Thecamoeba astrologa n. sp. – A new species of the genus Thecamoeba (Amoebozoa,Discosea, Thecamoebida) with an unusually polymorphic nuclear structure

Thecamoeba astrologa n. sp. was isolated from a plant litter sample. This species has a complex locomotive morphology, to a certain extent intermediate between the striate and the rugose morphotype. The shape of the actively moving cell is similar to that of striate thecamoebians, such as T. quadrilineata. However, in a slow movement, they can be easily confused with rugose species, like T. similis. Thecamoeba astrologa normally has peripheral asterisk-like nucleoli, which are unique among known thecamoebids. However, the structure of the nucleus is unusually variable during the growth of the culture and in some cases may become almost a vesicular one. The phylogenetic analysis based on the 18S rRNA gene sequence shows that this species belongs to the clade of thecamoebids, consisting of species possessing vesicular nuclei. This finding indicates that the structure of the nucleus derived from a single or few observations (as it usually happens in faunistic or ecological studies) may not be a reliable character of a thecamoebid amoeba. In some species nuclei may be highly polymorphic and dominating nuclear structure may depend on the age of the culture. Nuclei with constantly or temporarily peripheral nucleoli are now known in both major phylogenetic branches of the genus Thecamoeba.     

High-Frequency Rugose Exopolysaccharide Production by Vibrio cholerae Strains Isolated in Haiti

In October, 2010, epidemic cholera was reported for the first time in Haiti in over 100 years. Establishment of cholera endemicity in Haiti will be dependent in large part on the continued presence of toxigenic V. cholerae O1 in aquatic reservoirs. The rugose phenotype of V. cholerae, characterized by exopolysaccharide production that confers resistance to environmental stress, is a potential contributor to environmental persistence. Using a microbiologic medium promoting high-frequency conversion of smooth to rugose (S–R) phenotype, 80 (46.5%) of 172 V. cholerae strains isolated from clinical and environmental sources in Haiti were able to convert to a rugose phenotype. Toxigenic V. cholerae O1 strains isolated at the beginning of the epidemic (2010) were significantly less likely to shift to a rugose phenotype than clinical strains isolated in 2012/2013, or environmental strains. Frequency of rugose conversion was influenced by incubation temperature and time. Appearance of the biofilm produced by a Haitian clinical rugose strain (altered biotype El Tor HC16R) differed from that of a typical El Tor rugose strain (N16961R) by confocal microscopy. On whole-genome SNP analysis, there was no phylogenetic clustering of strains showing an ability to shift to a rugose phenotype. Our data confirm the ability of Haitian clinical (and environmental) strains to shift to a protective rugose phenotype, and suggest that factors such as temperature influence the frequency of transition to this phenotype.     

Aspergillus siamensis sp. nov. from soil in Thailand

A new species of Aspergillus section Fumigati, Aspergillus siamensis, isolated from coastal forest soil in Samaesarn island, Chonburi province, Eastern Thailand, is described and illustrated. This species is characterized by its broadly lenticular ascospores with two wide equatorial crests and finely spinulose and rugose convex surfaces and produced pale pinkish exudates after 14 d incubation on Czapek agar. The validation of this new species was supported further by analyses of the β-tubulin, calmodulin and actin gene sequences.     

Salmonella enterica Serovar Typhimurium DT104 Displays a Rugose Phenotype

Rugose phenotypes, such as those observed in Vibrio cholerae, have increased resistance to chlorine, oxidative stress, and complement-mediated killing. In this study we identified and defined a rugose phenotype in Salmonella enterica serovar Typhimurium DT104 and showed induction only on certain media at 25°C after 3 days of incubation. Incubation at 37°C resulted in the appearance of the smooth phenotype. Observation of the ultrastructure of the rugose form and a stable smooth variant (Stv), which was isolated following a series of passages of the rugose cells, revealed extracellular substances only in cells from the rugose colony. Observation of the extracellular substance by scanning electron microscopy (SEM) was correlated with the appearance of corrugation during development of rugose colony morphology over a 4-day incubation period at 25°C. In addition, the cells also formed a pellicle in liquid broth, which was associated with the appearance of interlacing slime and fibrillar structures, as observed by SEM. The pellicle-forming cells were completely surrounded by capsular material, which bound cationic ferritin, thus indicating the presence of an extracellular anionic component. The rugose cells, in contrast to Stv, showed resistance to low pH and hydrogen peroxide and an ability to form biofilms. Based on these results and analogy to the rugose phenotype in V. cholerae, we propose a possible role for the rugose phenotype in the survival of S. enterica serovar Typhimurium DT104.     

