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Optimization of isolation and PCR methods for the identification of CMS in onion
PETERKA, Václav
This bachelor thesis deals with the optimization of different methods of DNA isolation from different types of onion (Allium cepa L.) samples and the subsequent detection of the Ms locus and the detection of cytoplasmic male sterility (CMS). The different types of DNA extractions performed and their results were compared and evaluated. The best quality DNA was isolated from leaves using the CTAB-PVP. These test samples were also used for PCR for the detection of CMS and Ms locus using several different markers, which were compared with each other. The most suitable markers were then used in the PCR for the main samples. DNA isolation by CTAB-PVP was also used for further DNA isolation from the main samples of onion, which were two sterile lines and four fertile lines. The extracted DNA from these samples was first used to detect the Ms locus. Detection was performed using the marker AcSKP1. The samples were collected over several weeks. The Ms locus was detected in all samples (C1C6), regardless of the age of the plant. However, the Ms locus was detected in various forms of zygosity in samples from lines designated C2, C4 and C5. Furthermore, the DNA samples were used to detect CMS in plants and the type of CMS was determined. The markers cob and orfA501were used for this detection. Using these markers, it was found that the lines that were declared sterile (lines designated C1 and C4) were indeed sterile and had the CMS-T sterility type. The lines C2, C3 and C6 that were labelled as fertile were also detected as sterile samples. This means that these lines are not 100% fertile. In the line designated as C5, no sterility was detected in any of the samples.
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Relation locus of control on prevalence of depresivity by infertil person
Blašková, Ivana ; Loneková, Katarína (advisor) ; Hrachovinová, Tamara (referee)
This master thesis deals with infertility and its treatment. Both in the theoretical and empirical part, it focuses on the female aspects of the issue. The theoretical part summarizes an overview of key information important to understand the presented problems. We define the terms, describe the psychological and physiological causes of infertility, the process of treatment of infertility and aspects thereof. We focus on the depression rate related to infertility and its treatment, furthermore, we deal with type locus of control related to the experienced rate of depression. Our aim in the empirical part is to describe the depression rate, type of locus of control within the two samples of women acquired by us and capture the possible connections. The sample is made of 58 women who are currently undergoing a treatment for infertility and of the control group, which is made of 58 healthy women. To collect the data, we used the Beck Depression Inventory BDI-II, the Rotter's Internal- External Scale and the anamnestic questionnaire. Based on acquired data, we found out some specifity in depression rate and type of locus of control within our samples of women.
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Vliv polyploidizace a hybridizace na fyziologii reprodukce u ryb
TICHOPÁD, Tomáš
Sexual reproduction is one of the most common traits of eukaryotes and one of the most debated topics in biology. It involves complicated gene regulatory networks coordinating meiotic division, recombination, and the production of haploid gametes. Although these pathways are highly conserved, they have been repeatedly modified in many ways throughout the evolution of life forms. Interspecific hybridization and polyploidization are well known to alter the reproductive potential of an individual, and both phenomena are commonly linked with the so-called asexuality, i.e., the production of (partly) clonal gametes. Asexuality has attracted considerable attention as an excellent model to study the evolutionary dis/advantages of sex. Because of numerous associations with polyploidy, asexuals have also been considered as potential missing links in the formation of polyploid species. Nevertheless, asexual eukaryotic lineages are not an easily definable group. They are distributed throughout the tree of life and employ a broad spectrum of independently arisen mechanisms for gamete production. These mechanisms can noticeably differ even between closely related taxa, ranging from entirely ameiotic processes to those involving distorted meiotic divisions, which have different evolutionary consequences for each asexual lineage. It is therefore vital to understand whether there are any general rules followed in the processes leading to the abandonment of sexual reproduction or induction of sterility. This thesis investigates the effect of hybridization and polyploidy on reproductive modifications in several fish taxa. The first investigation was focused on the regulation of unreduced gamete production in asexual allotriploid females of Cobitis elongatoides-taenia-taenia. Naturally, it is accomplished via premeiotic endoreduplication (PMER) and followed by the pairing of sister chromatids. It has been discovered that the regulation of PMER is a female's exclusive trait, but the PMER execution depends on germ cell hybrid origin. Therefore, the asexuality phenotype in Cobitis is linked to phenotypic sex differentiation. Secondly, the spermatozoa characteristics of F1 hybrids of common carp and gibel carp were examined to evaluate morphological malfunctions. Males showed classical hybrid dysgenesis as their reproductive potential was negatively affected by the hybridization event, probably due to the high divergence between species. Although most of the spermatozoa were aneuploid or polyploid, a small fraction of spermatozoa was capable of fertilization and gave rise to F2 hybrid offspring. Thirdly, germ cells of ancestral hybrid Poecilia formosa were examined to confirm the PMER pathway; however, it was discovered that this species only initiates first meiotic genes, but meiosis itself is skipped. Therefore, P. formosa represents a rare exception to apomixis in fish taxa. Finally, a successful and effective protocol for triploid zebrafish production was established. Although the primary goal of this protocol was the establishment of sterile recipients for transplantation purposes, it represents an excellent opportunity for research of autopolyploidization effect on sex differentiation as all produced triploid zebrafish were confirmed as males.
