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: 9Citation - Scopus: 11Development of Simple Sequence Repeat Markers in Hazelnut (corylus Avellana L.) by Next-Generation Sequencing and Discrimination of Turkish Hazelnut Cultivars(Springer, 2018) Özturk, Süleyman Can; Göktay, Mehmet; Doğanlar, Sami; Allmer, Jens; Frary, AnneEuropean hazelnut (Corylus avellana) is a diploid tree species and is widely used in confections. Hazelnuts are, to a large part, produced in Turkey with the cultivar "Tombul" widely grown in the Black Sea region. In this work, the "Tombul" genome was partially sequenced by next-generation sequencing technology yielding 29.2% (111.85 Mb) of the similar to 385 Mb (1C). This sequence information was used to develop genetic markers in order to enable differentiation of material before the long maturation process and to facilitate future breeding strategies. A total of 90,142 simple sequence repeats (SSRs) were identified in the contigs giving a frequency of 1 SSR per 1240 nt in the assembly. Mononucleotides were the most abundant SSR marker type (60.9%) followed by di- and trinucleotides. Primer pairs were designed for 75,139 (83.3%) of the SSRs. Fifty SSR primers were applied to 47 hazelnut accessions from nine countries to test their effectiveness and polymorphism. The markers amplified an average of 3.2 fragments. The highest polymorphism information content value was for cavSSR11062 (0.97) and the lowest (0.04) was for cavSSR13386. Two markers were monomorphic: cavSSR12855 and cavSSR13267. Single-copy SSR primers were also assessed for their ability to discriminate 19 Turkish cultivars, and it was found that seven primer pairs (Cav4217, Cav14875, Cav14418, Cav2704, Cav12862, Cav3909, Cav1361) were sufficient for this task. Thus, this study developed new SSR markers for use in hazelnut breeding and genetic studies and also provide a method to distinguish and identify true-type Turkish cultivars.Article Citation - WoS: 7Citation - Scopus: 10Association Mapping of Agro-Morphological Traits in European Hazelnut (corylus Avellana)(Springer, 2019) Frary, Amy; Öztürk, Süleyman Can; Balık, Hüseyin İrfan; Balık, Selda Kayalak; Kızılcı, Gökhan; Doğanlar, Sami; Frary, AnneMore than half of European hazelnut (Corylus avellana) production occurs in Turkey. Despite this dominance, the yield of Turkish cultivars has remained stagnant over the past 10years with Italian yield nearly double that in Turkey. This difference is due to Turkey's unique cultivation system; hazelnuts are grown in bushy clusters (ocak system), not as single trees. Current hazelnut breeding efforts are shifting toward the development of materials for single plant orchards which are much higher yielding; thus, there is a need to explore germplasm for relevant agro-morphological traits and to determine their genetic control. The objectives of this study were to examine data for 44 such traits in 390 hazelnut accessions: 16 cultivars, 232 landraces and 142 wild individuals from nine provinces in Turkey and to map the loci associated with these characteristics using simple sequence repeat markers. Comparison of cultivars, landraces and wild hazelnut accessions revealed the effects of domestication and selection on the crop and indicated that useful alleles for traits such as cropping and reduced alternate bearing may exist in the wild germplasm. A total of 145 quantitative trait loci (QTL) were detected with the largest proportions identified for involucre (26%) and inflorescence (14%) morphology. Several markers co-localized with more than one trait including markers for male catkin abundance which were shared with plant vigor and height. Similarly, markers for female flower abundance co-localized with suckering and alternate bearing. Such markers and their linked QTL should be studied in more detail as they might help breeders select for plant vigor, decreased suckering and increased flower production: traits which will be extremely useful for Turkey's transition to single plant orchards.Article Citation - WoS: 10Citation - Scopus: 13Analysis of European Hazelnut (corylus Avellana) Reveals Loci for Cultivar Improvement and the Effects of Domestication and Selection on Nut and Kernel Traits(Springer Verlag, 2019) Frary, Amy; Öztürk, Süleyman Can; Balık, Hüseyin İrfan; Kayalak