Rugosity in Grimontia hollisae

Grimontia hollisae, formerly Vibrio hollisae, produces both smooth and rugose colonial variants. The rugose colony phenotype is characterized by wrinkled colonies producing copious amounts of exopolysaccharide. Cells from a rugose colony grown at 30°C form rugose colonies, while the same cells grown at 37°C form smooth colonies, which are characterized by a nonwrinkled, uncrannied appearance. Stress response studies revealed that after exposure to bleach for 30 min, rugose survivors outnumbered smooth survivors. Light scatter information obtained by flow cytometry indicated that rugose cells clumped into clusters of three or more cells (average, five cells) and formed two major clusters, while smooth cells formed only one cluster of single cells or doublets. Fluorescent lectin-binding flow cytometry studies revealed that the percentages of rugose cells that bound either wheat germ agglutinin (WGA) or Galanthus nivalis lectin (GNL) were greater than the percentages of smooth cells that bound the same lectins (WGA, 35% versus 3.5%; GNL, 67% versus 0.21%). These results indicate that the rugose exopolysaccharide consists partially of N-acetylglucosamine and mannose. Rugose colonies produced significantly more biofilm material than did smooth colonies, and rugose colonies grown at 30°C produced more biofilm material than rugose colonies grown at 37°C. Ultrastructurally, rugose colonies show regional cellular differentiation, with apical and lateral colonial regions containing cells embedded in a matrix stained by Alcian Blue. The cells touching the agar surface are packed tightly together in a palisade-like manner. The central region of the colony contains irregularly arranged, fluid-filled spaces and loosely packed chains or arrays of coccoid and vibrioid cells. Smooth colonies, in contrast, are flattened, composed of vibrioid cells, and lack distinct regional cellular differences. Results from suckling mouse studies showed that both orally fed rugose and smooth variants elicited significant, but similar, amounts of fluid accumulated in the stomach and intestines. These observations comprise the first report of expression and characterization of rugosity by G. hollisae and raise the possibility that expression of rugose exopolysaccharide in this organism is regulated at least in part by growth temperature.     

Rugose corals and associated carbonate microfossils from the Brigantian (Mississippian) of Castelsec (Montagne Noire, southern France)

The disused quarry east of Castelsec offers a view of shallow-marine carbonates of the poorly known Uppermost Mississippian of the Montagne Noire. At Castelsec, sections are studied in two characteristic facies types (bioclastic wackestone and microbial dominated boundstone) of the Upper Mississippian. The succession is rich in rugose corals and carbonate microfossils. Six genera with seven species belonging to a rugose coral fauna consisting of at least eight genera with several species are described herein; Dibunophyllum castelsecensis sp. nov. is described as new. Twenty-seven carbonate microfossils of different groups have been identified. The Castelsec succession is Brigantian in age, based on the stratigraphic occurrence of rugose corals, foraminifers, and calcareous algae observed in both sections. The rugose coral fauna shows relationships with the well-known fauna of northwestern Europe and the Ouralian-Asian Province. Typical elements of northwestern Europe are missing at Castelsec and vice versa. This differentiation between north and south is interpreted as responses to different palaeolatitudes and tectonic settings.     

Lower Emsian biostratigraphy and event stratigraphy of Ha Giang Province,northern Vietnam

A new species of rugose coral, Sanidophyllum dubium n. sp., and the typical Emsian (Early Devonian) rugose coral Xystriphylloides nobilis are described from the Mia Le Formation in northern Vietnam. The lower Emsian index conodonts ranging from the Polygnathus excavatus zone to the P. nothoperbonus zone are illustrated. The biostratigraphic correlation between northern Vietnam and South China shows that the Mia Le Formation in northern Vietnam is early Emsian in age, and its upper part can be correlated with the lower part of the Shizhou Member of the Yukiang Formation in Liujing, Guangxi and its equivalents in South China. Based on the study of the lower Emsian biostratigraphic sequence, the disappearance of Xystriphylloides nobilis fauna in the overlying bed of the uppermost Mia Le Formation and the extinction of the “tonkinensis fauna” (sensu lato) in the interval between the basal Si Phai Formation and the uppermost Mia Le Formation demonstrate the influence of the Yujiang Event in northern Vietnam.     

