Izabela Zgłobicka

New Insights into Plagiogrammaceae (Bacillariophyta) Based on Multigene Phylogenies and Morphological Characteristics with the Description of a New Genus and Three New Species

Plagiogrammaceae, a poorly described family of diatoms, are common inhabitants of the shallow marine littoral zone, occurring either in the sediments or as epiphytes. Previous molecular phylogenies of the Plagiogrammaceae were inferred but included only up to six genera: Plagiogramma, Dimeregramma, Neofragilaria, Talaroneis, Psammogramma and Psammoneis. In this paper, we describe a new plagiogrammoid genus, Orizaformis, obtained from Bohai Sea (China) and present molecular phylogenies of the family based on three and four genes (nuclear-encoded large and small subunit ribosomal RNAs and chloroplast-encoded rbcL and psbC). Also included in the new phylogenies is Glyphodesmis. The phylogenies suggest that the Plagiogrammaceae is composed of two major clades: one consisting of Talaroneis, Orizaformis and Psammoneis, and the second of Glyphodesmis, Psammogramma, Neofragilaria, Dimeregramma and Plagiogramma. In addition, we describe three new species within established genera: Psammoneis obaidii, which was collected from the Red Sea, Saudi Arabia; and Neofragilaria stilus and Talaroneis biacutifrons from the Mozambique Channel, Indian Ocean, and illustrate two new combination taxa: Neofragilaria anomala and Neofragilaria lineata. Our observations suggest that the biodiversity of the family is strongly needed to be researched, and the phylogenetic analyses provide a useful framework for future studies of Plagiogrammaceae.

A Description of Biremis panamae sp. nov., a New Diatom Species from the Marine Littoral, with an Account of the Phylogenetic Position of Biremis D.G. Mann et E.J. Cox (Bacillariophyceae)

Here we present a formal description of Biremis panamae Barka, Witkowski et Weisenborn sp. nov., which was isolated from the marine littoral environment of the Pacific Ocean coast of Panama. The description is based on morphology (light and electron microscopy) and the rbcL, psbC and SSU sequences of one clone of this species. The new species is included in Biremis due to its morphological features; i.e. two marginal rows of foramina, chambered striae, and girdle composed of numerous punctate copulae. The new species also possesses a striated valve face which is not seen in most known representatives of marine littoral Biremis species. In this study we also present the relationship of Biremis to other taxa using morphology, DNA sequence data and observations of auxosporulation. Our results based on these three sources point to an evolutionary relationship between Biremis, Neidium and Scoliopleura. The unusual silicified incunabular caps present in them are known otherwise only in Muelleria, which is probably related to the Neidiaceae and Scoliotropidaceae. We also discuss the relationship between Biremis and the recently described Labellicula and Olifantiella.

Microstructure and nanomechanical properties of single stalks from diatom Didymosphenia geminata and their change due to adsorption of selected metal ions

We present topographical and nanomechanical characterization of single Didymosphenia geminata stalk. We compared the samples before and after adsorption of metal ions from freshwater samples. Transmission electron microscopy studies of single stalk cross‐sections have shown three distinct layers and an additional thin extra coat on the external layer (called “EL”). Using scanning electron microscopy and atomic force microscopy (AFM), we found that topography of single stalks after ionic adsorption differed significantly from topography of pristine stalks. AFM nanoindentation studies in ambient conditions yielded elastic moduli of 214 ± 170 MPa for pristine stalks and 294 ± 108 MPa for stalks after ionic adsorption. Statistical tests showed that those results were significantly different. We conducted only preliminary comparisons between ionic adsorption of several stalks in air and in water. While the stalks with ions were on average stiffer than the pristine stalks in air, they became more compliant than the pristine stalks in water. We also heated the stalks and detected EL softening at 50°C ± 15°C. AFM nanoindentation in air on the softened samples yielded elastic moduli of 26 ± 9 MPa for pristine samples and 43 ± 22 MPa for stalks with absorbed metal ions. Substantial decrease of the EL elastic moduli after heating was expected. Significantly different elastic moduli for the samples after ionic adsorption in both cases (i.e., for heated and nonheated samples), as well as behavior of the stalks immersed in water, point to permanent structural EL changes due to ions.

