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Characterization of fungal community in spruce (Picea abies) litter using cultivation and T-RFLP
Kolářová, Zuzana ; Koukol, Ondřej (advisor) ; Brabcová, Vendula (referee)
Fungi have a key role in the decomposition of coniferous litter and affect nutrient cycling in forest ecosystems. Therefore, great emphasis is placed on exploring the diversity of these organisms. The aim of this thesis was to describe fungal diversity in spruce litter and revealed temporal development of this community in a forest regenerating after bark beetle outbreak. Another objective was to compare sites with different length of bark beetle damage. The study area was located in the Bohemian Forest mountain range. Litter bags with spruce needles were placed on the forest soil and several samplings were performed in the course of three years. Diversity of fungi and changes in the fungal community were assessed by two methods: cultivation of needles on 2ř MEA and fingerprinting method T-RFLP. In total 71 fungal species were obtained from needles during a three-year succession by cultivation approach. Using T-RFLP 122 different fragments were generally recorded. The dominant species were Scleroconidioma sphagnicola, Thysanophora penicillioides, Hormonema dematiodes, Ceuthospora pinastri, species of genus Chalara, Trichoderma polysporum, Mycena galopus and unknown species Helotiales sp. 1. Primary saprotrophs occured in the community mainly in first 8 months and then were replaced by basidiomycetes....
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Impact of river system structure on the genetic diversity of reed populations
Fuxová, Gabriela ; Fér, Tomáš (advisor) ; Kubátová, Barbora (referee)
Many plant species are closely related to river biotopes or to biotopes influenced by rivers. River systems create important linear corridors in ecosystems and directly or non-directly influence spatial spread of species in these environments. This offer many questions about species spread in this system. We can answer these questions by using molecular methods. Using 10 microsatellite (SSRs) primers, 202 individuals of Phragmites australis from 60 populations were analysed. Those analyses allowed reveal kinship of individuals, obtain information about spatial spread of populations and about spreading of common reed. Phragmites australis creates both - monoclonal and polyclonal - populations. Dependence of rate of clonality on environment was revealed. Populations from river banks are more monoclonal, populations from pond shores are more polyclonal. Populations are isolated. The highest percentage of variability was explained on among-population level. This is common for anemochoric species. Communication between populations is present, more on shorter distances. Evidence for vegetative spread was found on short distance. Generative spread is much more common. Long-distance spreading is mediated by generative diaspores - seeds. This spreading includes within-river spread, among-river spread and spreading...
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Application of molecular methods in population genetic studies
Šurinová, Mária ; Münzbergová, Zuzana (advisor) ; van Loo, Marcela (referee) ; Mandák, Bohumil (referee)
A vast range of factors shape the genetic structure of plant populations. In this thesis, I focus on two of them. The first factor, polyploidization, is a process of chromosome set multiplication through whole-genome duplication within a single species (autopolyploids) or hybridization of two different species (allopolyploids). It rapidly brings changes into genomes, allowing species to occupy distinct niches, adapt to new habitats, colonize them, or adapt to changing environment in their native range. But it comes at a certain cost - difficulties in mitosis and meiosis, changes in cellular architecture. Furthermore, after cytotype establishment, new individuals have to deal with cytotype exclusion effect, competition with parental individuals and higher nutrition requirements. In this thesis, I present the effect of polyploidization on populations of three species- Arabidopsis arenosa, Aster amellus and Festuca rubra. The second factor changing the population genetic structure presented in this thesis is the fragmentation of populations. Population fragmentation can be caused by natural or anthropogenic activities and often leads to overall reduction in population size and reduced connectivity among fragments. Restricted gene flow may threaten long-term population survival due to inbreeding...
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Study of genome of Metschnikowia yeasts by molecular methods
Schneiderwindová, Nicole ; Skoumalová, Petra (referee) ; Němcová, Andrea (advisor)
Yeasts of the genus Metschnikowia belonging to the family Metschnikowiacea are yeasts characterized by vegetative propagation through multilateral budding. These are yeasts widely distributed in nature. More than 35 species occurring have been defined in the wild. They most often occur on flowers, fruits, but also on insects or human skin. They have a wide range of uses due to their antifungal effects in agriculture and the cosmetics industry. This bachelor thesis deals with the study of usage of molecular methods to characterize selected species of yeasts of the genus Metschnikowia. It focuses on a detailed description of the yeast cell structure, karyotype and methods of reproduction in the theoretical part of the work. In the practical part on optimization and description of molecular methods including pulse gel electrophoresis methods used to separate the yeast genome and their subsequent observation of changes in individual parts of genome. First, the yeast was cultured under special conditions that are characteristic of Metschnikowia yeasts, then yeast DNA was isolated using methods suitable for DNA isolation, which was further examined by the PFGE molecular method. The DNA isolation procedure was first optimized for individual yeast strains, as it was necessary to verify the required ratio of low melting agarose to isolated DNA. That was because of it was important for the resulting gel blocks to be suitable for measurement by PFGE analysis. By optimizing the method was possible to create ideal blocks of isolated yeast DNA, which were subsequently subjected to PFGE analysis. Several measurements of PFGE analysis were performed at different time intervals in order to separate small and large yeast chromosomes. The CHEF standard of the yeast Hansenula wingei and the standard of the yeast Schizosaccharomyces pombe were used for the measurements. According to the measurement results, it can be determined that the yeast DNA isolation procedure and subsequent analysis by pulsed gel electrophoresis were successful, as the number of chromosomes of all used yeast species of the genus Metschnikowia was determined.