Early Silurian (Telychian) rugose coral fauna of Daguan area, northeast Yunnan province, China

Study on rugose coral fauna of the Sifengya Formation (early Telychian) and Daluzhai Formation (mid-late Telychian) in Daguan area, northeast Yunnan Province, China was carried out. Rugose coral fauna of the Sifengya Formation included 18 genera and 34 species, while Daluzhai Formation with nine genera, ten species. We described rugose coral fauna (12 genera, 19 species) including one new genus and five new species, i.e. Protoketophyllum daguanense gen. et sp. nov., Crassilasma huanggexiense sp. nov., Pseudophaulactis heae sp. nov., P. convolutus sp. nov., and Shensiphyllum minor sp. nov. The characteristics and geological significance of rugose coral fauna of Sifengya Formation and Daluzhai Formation were analyzed. Particularly, rugose coral fauna of the Sifengya Formation represent early Telychian rugosan fauna in the Upper Yangtze region and improve the sequences of early Silurian (Llandovery) rugose coral assemblages in Yangtze region. It is therefore very meaningful to further analyze radiation period of rugose coral fauna in such epoch. __________ Translated from Acta Palaeontologica Sinica, 2005, 44(2): 229–246 [译自: 古生物学报, 2005, 44(2): 229–246]     

Genetic Analysis and Prevalence Studies of the brp Exopolysaccharide Locus of Vibrio vulnificus

Phase variation in the Gram-negative human pathogen Vibrio vulnificus involves three colonial morphotypes- smooth opaque colonies due to production of capsular polysaccharide (CPS), smooth translucent colonies as the result of little or no CPS expression, and rugose colonies due to production of a separate extracellular polysaccharide (EPS), which greatly enhances biofilm formation. Previously, it was shown that the brp locus, which consists of nine genes arranged as an operon, is up-regulated in rugose strains in a c-di-GMP-dependent manner, and that plasmid insertions into the locus resulted in loss of rugosity and efficient biofilm production. Here, we have used non-polar mutagenesis to assess the involvement of individual brp genes in production of EPS and related phenotypes. Inactivation of genes predicted to be involved in various stages of EPS biosynthesis eliminated both the rugose colonial appearance and production of EPS, while knockout of a predicted flippase function involved in EPS transport resulted in a dry, lightly striated phenotype, which was associated with a reduction of brp-encoded EPS on the cell surface. All brp mutants retained the reduced motility characteristic of rugose strains. Lastly, we provide evidence that the brp locus is highly prevalent among strains of V. vulnificus.     

A new oligolectic bee species of the genus Rhophitulus Ducke (Hymenoptera,Andrenidae) from South Brazil

The genus Rhophitulus Ducke, 1907 is a large and complex group of bees of the tribe Protandrenini comprising small, slender, mostly black ground-nesting species that are restricted to South America. We describe a new species of Rhophitulus from Parque Nacional São Joaquim, Urubici, state of Santa Catarina, Brazil. Rophitulus ater sp. nov. is distinctive and easily distinguished from other species of the genus by a unique combination of morphological characters in both sexes, but especially by the following: dull black body, coarsely and densely punctate integument, basal area of metapostnotum depressed and areolate rugose, posterior margin of hind tibia and pygidial fimbria of female with blackish pilosity, and characters of the male genitalia with hidden sterna. The new species is closely associated with Blumenbachia catharinensis (Loasaceae), which is restricted to cloud forest of the southeastern rim of Serra Geral. Flowers of B. catharinensis are pollen and nectar resources and mating sites for the new species.     

Systematic implication of seed micromorphology in Arenaria (Caryophyllaceae) and allied genera

Variations in seed micromorphology of 64 taxa of Arenaria and allied genera were examined using scanning electron microscopy (SEM) in order to evaluate their diagnostic value for systematic studies in the genus and providing additional evidence on delimitation of natural groups. Significant diversity was found in seed coat morphology both among and within genera. Size, color, and shape of seeds appear to be of low importance at species level but provide useful evidence at generic rank. Cerastium and Stellaria are well distinguished from Arenaria by serrate seed margins and solid columellae on testa cells. The sculpturing pattern renders most characters potentially applicable in taxonomy and phylogeny assessment in Arenaria. The studied taxa show four basic types of sculpturing, viz. colliculate, rugose, papillate, and smooth. The colliculate type which is characteristic for the core group of Arenaria (A. subg. Arenaria) can be further divided into five subtypes. The smooth seed surface with indistinct cell boundaries represents a synapomorphy for A. subg. Leiosperma, whereas the winged seeds along with rugose surface characterize subg. Odontostemma. Otherwise seed micromorphology rarely provides adequate evidence for separation of natural groups, especially at sectional rank. We hypothesize that the seeds are highly polymorphic in certain groups of taxa showing recent adaptive radiation, such as the annual species of Arenaria, and do not provide strong support for natural groups within the genus.     

