Molecular Biology and Genetics / Moleküler Biyoloji ve Genetik
Permanent URI for this collectionhttps://hdl.handle.net/11147/9
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Article Citation - WoS: 8Citation - Scopus: 10Interaction With Fungi Promotes the Accumulation of Specific Defense Molecules in Orchid Tubers and May Increase the Value of Tubers for Biotechnological and Medicinal Applications: the Case Study of Interaction Between Dactylorhiza Sp. and Tulasnella Calospora(Frontiers Media S.A., 2022) Hampejsová, Romana; Berka, Miroslav; Berková, Veronika; Jersáková, Jana; Domkářová, Jaroslava; von Rundstedt, Friederike; Frary, Anne; Saiz-Fernández, Iñigo; Brzobohatý, Břetislav; Černý, MartinTerrestrial orchids can form tubers, organs modified to store energy reserves. Tubers are an attractive source of nutrients, and salep, a flour made from dried orchid tubers, is the source of traditional beverages. Tubers also contain valuable secondary metabolites and are used in traditional medicine. The extensive harvest of wild orchids is endangering their populations in nature; however, orchids can be cultivated and tubers mass-produced. This work illustrates the importance of plant-fungus interaction in shaping the content of orchid tubers in vitro. Orchid plants of Dactylorhiza sp. grown in asymbiotic culture were inoculated with a fungal isolate from Tulasnella calospora group and, after 3 months of co-cultivation, tubers were analyzed. The fungus adopted the saprotrophic mode of life, but no visible differences in the morphology and biomass of the tubers were detected compared to the mock-treated plants. To elucidate the mechanisms protecting the tubers against fungal infestation, proteome, metabolome, and lipidome of tubers were analyzed. In total, 1,526, 174, and 108 proteins, metabolites, and lipids were quantified, respectively, providing a detailed snapshot of the molecular process underlying plant-microbe interaction. The observed changes at the molecular level showed that the tubers of inoculated plants accumulated significantly higher amounts of antifungal compounds, including phenolics, alkaloid Calystegine B2, and dihydrophenanthrenes. The promoted antimicrobial effects were validated by observing transient inhibition of Phytophthora cactorum growth. The integration of omics data highlighted the promotion of flavonoid biosynthesis, the increase in the formation of lipid droplets and associated production of oxylipins, and the accumulation of auxin in response to T. calospora. Taken together, these results provide the first insights into the molecular mechanisms of defense priming in orchid tubers and highlight the possible use of fungal interactors in biotechnology for the production of orchid secondary metabolites.Article Citation - WoS: 4Citation - Scopus: 4Development of Liposomal Formulations of the Eggplant Glycoalkaloids Solasonine and Solamargine(Elsevier, 2022) Tatlıdil, Engin; Gürbüz, Nergiz; Doğanlar, Sami; Frary, AnneThe eggplant glycoalkaloids solasonine and solamargine are efficient biomacromolecules against skin diseases but are water-insoluble which results in inefficient treatment due to inadequate transdermal delivery. To address this problem, several liposomal formulations were prepared and evaluated for parameters including lecithin type, hydration temperature, and pH. The optimal formula with high physical and chemical stability included the lecithin Phospholipon 80H hydrated with 10 mM NaCl (pH 5.5). Solasonine - solamargine loaded liposomes were tested for their physical and chemical stability and drug leakage over a three-month period. Furthermore, the drug release profile of the loaded liposomes was evaluated with different release media. The glycoalkaloids and their liposomal formulations were assessed for their biological activity in culture using HaCaT and SCC-25 cell lines. This work resulted in a biologically effective liposomal formulation that was stable (size <220 nm, PDI [removed]80%) for at least three months.Article Citation - WoS: 36Citation - Scopus: 47Mapping of Quantitative Trait Loci for Antioxidant Molecules in Tomato Fruit: Carotenoids, Vitamins C and E, Glutathione and Phenolic