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Confirmation of \kur{Beauveria caledonica} occurence in Šumava National park by molecular markers
BINDER, Richard
Biological plant protection against insect pests is an important alternative to chemical protection. One of the most important group used in the biological plant protection against insect pests are the entomopathogenic fungi. Entomopathogenic fungi are microscopic fungi that are able to induce a primary disease to insect pests. It is a very heterogeneous group of species. Worldwide there were isolated and described more than 750 species of entomopathogenic fungi. Genus Beauveria is considered one of the most important genera of entomopathogenic fungi. In the Czech Republic there has been confirmed species B. bassiana, B. brongniartii and now, on the basis of this work, B. caledonica. This study is aimed to confirm the occurrence of B. caledonica in National Park Šumava. To confirm this occurrence, I used analyzes based on the methods of molecular markers. Molecular markers are an indispensable part of science in the field of mycology, for example the strain characterization, population genetics, detection and identification of fungi, phylogenetic studies and evolutionary biology. For this study there were used sequence analysis of ITS, EF1- and LSU regions. The output data of these analyzes were used to create phylogenetic trees. The result of my thesis is taxonomical classification of studied isolates on species level.
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Biosystematic studies in the family Cyperaceae
KOŠNAR, Jan
The thesis was focused on the microevolutionary mechanisms that contribute to morphological diversity in selected members of the sedge family (Cyperaceae). Natural hybridization, evidenced from both morphological characters and molecular markers, was revealed to be a potentially important source of diversification in the tropical spikerushes of Eleocharis subgenus Limnochloa. High levels of phenotypic plasticity of clonal growth but rare genetic (ecotypic) differentiation among contrasting morphotypes were found in the polymorphic species Carex nigra, which implied that taxonomic splitting of the species was unreasonable.
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Utilization of molecular markers in oil seed rape breeding programmes
KRISTINOVÁ, Helena
Current breeding of oilseed rape is focused on breeding of F1 hybrids and the male sterility and self-incompatibility could play the significant role in hybrid breeding programmes. Plant breeders also more widely utilize molecular genetic techniques for selection of desirable plants/genotypes. Molecular methods are faster, more reliable and more specific than conventional ones, which are based mainly on morphological descriptors. The aim of my thesis was to optimize PCR analysis methods and to develop specific molecular markers for target plant selection in oilseed rape hybrid breeding programmes. The new marker for detection of fertility restoration gene (Rf) in CMS Ogu-INRA plants was tested. New primer pair RsPPRF2/RsPPRR2 was designed in the coding region of PPR-B protein, which participates in the restoration of fertility in CMS Ogu-INRA. Also newly designed primers BrSLGIIF/BrSLGIIR were tested in SI plants. The optimal annealing temperaturse of these primers was 58 °C. But amplification in some SC plants was also observed. The optimization of the PCR reaction was performed for all designed primers. The set of F1 SI hybrids created by crossing of two lines with different S II haplotypes was tested by using of the PCR-RFLP technique for detection of polymorphism in amplified fragments.
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Possibilities of AFLP technique in study of \kur{Beauveria bassiana} genetic structure
BERANOVÁ, Kristina
Beauveria bassiana is widely used in biological control against the economically significant pests. Detection of morphological and genetic polymorphism among single species and strains in natural environment is important to study of the distribution and the effects in the ecosystem. The aim of this study was to determine the links of "local strains" with ecological aspects of biological control in protected zone of National park and assess the polymorphism of the population monitored in NP Šumava and exotic strains using molecular markers. There were analyzed 39 strains collected in the NP Šumava and the Krkonoše National Park in the Czech Republic and 11 strains from different countries and areas. The polymorphism of these strains was evaluated by different methods based on DNA analysis. In this study, the genetic variability is evaluated by AFLP markers. The results reveal that population from NP Šumava is very closed and the "local strains" can be very well characterized thanks these methods and compared with other strains from different parts of the Czech Republic and other countries. Thanks to these screening analyses, the strain Bba I101was determined for the preparation which will be used for bark beetle (Ips typographus) control in the National Park Šumava in the Czech Republic. This study was supported by grants GACR 521/08/H042, MSM 60076658-06, MZP SP/2d1/41/08.
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Analysis of low-copy sequences and its utilization for evaluation of genetic polymorphism in selected \kur{Beauveria bassiana} strains/isolates
JOZOVÁ, Eva
Beauveria bassiana is used in biological control against the economically significant pests. Detection of morphological and genetic polymorphism between different species and strains isolated in the natural environment is important to study the distribution and effects in the ecosystem. The aim of this study was to determine in which links are "local tribes" with ecological aspects of biological control in protected zone and assess the polymorphism of the sections by using molecular markers. Were analyzed 36 strains collected in the National Park Šumava in the Czech Republic. Polymorphism of these strains was determined according to a sequence of LSU (28S Large Subunit Ribosomal DNA). It is observed that the populations from National Park is closed, "the local tribes" because these methods can be very well characterized and compared with other strains from different parts of the Czech Republic and other countries. Populations from National Park showed no differences in polymorphism. Through this analysis, the sample Bba I101 was aligned. This preparation is re-used for bark beetle occurring in the National Park Šumava in the Czech Republic. This study was supported by grants GACR 521/08/H042, MSM 60076658-06, MGA SP/2d1/41/08.
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Diagnostics of species complex in family Adelgidae by molecular markers.
CHUNDELOVÁ, Daniela
Five molecular markers (mitochondrial AT-control region and COI, nuclear ITS and markers from primary enosymbiont Buchnera sp. argS and rrs) were tested for use in species identification of 2 selected genuses of the Adelgidae family. Markers COI, argS and rrs were succesfuly amplified and sequenced. Aligned sequences of those three markers were used for reconstruction of dendrogram by Neighbour-Joining metod with Kimura 2-Parameter model. These markers are suitable for resolution of taxonomic relationship of genuses within family Adelgidae, although further testing of these markers in conjunction with other markers (like ITS or AT-control region) will be required to delineate sister species with higher precision.
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Utilization of PCR-RFLP approach for evaluation of ITS polymorphism in selected \kur{Beauveria bassiana} strains/isolates
KOCKOVÁ, Lucie
The aim of this study was the evaluation of the suitability PCR-RFLP technique and ITS region analysis for determination the genetic variability of the entomopathogenic fungi - Beauveria bassiana, Beauveria brongniartii and Isaria fumosorosea. By the means of ITS-RFLP technique, it was possible to identify the uniformity of the B. bassiana isolates from NP Šumava, the higher level of polymorphism of B.bassiana isolates from different geographical areas was detected and it was possible to distinguish different entomopathogenic fungi used in this study.
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Identification of species within \kur{Adelgidae} by molecular markers
VĚCHTOVÁ, Pavlína
Four molecular markers (nuclear ITS, EF1 alpha and mitochondrial AT-Rich and COI) were tested for use in species identification of Adelgidae family. Markers EF1 alpha and COI were successfuly amplified and sequenced. Aligned sequences of those two markers were used for reconstruction of dendrogram by Minimum Evolution method with Kimura 2-Parameter model. Comparison of dendrogram with assumptions based on morfology and biology of Adelgids confirmed suitability of chosen molecular markers for reliable resolution of taxonomic relationship within Adelgidae, although further testing and markers will be required to delineate sister species with higher precision.
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