Search

The internal nanostructure of the diatoms Cyclotella meneghiniana, Seminavis robusta and Achnanthes subsessilis was investigated using small angle neutron scattering (SANS) to examine thin biosilica samples, consisting of isotropic (powder) from their isolated cell walls. The interpretation of SANS data was assisted by several other measurements. The N2 adsorption, interpreted within the Branuer-Emmet-Teller isotherm, yielded the specific surface area of the material. Fourier transform infrared (FTIR) and Raman spectroscopy indicates that the isolated material is amorphous silica with small amounts of organic cell wall materials acting as a filling material between the silica particles. A two-phase (air and amorphous silica) model was used to interpret small angle neutron scattering data. After correction for instrumental resolution, the measurements on two SANS instruments covered an extended range of scattering vectors 0.0011 nm(-1) < q < 5.6 nm(-1), giving an almost continuous SANS curve over a range of scattering vectors, q, on an absolute scale of intensity for each sample. Each of the samples gave a characteristic scattering curve where log (intensity) versus log (q) has a -4 dependence, with other features superimposed. In the high-q regime, departure from this behaviour was observed at a length-scales equivalent to the proposed unitary silica particle. The limiting Porod scattering law was used to determine the specific area per unit of volume of each sample illuminated by the neutron beam. The Porod behaviour, and divergence from this behaviour, is discussed in terms of various structural features and the proposed mechanisms for the bio-assembly of unitary silica particles in frustules.

The occurrence of cryptic and pseudocryptic species, often living in sympatry, is widespread among microalgae. This phenomenon raises important questions about niche partitioning between these closely related species. To date, however, few studies have addressed the ecological mechanisms underlying sympatry in cryptic and pseudocryptic species. As a result, we have only a limited understanding of the factors that govern their distribution along environmental gradients. Here, we used the ribosomal internal transcribed spacer (ITS), 18S rRNA gene, and the RUBISCO LSU (rbcL) chloroplast gene sequence data together with cell wall morphology to show that estuarine populations of the widespread and common benthic diatom Navicula phyllepta Kütz. consist of pseudocryptic species. Growth rate measurements in function of salinity showed that N. phyllepta strains assigned to the different species differed in their tolerance to low salinities (<5 practical salinity units, psu), which was reflected by their different (but widely overlapping) distribution in the Westerschelde estuary (the Netherlands). Multiple regression analyses of the factors determining the abundance of the different species in field samples revealed that, in addition to salinity, sediment type and ammonium concentrations were probably equally important. Our results show that N. phyllepta sensu lato comprises different species with specialized ecophysiological characteristics rather than generalists with a broad adaptability to different environmental conditions., (© 2009 Phycological Society of America.)

The diversities and distributions of diatoms are much more complex than was ever imagined. To understand the underlying mechanisms, research must focus on evolutionary processes occurring at a population level and employ sufficiently informative molecular markers. Using ten microsatellites and ITS rDNA sequence data, we investigated the genetic structure of populations of the benthic freshwater diatom Sellaphora capitata (until 2004 a cryptic entity within the S. pupula agg. species complex). This is the first time that microsatellites have been used to investigate the genetic structure of any freshwater or benthic microalga. Using an integrated approach (morphology, DNA barcoding and specificity of the microsatellite primers), we verified the identity of 70 S. capitata isolates obtained from lakes in the UK, Belgium and Australia. Standardized F'(ST) values were very high (>0.4) and in Bayesian analyses, isolates clustered according to their country of origin, with limited evidence of admixture. However, selected isolates from all countries were sexually compatible, a result consistent with limited ITS divergence. Considering the apparent absence of desiccation-resistant resting stages in most diatoms, we conclude that such levels of differentiation are likely to be a consequence of limited dispersal. With restricted dispersal, previously unacknowledged opportunities for allopatric speciation exist, which may help to explain the huge extant diversity of diatoms.

The major research questions addressed in this study were (i) whether green microalgae living in soft water (operationally defined water hardness<10mg CaCO(3)/L) are intrinsically more sensitive to Ni than green microalgae living in hard water (operationally defined water hardness >25mg CaCO(3)/L), and (ii) whether a single bioavailability model can be used to predict the effect of water hardness on the toxicity of Ni to green microalgae in both soft and hard water. Algal growth inhibition tests were conducted with clones of 10 different species collected in soft and hard water lakes in Sweden. Soft water algae were tested in a 'soft' and a 'moderately hard' test medium (nominal water hardness=6.25 and 16.3mg CaCO(3)/L, respectively), whereas hard water algae were tested in a 'moderately hard' and a 'hard' test medium (nominal water hardness=16.3 and 43.4 mg CaCO(3)/L, respectively). The results from the growth inhibition tests in the 'moderately hard' test medium revealed no significant sensitivity differences between the soft and the hard water algae used in this study. Increasing water hardness significantly reduced Ni toxicity to both soft and hard water algae. Because it has previously been demonstrated that Ca does not significantly protect the unicellular green alga Pseudokirchneriella subcapitata against Ni toxicity, it was assumed that the protective effect of water hardness can be ascribed to Mg alone. The logK(MgBL) (=5.5) was calculated to be identical for the soft and the hard water algae used in this study. A single bioavailability model can therefore be used to predict Ni toxicity to green microalgae in soft and hard surface waters as a function of water hardness.

