This work is aimed at the study of the effect of 17α-ethinylestradiol (EE2) on the production and characteristics of ligninolytic enzymes (laccase, Mn-dependent peroxidase and lignin peroxidase) in I. lacteus, T. versicolor, P. chrysosporium and P. ostreatus cultures grown on two types of liquid media. Enzyme activity production in fungal cultures was affected by the composition of culture medium. In the case of P. chrysosporium, the addition of EE2 to the complex- medium cultures led to a MnP activity stimulation and simultaneously LiP production was partially repressed in these cultures. In the mineral MM medium, no effect of EE2 on enzyme production by P. chrysosporium was observed. In EE2 treated MM cultures of P. ostreatus lower MnP activities were found when compared to biotic controls. In the case of T. versicolor cultures, the addition of EE2 to the complex medium caused laccase and LiP stimulation in the cultures. In the MM medium, however, only laccase production was affected by EE2. I. lacteus MnP production was partially repressed by EE2 in MM medium. In contrast to that, significantly higher MnP activities were detected in complex- medium I. lacteus cultures after the treatment with EE2. Further EE2 degradation by the fungal cultures was studied. The highest degradation effeciency was...

My diploma thesis deals with genetic variability of entomopathogenic fungi of the Isaria genus encountered in the Czech Republic. Individual representative of the genus can be found in soil where they attack all developmental stages of insects, giving preference to larvae and pupae. The Isaria fungi find application first and foremost where plants have to be provided biological protection. In case of mitosporic fungi is the precise identification very difficult, taxonomy is often unclear in many genera, including the genus Paecilomyces/Isaria to demonstrate their polyphyletic nature. The fungi are classified primarily with reliance on morphological studies. The most common markers used to identify fungi are the shapes and sizes of their conidia and the biological properties (germination of spores, tests of biological efficiency). Identification made in consideration of the morphological markers is inaccurate and very variable. To overcome those accuracies, there are very useful molecular DNA markers, which can be relevant in ecology, biology and in fungi genetics. This paper relies on applying the ITS region (Internal Transcribed Spacer) as a molecular marker. ITS regions are partial constituent rDNA carrying no code - that is why the regions are likely to accumulate evolutionary changes in the DNA sequence, which makes them suitable for extensive use in taxonomic analyses of many organisms. The study results in a phylogenetic trees constructed by comparing different sequences of ITS regions obtained from the samples of entomopathogenic fungi of the Isaria genus gathered in the Czech Republic during the monitoring stage 2013 to 2014. Thereunder detection of Isaria sp. occurring in the Czech Republic.

This diploma theses focuses on comparison of natural and intentionally inducated supressiveness of environment induced by application of entomopathogenic fungi Beauveria bassiana, Metarhizium anisopliae, Isaria fumosorosea a Lecanicillium muscarium. In tests were evaluated the in vitro parameters as well as the effectiveness in vivo biotests on insect host larva Tenebrio molitor. The species of entomopatoghenic fungi were applied in suspensions, single strains and also in combination of two strains. In the in vitro conditions the possibilities of objective evaluation of the supresivity level were tested by using the CFU test (Colony Forming Units) on three different nutrient media (PDA, PDA + A , PDA + D), as one of the basic evaluation parameters. Further the germination tests were evaluated according to GI (Germination Index), determination of radial growth (comparison of median cultures) and interaction of strain suspensions on nutrient media PDA. In the in vivo biotests were watched the epizooties from suspensions of these entomopathogenic fungi on insect larva Tenebrio molitor in competitive test of strains according to FDI (Fungl development index) evaluation scala. Chosen larva covered by fully sporulating mycelium from epizootie were further evaluated in CFU test. The result were dominant strain/s on the larva from applied suspensions.

The goals of the work are the evaluation of the pathogenic fungi occurence, detection of the occurence frequency, the strategy of losses estimates and the appreciation of the disease intensity. In the following I observe the infection with fungi pathogens from the tribe Fusarium including the interpretation of the proceeds items and the surface microflora on the seed of Avena nuda.

In laboratory bioassays, the efficacy of the entomopathogenic fungus Beauveria bassiana against the yellow mealworm (Tenebrio molitor) was tested under various temperature conditions. Six different strains of fungus B. bassiana was investigated. The evaluation was based on vitality bioassays including germination and growth index assessment and the bioassay of virulence based on target organism T. molitor was also assessed growth and yield of conidia different strains of fungus B. bassiana on natural substrates and artificial nutrient substrates.

The study of fungal diversity may lead to many fundamental discoveries and conclusions. Molecular genetics, and particularly high throughput sequencing methods using short DNA fragments as barcodes, has recently experienced a boom. The most frequently used marker for fungal research is the partial region of nuclear ribosomal DNA called ITS (Internal Transcribed Spacer). It occurs in the form of tandem repetitions of up to 200 copies. This fact greatly simplifies its amplification from the environment but also introduces some negatives. One of them can be an existence of intragenomic and intraspecific variability which confounds diversity estimates by exaggerating the real number of species. Using alternative low-copy markers can easily prevent these problems. In this study EF-1α and RPB2 protein- coding genes were compared with traditionally used ITS1 and ITS2 markers. An artificial mock community was created by blending genomic DNA of different fungal lineages. The community was sequenced for all markers and the data were processed according to guidelines commonly used in environmental studies. The results show that ITS2 is unequivocally a more suitable marker for environmental studies than other compared markers. The average coefficient of overestimation was deemed to be approximately two for ITS1, ITS2,...

