Atlas of foliar surface features in woody plants,IX. Betulaceae of eastern United States

Scanning electron microscopy was used to examine foliar surface features such as cuticular patterns, epicuticular wax, and trichome types in species ofAlnus, Betula, Carpinus, Corylus, andOstrya. Six trichome types are recognized in this survey, four non-glandular (acicular, filiform, aduncate, subulate) and two glandular (stipitate gland and peltate scale). The distribution of these trichomes among the genera supports the recognition of two tribes, Betuleae (Alnus andBetula) and Coryleae (Carpinus, Corylus, andOstrya). Trichome morphology can be an important supportive taxonomic character in determining evolutionary relationships.     

Atlas of foliar surface features in woody plants,V. fraxinus (Oleaceae) of eastern North America

Scanning electron microscopy of foliar surfaces of the seven species ofFraxinus native in eastern North America confirms the importance of cuticular patterns (smooth, striated, striate-papillose, and coronulate) as useful taxonomic character states. There are there trichome types (acicular hairs ornamented with micro-striae or-papillae, peltate glands, and scales) which are found in nearly all species and which vary only in density. No differences in surface features are found by which determination can be made of either sex or ploidy level inF. americana. Hybridization betweenF. americana andF. pennsylvanica is considered to be rare in nature and is not a probable cause for the overlap of character states and concomitant difficulty of identification based upon gross morphological features. Likewise, the putative hybrid origins ofF. biltmoreana andF. profunda are not supported by micro-morphological features. Illustrations are given for all species.     

The systematic significance of achene micromorphology in Carex retrorsa (Cyperaceae)

Scanning electron microscopy (SEM) was used to examine achene epidermal features of nine Carex species within sections Vesicariae and Lupulinae. Two major epidermal features are recognized in this study: the epidermal layers with one or two primary silicon bodies in each cell and the absence of any primary silicon body. Comparative data of achene morphology using SEM, and evidence compiled from previous investigations, support the placement of Carex retrorsa within section Vesicariae.     

Microbial attachment to particles in marine and freshwater ecosystems

Scanning electron microscopy observations ofin situ suspended marine and freshwater particles show diverse but similar modes of bacterial and fungal attachment. A survey of Sierra Nevada mountain lakes and pelagic and near-shore waters in the Pacific Ocean indicates that attachment is most noticeable in the near-surface waters where fresh dissolved and particulate input of carbon from phytoplankton and elevated temperatures favor microbial growth. The most common modes of attachment are: adhesive stalk formation, growth on adhesive webs, attachment by the use of pili-like appendages and slimy capsular secretions, and molecular or chemical sorption without the use of visualized structural appendages. Attached microbial growth is accelerated when particulate substrates are supplied, even when they are not rich in organic nutrients. This is the case in the Lake Tahoe basin, where microflora attached to eroded silts can significantly modify the organic carbon and nutrient content of such minerogenous particles.     

Prominent spermoderm patterns of Poaceae

Scanning electron microscopy studies of 60 genera and 118 species of the Poaceae revealed prominent spermoderm patterns. The most common patterns observed (with percent occurrence) were simple reticulate with straight walls (48.3%), simple reticulate with wavy walls (14.4%), and verrucate without pubescence (13.6%). No other pattern occurred in more than 10% of the species observed. Some genera possess common spermoderm patterns among species while others do not.     

Cytotoxicity of Fusarium T-2 in cultured Madin-Darby bovine kidney (MDBK) and primary fetal bovine kidney (PFBK) cells

Cytotoxicity of Fusarium T-2 toxin was evaluated in cultured Madin-Darby bovine kidney (MDBK) and primary fetal bovine kidney (PFBK) cells. The criteria for evaluation included number of adherent cells, phase contrast microscopy, and the scanning and transmission electron microscopy. The primary cells were more sensitive to the toxic effects of T-2 toxin than MDBK cultures. Cytotoxicity was observed when the cultures were exposed to the toxin for only 1 hour and then incubated with untreated media. Cell multiplication was decreased in both systems in 72 hour cultures. Scanning electron microscopy indicated loss of inter-cell contact and marked alterations in cell shape. Transmission electron microscopy indicated extensive proliferation of lysosomal bodies and proliferation of endoplasmic reticulum. The membrane system and mitochondria were not affected. Results indicated the kidney cells are highly sensitive to the toxic effects of T-2 toxin.     

