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Investigation of the genome sizes, karyotype and ploidy within the genus Monocercomonoides
Kornalíková, Martina ; Hampl, Vladimír (advisor) ; Čepička, Ivan (referee)
Oxymonads are a group of anaerobic flagellated protists living mainly in the gut of insects and vertebrates. Here, we invetigate the ploidy and karyotype of various strains of Monocercomonoides using Fluorescence in situ Hybridization (FISH). For ploidy we used probes against the SufDSU gene, which is known to be present in a single copy in the genome of M. exilis. For karyotype investigations we used FISH probes against the TTAGGG telomeric repeat sequence. The results showed that all the investigated strains are haploid with a single clear signal from our SufDSU probes. The karyotype analyses showed that the average number of signals per nucleus varied between 9 to 17 in most of the investigated strains, indicating number of chromosomes similar to M. exilis. A notable exception from this are the strains of M. mercovicensis, where we observed a much higher number of signals, suggesting much higher number of chromosomes. For estimating of the DNA content in the nuclei of these oxymonads we used the flow cytometry, with M. exilis as a standard. Our results indicate that two of the investigated strains, Monocercomonoides sp. strain OEV and Mural1, have smaller genome sizes than M. exilis, while the rest of the strains have genomes sizes larger than M. exilis. We observed an unusually large variation...
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Analyses of Monocercomonoides genome sizes, ploidies and karyotypes
Kornalíková, Martina ; Hampl, Vladimír (advisor) ; Krylov, Vladimír (referee)
Oxymonads are a group of flagellate protists living in low oxygen environments - mainly the guts of insects and vertebrates. In this study, we focus on the analysis of ploidy and karyotype of various species of oxymonads using Fluorescence In Situ Hybridization (FISH) with probes against single copy genes and telomeric repeats as well as estimating the DNA content in the nuclei of these oxymonads using flow cytometry. Using specific FISH probes against SufDSU gene, which is present in a single copy in the haploid genome, we showed that all studied strains are probably haploid. From the genome of Monocercomonoides exilis strain PA203 we know that oxymonads have the ancestral type of telomeric repeat (TTAGGG). Using a probe against these repeats we tried to label chromosome ends and estimate the number of chromosomes for seven strains (five species) of Monocercomonoides. With a single exception, the average number of signals per nucleus was below 20 indicating number of chromosomes below 10. In the strains of M. mercovicensis, we observed much higher number of signals suggesting that the cells have much higher number of chromosomes. Finally, we established the DNA content for several strains using flow cytometry. We used as a standard M. exilis strain PA203 knowing that the haploid genome size is...
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