Madinithidium gen. nov. (Bacillariophyceae), a new monoraphid diatom genus from the tropical marine coastal zone

Abstract: We studied a group of monoraphid diatom species (Bacillariophyceae, Achnanthidiaceae) found in the marine coastal environment of tropical islands of the Caribbean Sea, western Atlantic Ocean, Indian Ocean, South Pacific Ocean, the Adriatic Sea and the Black Sea. Based on light and electron microscope examination, Madinithidium was formally described as a genus new to science with the generitype defined as Madinithidium undulatum. The characteristic features of the valve structure are a strongly developed sternum and raphe sternum, transapical striae formed by a single areola (macroareolae) positioned in small depressions, elevated virgae and coaxial internal central raphe endings. Furthermore, the striae of both raphe and sternum valves are closed by finely perforated hymenes. Madinithidium and Achnanthidium species are difficult to identify correctly with light microscopy since their valves are small and finely structured. Four species recently described from the western Indian Ocean and assigned to Achnanthidium sensu lato – Achnanthidium capitatum, A. flexuistriatum, A. pseudodelicatissimum and A. scalariforme – possessed morphological features permitting their placement into Madinithidium gen. nov. The above-mentioned species were formally transferred to the new genus.

Ultrastructural and molecular characterization of diversity among small araphid diatoms all lacking rimoportulae. I. Five new genera, eight new species

Pennate diatoms are important contributors to primary production in freshwater and marine habitats. But the extent of their diversity, ecology, and evolution is still largely unknown. This is particularly evident among the clades of pennate diatoms without raphe slits, whose diversity is likely underestimated due to their small size and features that can be difficult to discern under light microscopy. In this study, we described five new araphid genera with eight new species based on morphological observations (light and electron microscopy) and molecular data (nuclear‐encoded small subunit ribosomal RNA and chloroplast‐encoded rbcL and psbC): Serratifera varisterna, Hendeyella rhombica, H. dimeregrammopsis, H. lineata, Psammotaenia lanceolata, Castoridens striata, C. hyalina, and Cratericulifera shandongensis. We also transferred Dimeregramma dubium to Hendeyella dubia. Phylogenetic analysis of the molecular data revealed that all the newly established taxa fell into a monophyletic group, with Fragilariforma virescens located at the base. The group was composed by two subclades: one comprising Castoridens, Cratericulifera, and Plagiostriata, and the larger including also the rest of the new genera plus some of the smallest known diatoms, such as Nanofrustulum, Opephora, Pseudostaurosira, Staurosirella, and Staurosira with a high level of support. This study enhances the general knowledge on the phylogeny and biodiversity of a group of small araphid diatoms that have been generally poorly described both by electron microscopy and DNA sequence data.

Multigene Assessment of Biodiversity of Diatom(Bacillariophyceae) Assemblages from the Littoral Zone of the Bohai and Yellow Seas in Yantai Region of Northeast China with some Remarks on Ubiquitous Taxa

ABSTRACT Witkowski, A.; Li, C. L.; Zg[lstrok]obicka, I.; Yu, S. X.; Ashworth, M.; Dąbek, P.; Qin, S.; Tang, C.; Krzywda, M.; Ruppel, M.; Theriot, E. C.; Jansen, R. K.; Car, A.; Płociński, T.; Wang, Y. C.; Sabir, J. S. M.; Daniszewska-Kowalczyk, G.; Kierzek, A., and Hajrah, N. H., 2016. Multigene assessment of biodiversity of diatom (Bacillariophyceae) assemblages from the littoral zone of the Bohai and Yellow Seas in Yantai region of Northeast China with some remarks on ubiquitous taxa. Diatoms are important contributors to the benthic microeukaryote flora. This manuscript lays the foundation for future metagenomic and environmental sequencing projects off coastal China by curating diatom DNA sequences from the Yantai region of the Bohai and Yellow Seas (Northeast China). These studies are based on cultures established from samples collected in different seasons from marine littoral and supralittoral zones in 2013 and 2014. Thirty-six diatom strains were cultured successfully and identification of these clones was determined by light and scanning electron microscopy(LM and SEM) and DNA sequencing of the nuclear-encoded small subunit ribosomal RNA (SSU)and chloroplast-encoded rbcL and psbC genes. The strains primarily represent raphid pennate genera, such as Amphora, Amphora (Oxyamphora), Caloneis, Diploneis, Halamphora, Navicula, Nitzschia, Parlibellus, Pleurosigma, Surirella and Tryblionella. When the DNA markers from these strains were analysed in a multi-gene phylogeny, we found that some clones-particularly within the genera Amphora, Navicula and Nitzschia—show greater than expected genetic diversity despite their very similar morphology and morphometrics. We also compared the molecular and morphological identities of several seemingly ubiquitous marine littoral taxa in the genera Amphora and Nitzschia from the Indian Ocean and Atlantic Ocean, the Red Sea and Adriatic Sea to their Yellow Sea counterparts.

