- Email: [email protected]
High-resolution melting analysis in sequence matching applications to identify similarities and dissimilarities in DNA
Abstracts / Journal of Biotechnology 161S (2012) 13–18
imately 32 ◦ C). The pH seemed to control both the maximal algal density in the pond and the productivity that were found to be maximum at pH values below 11. http://dx.doi.org/10.1016/j.jbiotec.2012.07.028
ORAL PRESENTATION Food and Nutritional Sciences Inhibitory activity of apigenin, quercetin, naringenin, (+)catechin, quercitrin, rutin, and naringin against purified NADH-cytochrome B5 reductase Haydar Celik 1,∗ , Muberra Kosar 2 1 Department of Pharmaceutical Basic Sciences, Faculty of Pharmacy, Erciyes University, Kayseri, 38039, Turkey 2 Department of Pharmacognosy, Faculty of Pharmacy, Erciyes University, Kayseri, 38039, Turkey
E-mail address: [email protected] (H. Celik).
17
and their effects on nutritional values, and the chemical reactions taking place in meal preparation. Information collected from literature combined with experimental findings related to commonly consumed dishes on food preservation, preparation, spoilage, and various cooking techniques like boiling, steaming, baking, frying and microwaving are assessed. Examples are given on various processes from boiling vegetables and roasting meat to baking cake and popping corn to emphasize the importance of chemical knowledge on meal preparation. It can be concluded that food processing can be regarded as an experimental science and the skill of meal making is developed only through continued special effort – it does not come naturally. http://dx.doi.org/10.1016/j.jbiotec.2012.07.030
ORAL PRESENTATION Food and Nutritional Sciences Determination of cytotoxic and genotoxic effects of benzaldehyde in vitro
Flavonoids are polyphenolic compounds exhibiting a broad spectrum of biological and pharmacological activities. The healthpromoting effect of these phytochemicals is attributed in part to their ability to modulate the activity of enzymes responsible for metabolic activation or detoxification of chemical carcinogens. Modulation of drug metabolizing enzymes by flavonoids is of great significance from a pharmacological and toxicological point of view concerning the potential flavonoid-drug and/or flavonoid/xenobiotic interactions and in guiding clinical use of drugs. In the present study, in an effort to identify the interaction of dietary flavonoids with NADH-cytochrome b5 reductase, we studied the modulatory effects of seven flavonoids with different chemical structures including apigenin, quercetin, naringenin, (+)catechin, quercitrin, rutin and naringin on the activity of purified bovine liver b5 reductase, which shares a high level of homology with human counterpart. The findings obtained using potassium ferricyanide as electron acceptor identified quercetin as potent inhibitor of b5 reductase with an IC50 value of 1.07 mol/L and suggest that inhibition is highly dependent on structure of individual flavonoids. The possible importance of this structure-dependent modulation in terms of altered therapeutic efficacy or toxicity of certain drugs and xenobiotics metabolized by cytochrome b5 reductase will be discussed in the presentation.
Benzaldeyde (BA) occurs naturally in a number of plants, including cherry, fig, peach fruits and carnation flowers at therapeutic doses. In addition it use in personal care products and food as a preservative. Although it has been reported that 50.3 g/mL is non toxic to different cell culture modals by National Toxicology Programme, we showed that this amount of BA is toxic to cultured lymphocytes with the help of recent cell culture assays. In this study, we aimed that determination of cytotoxic and apoptotic effects of different concentrations of BA on cultured human lymphocytes by using Lactate dehydrogenase (LDH) assay, cell proliferation (WST1) assay and TUNNEL test (apoptotic test) at 6th and 24th hours. The cytotoxicity increased treated with 10, 25 and 50 g/mL concentrations of BA (p < 0.001). Moreover, treatment with the same concentrations significantly decreased cell number (p < 0.05). Also TUNEL assay results show that the same concentration of BA caused DNA damage significantly (p < 0.001). According to our results toxic and genotoxic effects of BA have to evaluate with further investigations to use in cosmetic and food products.