Balık, Selda; Kızılcı, Gökhan; Doğanlar, Sami; Frary, AnneTurkey is a rich source of European hazelnut (Corylus avellana) germplasm with nearly 400 accessions in the national collection. This genetic material encompasses cultivars, landraces and wild genotypes which were characterized for 12 nut and 13 kernel traits over 2years in the 1990s. Analysis of these attributes revealed both the positive and negative impacts that human selection and breeding have had on hazelnut. Thus, while selection has resulted in larger nuts and kernels, cultivars have fewer nuts per cluster and kernels with larger internal cavities. Breeding has also resulted in a propensity for cultivars to have higher proportions of double kernels and empty nuts, two traits which reduce quality and yield. In addition, it is clear that while selection has successfully increased hazelnut fat content it has not impacted overall flavor, a much more complex trait. The nut and kernel phenotypic data were combined with genotypic data from 406 simple sequence repeat marker alleles for association mapping of the quantitative trait loci (QTL) for the traits. A total of 78 loci were detected in the population with the highest proportions for nut (24%) and kernel (26%) appearance parameters followed by quality (19%), shell thickness (16%) and yield-related (15%) traits. It is hoped that some of the identified QTL will be useful for future breeding of hazelnut for improved nut and kernel yield and quality.Article Citation - WoS: 17Citation - Scopus: 20Molecular Genetic Diversity of the Turkish National Hazelnut Collection and Selection of a Core Set(Springer Verlag, 2017) Öztürk, Süleyman Can; Balık, Hüseyin İrfan; Kayalak Balık, Selda; Kızılcı, Gökhan; Duyar, Ömür; Doğanlar, Sami; Frary, AnneEuropean hazelnut (Corylus avellana L.) is an economically and nutritionally important nut crop with wild and cultivated populations found throughout Europe and in parts of Asia. This study examined the molecular genetic diversity and population structure of 402 genotypes including 143 wild individuals, 239 landraces, and 20 cultivars from the Turkish national hazelnut collection using simple sequence repeat (SSR) markers. A total of 30 SSR markers yielded 407 polymorphic fragments. Diversity analysis of the Turkish hazelnut genotypes indicated that they fell into three subpopulations according to ad hoc statistics and neighbor-joining algorithm. Although all cultivars clustered together, they overlapped with the wild accessions and landraces. Thus, the dendrogram, principal coordinate, and population structure analyses suggest that they share the same gene pool. A total of 78 accessions were selected as a core set to encompass the molecular genetic and morphological diversity present in the national collection. This core set should have priority in preservation efforts and in trait characterization.Article Citation - WoS: 20Citation - Scopus: 24Newly Developed Ssr Markers Reveal Genetic Diversity and Geographical Clustering in Spinach (spinacia Oleracea)(Springer Verlag, 2017) Göl, Şurhan; Göktay, Mehmet; Allmer, Jens; Doğanlar, Sami; Frary, AnneSpinach is a popular leafy green vegetable due to its nutritional composition. It contains high concentrations of vitamins A, E, C, and K, and folic acid. Development of genetic markers for spinach is important for diversity and breeding studies. In this work, Next Generation Sequencing (NGS) technology was used to develop genomic simple sequence repeat (SSR) markers. After cleaning and contig assembly, the sequence encompassed 2.5% of the 980 Mb spinach genome. The contigs were mined for SSRs. A total of 3852 SSRs were detected. Of these, 100 primer pairs were tested and 85% were found to yield clear, reproducible amplicons. These 85 markers were then applied to 48 spinach accessions from worldwide origins, resulting in 389 alleles with 89% polymorphism. The average gene diversity (GD) value of the markers (based on a GD calculation that ranges from 0 to 0.5) was 0.25. Our results demonstrated that the newly developed SSR markers are suitable for assessing genetic diversity and population structure of spinach germplasm. The markers also revealed clustering of the accessions based on geographical origin with clear separation of Far Eastern accessions which had the