Late Moscovian (mid-Pennsylvanian) rugose corals from Wadi Araba (Egypt,Eastern Desert): Taxonomy,palaeoecology and palaeobiogeography

An important late Moscovian rugose coral association is described from the Rod El Hamal Formation of the Wadi Araba area (northern Eastern Desert, Egypt). The upper part of the formation yielded a moderately abundant but poorly diversified coral fauna composed of large dissepimented solitary rugose corals. In total, six species belong to the Bothrophyllidae and the Geyerophyllidae, including three new species: Bothrophyllum suezensis, Bothrophyllum cylindricum, and Amygdalophylloides omarai. Three undeterminable Bothrophyllidae taxa are left in open nomenclature. The corals were attached to the soft substrate by talons and rootlets, either rooted and growing upward as mud-stickers or as secondary recliners. They show frequent rejuvenations and constrictions. Encrustation and/or bioerosion are scarce. Carbonate microfacies indicate a habitat in littoral and inner neritic zones adjacent to a low terrigenous hinterland. Colonial taxa are missing and tabulate corals are rare. The corals from the Rod El Hamal Formation are the only example of a late Moscovian coral fauna on the southern margin of the Palaeotethys, that is to say along the northern margin of Gondwana. The association shows similarities with northwestern Spain (Cantabrian Mountains), the Donets and Moscow basins, and thus a general attribution to the Palaeotethys realm.     

Rugosa und Heterocorallia aus Obervisé-Geröllen der Marbella-Formation (Betische Kordillere,Südspanien)

Limestone pebbles of the post-lower Bashkirian Marbella Formation (Malaguides, Internal Zone of the Betic Cordillera) yielded sixteen species of rugose corals belonging to eleven genera and two species of heterocorals belonging to two genera. The fauna is of uppermost Viséan age (V3b-V3c). Its paleogeographic significance within the western Mediterranean is discussed. Lonsdaleia corbariensis, known up to now only from debris flow deposits of the Mouthoumet massif (Southern France), covers approximately half of the total fauna.Hexaphyllia mirabilis is shown to have juvenile stages with five septa.     

Erster Nachweis der GattungAmplexocarinia (Rugosa) im Eifelium der Nord-Eifel (Rheinisches Schiefergebirge)

The rugose coral genusAmplexocarinia Soshkina, 1928 is recorded for the first time from the Freilingen Formation (Upper Eifelian) of the Sotenich syncline (North Eifel, Rheinisches Schiefergebirge). A new species,Amplexocarinia freilingensis, is described, figured and compared toAmplexocarinia sp. from Eifelian/Givetian boundary beds of Moravia (Czechoslovakia) and to similar forms from different Devonian strata. Some remarks concerning the stratigraphy of the Freilingen Formation of the Sotenich syncline are given.     

A new species of the genus Anteon Jurine (Hymenoptera,Dryinidae) from Laos

A new species of Anteon Jurine, 1807 is described from Laos, Houaphanh Province: Anteon holzschuhi sp. n. Morphologically the new species is similar to Anteon semipolitum Olmi, 2008, but it is distinguished by the sculpture of the face partly reticulate rugose and partly with deep punctures similar to areolae; in Anteon semipolitum the face is completely punctate and unsculptured among punctures. In addition, in the new species the distance from the outer edge of a lateral ocellus to the compound eye (OOL) is about 3.3 times as long as the distance between the inner edges of a lateral ocellus and the median ocellus (OL); in Anteon semipolitum OOL is less than twice as long as OL. Published identification keys to the Oriental species of Anteon are modified to include the new species.     