Introgressive hybridization in two southern California species of Rhus (Anacardiaceae)

Five populations ofRhus integrifolia andR. ovata were studied and, based upon morphological characteristics and pollen viability, a number of putative hybrids were found in those areas where the two species are sympatric. Diagnostic characteristics most useful for distinguishing the species and their hybrids are leaf morphology, sepal pubescence, petiole length, and petal size. Introgression was evident in at least three populations, which suggested that hybridization between the two species is an ancient phenomenon. Evolutionarily, hybridization between the two taxa has probably been of little consequence.     

Seed morphology in New WorldAntirrhineae (Scrophulariaceae): Systematic and phylogenetic implications

Seeds of 39 species representing all native genera and sections of New WorldAntirrhineae were examined with the Scanning Electron Microscope and Light Microscope. Based on seed shape and surface ornamentation, seven morphological categories are recognized: cristate, tetracostate, foveolate, tumid tuberculate/cristate, circumalate, medusiform, and a miscellaneous category. Most sections and/or generic categories can be characterized by a single morphological type, althoughMaurandya s.l. has several distinctive seed types included within present generic boundaries. The cristate seed type is believed to be the “primitive type” from which the medusiform and tumid tuberculate/cristate types have developed. The tumid tuberculate/cristate type presumably gave rise to the circumalate, tetracostate, and foveolate patterns. The miscellaneous category (represented byLinaria andKickxia) may be phyletically remote from the other New World types. Seeds of the New WorldAntirrhineae mostly seem to be adapted for water dispersal although some adaptations for wind dispersal (e.g. wings, light weight) are evident. Free-hand sections and seed coat clearings indicate that circumalate seeds are of two different anatomical patterns; such structural diversity may be found in the other seed types.     

Copper tolerance in Frankia sp. strain EuI1c involves surface binding and copper transport

Several Frankia strains have been shown to be copper-tolerant. The mechanism of their copper tolerance was investigated for Frankia sp. strain EuI1c. Copper binding was shown by binding studies. Unusual globular structures were observed on the surface of the bacterium. These globular structures were composed of aggregates containing many relatively smaller “leaf-like” structures. Scanning electron microscopy with energy-dispersive X-ray (SEM-EDAX) analysis of these structures indicated elevated copper and phosphate levels compared to the control cells. Fourier transform infrared spectroscopy (FTIR) analysis indicated an increase in extracellular phosphate on the cell surface of copper-stressed cells. Bioinformatics’ analysis of the Frankia sp. strain EuI1c genome revealed five potential cop genes: copA, copZ, copC, copCD, and copD. Experiments with Frankia sp. strain EuI1c using qRT-PCR indicated an increase in messenger RNA (mRNA) levels of the five cop genes upon Cu²⁺ stress. After 5 days of Cu²⁺ stress, the copA, copZ, copC, copCD, and copD mRNA levels increased 25-, 8-, 18-, 18-, and 25-fold, respectively. The protein profile of Cu²⁺-stressed Frankia sp. strain EuI1c cells revealed the upregulation of a 36.7 kDa protein that was identified as FraEuI1c_1092 (sulfate-binding periplasmic transport protein). Homologues of this gene were only present in the genomes of the Cu²⁺-resistant Frankia strains (EuI1c, DC12, and CN3). These data indicate that copper tolerance by Frankia sp. strain EuI1c involved the binding of copper to the cell surface and transport proteins.     