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Možnosti molekulární a imunochemické detekce patogenů vybraných druhů rostlin
Sukharukava, Yulita
This thesis is focused on the techniques of classic, molecular and immunoenzymatic detection and identification of pathogens of winter wheat. The most important diseases, methods of detection and identification of pathogens of these diseases are described in detail. In the year 2016 I monitored the occurrence of Barley Yellow Dwarf Virus (BYDV) on small experimental plots in the strands of wheat species Hybnos and MvVekni at the site of the school farm Žabčice with the help of PCR (Polymerase Chain Reaction) method. On the base of molecular detection it was identified that 14% of plants were infected with this virus. BYDV-PAS virus strain was found. MvVekni species has shown resistance to Wheat Dwarf Virus (WDV).
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Molekulární metody vhodné pro identifikaci rostlinných patogenů
Prokešová, Kateřina
This thesis contains a literary research on the overview of molecular methods and immunochemical methods and their comparison. The next chapter is about wheat germs. The aim of the experimental part was to detect WDV in winter wheat (Triticum aestivum) in the primary phase of natural infection using the PCR method and evaluate the symptoms in infected plants. Finally, compare the nucleotide sequences of this virus available in GenBank in terms of their similarity in the area of the primers used by us.
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Testování genetické čistoty hybridního osiva zelí hlávkového pomocí molekulárních metod
O'Shea, Denisa
This diploma thesis deals with testing of hybrid seeds genetic purity in three varieties of head cabbage (Brassica oleracea var. capitata L.). The suitability of SSR primers chosen according to an earlier screening carried out as a part of a research project in the workplace of Mendeleum and hybrid seeds genetic purity in head cabbage by SSR molecular method was tested. Among the total of 8 SSR primers screened, only 5 could produce both parent specific markers and therefore they were able to differentiate hybrid offspring and its corresponding parental lines, by which crossbreeding they originated. For each variety, 70 samples of hybrid offspring and 2 samples of parental lines were tested always in two sets. By using these primers, the genetic purity of the variety D - F1 was determinated as 100% and of the varieties H - F1 and T - F1 as 98.57%. The results of this thesis have the potential to be used in practice by a commercial company that has provided the seed samples for testing.
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Conspecific brood parasitism: a case study on the common pochard and the barn swallow
Petrželková, Adéla ; Hořák, David (advisor) ; Musil, Petr (referee) ; Guillemain, Matthieu (referee)
Presented thesis is mainly concerned with the conspecific brood parasitism (CBP) in birds. CBP is an alternative reproductive strategy when a parasitic female lay egg or eggs to other (host) female's nest of the same species. Then the parasitic female leaves the host's nest and does not provide any energetic investment in a subsequent parental care. Distinguish parasitic eggs or young is problematic because there are no obvious morphological differences. Thus, the use of molecular method is crucial for proper determination of CBP. CBP can be used as 'a best-of-bad-job' when female does not have her own nest or lost her nest for example through predation. Other option is that female can increase her fecundity ('fecundity enhancement' hypothesis) when she lay parasitic egg/eggs and care about her own clutch afterward. Two different model species were studied. The first one was a precocial diving duck - the common pochard (Aythya ferina; Anseriformes; Anatidae). Protein fingerprinting was used for detection of CBP and for distinguishing between eggs of individual females. It was found that the rate of CBP was relatively high in this species (91%, 72%; Chapter 1, 4). Further results indicated that the host clutch size decreased with the number of parasitic eggs in the clutch. The study of individual...
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Impact of river system structure on the genetic diversity of reed populations
Fuxová, Gabriela ; Fér, Tomáš (advisor) ; Kubátová, Barbora (referee)
Many plant species are closely related to river biotopes or to biotopes influenced by rivers. River systems create important linear corridors in ecosystems and directly or non-directly influence spatial spread of species in these environments. This offer many questions about species spread in this system. We can answer these questions by using molecular methods. Using 10 microsatellite (SSRs) primers, 202 individuals of Phragmites australis from 60 populations were analysed. Those analyses allowed reveal kinship of individuals, obtain information about spatial spread of populations and about spreading of common reed. Phragmites australis creates both - monoclonal and polyclonal - populations. Dependence of rate of clonality on environment was revealed. Populations from river banks are more monoclonal, populations from pond shores are more polyclonal. Populations are isolated. The highest percentage of variability was explained on among-population level. This is common for anemochoric species. Communication between populations is present, more on shorter distances. Evidence for vegetative spread was found on short distance. Generative spread is much more common. Long-distance spreading is mediated by generative diaspores - seeds. This spreading includes within-river spread, among-river spread and spreading...
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Characterization of fungal community in spruce (Picea abies) litter using cultivation and T-RFLP
Kolářová, Zuzana ; Koukol, Ondřej (advisor) ; Brabcová, Vendula (referee)
Fungi have a key role in the decomposition of coniferous litter and affect nutrient cycling in forest ecosystems. Therefore, great emphasis is placed on exploring the diversity of these organisms. The aim of this thesis was to describe fungal diversity in spruce litter and revealed temporal development of this community in a forest regenerating after bark beetle outbreak. Another objective was to compare sites with different length of bark beetle damage. The study area was located in the Bohemian Forest mountain range. Litter bags with spruce needles were placed on the forest soil and several samplings were performed in the course of three years. Diversity of fungi and changes in the fungal community were assessed by two methods: cultivation of needles on 2ř MEA and fingerprinting method T-RFLP. In total 71 fungal species were obtained from needles during a three-year succession by cultivation approach. Using T-RFLP 122 different fragments were generally recorded. The dominant species were Scleroconidioma sphagnicola, Thysanophora penicillioides, Hormonema dematiodes, Ceuthospora pinastri, species of genus Chalara, Trichoderma polysporum, Mycena galopus and unknown species Helotiales sp. 1. Primary saprotrophs occured in the community mainly in first 8 months and then were replaced by basidiomycetes....
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