Scopus İndeksli Yayınlar Koleksiyonu / Scopus Indexed Publications Collection
Permanent URI for this collectionhttps://hdl.handle.net/11147/7148
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Article Citation - WoS: 3Citation - Scopus: 3Chromatographic Analysis for Targeted Metabolomics of Antioxidant and Flavor-Related Metabolites in Tomato(Bio-Protocol, 2021) Gürbüz Çolak, Nergiz; Tek, Neslihan; Frary, Anne; Doğanlar, SamiTargeted metabolomics is a useful approach to evaluate crop breeding studies. Antioxidant and flavor-related traits are of increasing interest and are considered quality traits in tomato breeding. The present study presents chromatographic methods to study antioxidants (carotenoids, vitamin C, vitamin E, phenolic compounds, and glutathione) and flavor -related characters (sugars and organic acids) in tomato. Two different extraction methods (for polar and apolar entities) were applied to isolate the targeted compounds. The extraction methods developed in this work were time and cost-effective since no further purification was needed. Carotenoids, vitamin C, glutathione, and phenolic acids were analyzed by HPLC-PDA using a RP C18 column at an appropriate wavelength for each compound. Vitamin E and sugars were analyzed by HPLC with RP C18 and NH2 columns and detected by FLD and RI detectors, respectively. In addition, organic acids were analyzed with GC-FID using a Rtx 5DA column after derivatization with MSTFA. As a result, sensitive analytical methods to quantify important plant metabolites were developed and are described herein. These methods are not only applicable in tomato but are also useful to characterize other species for flavor-related and antioxidant compounds. Thus, these protocols can be used to guide selection in crop breeding.Article Citation - Scopus: 18Organogenesis From Transformed Tomato Explants(Humana Press, 2005) Frary, Anne; Van Eck, JoyceTomato was one of the first crops for which a genetic transformation system was reported involving regeneration by organogenesis from Agrobacterium-transformed explants. Since the initial reports, various factors have been studied that affect the efficiency of tomato transformation and the technique has been useful for the isolation and identification of many genes involved in plant disease resistance, morphology and development. In this method, cotyledon explants from in vitro-grown seedlings are precultured overnight on a tobacco suspension feeder layer. The explants are then inoculated with Agrobacterium and returned to the feeder layer for a 2-d period of cocultivation. After cocultivation, the explants are transferred to an MS-based selective regeneration medium containing zeatin. Regenerated shoots are then rooted on a separate selective medium. This protocol has been used with several tomato cultivars and routinely yields transformation efficiencies of 10-15%.Article Citation - WoS: 3Citation - Scopus: 3Glycoalkaloid Isolation From Solanum Linnaeanum Berries(EDP Sciences, 2015) Gürbüz, Nergiz; Karabey, Fatih; Öztürk, Taylan Kurtuluş; Kılınç, Ali; Frary, Anne; Doğanlar, SamiGlycoalkaloids are plant secondary metabolites that can be both harmful and beneficial to human health. They cause gastroenteric symptoms, coma and even death at high concentrations. It is thought that glycoalkaloids are toxic to human health as a result of their effects on the nervous system and destruction of cell membranes. On the other hand, glycoalkaloids can be effective drugs. For example, solasodine is used to treat skin cancer and tomatidine is used in cancer chemotherapy. Solasodine is also used as a precursor of steroidal drugs. The goal of the work was to isolate and separate efficiently these similar compounds. Materials and methods. Glycoalkaloids from Solanum linnaeanum berries were isolated using column chromatography and confirmed via NMR spectroscopy and MS/MS spectrometry. Results and discussion. The chemical structures of glycoalkaloids are nearly identical. For example, solamargine differs from solasonine in having a methyl group instead of a hydroxyl group bound to the sugar residue of the compound. Thus, their molecular weight and polarity are quite similar. In this work, the process yielded 37.1 mg solasonine, 92.3 mg solasonine and solamargine mix and 56.2 mg solamargine from 81.67 g dried S. linnaeanum berries. Conclusion. Solanum linnaeanum berries are a good source of these glycoalkaloids and the developed protocol proved efficient for purification of solasonine and solamargine.Article