http://dx.doi.org/10.1016/j.jbiotec.2012.07.029
http://dx.doi.org/10.1016/j.jbiotec.2012.07.031
ORAL PRESENTATION
ORAL PRESENTATION
Food and Nutritional Sciences
Fundamental and Applied Aspects of Biotechnology
Meal chemistry: Science, skill and fun
High-resolution melting analysis in sequence matching applications to identify similarities and dissimilarities in DNA
Ömer Edip Kuzugudenli School of Sciences, Erciyes University, Kayseri, 38039, Turkey E-mail address: [email protected]. Overwhelming majority of the people preparing meals does not evaluate the chemical aspects of their actions – most people cannot understand and the remaining do not bother to think about the details. Foods usually go through a number of physical and chemical stages from their production to final eatable forms. This study was conducted for a better understanding of the chemistry of human body, food chemistry, uptake of food components in digestive systems, physical and chemical food processing techniques
Zeynep Ulker ∗ , Lokman Alpsoy Biology, Fatih University, Istanbul, 34500, Turkey E-mail address: [email protected] (Z. Ulker).
Irem Uzonur ∗ , Derya Sultan Karabulut, Fatih Koyuncu, Gamze Akdeniz, Zeynep Katmer Biology, Fatih University, Istanbul, 34500, Turkey E-mail address: [email protected] (I. Uzonur). High-resolution melting (HRM) analysis has applications in testing for known sequence variants (genotyping), screening for mutations (scanning) and identifying similarities or dissimilarities in DNA (sequence matching). It can discriminate nucleotide sequence differences by comparing DNA melting behaviour of different sample compositions with high accuracy. The aim is to show the power and
18
Abstracts / Journal of Biotechnology 161S (2012) 13–18
potential of the HRM method in identifying similarities and dissimilarities in DNA sequences of various sample types. The samples used in the work are chosen to be informative for the diverse applications of HRM in sequence matching. Samples tested are groups (human) with NAT2 SNP (rs1801280 341T > C), Urticadioica mighty ecotypes and bloom forming species in Golden Horn. All amplicons produced specifically have been analyzed for specific gene of interest in the above mentioned organisms using Qiagen’s Type-it HRM mix on Rotor-Gene 6000TM HRM platform with genotype autocalling function. High-resolution melting method which is a strong alternative to the conventional screening methods, because of its simplicity, accuracy, rapidity and cost-effectiveness can be very effective using Rotor-Gene 6000TM HRM platform and its genotype autocall function given the confidence intervals high enough with the aid of reliable positive controls for each case considered. http://dx.doi.org/10.1016/j.jbiotec.2012.07.032
resistance traits are coded by quantitative trait loci (QTLs) that cannot be transferred by genetic transformation, not to mention the consumer acceptance issue. We are going to discuss a strategy of combinatorial biotechnology where different tools should work together to deliver the resistance genes into crop gene pool. In the case of potato such a combination includes: genetic transformation of Solanum chacoense (chc resistant to Colorado potato beetle – CPB), for MMR deficiency to increase homeologous recombination, somatic hybridization of potato cultivars with chc by protoplast electrofusion, selection of resistant hybrids by using molecular markers (RAPD) linked to leptines biosynthesis, repellents for CPB, testing of resistance by the use of laboratory bioassay and finally integration into breeding new varieties resistant to CPB. Other examples for multiple resistance traits or resistance genes pyramiding will be presented. Acknowledgements for the funds to the project CNCS PNII-IDPCE-2011-3-0586 http://dx.doi.org/10.1016/j.jbiotec.2012.07.034
ORAL PRESENTATION Plant-Cell Biotechnology Gynogenesis induction in edible Alliums Arzu
Kaska ∗ ,
Fevziye Celebi-Toprak, Ali Ramazan Alan
Pamukkale University, Plant Genetics and Agricultural Biotechnology Application and Research Center (BIYOM), Kinikli, Denizli, 20070, Turkey E-mail address: [email protected] (A. Kaska). Genetic improvement of edible Alliums such as onion (Allium cepa L.; 2n = 2x = 16) and leek (A. ampeloprasum L.; 2n = 4x = 32) through conventional breeding is a time consuming process due to biennial nature and high levels of heterozygosity. Production of doubledhaploid (DH) plants via gynogenesis is an alternative to repeated self-pollination for recovery of genetically uniform onion and leek lines. We initiated work on gynogenesis utilizing a wide range of material developed in our edible Alliums improvement program. Our goal was to test the applicability of the DH technique to the development of improved onion and leek cultivars adapted to Turkey. In the first year of gynogenesis project, 30 onion and 7 leek plants were recovered from breeding lines maintained in our Allium germplasm. Flow cytometry analysis showed that gynogenic onion plants obtained were haploid, diploid, mixoploid, and tetraploid. All gynogenic leek plants analyzed were tetraploid. Our results clearly show that Turkish onion and leek germplasm materials are suitable for gynogenesis-based breeding programs. http://dx.doi.org/10.1016/j.jbiotec.2012.07.033
