The disertation thesis was focused on the analysis of variability of two nepoviruses genomes, Grapevine fanleaf virus -- GFLV and Arabis mosaic virus -- ArMV. The thesis described single isolates of these nepoviruses and their until recently non-explored genome portions coded by RNA1 strand, the coding regions for 1BHel and 1EPol proteins precisely. The main topic of this thesis was the identification of infected plants that were infected by nepoviruses GFLV and ArMV. Based on design of specific primers there was established sequence homology of coding sequences. Another topic was the development and optimalization of diagnostic system for the efficient GFLV and ArMV detection. This work was based on hypothesis of very high level GFLV and ArMV genomes variability. The results were recieved mainly from sequencing analyses performed by capillary automatic sequencer ABI-PRISM 310 (Applied Biosystems, Carlsbad, USA). Obtained results were assessed with CLC Main Workbench 5 software (CLC Bio, Aarhus, Denmark). Real-time PCR TaqMan multiplex was also designed using this software. This real-time tool is able to detect and quantify both nepoviruses within one PCR reaction simultaneously. There were sequenced and analysed the genetic codes at the nucleotide level for MP 2B , 2CCP, 1BHel and 1EPol proteins in this work. The first one was assessed because of the development of real-time PCR TaqMan multiplex. In this case were sequenced 290 bp from 14 GFLV and ArMV isolates, the variability level was established in frame of nucleotides 71.7-97.6%. The partial genetic code for 2CCP was assessed in frame of 6 GFLV isolates and the variability was established in 935 bp fragment to 83-86% at the nucleotide level and 81-91% at the amino acid level. The partial genetic code for 1BHel was sequenced in frame of 6 isolates GFLV and the variability was established in 379 bp in frame of all accessible GFLV and ArMV isolates, the variability was established 86.4-98.9% at the nucleotide level and 96-100% at the aminoacid level. The partial genetic code for 1EPol protein was sequenced in 6 GFLV isolates in frame of 365 bp and the variability was established for all accessible sequences in GenBank/NCBI. The variability was established at the nucleotide level 79.5-100% and at the aminoacid level 91.4-100%. All received sequences longer than 300 bp were submitted to GenBank/NCBI. The obtained partial genetic codes for 1BHel a 1EPol proteins coded in RNA1 were unique at the time of publication. This work provided the important results which were used subsequently like source information for construction of vectors intended for the preparation of transgenic plants. Then the incurred grapevine would show resistance against GFLV. Moreover, based on results of this work there was created certified methodology of the real-time PCR TaqMan multiplex method which can be standartly used as an efficient diagnostic tool for simultaneous detection of GFLV and ArMV. This real-time system was succesfully compared with ELISA what would be interesting mainly for diagnostic laboratories engaged in the certification processes of propagation material.

Abstract In order to examine the prevalence of Cryptosporidium in wild Arvicolinae in the Czech Republic and understand the role that wild rodents play in the transmission of this parasite to humans and livestock, 152 faecal samples from 129 common voles (Microtus arvalis) and 23 bank voles (Clethrionomys glareolus) were collected on 9 localities in 2012. All samples were examined for presence of Cryptosporidium sp. using both the aniline-carbol-methyl violet staining method and molecular tools. The age, sex and faecal consistency were noted at the time of sampling. Microscopical examination revealed the presence Cryptosporidium sp. oocysts in 2 samples originated from common voles and 2 samples from bank voles. Genotyping was done through PCR amplification and characterization of the SSU rRNA and actin loci. Cryptosporidium specific DNA was detected in 10 samples (4 from common voles and 6 from bank voles) including those microscopically positive. Cryptosporidium infection was not linked to diarrhoea. Sequence and following phylogeny analyses revealed two new Cryptosporidium genotypes originated from bank voles and two new genotype from common vole, phylogeneticaly distinct from known species and genotypes. The host specificity needs to be verified by experimental infection in the future.

In total 399 samples from three species of squirrels (Sciurus vulgaris, Sciurus carolinensis and Callosciurus finlaysonii) were collected from 2010 to 2013. All samples were examined for the presence of Cryptosporidium using by molecular methods, which demonstrated by presence of Cryptosporidium specific DNA in 18 samples (4.5%). Out of them 12 samples were positive onCryptosporidium ferret genotype, 3 on Cryptosporidium skunk genotype, 1 on Cryptosporidium chipmunk genotype I and 2 samples onCryptosporidium ubiquitum. Statistical analyses did not reveal any influence of gender on the occurrence of Cryptosporidium. Juvenile individuals of squirrels from the family Sciuridae are more often infected by Cryptosporidium than the adult animals.

Samples of different varieties grapevine from Małopolska and Podkarpacie collected on spring 2016 were tested for Grapevine Pinot gris virus (GPGV) by RT-PCR. 16 out of 65 plants were found positive for GPGV. Two set of primers were used for detection, targeting partial movement protein and coat protein, and RdRp domain, both of those regions were sequenced. Phylogenetic analyses settled the Polish isolates to described before asymptotic group. Also GPGV positive samples were tested by multiplex and simplex RT- PCR for multiple infections, and 18,5% of all samples were GPGV and GFkV positive. This was first survey of GPGV in Poland.