Vibrio cholerae O1 Strain TSI-4 Produces the Exopolysaccharide Materials That Determine Colony Morphology,Stress Resistance,and Biofilm Formation

Vibrio cholerae O1 strain TSI-4 (El Tor, Ogawa) can shift to a rugose colony morphology from its normal translucent colony morphology in response to nutrient starvation. We have investigated differences between the rugose and translucent forms of V. cholerae O1 strain TSI-4. Electron microscopic examination of the rugose form of TSI-4 (TSI-4/R) revealed thick, electron-dense exopolysaccharide materials surrounding polycationic ferritin-stained cells, while the ferritin-stained material was absent around the translucent form of TSI-4 (TSI-4/T). The exopolysaccharide produced by V. cholerae TSI-4/R was found to have a composition of N-acetyl-d-glucosamine, d-mannose, 6-deoxy-d-galactose, and d-galactose (7.4:10.2:2.4:3.0). The expression of an amorphous exopolysaccharide promotes biofilm development under static culture conditions. Biofilm formation by the rugose strain was determined by scanning electron microscopy, and most of the surface of the film was colonized by actively dividing rod cells. The corresponding rugose and translucent strains were compared for stress resistance. By having exopolysaccharide materials, the rugose strains acquired resistance to osmotic and oxidative stress. Our data indicated that an exopolysaccharide material on the surface of the rugose strain promoted biofilm formation and resistance to the effects of two stressing agents.Cholera is a serious epidemic disease that has killed millions of people and continues to be a major health problem worldwide. Vibrio cholerae, the bacterium that causes cholera, is a motile, gram-negative, curved rod with a single polar flagellum. The hypothesis that V. cholerae occupies an ecological niche in the estuarine environment requires that this organism be able to survive the dynamics of various physiochemical changes, including variations in nutrient concentrations. As a response to nutrient depletion, copiotrophic (31, 42), heterotrophic bacteria may undergo considerable morphological, physiological, and chemical changes (13, 22, 23, 26–28). In fact, to survive energy- and nutrient-deprived conditions, non-spore-forming, heterotrophic bacteria are known to undergo an active adaptation program (28). Brown and Williams have provided detailed experimental evidence that the molecular composition of the bacterial cell walls is essentially plastic and is remarkably responsive to the cell’s growth environment (5). Rice et al. (33) discovered that V. cholerae O1 from the Peru epidemic was able to shift to a phenotype having a wrinkled or rugose colony morphology. They also suggested that the V. cholerae rugose phenotype represents a fully virulent survival form of the organism that can persist in the presence of free chlorine. Morris et al. (29) reported that V. cholerae can shift to a rugose colony morphology associated with the expression of an amorphous exopolysaccharide (EPS) that promotes cell aggregation, and they also confirmed that rugose strains displayed resistance to killing by chlorine and complement-mediated serum bactericidal activity. They also indicated that these rugose strains cause human disease. However, the phenotypic characteristics associated with rugose morphology, relationships between these characteristics, and their relative importance in pathogenicity still remained to be identified.A large variety of EPSs are synthesized by gram-negative bacteria. While some have been implicated in the pathogenicity of plant and mammalian hosts, others have not been assigned a function, but many serve a structural role, benefiting the bacterium by enabling attachment to surfaces, improving nutrient acquisition, or providing protection from environmental stresses and host defenses (36). The EPSs cover the surfaces of many gram-negative and gram-positive bacteria. They may form a capsule composed of a high-molecular-weight polysaccharide attached to the cell surface, or they may produce slime either loosely attached to the cell surface or released to the culture fluid. Bacterial cells initiate the process of irreversible adhesion by binding to the surface by using EPS glycocalyx polymers and the development of microcolonies. The eventual production of a continuous biofilm on the colonized surface is a function of cell division within microcolonies and recruitment of bacteria from the planktonic phase. The biofilm concept has drawn attention to the bacterium’s ecological and biotechnological importance (8–11). We must now accept the unequivocal evidence that bacteria respond to changes in their environment by profound phenotypic variations in enzymatic activity, cell wall composition (34), and surface structure (2).In this study, we have isolated the rugose variants of V. cholerae O1 strain TSI-4 from starvation medium and determined EPS expression on the cell surface of the rugose strain by polycationic ferritin-labeled thin-section electron microscopy. While examining the morphological characteristics of these rugose strains, we found that they produced a continuous biofilm on the colonized surface and culture tube walls. Directly sampled, intact biofilms were subjected to electron microscopic analysis. We have also studied the role of the slime polysaccharide of V. cholerae TSI-4 in the bacterium’s resistance to osmotic and oxidative stress.     

