Hindcasting cyanobacterial communities in Lake Okaro with germination experiments and genetic analyses

Cyanobacterial blooms are becoming increasingly prevalent worldwide. Sparse historic phytoplankton records often result in uncertainty as to whether bloom-forming species have always been present and are proliferating in response to eutrophication or climate change, or if there has been a succession of new arrivals through recent history. This study evaluated the relative efficacies of germination experiments and automated rRNA intergenic spacer analysis (ARISA) assays in identifying cyanobacteria in a sediment core and thus reconstructing the historical composition of cyanobacterial communities. A core (360 mm in depth) was taken in the central, undisturbed basin of Lake Okaro, New Zealand, a lake with a rapid advance of eutrophication and increasing cyanobacteria populations. The core incorporated a tephra from an 1886 volcanic eruption that served to delineate recent sediment deposition. ARISA and germination experiments successfully detected akinete-forming nostocaleans in sediment dating 120 bp and showed little change in Nostocales species structure over this time scale. Species that had not previously been documented in the lake were identified including Aphanizomenon issatschenkoi , a potent anatoxin-a producer. The historic composition of Chrococcales and Oscillatoriales was more difficult to reconstruct, potentially due to the relatively rapid degradation of vegetative cells within sediment.     

Molecular detection of uncultured cyanobacteria and aminotransferase domains for cyanotoxin production in sediments of different Kenyan lakes

PCR-based denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene fragments was used to identify the cyanobacterial phylotypes in sediments and plankton of saline–alkaline and freshwater lakes of Kenya. The detection of the aminotransferase domain located on modules mcyE and ndaF using specific molecular markers confirmed the presence of potential toxin-producing cyanobacteria. The eight nucleotide sequences obtained from DGGE bands were placed in three divergent cyanobacterial clusters. Five nucleotide sequences were close to members of the genera Anabaenopsis and Umezakia ( Nostocales ), two sequences fell in the cluster with Arthrospira sp. ( Oscillatoriales ) and one sequence was related to Chroococcidiopsis sp. ( Pleurocapsales ). The presence of the latter taxon was demonstrated de novo in the investigated lakes. All nine attained nucleotide sequences of the aminotransferase region belonged to the mcyE module. Five sequences of the aminotransferase domain were included in the cluster having the nucleotide sequence of Anabaena sp. but showed a separate lineage. Other four aminotransferases were placed in the cluster represented by nucleotide sequence of Microcystis aeruginosa . To our knowledge, this is the first report on molecular detection of cyanobacterial phylotypes in sediments of African lakes and aminotransferase domains for cyanotoxin production from sediment samples in general.     

Effects of species traits on the genetic diversity of high-mountain plants: a multi-species study across the Alps and the Carpathians

Aim  To test the influence of various species traits, elevation and phylogeographical history on the genetic diversity of high-mountain plants in the Alps and Carpathians.
Location  The regular sampling grid comprised the whole range of the European Alps and the Carpathians.
Methods  Twenty-two high-mountain plant species were exhaustively sampled and their genetic diversity was assessed with amplified fragment length polymorphisms (AFLPs). ANOVAs were used to check for relationships between species traits and species genetic diversity, and to test whether genetic diversity was influenced by altitude and phylogeographical history (i.e. Alps versus Carpathians).
Results  In both mountain systems, species dispersed and pollinated by wind showed higher genetic diversity than species with self or insect pollination, and with animal- or gravity-dispersed seeds. Only in the Alps did altitudinal range size affect species genetic diversity significantly: species with narrow altitudinal ranges in the highest vegetation belts had significantly higher genetic diversity than those expanding over wide altitudinal ranges. Genetic diversity was species specific and significantly higher in the Alps than in the Carpathians, but it was not influenced by elevation.
Main conclusions  Wind pollination and wind dispersal seem to foster high genetic diversity. However, species traits are often associated and their effects on genetic diversity cannot be clearly disentangled. As genetic diversity is species specific, comparisons across species need to be interpreted with care. Genetic diversity was generally lower in the Carpathians than in the Alps, due to higher topographical isolation of alpine habitats in the Carpathians and this mountain massif's divergent phylogeographical history. Elevation did not influence genetic diversity, challenging the long-held view of decreasing genetic diversity with increasing elevation in mountain plants.     

Filling a gap in the phylogeny of flatworms: relationships within the Rhabdocoela (Platyhelminthes), inferred from 18S ribosomal DNA sequences

The phylogeny of the Rhabdocoela, a species-rich taxon of free-living flatworms, is reconstructed based on complete 18S rDNA sequences. The analysis includes 62 rhabdocoels and 102 representatives of all major flatworm taxa. In total, 46 new sequences are used, 41 of them from rhabdocoel species, five from proseriates. Phylogenetic analysis was performed using maximum parsimony and Bayesian inference. Clade support was evaluated with parsimony jackknifing, Bremer support indices and Bayesian posterior probabilities. The resulting cladogram corroborates that the Rhabdocoela is monophyletic, but its sister group remains uncertain. The 'Dalyellioida' and the 'Typhloplanoida', both former rhabdocoel subtaxa, are polyphyletic. Within the Rhabdocoela the monophyletic Kalyptorhynchia, characterized by a muscular proboscis, forms the sister group of all other rhabdocoels. The Schizorhynchia is a monophyletic subtaxon of the Kalyptorhynchia, with the split proboscis as a synapomorphy. Except for the Dalyelliidae and the Typhloplanidae, both freshwater taxa, none of the 'families' previously included in the 'Typhloplanoida' and the 'Dalyellioida' appears to be monophyletic. As a result of this analysis, three existing and four new taxon names are formally defined following the rules of the Phylocode.     

