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The genetic basis of Czech garlic "paličák" (A. sativum L. ssp. ophioscorodon) focusing on the biosynthetic pathways of the secondary metabolites
Čermák, Vladimír ; Ovesná, Jaroslava (advisor) ; Demnerová, Kateřina (referee)
Czech bolting garlic (Allium sativum L.) is the name for Czech varieties, which are categorized into subspecies sativum ssp. An inflorescence production and specific onion morphology is typical for this type of garlic. The genetic basis, that has been described by the analysis of microsatellites, are discussed in this thesis. Transcription analysis has revealed a large number of unigenes that have been assigned by genetic ontology to individual functions in the organism. Therefore, genetic differences from other varieties were confirmed. The impact of the environment and cultivation practices, including large- scale production, is being explored. The results can be used for further research or breeding. Consumers and experts have rated the Czech bolting garlic as a variety with a distinctly pungent taste and aroma. Substances causing this strong characteristic taste are secondary metabolites, alk(en)ylcysteine-S-oxides (ACSO), especially alliin and methiin. Chemical analysis did not confirm the exceptionally high value of these taste precursors or other substances that should distinguish Czech bolting garlic. Secondary metabolism includes other important substances that are used mainly in pharmacy and healthcare, where garlic has been used for thousands of years due to antibacterial effects,...
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Study of factors influencing efficiency of Canabis sativa transformation
Širl, Marek ; Ovesná, Jaroslava (advisor) ; Vondráková, Zuzana (referee)
Hemp (Cannabis sativa L.) is a multi-use crop, able to provide fibre celulose a hurds for industrial treatment seeds for oil preparation biomass for energy conversion and produces secondary metabolites useful for pharmaceutical application. For its resistence to stress ability to accumulate high concentration of heavy metals and low cultivations demands, it can also be used for phytoextractions. Current research is focused on establishment of cultivation protocol, which allows transformation of callus cultures, and their regeneration with high efficiency. In this thesis, several varieties of hemp were transferred to in vitro conditions and were tested for their ability to form callus. The best results were achieved using the hypocotyl segments in a nutrient medium supplemented with 1 mg/L of naphtylacetic acid and one of these two synthetic cytokinins 0,5 mg/L of thidiazuron or 5 mg/L of 6-benzylaminopurine. No significant difference in the use of these two cytokinins were observed. None of the explants on four different test media for regeneration of shoots were able to succesfully regenerate. Transformation of hemp was tested using two different methods. Transformed protoplasts from hemp leafs after agroinfiltration were isolated. This method turn out to be unsuitable for use with hemp due to its...
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A transcriptomic-based comparison of barley cultivars differing with respect to their low temperature acclimation capacity
Janská, Anna ; Ovesná, Jaroslava (advisor) ; Vaňková, Radomíra (referee) ; Honys, David (referee)
The PhD thesis is focused on a transcriptomics-based comparison of barley cultivars differing with respect to their low temperature acclimation capacity, with a particular focus on genes transcribed in the leaf and crown. The crown was of interest because of its importance for the winter survival of the plant. To involve both the first and the second phase of hardening, the test plants were exposed first to +3řC for 21 days, followed by - 3řC for one day. Freezing damage was assessed by measuring electrolyte leakage (Papers 2 and 3), using a modified version of a protocol developed by Prášil and Zámečník (1998). The same protocol was adapted to evaluate crown regrowth (Paper 2); for this purpose, the plants were cooled, then replanted and cut above the crown, and their survival rate calculated over the following week. Each RNA sample was queried by hybridization to an Affymetrix 22 K Barley1 GeneChip Genome Array (Close et al. 2004). The data were statistically analysed with the help of the software packages R, MAS 5.0 (Ihaka & Gentleman 1996) (Papers 2 and 3), Gene Spring GX 7.3 (Agilent Technologies, Santa Clara CA) and MapMan (Thimm et al. 2004; Usadel et al. 2005) (Paper 2), the "Self-Organizing Maps" algorithm (Kohonen et al. 1996) (Paper 3) and MIPS FunCat (Ruepp et al. 2004) (Paper 2). Paper...
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Development and validation of methods for GMO detection
Hodek, Jan ; Ovesná, Jaroslava (advisor) ; Konopásek, Ivo (referee) ; Pazlarová, Jarmila (referee)
The present work is focused on the issue of genetically modified organisms (GMOs) detection in the plants and derived products. Existing legislation in the European Union regulates monitoring of GMO and lays down rules for their labeling and traceability. The aim was to contribute to solving certain issues of critical points in GMO detection. The first point was DNA extraction. The problem was adressed to the example of DNA extraction from the fruit of papaya and candied papaya. For both food products the suitability of the selected extraction methods was verified and confirmed by amplification of a specific DNA sequence by PCR. The second critical point was related to PCR inhibitors. We observed the effect of real-time PCR inhibition due to the influence of the residual EDTA and the residual amount of heavy metal ions, which were present in laboratory plastic. In both examples, the inhibition affects the PCR efficiency. Another critical point in the GMO detection is use of screening elements. That work was focused on false positive results. Development of new methods for GMO detection was another goal of this work. We have developed the method for garden pea taxon-specific gene quantification by real-time PCR. The method was tested on 13 cultivars of garden pea and several related plants of...