ORAL PRESENTATION Plant-Cell Biotechnology Action of heavy metals on the function and state of photosystem II Jeyhuna Jafarova 1,∗ , Rena Ganiyeva 1 , Sona Bayramova 1 , Inna Kurbanova 1 , Ralphreed Gasanov 2 1
Biophysics Laboratory, Institute of Botany, National Academy of Sciences, Patamdart shosse, 40, AZ 1073, Azerbaijan 2 Department of Biophysics and Molecular biology, Z. Khalilov str. 23, Baku State University, Baku, AZ 1148, Azerbaijan E-mail address: [email protected] (J. Jafarova). Action sites of different metals in the electron transport reactions of Photosystem II (PS II) evaluated by delayed fluorescence in ms range (ms DF) and pigment-protein and protein-protein interaction by electrophoretic measurements are presented. The metal ions had multiple actions on PS II. On the donor side the presence of Cd2+ inhibits components of ms DF reflecting oxygen evolution. On the same side Co2+ and Ni2+ display toxic effects at the pH dependent form and interact with acceptor side also. Al3+ , Zn2+ and Mn2+ ions influence components on the acceptor side of PS II (possibly interact and modify the QB site). Herewith Al3+ and Mn2+ toxic effects are manifested only in acidic pH. Investigated metals bring about monomerization of oligomeric and dimeric chlorophyll–protein complexes (CPC) and destabilization a protein-protein interactions. Molecular mechanisms of metals action are discussed. http://dx.doi.org/10.1016/j.jbiotec.2012.07.035
ORAL PRESENTATION Plant-Cell Biotechnology Combining different biotechnological tools for better introgression of resistance genes into crops: The case of potato Elena Rakosy-Tican Department of Molecular Biology and Biotechnology, Babes-Bolyai, University, Cluj-Napoca, 400006 Romania E-mail address: [email protected]. Many crop plants wild relatives represent a rich source of resistance genes toward biotic or abiotic stress. Such genes can be faster introgressed into crops by using biotechnological tools. Usually,
imately 32 ◦ C). The pH seemed to control both the maximal algal density in the pond and the productivity that were found to be maximum at pH values below 11. http://dx.doi.org/10.1016/j.jbiotec.2012.07.028
ORAL PRESENTATION Food and Nutritional Sciences Inhibitory activity of apigenin, quercetin, naringenin, (+)catechin, quercitrin, rutin, and naringin against purified NADH-cytochrome B5 reductase Haydar Celik 1,∗ , Muberra Kosar 2 1 Department of Pharmaceutical Basic Sciences, Faculty of Pharmacy, Erciyes University, Kayseri, 38039, Turkey 2 Department of Pharmacognosy, Faculty of Pharmacy, Erciyes University, Kayseri, 38039, Turkey
E-mail address: [email protected] (H. Celik).
17
and their effects on nutritional values, and the chemical reactions taking place in meal preparation. Information collected from literature combined with experimental findings related to commonly consumed dishes on food preservation, preparation, spoilage, and various cooking techniques like boiling, steaming, baking, frying and microwaving are assessed. Examples are given on various processes from boiling vegetables and roasting meat to baking cake and popping corn to emphasize the importance of chemical knowledge on meal preparation. It can be concluded that food processing can be regarded as an experimental science and the skill of meal making is developed only through continued special effort – it does not come naturally. http://dx.doi.org/10.1016/j.jbiotec.2012.07.030
ORAL PRESENTATION Food and Nutritional Sciences Determination of cytotoxic and genotoxic effects of benzaldehyde in vitro
Flavonoids are polyphenolic compounds exhibiting a broad spectrum of biological and pharmacological activities. The healthpromoting effect of these phytochemicals is attributed in part to their ability to modulate the activity of enzymes responsible for metabolic activation or detoxification of chemical carcinogens. Modulation of drug metabolizing enzymes by flavonoids is of great significance from a pharmacological and toxicological point of view concerning the potential flavonoid-drug and/or flavonoid/xenobiotic interactions and in guiding clinical use of drugs. In the present study, in an effort to identify the interaction of dietary flavonoids with NADH-cytochrome b5 reductase, we studied the modulatory effects of seven flavonoids with different chemical structures including apigenin, quercetin, naringenin, (+)catechin, quercitrin, rutin and naringin on the activity of purified bovine liver b5 reductase, which shares a high level of homology with human counterpart. The findings obtained using potassium ferricyanide as electron acceptor identified quercetin as potent inhibitor of b5 reductase with an IC50 value of 1.07 mol/L and suggest that inhibition is highly dependent on structure of individual flavonoids. The possible importance of this structure-dependent modulation in terms of altered therapeutic efficacy or toxicity of certain drugs and xenobiotics metabolized by cytochrome b5 reductase will be discussed in the presentation.
