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Anabaena - Phenotypic and genotypic diversity of planktonic strains in fishponds and reservoirs of the Czech Republic
ZAPOMĚLOVÁ, Eliška
Morphological diversity of 61 Anabaena populations of 13 morphospecies was described under the field conditions of Czech fishponds and reservoirs. Polyphasic approach was then applied in classification of 45 clonal strains isolated from those populations. Detailed morphological analyses were performed and partial 16S rRNA gene sequences were obtained for 33 of the strains, and secondary metabolite production was evaluated in 20 strains. Plasticity of morphological characteristics under varied conditions of light, temperature, nitrogen and phosphorus was studied in selected strains, as well as their temperature and light growth requirements. The results were then discussed with respect to the delimitation of single Anabaena morphospecies. A new genus Sphaerospermum was defined for the morphospecies Anabaena kisseleviana, A. reniformis and Aphanizomenon aphanizomenoides, whose phenotypic and genotypic features differed considerably from all other Anabaena morphospecies. Unique information was provided on the occurrence and distribution of A. reniformis and Aph. aphanizomenoides in the Czech Republic.
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Floristic study of cyanobacteria and algae in south-east part of Vysočina region
MELICHAR, Antonín
The phytoplankton samples from 45 localities in Vysočina region were collected in 2009. Species composition and relative abundance of species were studied; ecological factors like pH, conductivity, temperature, water transparency and dissolved ions were measured. Ecological indices (Shannon and Simpson index of diversity) were calculated and compared with other ecological factors. Checklist of all species present in Vysočina region was made. There were found 262 species and 4 species new for the Czech Republic.
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Metabolic rhythms of unicellular, nitrogen fixing cyanobacteria and possible interplay with modeled KaiABC circadian oscillator
Červený, Jan ; Nedbal, Ladislav
Circadian clocks of living organisms provide evolutionary advantage and make living in dynamic environment more efficient. The clocks affect human lives and health as well as control the simplest organisms such as prokaryotic cyanobacteria. The cyanobacteria represent an excellent model that is amendable to a multitude of genetic, biochemical, and biophysical methods. The cyanobacterium, Cyanothece sp. ATCC 51142 relies on the circadian clock to permit, in the same cell, anoxygenic nitrogen fixation at night and oxygenic photosynthesis during solar day. We measured real-time, in-situ photosynthetic and respiratory activities as well as the culture growth under light forcing conditions and also under constant light, i.e. free-running mode. Interestingly, the experiments show a strong 24h-period dynamic pattern that is replaced by apparent 12h-period in free running mode. The 24h-pattern does not change significantly when changing the light/dark ratio from 16hL/8hD to 12hL/12hD and 8hL/16hD. Furthermore, we tried to elucidate connection between these metabolic rhythms and known structure of KaiABC circadian oscillator by means of mathematical modeling. One of simulation results show a strong correlation between the presumed catabolic event indicated by significant peak in respiratory activity, and simulated dynamics of KaiB4 complex in modeled circadian pacemaker. A causal relationship between these 2 events is suggested because KaiB4 facilitates dephosphorylation of KaiC6 hexamer, which is known to signal upcoming dark period.
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Changes in water quality in outdoor swimming pools and swimming areas during the summer season
MARŠÁLKOVÁ, Alena
Good quality of bathing water is an increasingly important factor taken into account when we choose summer vacations, a place to stay for a weekend or a weekend trip destination. In the Czech Republic, bathing waters may be divided according to the legal status into outdoor swimming pools, surface waters used for bathing (swimming areas), other water surfaces or artificial pools. In my thesis I dealt only with outdoor swimming pools and swimming areas. The difference between them lies in the fact that outdoor swimming pools have their own operators who monitor water quality and provide other services in the waterside. Usually there is a fee charged. Swimming areas have no operators and water quality monitoring is placed under the responsibility of regional public health authorities. In the Czech Republic, at present, 188 outdoor swimming pools and swimming areas have been monitored on a regular basis each bathing season. For my research I chose four sites located in the South Bohemian region and four sites in the South Moravian region. For each of these eight sites, I collected results of laboratory analyses, including categories of water quality in bathing seasons of the past five years, it means from the year 2005. With these obtained data I assessed the water quality development both during the bathing season of 2009 and its evolution over the past five years. On the basis of particular categories of water quality I also tried to compare water quality in the South Bohemian and the South Moravian region and also to compare water quality between individual sampling points in one locality. Another aim was to evaluate some measures and interventions made in order to improve water quality. I focused particularly on the pond Olšovec in the South Moravian region and the VN (water reservoir) Orlík in the South Bohemian region. The results do not show any prominent changes in water quality both during the bathing season 2009 and over the past five years. Pronounced fluctuations in water quality were recorded only at certain locations during the 2007 bathing season. Not even between individual regions significant differences in water quality were found. The proportion of individual water quality categories in the two regions did not differ by more than 4%. We can therefore say that water quality in selected localities in the South Bohemian and South Moravian regions is comparable. As for certain measures and interventions implemented to improve water quality, it is important to focus also on the wider neighborhood of the locality, because it could also be a source of substances worsening water quality.
