this endangered species in China.     

Microvasculature of the bear heart demonstrated by scanning electron microscopy

Morphological features of the coronary microvasculature were examined in ten Black bears (Ursus americanus). Tissue samples were collected for scanning electron microscopy from five animals, and vascular casts of the coronary vessels were prepared from five. Each portion of the microvasculature examined had specific morphological characteristics which may contribute to the control of blood flow. All indications from the present study suggest that the structure of the coronary microvasculature in the Black bear is similar to that of mammalian species previously examined.     

A protocol for isolating Xenopus oocyte nuclear envelope for visualization and characterization by scanning electron microscopy (SEM) or transmission electron microscopy (TEM)

This protocol details methods for the isolation of oocyte nuclear envelopes (NEs) from the African clawed toad Xenopus laevis, immunogold labeling of component proteins and subsequent visualization by field-emission scanning electron microscopy (FESEM) and transmission electron microscopy (TEM). This procedure involves the initial removal of the ovaries from mature female X. laevis, the dissection of individual oocytes, then the manual isolation of the giant nucleus and subsequent preparation for high-resolution visualization. Unlike light microscopy, and its derivative technologies, electron microscopy enables 3-5 nm resolution of nuclear structures, thereby giving unrivalled opportunities for investigation and immunological characterization in situ of nuclear structures and their structural associations. There are a number of stages where samples can be stored, although we recommend that this protocol take no longer than 2 d. Samples processed for FESEM can be stored for weeks under vacuum, allowing considerable time for image acquisition.     

Characterization of theGeosiphon pyriforme symbiosome by affinity techniques: confocal laser scanning microscopy (CLSM) and electron microscopy

Summary Geosiphon pyriforme represents a photoautotrophic endosymbiosis of aGlomus-like fungus with the cyanobacteriumNostoc punctiforme. The fungus forms unicellular bladders of up to 2 mm in length and 0.5 mm in diameter growing on the soil surface and harboring the endosymbioticNostoc filaments. The cyanobacteria are located in a compartment (the symbiosome) bordered by a host membrane. The space between this symbiosome membrane (SM) and theNostoc cell wall is filled with an about 30–40 nm thick layer of amorphous material, which is present also in the regions of the symbiosome where noNostoc filaments are located. At these sites the amorphous material consists of a 20–30 nm thick layer separating the SM. The region between the SM and the cyanobacterium is defined as symbiosome space (SS). Fungal bladders, hyphae and free livingNostoc were analyzed by affinity techniques as well as the material occurring in the SS. FITC-coupled lectins with sugar specificity to -D-mannosyl/-D-glucosyl (Con A), N-acetyl--D-glucosamine oligomers (WGA), -L-fucosyl (UEA-I), -D-galactosyl (RCA-120), -D-galactosyl (BS-I-B₄), N-acetyl--D-galactosamine (HPA), and sialic acid (EBL) residues were tested. WGA binding and calcofluor white staining demonstrated that the bladder wall as well as the SS contain fibrillar chitin. Of the other lectins only Con A clearly labeled the symbiosome. On the contrary, the lectin binding properties of the slime produced by free livingNostoc-colonies indicate the presence of mannose, fucose, GalNAc, sialic acid, and galactose, while chitin or GlucNAc-oligomers could not be detected. The symbiosome was also investigated electron microscopically. WGA-gold binding confirmed the presence of chitin, while a slight PATAg reaction indicated some polysaccharidic molecules within the SS. Our results show that the amorphous material within the SS contains molecules typical of the fungal cell wall and suggest that the SM is related to the fungal plasma membrane. The applied lectins all bind to the hyphal surface, indicating a high molecular complexity. Mannosyl, -galactosyl, and sialic acid residues are strongly exposed at the outer cell wall layer, whereas GlucNAc, GalNAc, and -galactosyl residues seem to be present in smaller amounts. The symbiotic interface established between the fungus andNostoc inGeosiphon shows many similarities to that occurring between fungi and root cells in arbuscular mycorrhizas.Abbreviations AM arbuscular mycorrhiza - BS-I-B₄ Bandeiraea simplicifolia lectin I isolectin B₄ - CLSM confocal laser scanning microscopy - Con A Concanavalin A - EBL elderberry bark lectin I - FITC fluorescein isothiocyanate - HPA Helix pomatia agglutinin - PATAg periodic acid-thiocarbohydrazide-Ag proteinate - SM symbiosome membrane - SS symbiosome space - RCA-120 Ricinus communis agglutinin 120 - UEA-I Ulex europaeus agglutinin I - WGA wheat germ agglutinin Dedicated to Professor Dr. Peter Sitte at the occasion of his 65th birthday