Acids(Elsevier, 2020) Gürbüz Çolak, Nergiz; Tek, Neslihan; Ülger, Mehmet; Frary, Anne; Doğanlar, SamiThe nutritional value of a crop lies not only in its protein, lipid, and sugar content but also involves compounds such as the antioxidants lycopene, beta-carotene and vitamin C. In the present study, wild tomato Solanum pimpinellifolium LA 1589 was assessed for its potential to improve antioxidant content. This wild species was found to be a good source of alleles for increasing beta-carotene, lycopene, vitamin C and vitamin E contents in cultivated tomato. Characterization of an LA 1589 interspecific inbred backcross line (IBL) mapping population revealed many individuals with transgressive segregation for the antioxidants confirming the usefulness of this wild species for breeding of these traits. Molecular markers were used to identify QTLs for the metabolites in the IBL population. In total, 64 QTLs were identified for the antioxidants and their locations were compared to the map positions of previously identified QTLs for confirmation. Four (57 %) of the carotenoid QTLs, four (36 %) of the vitamin QTLs, and 11 (25 %) of the phenolic acid QTLs were supported by previous studies. Furthermore, several potential candidate genes were identified for vitamins C and E and phenolic acids loci. These candidate genes might be used as markers in breeding programs to increase tomato's antioxidant content.Article Citation - WoS: 12Citation - Scopus: 14Transcriptomic Analysis of Selenium Accumulation in Puccinellia Distans (jacq.) Parl., a Boron Hyperaccumulator(Elsevier, 2020) Kök, Aysu Başak; Mungan, Mehmet Direnç; Doğanlar, Sami; Frary, AnneSelenium (Se) is present in a wide variety of natural and man-made materials on Earth. Plants are able to tolerate and (hyper)accumulate Se to different extents. In fact, some species can tolerate and accumulate multiple elements. Puccinellia distans (P. distans), weeping alkali grass, is known to hyperaccumulate extreme concentrations of boron and tolerate high levels of salinity, therefore, we investigated the Se accumulation and tolerance capacities of this species. In addition, P. distans' Se tolerance mechanism was studied using a transcriptomic approach. The results of this study indicated that, when grown in a hydroponic system containing 80 or 120 mu M Se, P. distans shoots accumulated from 1500 to 2500-fold more Se than plants grown without the element. Thus, P. distans was discovered to be a novel Se accumulator plant. RNA sequencing results and biochemical analyses helped to shed light on the Se tolerance and accumulation mechanism of P. distans. Here, we suggest that upregulation of Se assimilation and stress response genes may be due to induction of jasmonic acid signaling. In addition, we propose that the cell wall may play an important role in restriction of Se movement to the cytoplasm. Also, we hypothesize that Se accumulates in cells by sequestration of selenate in the vacuole. (C) 2019 Elsevier Ltd. All rights reserved.Article Citation - WoS: 18Citation - Scopus: 21Exploring Wild Alleles From Solanum Pimpinellifolium With the Potential To Improve Tomato Flavor Compounds(Elsevier, 2020) Gürbüz Çolak, Nergiz; Tek, Neslihan; Ülger, Mehmet; Frary, Anne; Doğanlar, SamiMost consumers complain about the flavor of current tomato cultivars and many pay a premium for alternatives such as heirloom varieties. Breeding for fruit flavor is difficult because it is a quantitatively inherited trait influenced by taste, aroma and environmental factors. A lack of genetic diversity in modern tomato cultivars also necessitates exploration of new sources for flavor alleles. Wild tomato S. pimpinellifolium and inbred backcross lines were assessed for individual sugars and organic acids which are two of the main components of tomato flavor. S. pimpinellifolium was found to harbor alleles that could be used to increase glucose and fructose content and adjust acidity by altering malic and citric acid levels. Single nucleotide polymorphism markers were used to detect 14 quantitative trait loci (QTLs) for sugars and 71 for organic acids. Confirmation was provided by comparing map locations with previously identified