Master Degree / Yüksek Lisans Tezleri
Permanent URI for this collectionhttps://hdl.handle.net/11147/3008
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Master Thesis Characterization of Pollen-E1 Gene Might Play Role in Salt Tolerance in Beta Maritima(Izmir Institute of Technology, 2015) Uysal, Özge; Karakaya, Hüseyin Çağlar; Karakaya, Hüseyin Çağlar; 04.03. Department of Molecular Biology and Genetics; 04. Faculty of Science; 01. Izmir Institute of TechnologySalinity stress has a negative impact on the growth of plants, which affects homeostasis and productivity. The uptake of non-essential salt ions change the osmotic balance of the cell and cause dehydration. Higher plants develop salt tolerance mechanisms to avoid dehydration. In this project, we isolated and characterized salt tolerance genes in Beta maritima plant. For this purpose, functional genomics technique was used by over expressing cDNAs in yeast and colonies can grow toxic salt media isolated and characterized. We found several colonies and we focused on uncharacterized Pollen-E1 gene with an unknown function. Pollen-E1 cDNA confers salt tolerance to yeast cells. Intracellular sodium measurements of Pollen-E1 overexpressed in yeast cells showed decreased salt levels as compared to wild type suggesting that sodium was transported out of the cell. Pollen-E1 protein localized in endomembrane systems in the yeast cells. In mRNA expression analysis, Pollen-E1 mRNA levels induced immediately in leaves and later stages in root systems under salt stress. Our results showed that is the uncharacterized and unknown function Pollen-E1 gene might have some role of regulating salt tolerance in Beta maritima.Master Thesis Identification and Characaterization of Nickel Tolerance Genes in Beta Maritima(Izmir Institute of Technology, 2009) Bozdağ, Gönensin Ozan; Karakaya, Hüseyin Çağlar; Karakaya, Hüseyin Çağlar; 04.03. Department of Molecular Biology and Genetics; 04. Faculty of Science; 01. Izmir Institute of TechnologyNickel is an essential micronutrient for plant growth. However, an excess amount of nickel is toxic for plants and animals. The Beta maritima plant which is known crop as salt and manganese tolerant nature. In this study, we aimed to identify nickel detoxification metabolism genes of Beta maritima by screening plant.s overexpressed cDNA library in yeast cells. After initial screening, three transformants were isolated that were able to grow in the presence of toxic nickel concentration. After the sequence analyses, three genes were named as NIC3, NIC6 and NIC8. We tested the growth rates of the yeast cells those were overexpressing cDNAs in solid mediums and showed similar nickel detoxification patterns. Then intracellular nickel concentrations were measured to see whether mentioned cDNAs pump nickel out of the cell or not. We also analysed the gene expression levels of NIC3 and NIC6 genes after appllying nickel stress to the plants to see the relations between nickel treatment and transcription changes of these two genes. Our data suggest that NIC3, NIC6 and NIC8 genes, besides their other functions, confer yeast cells nickel tolerance and may play roles in heavy metal detoxification for plant cells.Master Thesis Identification and Characterization of Manganese Tolerance Genes in Beta Vulgaris Subsp. Maritima(Izmir Institute of Technology, 2011) Erbaşol, Işıl; Karakaya, Hüseyin Çağlar; Karakaya, Hüseyin Çağlar; 04.03. Department of Molecular Biology and Genetics; 04. Faculty of Science; 01. Izmir Institute of TechnologyManganese is an essential element for higher organisms however uptake of excess amount of manganese causes toxicity. Beta vulgaris subsp. maritima, the member of Chenopodiaceae family, is known to tolerate high concentration of sodium. Due to its living conditions, Beta vulgaris subsp. maritima adapted many different stress conditions. Therefore it is an ideal plant for studying plant tolerance mechanisms. In this study, we aimed to identify the genes which are responsible for manganese tolerance in Beta vulgaris subsp. maritima by screening its cDNA library in Saccharomyces cerevisiae cells. After initial screening in the presence of toxic manganese concentration; 2,7mM MnCl2, three resistant yeast colonies were selected. After the sequence and similarity analysis, two genes which might involve in manganese tolerance were identified and named as BmMn1 and BmMn2. The results of solid media tests with different yeast strains which transformed with the genes revealed that BmMn1 provides a remarkable manganese tolerance like BmMn2 and slightly nickel tolerance. They do not show tolerance to other metals such as zinc, cadmium, boron and cobalt. Identified manganese concentrations in pmr1 yeast strainstransformed with BmMn1, BmMn2 or empty vector pointed that BmMn1 and BmMn2 transport excess manganese out of the cell. In addition, GFP localization in the yeast cell proved that the BmMn1 and BmMn2 are located in Golgi apparatus. qRT-PCR analyses of Beta vulgaris subsp. maritima which was exposed to 2mM Mn2+ suggested a dynamic regulation for the expression of these two genes. The results indicate that BmMn1 and BmMn2 have a role in detoxification of excess amount of manganese in Beta vulgaris subsp. maritima.