Antagonistic interactions between Aureobasidium pullulans and Fusarium culmorum,a fungal pathogen of winter wheat

The effect of the biological control agent Aureobasidium pullulans (de Bary) G. Arnaud on the development of Fusarium head blight (FHB) on winter wheat and kernel contamination with fungi of the genera Fusarium, Acremonium, Cladosporium and Penicillium was analyzed in a greenhouse experiment. Scanning electron microscopy was used to evaluate the distribution of A. pullulans cells and aggregates on wheat kernels, infection structures of Fusarium culmorum (W.G. Smith) Sacc and the antagonist-pathogen interactions. Biological control with A. pullulans reduced FHB severity by 21.67 % and improved grain filling by 5.02 %, compared with the control treatment. The survival of A. pullulans was good (to 31 cells per kernel), in particular on the surface and in the crease of kernels, including in pathogen-inoculated wheat plants. A. pullulans cells firmly adhered to F. culmorum hyphae, and damaged them. In most cases, autochthonous communities of filamentous fungi of the genera Acremonium and Penicillium developed at a slower rate after kernel inoculation with the pathogen.     

Taxonomic and phylogenetic significance of seed coat microsculpturing in Mentzelia (Loasaceae) in Wyoming and adjacent western states

Six hundred seeds belonging to the genusMentzelia (Loasaceae) were examined using the Scanning Electron Microscope. The seeds represented all six sections of the genus and approximately 76% of the total number of species from northern Mexico and western United States. Emphasis was placed on seed material from Wyoming and adjoining states. Ovule serial sectioning was done to determine testa ontogeny. In all cases seed surface structures could be related to excrescences developing from the integument cells. Six basic seed coat relief features could be distinguished, corresponding to the six sections in the genus. Minor variations in the basic types are constant and characteristic for species or, in very few instances, for species groups. A dendrogram was constructed from the seed testa data which supports the phylogeny established from chromosomal and morphological data. Practical application of seed coat microcharacter specificity to identification ofMentzelia seeds from prehistoric sites is suggested.     

Initial attachment ofCandida albicans cells to buccal epithelial cells

The rapid-freezing technique was applied in association with scanning and transmission electron microscopy to observe the initial attachment (or contact) ofCandida albicans cells to exfoliated human buccal epithelial cells. Low temperature scanning electron microscopy provided detailed three-dimensional morphological features of the yeast-epithelial cell association; adhesion ofC. albicans cells to host cells was primarily owing to an interaction between fibrillar layer of the yeast cell wall and the membrane interdigitations of the epithelial cells. Such a particular interconnection between the two cells was confirmed by the freeze-substitution fixation for transmission electron microscopy. These results clearly demonstrate the outermost fibrillar cell wall layer ofC. albicans responsible for adhesion to host cells.     

Is the insular endemic Psidium socorrense (Myrtaceae) at risk of extinction through hybridization?

Natural hybridization between an insular endemic species and a widely distributed congener may endanger the endemic through genetic assimilation or outbreeding depression. Furthermore, hybrids can exhibit complex morphological variation, causing taxonomic problems in the identification of the involved taxa. In this work, we used a combination of leaf morphological and molecular markers (RAPD) to establish the differentiation between Psidium sp. aff. sartorianum and the insular endemic P. socorrense. It was also determined if hybridization between these taxa occurs in the southern slope of Isla Socorro, Mexico. Plant collection was carried along an altitudinal gradient (100–800 m). We collected eight populations separated 100 m a.s.l. apart from each other; 25 individuals were collected per population. Psidium socorrense and P. sp. aff. sartorianum differed significantly in all but two morphological characters measured. Also, a high number of diagnostic RAPD markers were found for each taxon. These results suggest that two Psidium species occur at Isla Socorro. Furthermore, both morphological and RAPD markers revealed a hybrid zone located in the southern slope of Isla Socorro (400–700 m a.s.l.) with an asymmetrical pattern of gene flow towards P. socorrense. We suggest that the disturbance caused by the sheep population in the mixed stand favors the establishment of hybrids. We further discuss whether hybridization represents a threat to the insular endemic P. socorrense.     