Simonsenia aveniformis sp. nov. (Bacillariophyceae), molecular phylogeny and systematics of the genus, and a new type of canal raphe system.

The genus Simonsenia is reviewed and S. aveniformis described as new for science by light and electron microscopy. The new species originated from estuarine environments in southern Iberia (Atlantic coast) and was isolated into culture. In LM, Simonsenia resembles Nitzschia, with bridges (fibulae) beneath the raphe, which is marginal. It is only electron microscope (EM) examination that reveals the true structure of the raphe system, which consists of a raphe canal raised on a keel (wing), supported by rib like braces (fenestral bars) and tube-like portulae; between the portulae the keel is perforated by open windows (fenestrae). Based on the presence of portulae and a fenestrated keel, Simonsenia has been proposed to be intermediate between Bacillariaceae and Surirellaceae. However, an rbcL phylogeny revealed that Simonsenia belongs firmly in the Bacillariaceae, with which it shares a similar chloroplast arrangement, rather than in the Surirellaceae. Lack of homology between the surirelloid and simonsenioid keels is reflected in subtle differences in the morphology and ontogeny of the portulae and fenestrae. The diversity of Simonsenia has probably been underestimated, particularly in the marine environment.

Sexual reproduction in Schizostauron (Bacillariophyta) and a preliminary phylogeny of the genus

Abstract: The focus of this paper is the sexual reproduction and phylogeny of Schizostauron Grunow, which were studied using clonal cultures isolated from the Indian Ocean coast of Mozambique near Tofo and Bazaruto. Taxa of Schizostauron were characterized by light and electron microscopy and a phylogeny derived from three genes (rbcL, psbC and short subunit). Schizostauron was established in the second half of the 19th century, then forgotten, with its taxa included in Cocconeis or Achnanthes sensu lato. Schizostauron, together with Astartiella and perhaps also Kolbesia and Karayevia, seems to form a third lineage of monoraphid diatoms, which are related to biraphid diatoms belonging to the Stauroneidaceae and Parlibellus, but not to the two other lineages of monoraphids (Achnanthes and the Achnanthidiaceae–Cocconeidaceae group). In culture, size reduction was followed by release of gametes and auxosporulation in mixtures of clones. Despite some morphological differences, four clones from the Bazaruto population proved to be sexually compatible, one of the clones being sexually compatible with the three others. Before gametogenesis, cells gathered in groups of two to eight through active movement. Some groups of cells surrounded themselves by weakly visible mucilage. Reproduction was isogamous morphologically, and apparently also behaviourally. Growing auxospores were surrounded by the empty irregularly arranged frustules of parental cells. In numerous aspects the pattern of sexual reproduction of Schizostauron is similar to that of Achnanthes sensu stricto. The rbcL identity matrix (IDM) for sexually compatible clones ranged between 0.998 and 0.984. One clone derived from the Tofo population had an IDM below 0.900 and was sexually isolated from the remaining clones. A description of Schizostauron davidovichiorum sp. nov. is provided.

Visualization of the internal structure of Didymosphenia geminata frustules using nano X-ray tomography

For the first time, the three-dimensional (3D) internal structure of naturally produced Didymosphenia geminata frustules were nondestructively visualized at sub-100 nm resolution. The well-optimized hierarchical structures of these natural organisms provide insight that is needed to design novel, environmentally friendly functional materials. Diatoms, which are widely distributed in freshwater, seawater and wet soils, are well known for their intricate, siliceous cell walls called ‘frustules’. Each type of diatom has a specific morphology with various pores, ribs, minute spines, marginal ridges and elevations. In this paper, the visualization is performed using nondestructive nano X-ray computed tomography (nano-XCT). Arbitrary cross-sections through the frustules, which can be extracted from the nano-XCT 3D data set for each direction, are validated via the destructive focused ion beam (FIB) cross-sectioning of regions of interest (ROIs) and subsequent observation by scanning electron microscopy (SEM). These 3D data are essential for understanding the functionality and potential applications of diatom cells.