Benzaldeyde (BA) occurs naturally in a number of plants, including cherry, fig, peach fruits and carnation flowers at therapeutic doses. In addition it use in personal care products and food as a preservative. Although it has been reported that 50.3 g/mL is non toxic to different cell culture modals by National Toxicology Programme, we showed that this amount of BA is toxic to cultured lymphocytes with the help of recent cell culture assays. In this study, we aimed that determination of cytotoxic and apoptotic effects of different concentrations of BA on cultured human lymphocytes by using Lactate dehydrogenase (LDH) assay, cell proliferation (WST1) assay and TUNNEL test (apoptotic test) at 6th and 24th hours. The cytotoxicity increased treated with 10, 25 and 50 g/mL concentrations of BA (p < 0.001). Moreover, treatment with the same concentrations significantly decreased cell number (p < 0.05). Also TUNEL assay results show that the same concentration of BA caused DNA damage significantly (p < 0.001). According to our results toxic and genotoxic effects of BA have to evaluate with further investigations to use in cosmetic and food products.
http://dx.doi.org/10.1016/j.jbiotec.2012.07.029
http://dx.doi.org/10.1016/j.jbiotec.2012.07.031
ORAL PRESENTATION
ORAL PRESENTATION
Food and Nutritional Sciences
Fundamental and Applied Aspects of Biotechnology
Meal chemistry: Science, skill and fun
High-resolution melting analysis in sequence matching applications to identify similarities and dissimilarities in DNA
Ömer Edip Kuzugudenli School of Sciences, Erciyes University, Kayseri, 38039, Turkey E-mail address: [email protected]. Overwhelming majority of the people preparing meals does not evaluate the chemical aspects of their actions – most people cannot understand and the remaining do not bother to think about the details. Foods usually go through a number of physical and chemical stages from their production to final eatable forms. This study was conducted for a better understanding of the chemistry of human body, food chemistry, uptake of food components in digestive systems, physical and chemical food processing techniques
Zeynep Ulker ∗ , Lokman Alpsoy Biology, Fatih University, Istanbul, 34500, Turkey E-mail address: [email protected] (Z. Ulker).
Irem Uzonur ∗ , Derya Sultan Karabulut, Fatih Koyuncu, Gamze Akdeniz, Zeynep Katmer Biology, Fatih University, Istanbul, 34500, Turkey E-mail address: [email protected] (I. Uzonur). High-resolution melting (HRM) analysis has applications in testing for known sequence variants (genotyping), screening for mutations (scanning) and identifying similarities or dissimilarities in DNA (sequence matching). It can discriminate nucleotide sequence differences by comparing DNA melting behaviour of different sample compositions with high accuracy. The aim is to show the power and
18
Abstracts / Journal of Biotechnology 161S (2012) 13–18
potential of the HRM method in identifying similarities and dissimilarities in DNA sequences of various sample types. The samples used in the work are chosen to be informative for the diverse applications of HRM in sequence matching. Samples tested are groups (human) with NAT2 SNP (rs1801280 341T > C), Urticadioica mighty ecotypes and bloom forming species in Golden Horn. All amplicons produced specifically have been analyzed for specific gene of interest in the above mentioned organisms using Qiagen’s Type-it HRM mix on Rotor-Gene 6000TM HRM platform with genotype autocalling function. High-resolution melting method which is a strong alternative to the conventional screening methods, because of its simplicity, accuracy, rapidity and cost-effectiveness can be very effective using Rotor-Gene 6000TM HRM platform and its genotype autocall function given the confidence intervals high enough with the aid of reliable positive controls for each case considered. http://dx.doi.org/10.1016/j.jbiotec.2012.07.032