Ten polymorphic microsatellite markers were developed for the benthic freshwater diatom Sellaphora capitata and tested on 40 isolates from a Belgian pond. Genotyping was very successful (95%). The number of alleles per locus ranged from three to 12 (mean 6.6) and expected heterozygosities from 0.2 to 0.86 (mean 0.67). This is the first time that microsatellite markers have been developed for a freshwater or benthic diatom., (© 2008 The Authors. Journal compilation © 2008 Blackwell Publishing Ltd.)

Despite the growing interest in diatom genomics, detailed time series of gene expression in relation to key cellular processes are still lacking. Here, we investigated the relationships between the cell cycle and chloroplast development in the pennate diatom Seminavis robusta. This diatom possesses two chloroplasts with a well-orchestrated developmental cycle, common to many pennate diatoms. By assessing the effects of induced cell cycle arrest with microscopy and flow cytometry, we found that division and reorganization of the chloroplasts are initiated only after S-phase progression. Next, we quantified the expression of the S. robusta FtsZ homolog to address the division status of chloroplasts during synchronized growth and monitored microscopically their dynamics in relation to nuclear division and silicon deposition. We show that chloroplasts divide and relocate during the S/G2 phase, after which a girdle band is deposited to accommodate cell growth. Synchronized cultures of two genotypes were subsequently used for a cDNA-amplified fragment length polymorphism-based genome-wide transcript profiling, in which 917 reproducibly modulated transcripts were identified. We observed that genes involved in pigment biosynthesis and coding for light-harvesting proteins were up-regulated during G2/M phase and cell separation. Light and cell cycle progression were both found to affect fucoxanthin-chlorophyll a/c-binding protein expression and accumulation of fucoxanthin cell content. Because chloroplasts elongate at the stage of cytokinesis, cell cycle-modulated photosynthetic gene expression and synthesis of pigments in concert with cell division might balance chloroplast growth, which confirms that chloroplast biogenesis in S. robusta is tightly regulated.

Diatoms are a species-rich group of photosynthetic eukaryotes, with enormous ecological significance and great potential for biotechnology. During the last decade, diatoms have begun to be studied intensively using modern molecular techniques and the genomes of four diatoms have been wholly or partially sequenced. Although new insights into the biology and evolution of diatoms are accumulating rapidly due to the availability of reverse genetic tools, the full potential of these molecular biological approaches can only be fully realized if experimental control of sexual crosses becomes firmly established and widely accessible to experimental biologists. Here we discuss the issue of choosing new models for diatom research, by taking into account the broader context of diatom mating systems and the place of sex in relation to the intricate cycle of cell size reduction and restitution that is characteristic of most diatoms. We illustrate the results of our efforts to select and develop experimental systems in diatoms, using species with typical life cycle attributes, which could be used as future model organisms to complement existing ones., ((c) 2008 Wiley Periodicals, Inc.)

The enormous species diversity of diatoms correlates with the remarkable range of cell sizes in this group. Nuclear DNA content relates fundamentally to cell volume in other eukaryotic cells. The relationship of cell volume to G1 DNA content was determined among selected members of the genus Thalassiosira, one of the most species-rich and well-studied centric diatom genera. Both minimum and maximum species-specific cell volume correlated positively with G1 DNA content. Phylogeny based on 5.8 S and ITS rDNA sequences indicated that multiple changes in G1 DNA content and cell volume occurred in Thalassiosira evolution, leading to a 1,000-fold range in both parameters in the group. Within the Thalassiosira weissflogii (Grunow) G. A. Fryxell et Grunow species complex, G1 DNA content varied 3-fold: differences related to geographic origin and time since isolation; doubling and tripling of G1 DNA content occurred since isolation in certain T. weissflogii isolates; and subcultures of T. weissflogii CCMP 1336 diverged in DNA content by 50% within 7 years of separation. Actin, β-tubulin, and Spo11/TopVIA genes were selected for quantitative PCR estimation of haploid genome size in subclones of selected T. weissflogii isolates because they occur only once in the T. pseudonana Hasle et Heimdal genome. Comparison of haploid genome size estimates with G1 DNA content suggested that the most recent T. weissflogii isolate was diploid, whereas other T. weissflogii isolates appeared to be polyploid and/or aneuploid. Aberrant meiotic and mitotic cell divisions were observed, which might relate to polyploidization. The structural flexibility of diatom genomes has important implications for their evolutionary diversification and stability during laboratory maintenance., (© 2008 Phycological Society of America.)