In 1971, the algal class Eustigmatophyceae, Stramenopiles, was detached by Hibberd and Leedale from the class Xanthophyceae, Stramenopiles, on the basis of prominent ultrastructural differences in vegetative cells and more importantly in zoospores, such as the presence of big extraplastidial stigma. The class was divided into four families, six genera and twelve species. It seems so far, that there is a deep dichotomy within the class. Eliáš et al. (unpublished) recom- mend to establish two orders based on molecular sequencing of 18S rDNA and rbcL: Eustigmatales (sen- su Eustigmatophyceae described by Hibberd) and Goniochloridales (order containing Goniochloris, Pseu- dostaurastrum and undescribed relatives). The order Eustigmatales is divided into three separate lineages, most likely families: Eustigmataceae (A1), Monodopsidaceae (A2) and a new family Pseudellipsoidiona- ceae (A3). In this study the ultrastructure of 10 eustigmatophycean strains from three separate lineages was investigated. The typical characteristics of this class as chloroplast without a girdle lamella, a reddish glo- bule and lamellate vesicles were found in all strains studied. The appearance of the lamellate vesicles was found to change during the life cycle. My research indicates that other characteristics, formerly...

In this Master's thesis I engage in the problematics of the species concept of green subaerial alga Trentepohlia (Ulvophyceae, Chlorophyta). This genus has been studied intensively since the 18th century. For more than 200 years, Trentepohlia species have been delimited using various morphological criteria. However, recent molecular studies showed inadequacies in this approach Therefore, my goal was a precise morphological study of the European Trentepohlia species and the re-evaluation of their phylogenetic position using rbcL gene sequencing. During the period 2013-2015, I carried out a detailed study of the oldest references of Trentepohlia species and compiled a delimitation key of the European species. Based on this research I concluded that it would be desirable to synonymize T. odorata with T. jolithus, as well as T. uncinata with T. arborum. I also sampled Trentepohlia species within Europe and carefully studied and described their morphology. Fresh Trentepohlia thalli were molecularly characterized by cloning, which revealed a common mixture of Trentepohlia species in what on a first sight appears to be a homogenous crust. Phylogenetic analyses based on the rbcL confirmed the ongoing inconsistencies among morphologically and molecularly delimited species and also the existence of cryptic...

This study was aimed at discovering the taxonomical identity of freshwater populations of a prevailably marine green-algal genus Ulva in the Czech Republic. Furtherly, collected samples were compared to available relevant material from European herbaria. The identity of collected specimens was successfully resolved by a combined methodology that involved classical and modern molecular techniques, and discussed in the framework of distribution and ecology of close European taxa.

The Phylogenetic reconstruction has seen great development in the last 30 years. Computers have become more powerful and more generally accessible, and computer algorithms more sophisticated. It comes the effort of scientists to reconstruct the entire tree of life from a large amount of phylogenetic data. Just for this purpose are formed phylogenetic supertrees that allow the combination of all information gathered so far. The aim of this work is to find a method to construct supertree that will give correct results.

Tato diplomová práce se zabývá možností využití bezeztrátových kompresních algoritmů ke klasifikaci biologických sekvencí. Nejdříve je představena literární rešerše o bezeztrátových kompresních algoritmech, která byla využita k výběru slovníkového algoritmu vytvořeného A. Lempelem a J. Zivem v roce 1976 (LZ77). Tento algoritmus je běžně používán k datové kompresi a v předkládané práci byl modifikován tak, aby umožnil klasifikaci biologických sekvencí. K algoritmu byly navrženy další modifikace, které rozvíjí jeho klasifikační možnosti. V průběhu práce byla sestavena sada datasetů biologických sekvencí, která umožnila podrobné testování algoritmu. Algoritmus byl porovnán s klasickými zarovnávacími metodami: Jukes-Cantor, Tamura a Kimura. Bylo ukázáno, že algoritmus dosahuje srovnatelných výsledků v oblasti klasifikace biologických sekvencí a dokonce je u 20% datasetů překonává. Lepší výsledky dosahuje zejména u sekvencí, jež jsou si vzájemně vzdálené.

Phylogenetic relationships of a sample comprising 248 bats belonging to 19 species and four families (Hipposideridae, Rhinolophidae, Molossidae and Vespertilionidae) from Senegal (Western Africa) were investigated with the use of multi-locus sequence data and non- differentially stained chromosomes. The karyotypes of Hipposideros ruber, H. tephrus, H. jonesi and H. cyclops were described for the first time. The standard Hipposideros formula was recorded in H. tephrus, H. jonesi and H. ruber (2n = 32, FNa = 60, FN = 64). The karyotypes of H. cyclops (2n = 36, FN = 66) and H. gigas (2n = 52, FN = 64) substantially diverged from this typical chromosomal complement. Rhinolophus landeri and R. fumigatus shared the same diploid number (2n = 58), but differed in the chromosome morphology (R. fumigatus - FNa = 60, FN = 64; R. landeri - FNa = 64, FN = 68). Rhinolophus landeri was found karyotypically distinct to other African populations, thus signalling a possible presence of cryptic forms within this species. The karyotypes of Chaerephon pumilus and Mops condylurus had a 2n = 48, FN = 54 and were similar to other previously studied species of this chromosomally conservative family. Chromosomal, Bayesian, maximum likelihood and genetic distance analyses revealed an indication for the existence of cryptic...