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The sampling methodology for GM plants unintentionally present in the environment
Ovesná, Jaroslava ; Kučera, Ladislav ; Sovová, Tereza ; Mitrová, Katarína ; Pouchová, Vladimíra
In this methodology, sampling of plants from agricultural crops(corn, soybean, canola and potato) is defined to assess the presence of unauthorized GM plants in the environment.The methodology is applied mainly in the field of inspection controls of the compliance with Act no.78/2004. The sampling and collected samples must represent to a maximum extent the crops of the controlled part of the field (land parcel or land parcel portion). The basic applications of the methodology include:(a) low portion of unauthorized GM plants(seed contaminated with GM plants unauthorized for cultivation), (b) monitoring of the presence of unauthorized GM plants on land parcel, where there was such an occurence recorded in previous years (for example, plants growing on the field from seeds or tubers from the so called "soil´s seed bank").
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The use of microsatellite analysis for the characterisation of onion
Mitrová, Katarína ; Ovesná, Jaroslava
The object of the methodology is the process characterizing the autenticity of varieties of onion put through microsatellite analysis (SSR- Single Sequence Repeats) for the characterization of varieties of garlic into the practice. This method allows to identify varieties of onion and characterize genetic resources using DNA markers and determine their genetic similarity based on the length variability of short repeating sequences (microsatellites) in the characterization of varieties of onion, Allium cepa L. The principle of the method is amplification of the genome containing the microsatelite locus by polymerase chain reaction (PCR) using specific primers and subsequent analysis of the lengtht of the products and the identification of the DNA profile to the declared variety.
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The use of microsatellite analysis for the characterisation of garlic
Mitrová, Katarína ; Ovesná, Jaroslava ; Svobodová, Leona
The object of the methodology is the process characterizing the autenticity of varieties of garlic put through microsatellite analysis (SSR- Single Sequence Repeats) for the characterization of varieties of garlic into the practice. This method allows to identify varieties of garlic and characterize genetic resources using DNA markers and determine their genetic similarity based on the length variability of short repeating sequences (microsatellites) in the characterization of varieties of garlic, Allium sativum L. The principle of the method is amplification of the genome containing the microsatelite locus by polymerase chain reaction (PCR) using specific primers and subsequent analysis of the lengtht of the products and the identification of the DNA profile to the declared variety.
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Methodology for analysis of dairy products and their soy analogues for the presence of RoundUp Ready soybean
Tesařová, Zuzana ; Šídová, Dana ; Vráblík, Aleš ; Hodek, Jan ; Ovesná, Jaroslava
Dairy products may be subject to adulteration, where there is a relatively expensive raw material (e.g. cream) replaced by the addition of vegetable fats. Milk can be replaced in dairy products for example by soybean extract. Because soy belongs among strong allergens and the incidence of allergy is rather high. Therefore it is necessary to have a method for its detection. This work introduces the method for detection of soy, including analysis of genetic modified RoundUp Ready soybean. Detection of soybean in dairy products is based on analysis of amplification products melting profiles by High Resolution Melting (HRM) method. Analysis of geneticaly modified RoundUp Ready soybean is based on DNA detection via Polymerase Chain Reaction(PCR).
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Zabezpečení laboratorní detekce geneticky modifikovaných organismů
Výzkumný ústav rostlinné výroby‚ Praha-Ruzyně ; Koucká, Petra ; Hodek, Jan ; Pouchová, Vladimíra ; Kučera, Ladislav ; Ovesná, Jaroslava
Podklady pro metodiku vzorkování rostlinných materiálů pro potřeby správních úřadů. Zpracování informací z ENGL pro potřeby ČIŽP. Shromažďování referenčních materiálů. Metodiky stanovení GMO. Svazek obsahuje sborník na téma Geneticky modifikované organismy - současnost, rozšíření a možné interakce s životním prostředím. Jednotlivé příspěvky ve sborníku: geneticky modifikované organismy - základní informace, Možnosti stanovení GMO, Křižitelnost plodin a planých druhů jako potenciál pro únik genetických modifikací, Šíření pylu v porostech lnu setého, Problematika GM hrachu v ČR, Perspektivy využití GMO, Problematika GM bramboru v ČR.
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