loci. Thus, seven (50 %) of the sugar QTLs and 22 (31 %) of the organic acids loci were supported by analyses in other tomato populations. Examination of the genomic sequence containing the QTLs allowed identification of potential candidate genes for several flavor components. © 2020 Elsevier B.V.Article Citation - WoS: 179Citation - Scopus: 229Health Benefits and Bioactive Compounds of Eggplant(Elsevier, 2018) Gürbüz, Nergiz; Uluışık, Selman; Frary, Anne; Frary, Amy; Doğanlar, SamiEggplant is a vegetable crop that is grown around the world and can provide significant nutritive benefits thanks to its abundance of vitamins, phenolics and antioxidants. In addition, eggplant has potential pharmaceutical uses that are just now becoming recognized. As compared to other crops in the Solanaceae, few studies have investigated eggplant's metabolic profile. Metabolomics and metabolic profiling are important platforms for assessing the chemical composition of plants and breeders are increasingly concerned about the nutritional and health benefits of crops. In this review, the historical background and classification of eggplant are shortly explained; then the beneficial phytochemicals, antioxidant activity and health effects of eggplant are discussed in detail.Article Citation - WoS: 15Citation - Scopus: 15Identification of Stable Qtls for Fiber Quality and Plant Structure in Upland Cotton (g. Hirsutum L.) Under Drought Stress(Elsevier Ltd., 2018) Akköse Baytar, Asena; Peynircioğlu, Ceng; Sezener, Volkan; Basal, Hüseyin; Frary, Anne; Frary, Amy; Doğanlar, SamiCotton is an economically important commodity for nearly fifty industries including the textile sector which is largely based on cotton fiber. Identification of markers linked to loci for fiber traits under drought stress may be particularly beneficial because such loci could provide the genetic adaptability needed to produce good fiber under water limitation. In the present study, 177 simple sequence repeat (SSR) markers were used to detect significant quantitative trait loci (QTLs) linked to 11 fiber quality and plant structure traits in a panel of 99 Upland cotton (Gossypium hirsutum L.) genotypes using GLM and MLM analysis. The fiber quality traits, including fiber length (FL), fiber fineness (FF), fiber strength (FS), fiber elasticity (FE), fiber uniformity (FU), spinning conversion index (SCI), earliness (EAR), 1st position boll retention (1st PBR), 2nd position boll retention (2nd PBR), total boll number (TBN) and plant height (PH), were tested under both well-watered and water-limited irrigations in two locations. At both locations, GLM identified a total of 74 and 70 QTLs under well-watered and water limited conditions, respectively, at p ≤ 0.005. MLM detected seven and 23 QTLs under well-watered and water-limited conditions, respectively. Of the identified QTLs, some QTLs were detected in both locations: three for well-watered and two for water-stress conditions. Moreover, a total of 19 QTLs were stable under both watering-regimes. The QTLs identified herein could be useful in the development of cotton cultivars that have adaptability to drought conditions worldwide.Article Citation - WoS: 18Citation - Scopus: 21Molecular Genetic Diversity in the Origanum Genus: Est-Ssr and Srap Marker Analyses of the 22 Species in Eight Sections That Naturally Occur in Turkey(Elsevier, 2018) Taşcıoğlu, Tülin; Sadıkoğlu, Narin; Doğanlar, Sami; Frary, AnneOriganum (L.) is a genus of herbaceous perennials with culinary and medicinal uses with many species found in the Mediterranean region. The taxonomic classification of species belonging to this genus has been studied using morphological, biochemical and genetic diversity analyses. In this study, the genetic diversity of 22 Turkish Origanum species (including 24 taxa from eight sections) was examined with 46 herbarium specimens from the Mediterranean, Eastern Anatolian, Central Anatolian, and Black Sea regions of Turkey. Molecular marker data were generated from 25 SRAP primer pairs and six EST-SSR primers which produced 325 alleles. Dendrogram, principal coordinate and population structure analyses revealed the relationships among Origanum sections, species