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Determination of athenticity of plant foods by molecular techniques
Plášková, Anna ; Mikulíková, Renata (referee) ; Němcová, Andrea (advisor)
The aim of presented diploma thesis was to determination of authenticity of fruit baby foods for early infant feeding using molecular methods. In the experimental part, isolation kit was used for isolation of plant DNA from fruits (strawberry, apricot, raspberry, apple) and from six commercial fruit products for children. Isolated DNA was characterized and verified using PCR methods with primers specific for plant rDNA (ITS2). Specific primer pairs were designed to amplify DNA for the detection of one fruit species. Primer specificity was assessed with four fruit species. A mixture of fruit puree from the two fruits was used to determine the sensitivity of the multiplex PCR assay. Six commercial fruit products were evaluated to verify the applicability of the multiplex PCR assay. The methodology of molecular detection of fruit DNA by qPCR and multiplex qPCR (duplex) includes approaches, which enable to detect two fruits (strawberry-raspberry, apricot-apple) in one reaction and thus reduces time and money requirements.
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Analysis of authenticity of some food products with fruit component by molecular and instrumental techniques
Pecháček, Michal ; Němcová, Andrea (referee) ; Márová, Ivana (advisor)
This master’s thesis was focused on determining the authenticity of some food products with fruit component by molecular and instrumental techniques. The thesis was divided into theoretical and practical part. The theoretical part of the work was focused on food adulteration, methods of determining the food authenticity, analysed types or technologies of production of individual food products, which were analysed in this work. This part was also focused on molecular and instrumental techniques, which are currently used to determine the food authenticity. The practical part of the thesis was divided into molecular and instrumental part. Within the molecular part, DNA isolation was performed using the EliGene Plant DNA Isolation kit. Inhibitors such as polysaccharides seemed to be the biggest problem during the DNA isolation. Therefore, an isolation method using pectinase incubation was performed. The isolated DNA was subsequently subjected to PCR and the resulting PCR products were analysed by a melting curve analysis. This method was used for ITS2 primers, which were used for the plant DNA detection. In the case of species-specific primers BAS1 and Pa3LTP that were used for the peach and apricot detection, high resolution melting (HRM) analysis was performed. During HRM, the focus was on the melting temperature of the specific PCR products. The melting temperature of the BAS1 specific product was set at 78,4 °C and at 86,43 °C for the specific product of Pa3LTP primers. Finally, specific PCR products were subjected to agarose gel electrophoresis. In the case of ITS2 primers, which served to verify the amplifiability of plant DNA, a band of 500 bp was detected. In the case of species-specific primers BAS1 and Pa3LTP, bands from 100 to 150 bp were detected. In the instrumental part of this thesis the HPLC/PDA analysis of the phenolic compounds was performed. The most suitable procedure for the analysis of phenolic substances was the purification procedure using ethanol without further concentration At the end of the work, the individual methods were compared with each other. At the same time, the influence of the matrix on the overall determination of food authenticity was monitored. While molecular techniques could be used to determine the food authenticity by determining the presence of specific DNA, instrumental techniques would be more suitable for detecting food adulteration and detecting specific substances.
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Jaroslavice – place in the landscape
Galková, Ivana ; Eichler, Zdeněk (referee) ; Ponešová, Barbora (advisor)
The main aim of my thesis is to create sustainable local food production in Jaroslavice region. This region is situated near the Austria – Czechia border. The political changes of previous century influenced this rural location a lot. Changes resulted in socioeconomic degradation of this area. My project tends to shorten production connections between producers and consumers. It also takes the part with local farmers and farmer´s market. My architecture proposal is a building of regional food center focused on fruit production. The main fruits are apples, apricots and pitches according to the local climate conditions. The center has three main parts – processing, storage and retail.
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Cultivation of stone fruits in the conditions of occurrence of the causal agents of cork necrosis and premature death of apricots and peaches in production orchards
Pánková, Iveta ; Krejzar, Václav ; Krejzarová, Radka
The methodology monitors the vegetation season in the orchards of stone fruits with regard to the possible influence of performed agrotechnical measurements on the occurrence of the pests, especially bacterial causal agents from the complex Pseudomonas syringae (Ps), Ps. pv. syringae and Ps. pv. morsprunorum races 1. and 3. Regulation of bacterial pathogens is becoming increasingly difficult due to the current possibilities of fruit tree protection, the range of cultivated genotypes and the complex life cycle of pathogens. Manifestations of the disease vary in individual growing seasons depending on the weather. However, the effects and severity of the disease are cumulative. Caring for the health of the tree begins with the production of nursery sprouts and must continue throughout the life of the tree. The methodology describes the individual standard agrotechnical measurements in production orchards in the context of the pathogen life cycle. Based on the results of the project, possible labor and economically acceptable modifications of agricultural practices including timing are proposed in order to reduce the inoculum of cork necrosis agents and reduce economic losses due to premature death of apricot trees.
