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Genotyping of bacterial genus \kur{Xanthomonas} pathogenic for tomato and pepper by multilocus sequencing.
STEHLÍKOVÁ, Dagmar
The subject of this bachelor thesis is to estimate the degree of relationship of 20 collected isolated samples of Xanthomonas genus causing bacterial spot of tomatoes and peppers. To assess the relationship, the method of multilocus sequence analysis has been used (MLSA). Sequences of genes atpD, rpoD, gyrB, glnA and efP were found and analyzed using MEGA software version 5.1. The result was evaluated using a dendrogram according to which the isolates were divided into two clusters. The first cluster includes isolated of Xanthomonas euvesicatoria belonging to group A. The second cluster includes isolates of the species Xanthomonas vesicatoria belonging to group B. The results obtained in this work are consistent with the present classification of the xanhtomonads causing bacterial spot of tomatoes and peppers.
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Isolation of the antimicrobial peptide gene (defensin) from the hard tick \kur{Ixodes ricinus}, challenged by the pathogen infection
SKLADANÁ, Veronika
Antimicrobial peptides are major components of the innate immune response of epithelial cells. In hematophagous organisms, which acts as vectors of parasitic diseases, in particular Lyme borreliosis, the gut pathogen induce the expression of defensin, that provide the first barier of host defence. This raises the posibility that defensin may play a key role in the development of parasitic infection. The gene expressed in midgut was isolated from cDNA of the hard tick, Ixodes ricinus. The gene is coding the protein, that is produced and secreted during tick infection, and is known as defensin. Expression of the gene for defensin (224 bp) was induced by the pathogen. The gene was cloned into bacterial expression system.
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Identification and sequencing genom of a new virus infecting lucerne
BEČKOVÁ, Martina
Samples of lucerne plants characteristic with local necrosic lesions, leave malformation and yellow spots on leaves were investigated with transmission electron microscopy. Virus particles observed there were filamentous ones of 600 to 700 nm long. Nucleic acid was isolated, transcribed and amplified using PCR. Genus-specific primers were designed based on reverse genetics from the highly conserved genes for carlaviruses, potexviruses and potyviruses. Successful amplification with carlavirus-specific primers, sequencing and comparison with sequences in GenBank database revealed presence of a carlavirus. This was later identified by nucleotide sequence comparison as a new isolate V4 of Alfalfa latent virus. Specific primers for isolate V4 were designed in a coat protein position. Half of the genom of this virus was obtained with PCR and PCR modified amplifications and compared with sequences of Alfalfa latent virus and Pea streak virus from GenBank.
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Fylogeography and genetics variability of \kur{Diuraphis noxia} (\kur{Aphididae})
PAŠÍKOVSKÝ, Jiří
The aim of this work was a research of the genetic variability of natural populations of Russian wheat aphid Diuraphis noxia (Aphididae) by means of microsatellite markers and markers for EPIC-PCR. First goal was to introduce the methods and optimise them for Diuraphis noxia. In the follow-up pilot study, specimens from 47 lines representing 12 populations from all over the world were analysed. Having used microsatellite markers, I proved expected variability among individual populations and within them. The highest genetic variability was detected between Chile and Algeria using markers for cytochrome C in EPIC-PCR. These findings can be used for further studies of the genetic variability of the aphid Diuraphis noxia.
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Application of DNA barcoding on genus \kur{Folsomia} (Collembola) and mitochondrial geonome of \kur{F. candida}
SLÁMOVÁ, Martina
Mitochondrial molecular marker COI was tested for use in species identification of selected species of genus Folsomia (Collembola). Marker was succesfuly amplified and sequenced. Dendrogram constructed by Neighbor-Joining method with Kimura-2-Parameter model grouped all individuals into presumed species clusters and high intraspecific variability of F. quadrioculata suggests the existence of cryptic species. Furthermore, 65 % of mitochondrial geonome of F. candida was obtained with 16 tRNA genes, 9 proteincoding genes and 2 rRNA genes identified. So far the genome characteristics correspond to the one described in G. hodgsoni.
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