Master Degree / Yüksek Lisans Tezleri
Permanent URI for this collectionhttps://hdl.handle.net/11147/3008
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Master Thesis A Proteomic Approach for Indentifying Boron-Stress Tolerant Proteins in Barley Genotpes(Izmir Institute of Technology, 2008) Atik, Ahmet Emin; Yalçın, TalatBoron is an essential micro-nutrient for plants. However, when boron is present at high concentrations in the soil or ground water, healthy plant growth and development can be affected by boron toxicity. Turkey constitutes about 72 % of the total boron reserves in the world. In Turkey, barley (Hordeum vulgare) is the second widely grown cereal after wheat. Boron toxicity is one of the major factors limiting the yield of barley in Central Anatolia of Turkey. In the present study, proteomic approach was used to investigate the boronstress tolerant proteins in Anadolu barley genotype (boron-tolerant). Eight-day-old barley plants were treated with 10 mM H3BO3 for seven days. Control plants received no boron treatment during this period. Total proteins of leaves were extracted and separated by two-dimensional polyacrylamide gel electrophoresis. Control and borontreated Anadolu genotype.s proteome maps were compared and the novel proteins were fragmented into peptides using in-gel digestion technique. Liquid chromatographytandem mass spectrometry (LC-MS/MS) analysis and database searching gave way to identify ten spots representing seven different proteins. Two spots were identified as the same protein and one protein could not identified. The identified seven proteins are namely, ribulose 1,5-bisphosphate carboxylase/oxygenase large chain (RuBisCo largechain), thaumatin-like protein TLP5, basic pathogenesis-related protein PR5, RNase Slikeprotein, vacuolar proton-translocating ATPase subunit E, PSI type III chlorophyll a/b-binding protein, and light-harvesting complex I; LHC I. Among the identified seven proteins, vacuolar proton-translocating ATPase (VATPase) subunit E is the important one for boron tolerance in tolerant barley genotype. It is shown that the accumulation of excess boron in the vacuolar compartment of the plant cell by the help of V-ATPase subunit E protein. This is the known as internal tolerance mechanisms for Anadolu genotype of barley to survive under boron stress. It was proposed that, this might be the defense mechanism in boron-tolerant barley genotype under toxic boron concentrations.Master Thesis Identification of Genes That Play Roles in Boron Metabolism/(Izmir Institute of Technology, 2008) Kaya, Alaattin; Koç, AhmetBoron, is an essential microelement that plays a role in plant and animal development. However, an excess amount of boron is toxic for both types of organisms.The yeast Saccharomyces cerevisiae is an ideal model organism for describing the mechanism and regulation of metal ion transport in eukaryotes. We screened a yeast genomic DNA library to find genes that confer boron resistance to wild-type cells.Thirty transformants were isolated that were able to grow in the presence of a toxic amount of boron and all of them contained the multidrug efflux transporter gene ATR1 (YML116w) in the expression cassette. Our subsequent analysis revealed that ATR1 deletion mutants (.atr1) were sensitive to boron treatment and hyper-accumulated boron inside cells, whereas wild-type cells overexpressing the ATR1 gene were resistant to boron and hypoaccumulated boron in cells. We also analysed the global gene expression pattern in response to boron treatment and found that expression of ATR1 is upregulated along with many other transporter and amino acid biosynthesis genes. Our data suggest that the ATR1 gene functions as a boron efflux pump and is reqired for boron tolerance in yeast cells.Master Thesis Identification and Characterization of Boron Tolerant Genes in Hordeum Vulgare (barley) by Using Mrna Differential Display and Rt-Pcr Techniques(Izmir Institute of Technology, 2007) Akıncı, Ersin; Karakaya, Hüseyin ÇağlarBoron, is a microelement that plays a role in plant development. In contrast, excess amount of boron is toxic for plants. Turkey is the second country that has the largest boron reserve in the world, thus boron is one of the major problems in agriculture in Turkey. Barley (Hordeum vulgare) is the second widely produced cereal after wheat. Because barley is used in human diet, animal feeding and beer industry it is an economically valuable crop. There are ten different barley varieties in Turkey and these varieties show different genetic variations against boron toxicity. In this study, Hamidiye (boron sensitive) and Anadolu (boron tolerant) varieties were used to identify genes responsible for boron tolerance. RT-PCR and mRNA Differential Display techniques were used from root and leaf samples of Hamidiye and Anadolu varieties grown in laboratory with or without boron conditions. Eight differentially expressed genes identified by using mRNA Differential Display technique. Sequence of these genes gave homology to an eukaryotic translation initiation factor in Arabidopsis thaliana, a chlorophyll a/b binding protein precursor in Triticum aestivum, an elongation factor in Oryza sativa, short-chain dehydrogenase/reductase family protein in Arabidopsis thaliana, a thioredoxin h isoform in Triticum aestivum, a shaggy-like kinase protein in Triticum aestivum, chloroplast genome in Hordeum vulgare, a hypothetical protein in Arabidopsis thaliana. Expression level of six of forty three antiporter genes showed differences between Anadolu and Hamidiye cultivars in Real Time PCR.