Fulltext:  PDF
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Analysis of authenticity of some food products with fruit component by molecular and instrumental techniques
Pecháček, Michal ; Němcová, Andrea (referee) ; Márová, Ivana (advisor)
This master’s thesis was focused on determining the authenticity of some food products with fruit component by molecular and instrumental techniques. The thesis was divided into theoretical and practical part. The theoretical part of the work was focused on food adulteration, methods of determining the food authenticity, analysed types or technologies of production of individual food products, which were analysed in this work. This part was also focused on molecular and instrumental techniques, which are currently used to determine the food authenticity. The practical part of the thesis was divided into molecular and instrumental part. Within the molecular part, DNA isolation was performed using the EliGene Plant DNA Isolation kit. Inhibitors such as polysaccharides seemed to be the biggest problem during the DNA isolation. Therefore, an isolation method using pectinase incubation was performed. The isolated DNA was subsequently subjected to PCR and the resulting PCR products were analysed by a melting curve analysis. This method was used for ITS2 primers, which were used for the plant DNA detection. In the case of species-specific primers BAS1 and Pa3LTP that were used for the peach and apricot detection, high resolution melting (HRM) analysis was performed. During HRM, the focus was on the melting temperature of the specific PCR products. The melting temperature of the BAS1 specific product was set at 78,4 °C and at 86,43 °C for the specific product of Pa3LTP primers. Finally, specific PCR products were subjected to agarose gel electrophoresis. In the case of ITS2 primers, which served to verify the amplifiability of plant DNA, a band of 500 bp was detected. In the case of species-specific primers BAS1 and Pa3LTP, bands from 100 to 150 bp were detected. In the instrumental part of this thesis the HPLC/PDA analysis of the phenolic compounds was performed. The most suitable procedure for the analysis of phenolic substances was the purification procedure using ethanol without further concentration At the end of the work, the individual methods were compared with each other. At the same time, the influence of the matrix on the overall determination of food authenticity was monitored. While molecular techniques could be used to determine the food authenticity by determining the presence of specific DNA, instrumental techniques would be more suitable for detecting food adulteration and detecting specific substances.
|
|
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Determination of athenticity of plant foods by molecular techniques
Plášková, Anna ; Mikulíková, Renata (referee) ; Němcová, Andrea (advisor)
The aim of presented diploma thesis was to determination of authenticity of fruit baby foods for early infant feeding using molecular methods. In the experimental part, isolation kit was used for isolation of plant DNA from fruits (strawberry, apricot, raspberry, apple) and from six commercial fruit products for children. Isolated DNA was characterized and verified using PCR methods with primers specific for plant rDNA (ITS2). Specific primer pairs were designed to amplify DNA for the detection of one fruit species. Primer specificity was assessed with four fruit species. A mixture of fruit puree from the two fruits was used to determine the sensitivity of the multiplex PCR assay. Six commercial fruit products were evaluated to verify the applicability of the multiplex PCR assay. The methodology of molecular detection of fruit DNA by qPCR and multiplex qPCR (duplex) includes approaches, which enable to detect two fruits (strawberry-raspberry, apricot-apple) in one reaction and thus reduces time and money requirements.
|
|
|
Jaroslavice – place in the landscape
Galková, Ivana ; Eichler, Zdeněk (referee) ; Ponešová, Barbora (advisor)
The main aim of my thesis is to create sustainable local food production in Jaroslavice region. This region is situated near the Austria – Czechia border. The political changes of previous century influenced this rural location a lot. Changes resulted in socioeconomic degradation of this area. My project tends to shorten production connections between producers and consumers. It also takes the part with local farmers and farmer´s market. My architecture proposal is a building of regional food center focused on fruit production. The main fruits are apples, apricots and pitches according to the local climate conditions. The center has three main parts – processing, storage and retail.
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