The decomposition of organic substrates represents an important part of the global carbon cycle and affects its global change through CO2 release. In temperate forests, deadwood represents a large carbon stock, its amount and decomposition is crucial for ecosystem biodiversity and functioning. The fungi are omnipresent powerful decayers in all terrestrial ecosystems. Their ability to decompose all deadwood compounds, mainly lignocellulose, is highly important. Without fungi, the wood decompositions and the release of withheld nutrients back to nutrient cycles couldn't be performed. While many studies were concerned with the estimation of decomposition rates of deadwood, still deeper knowledge about microbial decomposition processes and the diversity of saproxylic species and their interaction is needed. The fungi are still underrepresented in dead wood studies. This study had two main objectives. First was to describe the fungal community on downed deadwood of Fagus sylvatica and Abies alba in natural forest of Salajka in the Czech Republic, to reflect the substrate changes during the different decay stages, and to link the enzyme activities to fungal community composition and their described ecophysiologies. Second aim was to describe the fungal communities on standing and downed dead logs of...

The development of the chemical and pharmaceutical industry and the massive use of synthetic substances in various industries has become indispensable for our society. Unfortunately, this progress has brought a whole new kind of pollution. It is a group of substances which imagine for living organisms’ greater risk than 'normal' organic pollution. Because they have extraneous origin compared with naturally occurring compounds, they are called "Xenobiotics." It is a group comprising a large number of different substances, with dissimilar physical, chemical and biological properties such as pharmaceuticals, pesticides, detergents, paints, varnishes, plastic packaging, food additives, cosmetic products, drugs, and many others. Although these substances began to enter the water cycle as early as 30 years of the 20th century, their concentration often moves to the limit of the detection limit, so their presence in the water began to be examined in more detail in the 1980s, following the development of analytical technologies chemical composition of water. The main danger of these substances is that doesn’t primarily effect on water quality, but directly affect on the organisms by confusing of the endocrine system or by directly damaging RNA or DNA, causing mutagenic changes, cancer-causing proliferation, birth defects, metabolic disorders, gender changes, the degeneration of reproductive capacity of entire populations or the disappearance of self-preservation instinct. The source of xenobiotics in the water cycle is predominantly wastewater, which generated through the wastewater treatment, which not designed to eliminate this type of pollution. he dissertation thesis deals with the issue of xenobiotics in wastewater and is divided into the theoretical and experimental part. The theoretical part summarizes the xenobiotics current state of knowledge in wastewater from their history, origin, classification, possibilities of elimination to impacts on the environ

Biogeochemical cycle of chlorine, particularly the formation of organically bound chlorine is still not well understood. In continental ecosystems chlorides act as source of chlorine, and also as a stress factor. Chlorides originate from precipitation of marine cloud masses. Organically bound chlorine in the environment is formed naturally by biotical and abiotical way. The biotical factors are microorganisms, plants, soil enzymes and animals. Volatile chlorinated hydrocarbons (VOCl) represent one group of organically bound chlorines. Several volatile chlorinated hydrocarbons reacts with atmospheric ozone, consequently causing depletion of the ozone layer. The most important known terrestrial source of volatile chlorinated hydrocarbons is the spruce forest ecosystem. Chlorine in the soil can be transformed by microorganisms into organically bound chlorine or translocated by transpiration stream in plants, where they are also transformed enzymatically into organically bound chlorine, and both of them can be emitted into the atmosphere. Too large amounts of chloride can affect the physiological functions of plants. In this thesis experiments were designed for measuring the natural emissions of volatile halogenated hydrocarbons from plants and fungi, with various periods of incubation, and also to...

Deadwood is a hotspot of microbial diversity and its decomposition contributes to carbon and nitrogen cycling in temperate forests. The historically recognized importance of fungi in the decomposition of deadwood has recently been complemented by the description of bacterial functions thanks to the rapid progress of culture-independent methods based on the analysis of nucleic acids. To study different aspects of deadwood decomposition, a temperate mixed forest in Zofinsky prales National Nature Reserve was selected as a site with rich historical forestry data where deadwood decomposition represents an important process in wood turnover. The aim of this thesis is to describe role of bacteria in deadwood decomposition at fine scale resolution with respect to community composition, enzyme transcription, and metabolic potential of dominant species. Effects of deadwood age together with pH and water content on the bacterial community composition proved to be more important than tree species identity. Bacteria showed distinct composition between early and late community in decomposing deadwood. The bacterial community was also under a significant influence of fungal community composition. Despite being in a close contact, bacterial and fungal communities differed significantly between deadwood and the...