The common and cosmopolitan freshwater benthic diatom Sellaphora pupula (Kütz.) Mereschk. is a model system for studying the nature of species in microalgae; the biological significance of morphological variation in this species complex has been widely demonstrated. The aim of this study was to establish a two-gene phylogeny (18S rDNA and rbcL) for 23 Sellaphora taxa, including 19 S. pupula aggregate (agg.) demes or species, S. bacillum (Ehrenb.) D. G. Mann, and S. laevissima (Kütz.) D. G. Mann. A range of analyses on separate and combined data sets indicated that Sellaphora is a monophyletic group containing four major clades. Of the traditionally recognized species, S. bacillum and S. laevissima are natural groups, but S. pupula is paraphyletic or polyphyletic because S. bacillum groups with S. pupula"small lanceolate" and S. lanceolata. Thickened bars at the poles of valves within the core "pupula-bacillum" group may be a morphological synapomorphy; the fossil record suggests that this group is at least 12 million years old. Otherwise, there was no clear pattern in the distribution of different morphologies among the major clades; each clade was also heterogeneous with respect to mating system. More intensive and extensive sampling will doubtless uncover even greater diversity; the challenge lies in its interpretation. Our results demonstrate the limitations of paleoecological, ecological, and biogeographical research based on morphospecies., (© 2008 Phycological Society of America.)

The study of reproductive isolation between populations, combined with estimates of genetic divergence, provides important insights into mechanisms of speciation. In this study, sixteen morphologically heterogeneous sympatric clones of Eunotia bilunaris sensu lato (Bacillariophyta) were brought into culture to study their phylogenetic relationships and pre- and postzygotic reproductive barriers. An ITS rDNA phylogeny was congruent with morphology and divided the clones into three groups ('slender', 'robust' and 'labile'), pointing to the presence of several species in E. bilunaris. Whereas most strains had a heterothallic mating system, four 'labile' clones displayed apomictic behaviour. A further 'labile' clone had a heterothallic mating behaviour, however, suggesting a very recent origin for apomixis. Despite high sequence divergence, hybridization occurred between clones belonging to different groups, but was 20-400 times less frequent than in intra-group matings. F1 hybrids had an intermediate morphology and were almost completely sterile; gamete formation was generally arrested in the early stages of meiosis I. The ITS divergence of 11.5-12.3% between the 'robust' and 'slender' clones seems to represent an upper limit of divergence in which cell pairing, gamete formation and auxosporulation are still possible but heavily reduced, and where hybrid sterility has already evolved.

There is a long-standing belief that microbial organisms have unlimited dispersal capabilities, are therefore ubiquitous, and show weak or absent latitudinal diversity gradients. In contrast, using a global freshwater diatom data set, we show that latitudinal gradients in local and regional genus richness are present and highly asymmetric between both hemispheres. Patterns in regional richness are explained by the degree of isolation of lake districts, while the number of locally coexisting diatom genera is highly constrained by the size of the regional diatom pool, habitat availability, and the connectivity between habitats within lake districts. At regional to global scales, historical factors explain significantly more of the observed geographic patterns in genus richness than do contemporary environmental conditions. Together, these results stress the importance of dispersal and migration in structuring diatom communities at regional to global scales. Our results are consistent with predictions from the theory of island biogeography and metacommunity concepts and likely underlie the strong provinciality and endemism observed in the relatively isolated diatom floras in the Southern Hemisphere.

The diatoms are the most speciose group of algae, having global ecological significance in the carbon and silicon cycles. They are almost unique among algae in being diplontic, and sexual reproduction is an obligate stage in the life cycle of most diatom species. It is unclear which are the principal factors that have fostered the evolutionary success of diatoms, but the unique life cycle (which is correlated with a curious wall structure and cell division mechanism) and size-dependent control of sexuality must have played an important part. Progress in understanding life cycle dynamics and their interrelationships with population biology and evolution will depend on how successfully sex can be initiated and manipulated experimentally, and our review provides a foundation for such work. Relevant data are scattered in time and come mostly from non-English publications, producing a false impression of diatoms as recalcitrant with respect to sexualization. Recent advances dependent on experimental cultures include the discovery of widespread heterothallism (including some complex types of behavior) in pennate diatoms, sexual diversity among clones of centric diatoms, more flexible size restitution strategies in centric diatoms than had been suspected, and use of reproductive isolation as a criterion in diatom taxonomy. We identify unsolved problems in the life history of diatoms, including aspects of sexualization, cell-cell recognition, sexual reproduction, and the development of the special expanding cell (the auxospore), which is crucial to morphogenesis in this group. Some of these problems are being addressed using modern molecular genetic tools, and progress will be facilitated when whole-genome sequences are published (e.g., for Thalassiosira pseudonana). Problems of culture maintenance and methods for manipulating the life cycle are discussed.