overall highest genetic diversity when compared with accessions from Persia, Turkey, Europe, and the USA. Thus, the SSR markers have good potential to provide valuable information for spinach breeding and germplasm management. Also they will be helpful for genome mapping and core collection establishment.Article Citation - WoS: 1Citation - Scopus: 1Genetic Diversity and Population Structure of Faba Bean (vicia Faba) From Turkey(American Society for Horticultural Science, 2016) Göl, Şurhan; Doğanlar, Sami; Frary, AnneFaba bean (Vicia faba L.) is an important food and feed legume because of the nutritional value of its seed protein and starch content, good biomass, and high efficiency nitrogen fixation.This study analyzed the molecular genetic diversity and population structure of 101 Turkish faba bean accessions using simple sequence repeat (SSR) markers.A total of 32 SSR markers yielded 281 loci of which 277 (98.6%) were polymorphic.Mean dissimilarity between genotypes was 0.355, a relatively high value which was expected given faba bean’s large genome and limited breeding history.According to the results, faba bean individuals grouped into three main clusters based on both distance matrix (neighbor-joining algorithm) and model-based (population structure) clustering analyses.Clustering was not correlated with seed size or origin within Turkey.Six cultivars were included in the analysis and showed high genetic diversity compared with the landraces, as expected, given the fact that at least some of the cultivars were bred by hybridization.A total of 47 individuals were selected for the core collection to represent the diversity of Turkish faba bean germplasm.This core set encompasses material adapted to all growing regions and should be a priority for morphological characterization. © 2016, American Society for Horticultural Science.All rights reserved.Article Citation - WoS: 35Citation - Scopus: 37Molecular Genetic Diversity and Association Mapping of Morphine Content and Agronomic Traits in Turkish Opium Poppy (papaver Somniferum) Germplasm(Springer Verlag, 2016) Çelik, İbrahim; Camcı, Hüseyin; Köse, Arzu; Koşar, Ferda Çelikoğlu; Doğanlar, Sami; Frary, AnneAs the sole plant source of many potent alkaloids, opium poppy (Papaver somniferum L.) is an important medicinal crop. Nevertheless, few studies have characterized opium poppy germplasm with crop-specific molecular markers. Because Turkey is a diversity center for opium poppy, Turkish germplasm is a valuable genetic resource for association mapping studies aimed at identifying QTLs controlling morphine content and agronomic traits. In this study, the morphological diversity and molecular diversity of 103 Turkish opium poppy landraces and 15 cultivars were analyzed. Potentially useful morphological variation was observed for morphine content, plant height, and capsule index. However, the landraces exhibited limited breeding potential for stigma number, and seed and straw yields. Both morphological and molecular analyses showed distinct clustering of cultivars and landraces. In addition, a total of 164 SSR and 367 AFLP polymorphic loci were applied to an opium poppy association mapping panel composed of 95 opium poppy landraces which were grown for two seasons. One SSR and three AFLP loci were found to be significantly associated with morphine content (P < 0.01 and LD value (r2) = 0.10–0.32), and six SSR and 14 AFLP loci were significantly associated with five agronomic traits (plant height, stigma number, capsule index, and seed and straw yields) (P < 0.01 and LD value (r2) = 0.08–0.35). This is the first report of association mapping in this crop. The identified markers provide initial information for marker-assisted selection of important traits in opium poppy breeding.Article Citation - WoS: 27Citation - Scopus: 31Development of Genomic Simple Sequence Repeat Markers in Opium Poppy by Next-Generation Sequencing(Springer Verlag, 2014) Çelik, İbrahim; Gültekin, Visam; Allmer, Jens; Doğanlar, Sami; Frary, AnneOpium poppy (Papaver somniferum L.) is an important pharmaceutical crop with very few genetic marker resources. To expand these resources, we sequenced genomic DNA using pyrosequencing technology and examined the DNA sequences for simple sequence repeats (SSRs). A total of 1,244,412 sequence reads