Unveiling Gymnosporangium corniforme, G. unicorne, and G. niitakayamense sp. nov. in Taiwan

Three rust fungi from high mountains and pear-producing areas in Taiwan were described using morphological and molecular data based on 34 specimens. Gymnosporangium corniforme was demonstrated to produce spermogonia and aecia on Sorbus randaiensis based on molecular analyses and inoculation experiments. The pear rust pathogen G. unicorne was discovered in Taiwan for the first time. Gymnosporangium niitakayamense sp. nov. was observed on the leaves of Photinia niitakayamensis. It was distinct from other species in peridial cell wall structures, i.e., smooth outer wall, rugose side wall, and coralloid projections on the inner wall, and in having echinulate aeciospores.     

Seed coat sculpture in Comanthera (Eriocaulaceae) and its implications on taxonomy and phylogenetics

Eriocaulaceae occurs primarily in the Neotropics, diversifying mostly in mountainous areas in southeastern Brazil. They are popularly known as everlasting plants, because once dried, they retain the appearance of living structures, being widely used as ornamentals. Most of the commercialized species belong to the genus Comanthera L.B.Sm., which was reestablished from Syngonanthus Ruhland. Comanthera groups 41 species, 26 in Comanthera subg. Comanthera and 15 in C. subg. Thysanocephalus, all restricted to South America. Phylogenetic analyses show Syngonanthus and Comanthera, as well as both Comanthera subgenera, as monophyletic and sister groups. Seed coat ornamentation proved to be useful and informative to help in determining taxonomic relationships in many plant groups. This study aims to characterize the external morphology of the seed coat of Comanthera and its subgenera using scanning electron microscopy, and discuss its taxonomic and phylogenetic implications. Seeds from nine species of Comanthera were analyzed, seven of which belong to C. subg. Comanthera and two to C. subg. Thysanocephalus. The results were compared with four species of Syngonanthus and one of Leiothrix. In Comanthera and Leiothrix the external periclinal walls remain intact, whereas they degenerate in the remaining genera. However, seed coat ornamentation in Leiothrix and Comanthera has distinct origins, as in the former, it comes from projections of the external layer of the outer integument, forming a striate pattern, and in the latter, from projections of the inner layer, conferring a rough pattern. Comanthera subg. Comanthera shows a rugose undulatory pattern, whilst C. subg. Thysanocephalus has a rugose micropapillate pattern. The newly reported features of the seed coat provide support for the Comanthera subgenera, thus presenting taxonomic and phylogenetic value.     

Seed Coat Microsculpturing Is Related to Genomic Components in Wild Brassica juncea and Sinapis arvensis

It has been reported that wild Brassica and related species are widely distributed across Xinjiang, China, and there has been an argument for species identification. Seed coat microsculpturing (SCM) is known to be an excellent character for taxonomic and evolutionary studies. By identifying collections from Xinjiang, China, and combining SCM pattern, flow cytometry, and genome-specific DNA markers as well as sexual compatibility with known species, this study aimed to detect potential relationships between SCM and genomic types in wild Brassica and related species. Three wild collections were found to be tetraploid with a SCM reticulate pattern similar to B. juncea, and containing A and B genome-specific loci, indicating relatively high sexual compatibility with B. juncea. The others were diploid, carrying S-genome-specific DNA markers, and having relatively high sexual compatibility with Sinapis arvensis. Moreover, their SCM was in a rugose pattern similar to that of S. arvensis. It was suggested that SCM, as a morphological characteristic, can reflect genomic type, and be used to distinguish B-genome species such as B. juncea from the related S. arvensis. The relationship between SCM and genomic type can support taxonomic studies of the wild Brassica species and related species.     

Systematic implications of flavonoid pigments in the fern genus hemionitis (adiantiaceae)

The closely related fern generaHemionitis L. andGymnopteris Bernhardi are separated primarily on differences in leaf architecture and venation. Studies indicate that these characters are highly variable and unreliably diagnostic. Further, the type species of the two genera readily hybridize with each other. Spore morphology, as exhibited by SEM, does not support the traditional alignment of the species in these two genera: some species ofHemionitis andGymnopteris have the same rugose to papillate spores, while other species from both genera possess crested spores. The flavonoid chemistry of these taxa coincides with spore type, i.e., taxa from both genera which possess crested spores produce kaempferol and quercetin 3-0-glycosides, while species with tuberculate spores produce only quercetin 3,4′-0-glycosides. The spore and chemical data suggest a realignment of these taxa within a single genus, which would avoid the rather tenuous dependence on a single vegetative character for generic distinctions.