resistance traits are coded by quantitative trait loci (QTLs) that cannot be transferred by genetic transformation, not to mention the consumer acceptance issue. We are going to discuss a strategy of combinatorial biotechnology where different tools should work together to deliver the resistance genes into crop gene pool. In the case of potato such a combination includes: genetic transformation of Solanum chacoense (chc resistant to Colorado potato beetle – CPB), for MMR deficiency to increase homeologous recombination, somatic hybridization of potato cultivars with chc by protoplast electrofusion, selection of resistant hybrids by using molecular markers (RAPD) linked to leptines biosynthesis, repellents for CPB, testing of resistance by the use of laboratory bioassay and finally integration into breeding new varieties resistant to CPB. Other examples for multiple resistance traits or resistance genes pyramiding will be presented. Acknowledgements for the funds to the project CNCS PNII-IDPCE-2011-3-0586 http://dx.doi.org/10.1016/j.jbiotec.2012.07.034
ORAL PRESENTATION Plant-Cell Biotechnology Gynogenesis induction in edible Alliums Arzu
Kaska ∗ ,
Fevziye Celebi-Toprak, Ali Ramazan Alan
Pamukkale University, Plant Genetics and Agricultural Biotechnology Application and Research Center (BIYOM), Kinikli, Denizli, 20070, Turkey E-mail address: [email protected] (A. Kaska). Genetic improvement of edible Alliums such as onion (Allium cepa L.; 2n = 2x = 16) and leek (A. ampeloprasum L.; 2n = 4x = 32) through conventional breeding is a time consuming process due to biennial nature and high levels of heterozygosity. Production of doubledhaploid (DH) plants via gynogenesis is an alternative to repeated self-pollination for recovery of genetically uniform onion and leek lines. We initiated work on gynogenesis utilizing a wide range of material developed in our edible Alliums improvement program. Our goal was to test the applicability of the DH technique to the development of improved onion and leek cultivars adapted to Turkey. In the first year of gynogenesis project, 30 onion and 7 leek plants were recovered from breeding lines maintained in our Allium germplasm. Flow cytometry analysis showed that gynogenic onion plants obtained were haploid, diploid, mixoploid, and tetraploid. All gynogenic leek plants analyzed were tetraploid. Our results clearly show that Turkish onion and leek germplasm materials are suitable for gynogenesis-based breeding programs. http://dx.doi.org/10.1016/j.jbiotec.2012.07.033
ORAL PRESENTATION Plant-Cell Biotechnology Action of heavy metals on the function and state of photosystem II Jeyhuna Jafarova 1,∗ , Rena Ganiyeva 1 , Sona Bayramova 1 , Inna Kurbanova 1 , Ralphreed Gasanov 2 1
Biophysics Laboratory, Institute of Botany, National Academy of Sciences, Patamdart shosse, 40, AZ 1073, Azerbaijan 2 Department of Biophysics and Molecular biology, Z. Khalilov str. 23, Baku State University, Baku, AZ 1148, Azerbaijan E-mail address: [email protected] (J. Jafarova). Action sites of different metals in the electron transport reactions of Photosystem II (PS II) evaluated by delayed fluorescence in ms range (ms DF) and pigment-protein and protein-protein interaction by electrophoretic measurements are presented. The metal ions had multiple actions on PS II. On the donor side the presence of Cd2+ inhibits components of ms DF reflecting oxygen evolution. On the same side Co2+ and Ni2+ display toxic effects at the pH dependent form and interact with acceptor side also. Al3+ , Zn2+ and Mn2+ ions influence components on the acceptor side of PS II (possibly interact and modify the QB site). Herewith Al3+ and Mn2+ toxic effects are manifested only in acidic pH. Investigated metals bring about monomerization of oligomeric and dimeric chlorophyll–protein complexes (CPC) and destabilization a protein-protein interactions. Molecular mechanisms of metals action are discussed. http://dx.doi.org/10.1016/j.jbiotec.2012.07.035
ORAL PRESENTATION Plant-Cell Biotechnology Combining different biotechnological tools for better introgression of resistance genes into crops: The case of potato Elena Rakosy-Tican Department of Molecular Biology and Biotechnology, Babes-Bolyai, University, Cluj-Napoca, 400006 Romania E-mail address: [email protected]. Many crop plants wild relatives represent a rich source of resistance genes toward biotic or abiotic stress. Such genes can be faster introgressed into crops by using biotechnological tools. Usually,