were obtained covering 474 Mb. Approximately half of the reads (52 %) were assembled into 166,724 contigs representing 105 Mb of the opium poppy genome. A total of 23,283 non-redundant SSRs were identified in 18,944 contigs (11.3 % of total contigs). Trinucleotide and tetranucleotide repeats were the most abundant SSR repeats, accounting for 49.0 and 27.9 % of all SSRs, respectively. The AAG/TTC repeat was the most abundant trinucleotide repeat, representing 19.7 % of trinucleotide repeats. Other SSR repeat types were AT-rich. A total of 23,126 primer pairs (98.7 % of total SSRs) were designed to amplify SSRs. Fifty-three genomic SSR markers were tested in 37 opium poppy accessions and seven Papaver species for determination of polymorphism and transferability. Intraspecific polymorphism information content (PIC) values of the genomic SSR markers were intermediate, with an average 0.17, while the interspecific average PIC value was slightly higher, 0.19. All markers showed at least 88 % transferability among related species. This study increases sequence coverage of the opium poppy genome by sevenfold and the number of opium poppy-specific SSR markers by sixfold. This is the first report of the development of genomic SSR markers in opium poppy, and the genomic SSR markers developed in this study will be useful in diversity, identification, mapping and breeding studies in opium poppy.Article Citation - WoS: 6Citation - Scopus: 8Molecular Genetic Diversity in the Turkish National Melon Collection and Selection of a Preliminary Core Set(American Society for Horticultural Science, 2013) Frary, Anne; Şığva, Hasan Özgür; Tan, Ayfer; Taşkın, Tuncer; İnal, Abdullah; Mutlu, Sevgi; Haytaoğlu, Mehmet; Doğanlar, SamiTurkey is a secondary center of diversity for melon (Cucumis melo) and is home to a variety of regional morphotypes. This diversity is housed in a national germplasm repository with more than 500 accessions. Molecular genetic variability of 209 melon genotypes from 115 accessions of this collection was characterized using amplified fragment length polymorphisms (AFLPs). Ten AFLP primer combinations yielded 279 reproducible fragments, which were used for dendrogram and principal coordinate analyses. These analyses showed two major clusters of Turkish melons: one group contained highly similar genotypes (maximum Dice dissimilarity coefficient of 0.18), whereas the other group was genetically more diverse (maximum dissimilarity 0.41). Although average dissimilarity was low (0.13), a broad range of genetic diversity was observed in the collection. A marker allele richness strategy was used to select a core set of 20 genotypes representing the allelic diversity of the AFLP data. The core set had double the average diversity (0.26) of the entire set and represented the major morphotypes present in the collection. Molecular genetic diversity of the core set was further validated using simple sequence repeat marker data (116 polymorphic fragments), which confirmed that the selected core set retained high levels of molecular genetic diversity.Article Citation - WoS: 24Citation - Scopus: 26Development of Est-Ssr Markers for Diversity and Breeding Studies in Opium Poppy(John Wiley and Sons Inc., 2013) Şelale, Hatice; Çelik, İbrahim; Gültekin, Visam; Allmer, Jens; Doğanlar, Sami; Frary, AnneAll publicly available opium poppy expressed sequence tag (EST) sequences, totalling 20 885, were assembled into unigenes and examined for simple sequence repeats (SSRs). Nearly 19% of the 14 957 unigenes contained SSRs with 4% harbouring more than one SSR. Average density of the SSRs was 1 SSR per 3.6 kb of non-redundant EST sequence. Trinucleotide SSRs were most frequently identified (39%), and many of the most prevalent motifs were AT-rich. Flanking primers were designed for 86% of the SSRs and 67 primer pairs were tested on 37 opium poppy accessions and seven related species. All markers were transferable to the related species. Polymorphism information content (PIC) values for the markers were intermediate for comparisons within opium poppy (average of 0.27) and slightly higher for comparisons across species (average of 0.29). The markers were found to be useful for diversity analysis as they successfully distinguished among Turkish opium poppy accessions and land races.