and individuals. Gene flow (PhiPT and Nm) was also studied for a deeper understanding of the relationships and hybridization patterns between sections and species. Molecular separation of the Origanums roughly corresponded to the taxonomy that Ietswaart proposed in 1980 but also suggested that hybridization among sections and species may result in convergence and/or divergence of different sections and species.Article Citation - WoS: 14Citation - Scopus: 16Transcriptomic Analysis of Boron Hyperaccumulation Mechanisms in Puccinellia Distans(Elsevier Ltd., 2018) Öztürk, Saniye Elvan; Göktay, Mehmet; Has, Canan; Babaoğlu, Mehmet; Allmer, Jens; Doğanlar, Sami; Frary, AnnePuccinellia distans, common alkali grass, is found throughout the world and can survive in soils with boron concentrations that are lethal for other plant species. Indeed, P. distans accumulates very high levels of this element. Despite these interesting features, very little research has been performed to elucidate the boron tolerance mechanism in this species. In this study, P. distans samples were treated for three weeks with normal (0.5 mg L−1) and elevated (500 mg L−1) boron levels in hydroponic solution. Expressed sequence tags (ESTs) derived from shoot tissue were analyzed by RNA sequencing to identify genes up and down-regulated under boron stress. In this way, 3312 differentially expressed transcripts were detected, 67.7% of which were up-regulated and 32.3% of which were down-regulated in boron-treated plants. To partially confirm the RNA sequencing results, 32 randomly selected transcripts were analyzed for their expression levels in boron-treated plants. The results agreed with the expected direction of change (up or down-regulation). A total of 1652 transcripts had homologs in A. thaliana and/or O. sativa and mapped to 1107 different proteins. Functional annotation of these proteins indicated that the boron tolerance and hyperaccumulation mechanisms of P. distans involve many transcriptomic changes including: alterations in the malate pathway, changes in cell wall components that may allow sequestration of excess boron without toxic effects, and increased expression of at least one putative boron transporter and two putative aquaporins. Elucidation of the boron accumulation mechanism is important in developing approaches for bioremediation of boron contaminated soils.Article Citation - WoS: 18Citation - Scopus: 29Cry1ac-Mediated Resistance To Tomato Leaf Miner (tuta Absoluta) in Tomato(Springer Verlag, 2017) Şelale, Hatice; Dağlı, Fatih; Mutlu, Nedim; Doğanlar, Sami; Frary, AnneTomato leaf miner (Tuta absoluta) is a major pest of tomato in regions with hot climates such as South America and the Mediterranean. This insect feeds on almost every part of the plant and forms galleries while feeding on the plant’s inner tissues. Thus, it can cause plant death and is costly and difficult to control with chemical sprays. In this study, we transferred a modified Bacillus thuringiensis cry1Ac gene to tomato plants via Agrobacterium tumefaciens mediated transformation. Introduction of the cry1Ac gene to the tomato genome was confirmed with PCR and Southern blot analysis in 12 independent events. Insertion sites of the transgene in the tomato genome were determined with TAIL-PCR (thermal asymmetric interlaced polymerase chain reaction) for four selected transgenic lines. Cry1Ac gene expression was verified at both the transcriptional and translational levels, with RT-qPCR and Western blot analyses, respectively. Expression of the Cry1Ac protein in tomato resulted in T. absoluta mortality rates of 38–100% depending on transgenic line. In addition, gallery formation was reduced in 57–100% of the transgenic plants. Moreover, it was found that a single copy of the gene in the hemizygous condition is sufficient to confer tolerance to leaf miner. This is the first reported development of tomato plants resistant to T. absoluta. These transgenic plants are promising for development of commercial tomato cultivars resistant to leaf miner, which will limit the use of environmentally harmful chemicals for control of this pest.
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