Centric diatoms from Lake Frolikha (Transbaikal area) and peculiarities of distribution of some taxa in Asia

The species composition of centric diatoms from Lake Frolikha (Transbaikal area) has been studied. The lake is located on the eastern shore of Lake Baikal and connected with it by a river. Twenty-three species of Centrophyceae from 7 genera (Aulacoseira, Cyclostephanos, Cyclotella, Discostella, Handmania, Pliocaenicus, and Stephanodiscus) have been found. The most represented genus is Aulacoseira (11 species). Fifteen species and 4 genera are new for the lake record. All revealed species are known in other lakes of the Baikal region; however Baikal endemics are absent in Lake Frolikha. The flora of centric diatoms in Lake Frolikha can be divided into two groups. The first group includes taxa common in Lake Baikal, and the second group includes taxa not typical for Lake Baikal. The level of differences between flora in Lake Baikal and Lake Frolikha is high (43%) despite the close location and connection to the river. An analysis of distribution of interesting species in Asia is presented.     

Expression Analysis of Bombyx mori Bidensovirus Structural Proteins and Assembly of Virus-like Particles in Insect Cells

Bombyx mori bidensovirus (BmBDV) is a new designated species of the new genus Bidensovirus in the new family Bidnaviridae, which contains two single-stranded linear DNAs (VD1 and VD2) and causes the chronic densonucleosis disease of silkworm. Previous researches revealed that VD1-ORF3 encodes the major structural proteins VPs. In this work, through western blot, we found that VPs expressed from 48 h post-inoculation and kept increasing until 120 h post-inoculation in midgut of Bombyx mori. In order to further investigate the translation of vp gene, the ORFs (vp1 and vp2) of the VP started just up-stream of the first two candidate initiation codons were expressed in Sf9 cells by a baculovirus expression system. The expression products were purified by gradient density centrifugation and analyzed by Western blot and electron microscopy. The results showed that the expressions of vp1 yielded three proteins (VP1, VP1′, and VP2), which are the same with the viral VPs expression in midgut of Bombyx mori, and vp2 generated two VPs with the molecular weights of about 51 kDa (VP2) and 37 kDa. The observation by electron microscopy indicated that these VPs can auto-assemble into virus-like particles that could not be distinguished from virus particles. These findings will provide materials for studying the structure of BmBDV and be helpful in the studies on BmBDV-based disease in silkworms.     

Evaluation of an immune-privileged scaffold for In vivo implantation of tissue-engineered trachea

Forty tracheas were harvested from donor New Zealand rabbits. Thirty of the tracheas were randomly divided into four treatment groups corresponding to 4, 5, 6, or 7% NaClO₄ and one untreated group (n = 6 each group). Scanning electron microscopy distinctly revealed the cilium of epithelial cells in the fresh trachea. The internal surface of the trachea was rough in the 4% treatment group and smooth in the 5% treatment group, whereas the matrix was fractured in the 6% treatment group and highly fractured in the 7% treatment group. We observed that the number of nuclei in the cells of the 4, 5, 6, and 7% treatment groups decreased compared to the cells of the untreated group (p < 0.05). Although there was a significant decrease in maximum tensile strength, tensile strain at fracture and the elastic modulus (p < 0.05) with increasing concentrations of NaClO₄, the content of glycosaminoglycans (GAGs) did not significantly decline (p > 0.05) in the 5% treatment group. In addition, histopathological analysis showed that the fiber component and basement membrane of the matrix in the 5% treatment group were retained after optimal decellularization. Despite the preserved cartilage, in vitro immunohistochemical analysis revealed that the matrix did not show the presence of major histocompatibility complex (MHC) antigens. The remaining ten donor tracheas, which were divided into a positive control group and an optimal decellularized group, were used for allogeneic transplantation. Blood samples were taken regularly, and histologic examinations were performed at 30 days postimplantation, which showed no significant immune rejection. In conclusion, we surveyed the structural integrity through morphological observation and compared the biomechanical and immunogenic changes in the tracheal matrix under the different treatments. The optimal decellularized tracheal matrix with preserved cartilage, which was acquired via 5% NaClO4 treatment, exhibited structural integrity, antigen cell removal and immune privilege and would be suitable for use as a tissue-engineered trachea for in vivo transplantation in rabbit models.