|
|
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.
|
|
|
Studz of direct and indirect plasma application on onion seeding bulbs
Krejsová, Lenka ; Enev, Vojtěch (referee) ; Krčma, František (advisor)
Nowadays, agriculture is based on conventional methods that involve the application of pesticides, agricultural chemicals and fertilizers. These substances have a negative impact on nature and human health, so more environmentally friendly methods are being sought. This thesis deals with the analysis of fragrances in onions treated with plasma before planting. The treatment was carried out to see if it would improve onion growth while maintaining or increasing the fragrance content. The theoretical part deals with the soil and it is characterization methods, plasma, quantitative and qualitative methods for the determination of volatile substances. It contains the principles of UV-VIS spectrometry, atomic absorption spectrometry (AAS), mass spectrometry (MS) as well as tandem connection of gas chromatography with mass spectrometry (GC–MS). The experimental part was carried out over a period of three years at 22 sites. The preparation and analysis of soil samples are described, which was also evaluated. Furthermore, the procedures for treating the bulbs with corona discharge and plasma-activated water before planting were described. In total, four treatment variants were carried out and bulbs from the fifth variant were not additionally treated. Each treatment variant was planted in four replications of ten bulbs each. Twenty selected grown onions from one site were individually converted to liquid samples and the saturated vapor was analysed by proton transfer reaction time of flight mass spectrometry (PTR–TOF–MS). Thanks to this analysis, the concentrations of volatile substances were determined. Some samples were analyzed by gas chromatography with mass spectrometer as detector to identify the compounds. Subsequently, the data were processed and evaluated. From the results obtained, it is evident that plasma treatment can be useful for the treatment of onions in agriculture, because it confirmed harvest increase without decrease of fragrances concentrations. Thus, it is necessary to carry out experiments on a larger scale in the future.
|
|
|
Analysis of aromatic compounds in plasma treated onion
Krejsová, Lenka ; Kozáková, Zdenka (referee) ; Krčma, František (advisor)
This bachelor thesis deals with the analysis of fragrances in onions, which were exposed to plasma before planting. The adjustment was made to see if onion growth and yields would improve. The theoretical part deals with the quantitative and qualitative methods for the determination of volatile substances. It contains the principle and instrumentation of mass spectrometry as well as the tandem connection of gas chromatography with mass spectrometry. In the experimental part, a liquid sample was obtained by pressing. After 60 minutes, saturated vapors were analyzed by reactive ionization mass spectrometry (PTR-MS). Thanks to this analysis, the concentration of volatile substances was determined. Some samples were analyzed by gas chromatography with mass spectrometer as detector to identify the compounds, because PTR-MS does not allow distinguishing of isomers. Subsequently, the data were processed and evaluated. From the gathered obtained it is clear that the plasma treatment has an effect on the fragrances concentrations. After six months of storage, the presence of some volatile substances increased due to increased biological activity.
|
|
|
Analysis of onion and garlic samples of different geographical origin
Korček, Jakub ; Mikulíková, Renata (referee) ; Diviš, Pavel (advisor)
Presented master thesis dealt with the analysis of garlic and onion samples and tried to find correlations between chemical composition and country of origin. The parameters examined were dry content, crude protein content, concentration of phenolic compunds, carbohydrate content (fructose, glucose), alliin content and concentration of selected elements (P, Mg, Ca, Na, K, Fe, Zn). Average content of dry matter of fresh samples was 35,84 ±2,12 g/100 g and of dehydrated samples was 90,61 ±2,90 g/100 g. Concentration of phenolic compounds was measured spectrophotometrically with Folin-Ciocault reagent. Average phenolic compounds content of samples was 0,1840 ±0,1286 GAE g/100 g. Crude protein content was measured by Kjeldahl method, and calculated from total nitrogen content. Carbohydrates were measured by HPLC-ELSD after hydrolysis of fructans. Average content of fructose of garlic samples was 57,014 ±0,863 g/100 g, of onion samples was 33,718 ±1,168 g/100 g. Average content of glucose of onion samples was 22,633 ±0,405 g/100 g. Alliin content was measured by HPLC-DAD method. Average alliin content of fresh samples was 4,644 ±0,446 g/100 g and of dehydrated samples was 1,962 ±0,180 g/100 g. Elemental analysis was conducted by ICP-OES method. Average concentration of selected elements was: P 2,15 ±0,11 mg/g, Mg 0,638 ±0,03 mg/g, Ca 1,246 ±0,05 mg/g, Na 0,550 ±0,08 mg/g, K 7,49 ±0,41 mg/g, Fe 79,3 ±6,16 mg/kg, Zn 11,4 ±3,33 mg/kg. Obtained data were statistically processed on significance level 0,05. Based on the principal components analysis, it was found, that the best parameters to differentiate samples from Czechia, Poland and Ukraine from other countries were content of nitrogen, phosphorus, magnesium, sodium and calcium. It was also discovered, that genus differences between onion and garlic have greater significance than geographical differences.