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Genes of early meiotic prophase I of spermatogenesis in house mouse
Škaloudová, Eliška ; Trachtulec, Zdeněk (advisor) ; Forman, Martin (referee)
Meiosis is an essential cellular process that is necessary for gamete formation in all sexually reproducing organisms. This work is focused on the description of the genes of early stages of meiotic division in males of a mammalian model, the house mouse. The first part summarizes meiosis focusing on prophase I, which is longer than prophase II. Prophase I is divided into five stages, namely leptotene, zygotene, pachytene, diplotene, and diakinesis. Mouse spermatogenesis and its differences from oogenesis are also briefly described. The second part provides a list of genes encoding proteins required for initiation of meiotic division, pairing and synapses of chromosomes, and initiation of the catalysis of double-strand breaks. Double-strand breaks are repaired by homologous recombination, which may result in so-called crossing-over, the major source of genetic variability. The work deals with the early stage of homologous recombination and components required for this process. Localization of meiotic double-strand breaks in the genome is not random and is under the control of the Prdm9 gene, which seems to take multiple roles, such as the formation of new subspecies of the house mouse. Knowledge of the genes controlling the early stages of meiotic division is a prerequisite to understanding some of...
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Womenś awareness of methods of assisted reproduction in the Czech Republic
Dolejšková, Michaela ; Černý, Andrej (advisor) ; Šimják, Patrik (referee)
The theme of my thesis is women's awareness about the methods of assisted reproduction in the Czech Republic. The work is devided into a theoretical part and practical part. In the theoretical part I deal with the physiology of reproductive organs of women and men, further I describe the term infertility and its causes. In the following chapters I focus on the process of finding a specialist, subsequent examinations and possible methods of assisted reproduction. In the practical part I examine the awareness of women in this area. As a data source I use an anonymous quantitative questionnaire, shared on social networks, filled in by women of different ages. 193 women completed the questionnaire. I made several hypotheses, which I confirmed or refuted thanks to the answers. Research has confirmed the hypothesis that more than half of women know when and to which doctor to address fertility problems. Subsequently, the second hypothesis was confirmed-most women know at least some of the risk factors affecting fertility. Furthermore, I assumed that most women are aware of the importance of age in the pursuit of conception. Although I confirmed the hypothesis thanks to a questionnaire survey, but not every woman knows how low the age at which fertility reduction begins to manifest. I refute the...
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Womenś awareness of methods of assisted reproduction in the Czech Republic
Dolejšková, Michaela ; Černý, Andrej (advisor) ; Šimják, Patrik (referee)
The theme of my thesis is women's awareness about the methods of assisted reproduction in the Czech Republic. The work is devided into a theoretical part and practical part. In the theoretical part I deal with the physiology of reproductive organs of women and men, further I describe the term infertility and its causes. In the following chapters I focus on the process of finding a specialist, subsequent examinations and possible methods of assisted reproduction. In the practical part I examine the awareness of women in this area. As a data source I use an anonymous quantitative questionnaire, shared on social networks, filled in by women of different ages. 193 women completed the questionnaire. I made several hypotheses, which I confirmed or refuted thanks to the answers. Research has confirmed the hypothesis that more than half of women know when and to which doctor to address fertility problems. Subsequently, the second hypothesis was confirmed-most women know at least some of the risk factors affecting fertility. Furthermore, I assumed that most women are aware of the importance of age in the pursuit of conception. Although I confirmed the hypothesis thanks to a questionnaire survey, but not every woman knows how low the age at which fertility reduction begins to manifest. I refute the...
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Relation locus of control on prevalence of depresivity by infertil person
Blašková, Ivana ; Loneková, Katarína (advisor) ; Hrachovinová, Tamara (referee)
This master thesis deals with infertility and its treatment. Both in the theoretical and empirical part, it focuses on the female aspects of the issue. The theoretical part summarizes an overview of key information important to understand the presented problems. We define the terms, describe the psychological and physiological causes of infertility, the process of treatment of infertility and aspects thereof. We focus on the depression rate related to infertility and its treatment, furthermore, we deal with type locus of control related to the experienced rate of depression. Our aim in the empirical part is to describe the depression rate, type of locus of control within the two samples of women acquired by us and capture the possible connections. The sample is made of 58 women who are currently undergoing a treatment for infertility and of the control group, which is made of 58 healthy women. To collect the data, we used the Beck Depression Inventory BDI-II, the Rotter's Internal- External Scale and the anamnestic questionnaire. Based on acquired data, we found out some specifity in depression rate and type of locus of control within our samples of women.
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