Dkk-3 is elevated in CSF and plasma of Alzheimer's disease patients

Biomarkers in CSF can offer improved diagnostic accuracy for Alzheimer's disease (AD). The present study investigated whether the glycoprotein and putative tumor suppressor Dickkopf homolog 3 (Dkk-3) is secreted into CSF and evaluated its applicability as a diagnostic marker for AD. Using our highly specific immunoenzymometric assay, Dkk-3 levels were measured in plasma and/or CSF of patients suffering from depression, mild cognitive impairment (MCI), or AD and compared with healthy subjects. Dkk-3 identity was verified by western blot and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS)/MS. High concentrations of Dkk-3 were detected in CSF compared with plasma (28.2 ± 1.3 vs. 1.22 ± 0.04 nmol/L, respectively). Consistently Dkk-3 expression was demonstrated in neurons of the cortex and epithelial cells of the choroid plexus, the major source of CSF. Significantly increased Dkk-3 levels in plasma and CSF were observed for AD patients compared with healthy subjects but not patients suffering from MCI or depression. In summary, our data indicate that elevated Dkk-3 levels are specifically associated with AD and might serve as a potential non-invasive AD biomarker in plasma.     

Morphological variation, photobiont association and ITS phylogeny of Chaenotheca phaeocephala and C. subroscida (Coniocybaceae, lichenized ascomycetes)

Chaenotheca phaeocephala and C. subroscida have been characterized by differences in their morphology and ecology. They are, however, morphologically plastic and sometimes seem to intergrade. It has been suspected that the differences in morphology between these two taxa could possibly be phenotypic and caused by their occurrence in different habitats and/or by the association with different photobionts. To test if there is a genetic difference between the species ITS1–5.8S-ITS2 rDNA sequences of six specimens of the mycobiont of C. phaeocephala and five of C. subroscida were compared. Further the ITS1–5.8S-ITS2 rDNA sequences of the photobiont of one specimens of C. phaeocephala and six of C. subroscida were compared to each other and to other species of Trebouxia. The mycobiont sequences of both C. phaeocephala and C. subroscida were monophyletic, substantially different and the groups had strong supports. They also proved to be associated with different Trebouxia species, C. phaeocephala with T. arboricola and Chaenotheca subroscida with T. jamesii. The morphologically based concepts of C. phaeocephala and C. subroscida thus not merely represent phenotypic variation but also are associated with considerable genetic differences in the extremes of the ITS1–5.8S-ITS2 rDNA region.     

Coordination of export and glycosylation of landomycins in Streptomyces cyanogenus S136

In Streptomyces cyanogenus S136 gene cluster for biosynthesis of polyglycosylated angucycline landomycin A (LaA), a divergently oriented gene pair for a TetR-family regulator ( lanK ) and an efflux protein ( lanJ ) is located, whose functions remained obscure. Overexpression and disruption studies showed that lanK and lanJ genes control LaA resistance. Also, a constitutive lanK overexpression led to predominant accumulation of LaA precursors bearing shorter glycoside chains. These data as well as the results of in vitro and in vivo assays of LanK activity are consistent with the idea that LanK represses lanJ and some downstream genes involved in conversion of landomycin D (a disaccharide LaA precursor) into LaA. LaA and some of its precursors accumulate in the producing cell and relieve repression by LanK, thus amplifying the biosynthesis and export of landomycins with long glycoside chains. Therefore, the main biological role of LanK appears to be the inhibition of premature extrusion of early LaA precursors from the cells, which in turn creates the optimal conditions for accumulation of LaA as the major landomycin in S. cyanogenus S136.     

Activation of Myelin Protein Kinase by Diacylglycerol and 4β-Phorbol 12-Myristate 13-Acetate

In an effort to gain a more complete understanding of the regulation of myelin basic protein phosphorylation, we have been interested in defining further the mode of regulation of the myelin protein kinase involved in this posttranslational modification. Here we report the partial purification of a protein kinase from rat brain myelin. By gel filtration, it was determined that the molecular weight of this enzyme was in the range of 70-80 X 10(3) daltons Furthermore, it was established that at low calcium concentrations, this enzyme was markedly activated by phosphatidylserine in combination with either 4 beta-phorbol 12-myristate 13-acetate or diolein. The enzyme was not affected by cyclic AMP or by calcium, alone or in combination with calmodulin. On the basis of these findings this enzyme can be identified as a protein kinase C-like enzyme.