|
|
|
Determination of volatile substances of natural extracts intended for cosmetic application
Paskevych, Olha ; Řezáčová, Veronika (referee) ; Mravcová, Ludmila (advisor)
Tato bakalářská práce je soustředěná na zkoumaní přírodních komerčně dostupných extraktů, které se používají do přírodní kosmetiky. Literární rešerše byla zaměřena na téma obsahu těkavých látek v přírodních extraktech používaných v kosmetice, jejich charakterizace a možnosti stanovení, problematika výskytu alergenů v přírodní kosmetice. Na základě tohoto přehledu byla navržena optimální metoda pro analýzu vzorků přírodních extraktů s přihlédnutím k dostupné analytické instrumentaci. Následující výtažky byly studovány se zaměřením na kvalitativní stanovení: mandle, aloe vera, hřebíček, Etiopská káva, zázvor, cibule.
|
|
|
Acyklické organosirné sloučeniny v cibuli kuchyňské (\kur{Allium cepa})
ŠUSTR, Ondřej
This Bachelor thesis deals with acyclic organosulfur compounds of onion (Allium cepa). The aim of the work was to isolate and structurally characterize some of these compounds using a series of preparative HPLC fractionations. The theoretical part deals with major groups of currently known organosulfur compounds of onion, their formation pathways and biological properties. The experimental part describes the process of isolation of cepaenes.
|
|
| |
|
|
Analysis of aromatic compounds in plasma treated onion
Krejsová, Lenka ; Kozáková, Zdenka (referee) ; Krčma, František (advisor)
This bachelor thesis deals with the analysis of fragrances in onions, which were exposed to plasma before planting. The adjustment was made to see if onion growth and yields would improve. The theoretical part deals with the quantitative and qualitative methods for the determination of volatile substances. It contains the principle and instrumentation of mass spectrometry as well as the tandem connection of gas chromatography with mass spectrometry. In the experimental part, a liquid sample was obtained by pressing. After 60 minutes, saturated vapors were analyzed by reactive ionization mass spectrometry (PTR-MS). Thanks to this analysis, the concentration of volatile substances was determined. Some samples were analyzed by gas chromatography with mass spectrometer as detector to identify the compounds, because PTR-MS does not allow distinguishing of isomers. Subsequently, the data were processed and evaluated. From the gathered obtained it is clear that the plasma treatment has an effect on the fragrances concentrations. After six months of storage, the presence of some volatile substances increased due to increased biological activity.
|
|
|
Determination of volatile substances of natural extracts intended for cosmetic application
Paskevych, Olha ; Řezáčová, Veronika (referee) ; Mravcová, Ludmila (advisor)
Tato bakalářská práce je soustředěná na zkoumaní přírodních komerčně dostupných extraktů, které se používají do přírodní kosmetiky. Literární rešerše byla zaměřena na téma obsahu těkavých látek v přírodních extraktech používaných v kosmetice, jejich charakterizace a možnosti stanovení, problematika výskytu alergenů v přírodní kosmetice. Na základě tohoto přehledu byla navržena optimální metoda pro analýzu vzorků přírodních extraktů s přihlédnutím k dostupné analytické instrumentaci. Následující výtažky byly studovány se zaměřením na kvalitativní stanovení: mandle, aloe vera, hřebíček, Etiopská káva, zázvor, cibule.
|
|
|
Analysis of onion and garlic samples of different geographical origin
Korček, Jakub ; Mikulíková, Renata (referee) ; Diviš, Pavel (advisor)
Presented master thesis dealt with the analysis of garlic and onion samples and tried to find correlations between chemical composition and country of origin. The parameters examined were dry content, crude protein content, concentration of phenolic compunds, carbohydrate content (fructose, glucose), alliin content and concentration of selected elements (P, Mg, Ca, Na, K, Fe, Zn). Average content of dry matter of fresh samples was 35,84 ±2,12 g/100 g and of dehydrated samples was 90,61 ±2,90 g/100 g. Concentration of phenolic compounds was measured spectrophotometrically with Folin-Ciocault reagent. Average phenolic compounds content of samples was 0,1840 ±0,1286 GAE g/100 g. Crude protein content was measured by Kjeldahl method, and calculated from total nitrogen content. Carbohydrates were measured by HPLC-ELSD after hydrolysis of fructans. Average content of fructose of garlic samples was 57,014 ±0,863 g/100 g, of onion samples was 33,718 ±1,168 g/100 g. Average content of glucose of onion samples was 22,633 ±0,405 g/100 g. Alliin content was measured by HPLC-DAD method. Average alliin content of fresh samples was 4,644 ±0,446 g/100 g and of dehydrated samples was 1,962 ±0,180 g/100 g. Elemental analysis was conducted by ICP-OES method. Average concentration of selected elements was: P 2,15 ±0,11 mg/g, Mg 0,638 ±0,03 mg/g, Ca 1,246 ±0,05 mg/g, Na 0,550 ±0,08 mg/g, K 7,49 ±0,41 mg/g, Fe 79,3 ±6,16 mg/kg, Zn 11,4 ±3,33 mg/kg. Obtained data were statistically processed on significance level 0,05. Based on the principal components analysis, it was found, that the best parameters to differentiate samples from Czechia, Poland and Ukraine from other countries were content of nitrogen, phosphorus, magnesium, sodium and calcium. It was also discovered, that genus differences between onion and garlic have greater significance than geographical differences.
|
guest ::