Citation - WoS: 25Citation - Scopus: 25Application of Est-Ssrs To Examine Genetic Diversity in Eggplant and Its Close Relatives(TUBITAK, 2011) Tümbilen, Yeliz; Frary, Anne; Daunay, Marie Christine; Doğanlar, SamiWithin the genus Solanum, the term 'eggplant' encompasses several cultivated species that are used for food and, to a lesser extent, for medicine. Th e use of one common name to describe more than one species and the existence of many related wild species have led to taxonomic confusion which, in turn, have complicated analyses of evolutionary relationships and genetic diversity within this groups of species. A further challenge for eggplant research is that, despite the fact that the use of molecular markers for phylogenetic studies is well-established, very few studies have described the development of new markers for eggplant. In our work, genic microsatellite (SSR) markers were identified from an expressed sequence tag library of S. melongena and used for analysis of 47 accessions of eggplant and closely related species. Th e markers had very good polymorphism in the 18 species tested including 8 S. melongena accessions. Moreover, genetic analysis performed with these markers showed concordance with previous research and knowledge of eggplant domestication. Th ese markers are expected to be a valuable resource for studies of genetic relationships, fingerprinting, and gene mapping in eggplant.Article Citation - WoS: 9Citation - Scopus: 13Determination of Genetic Diversity in Watermelon [citrullus Lanatus (thunb.) Matsum & Nakai] Germplasm(Southern Cross Publishing and Printing Pty Ltd, 2011) Ulutürk, Zehra İpek; Frary, Anne; Doğanlar, SamiThe genetic diversity and relatedness of 90 watermelon (Citrullus lanatus var. lanthus and Citrullus. lanatus var. citroides) accessions were molecularly characterized using 30 sequence-related amplified polymorphism (SRAP) marker combinations. The accessions were collected from Turkey (59 accessions) and other regions of the world by the United States Department of Agriculture (USDA) and the molecular data were analysed using UPGMA (Unweighted Pair Group Method Analysis). The SRAP combinations were highly polymorphic (97%) with 87 polymorphic bands determined among 201 amplified fragments (43%). The UPGMA analysis characterized five major clusters (A, B, C, D and E). While minimum genetic similarity among groups A, B and C was high (83%), group E was the most distant with 63% genetic similarity. Principal component analysis (PCA) was performed and used to produce a two-dimensional plot from which two main groups could be distinguished. Based on the analyses, genetic diversity of watermelons was very low and Turkish watermelons were not distinct from other countries' accessions. These results could be used for generation of a core collection of watermelon by elimination of redundant accessions and for watermelon breeding by helping to identify useful, genetically distinct lines. In addition, the study indicated that SRAP markers are useful for analysing genetic diversity in crops like watermelon which have low variability.Article Citation - WoS: 23Citation - Scopus: 26Development of Practical Hplc Methods for the Separation and Determination of Eggplant Steroidal Glycoalkaloids and Their Aglycones(Taylor and Francis Ltd., 2008) Eanes, Ritchie C.; Tek, Neslihan; Kırsoy, Öyküm; Frary, Anne; Doğanlar, Sami; Almeida, Adelia E.A practical set of HPLC methods was developed for the separation and determination of the eggplant steroidal glycoalkaloids, solanine, chaconine, solasonine, solamargine, and their aglycones, solasodine and solanidine. A gradient method was initially developed, but proved to be neither robust nor practical. Three separate isocratic methods using acetonitrile and ammonium dihydrogen phosphate were developed and shown to be more repeatable, less subject to fluctuations in mobile phase composition, and less time consuming. The effect of adjusting buffer pH, column temperature, and buffer type (triethylammonium phosphate vs. ammonium dihydrogen phosphate) were evaluated. It was also discovered that, by addition of 10% methanol to the acetonitrile portion of the mobile phase, more control over the separations was possible. The use of methanol as a mobile phase entrainer greatly improved separations in some cases and its effectiveness was also dependent upon column temperature. Assessments of the method recovery, limit of detection, and limit of quantitation were made using extracts from S. melongena and S. linnaeanum.