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Cytotoxicity and secondary metabolites of cyanobacteria
HROUZEK, Pavel
Cyanobacteria are well-known producers of secondary metabolites of different chemical structures and a wide range of biological functions. In the present thesis, the cytotoxic activity of cyanobacteria, originating from different habitats, was studied in order to reveal whether cytotoxicity is an environmentally dependent characteristic. In addition, the data were compared with the toxicity of these extracts to the model invertebrate Artemia salina. The obtained data suggest that cytotoxic cyanobacteria are favoured under some conditions and thus more frequent in particular localities. The majority of the studied extracts and fractions exhibited cytotoxitity to the Sp/2 cell line not accompanied by toxicity to A. salina. Moreover, in most of the strains with both activities to A. salina and Sp/2 cells the toxic effect was caused by an identical fraction. This result suggests that the toxic effect of the cyanobacterial secondary metabolites is mostly affecting basal cell metabolism rather than targeting specific organisms. In one of studied strains, Cylindrospermum sp. C24/1989, novel lipopeptides puwainaphycin F and G have been detected, isolated and their structure and biological effect have been characterized. Both of these structures interfere with eucaryotic membranes and cause a Ca2+ leakage into the cell. Subsequently, an enhanced tyrosine phosohorylation and relocalization of f-actin in the cell was observed. Lastly, the correlation between metabolite production and the reconstructed phylogeny was studied in planktonic Dolichospermopsis strains. Most of the detected compounds were found to be randomly disspersed across the reconstructed phylogeny and thus cannot be considered as good chemotaxonomic markers. This result also hampers the possible detection of toxic cyanobacteria by morphological methods or molecular detection based on the 16SrDNA gene.
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Analysis of the role of PilA proteins in the cyanobacterium \kur{Synechocystis} sp. PCC 6803.
LINHARTOVÁ, Markéta
The PilA proteins are major components of pili fibres which are essential for motility in bacteria. This project was focused on the role of the PilA proteins in cyanobacteria, specifically in the alternative functions that are not directly related to motility. The major task of this project was also to develop a method for the purification of the PilA1 protein from Synechocystis under native conditions and to analyze purified PilA1 protein in detail.
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Cytotoxicity of bentic and periphyton cyanobacteria
SKÁCELOVÁ, Kateřina
This study is focused on assessment of cytotoxic activity of 14 strains of benthic cyanobacteria against HeLa cell line. Extracts of the strains generating strong cytotoxicity were analyzed and fractionated with HPLC-MS, in order to determine active fractions. Mechanisms of effect on the cells were investigated using immunofluorescent staining. Two out of 14 studied strains exhibited strong cytotoxic activity. Extract of Cylindrospermum sp. CY OM contains active fraction inducting apoptosis and also a fraction affecting adhesion of cells. Activity of Anabaena oscillarioides HINDAK 1984/43 strain was caused by two cytotoxic fractions, in one of them a new linear peptide was discovered.
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Cytotoxicity of the filamentous cyanobacteria in relation to its ecology
TOMEK, Petr
The aims of this study was to screen a cytotoxic activity in 53 strains of the genera Nostoc and Anabaena and to compare occurence of active isolates within different ecological groups. Selected cytotoxic strains were subjected to HPLC/MS analysis to identify their extracts composition. Isolates with strong cytotoxic effect containing unknown substances were fractionated to obtain pure fraction of this compound. Finally, the selected strains toxicity dependence on time and concentration was evaluated.
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Construction of expression vector for cyanobacterium \kur{Synechocystis} sp. PCC 6803
RICHTOVÁ, Jitka
The cyanobacterium Synechocystis sp. PCC 6803 is an important model organism used in the study of photosynthesis and related processes. The only disadvantage of Synechocystis for molecular biology studies is that it lacks a cloning/expression system. The aim of this work was to evaluate whether it is possible to use a mutated high copy version of the native Synechocystis plasmid for expression of a chosen gene in this organism.
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