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Testing of primers for real-time PCR-HRM analysis of fruit products containing one fruit species
Boháčová, Barbora ; Dzurendová, Simona (referee) ; Fialová, Lenka (advisor)
Determining the authenticity of fruit products, which are often counterfeited by substituting part of a more expensive fruit with a cheaper but botanically similar fruit, is a current topic in the food industry. A prime example is the dilution of apricot products with peach puree. This study focuses on testing specific primers AGS18 and PdCass to reveal the true proportion of apricot content in model products mixed with peach puree using PCR analysis followed by HRM analysis. Testing of these primers for authenticity determination revealed limited utility of AGS18 primers due to the formation of small amount or no specific products during reactions with fruit DNA. PdCass primers required PCR condition optimization, but subsequently, specific DNA sequences from fruit leaves could be amplified in sufficient quantities. PCR analysis of DNA from model products with PdCass primers provided specific products in samples with apricot content of 70% or less. HRM analysis of samples and calculation of GCP did not distinguish purees with 0%, 10%, and 30% apricot content. However, purees with 50% and 70% apricot content exhibited significantly different melting curves compared to other samples.
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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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Moderní odrůdový sortiment broskvoně obecné a možnosti uplatnění v pěstitelské praxi
Grund, Martin
The purpose of this bachelor thesis is to theoretically study floricultural and pomological features of peach. The peach’s traditional cultivars used in Czech orchards and modern cultivars, which might enrich our range, were examined. In the experimental part of this thesis the dates of beginning and full flowering, flowering richness, and the shape of flower were observed in 10 cultivars in gene pool in Horticultural faculty in Lednice. The observing ran through the years 2020 and 2021, and every time there were chosen 3 trees from each cultivar which were evaluated and from which the average value was calculated. The gained values from both years were then compared.
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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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Hodnocení pevnosti ovoce pomocí měření odezvy při impulzivním zatěžování
Dvořáková, Pavla ; Nedomová, Š. ; Severa, L. ; Trnka, Jan ; Buchar, J.
Dynamic resonance frequency based on dynamic frequency response of excited peach was determined by peach physical properties,such as, peach firmness, peach mass, and day of harvesting. Analysis showed that dominant frequency increased with peach firmness increasing, and dominant frequency decreased with increasing of the peach mass.
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