Bahattin Tanyolac

Inter-simple sequence repeat (ISSR) and RAPD variation among wild barley (Hordeum. vulgare subsp. spontaneum) populations from west Turkey

Inter-Simple Sequence Repeat (ISSR) and Randomly Amplified Polymorphic DNA (RAPD) markers were used to analyze genetic distance among H. vulgare subsp. spontaneum populations from west Turkey. Fifty-five RAPD and 10 ISSR primers were used to detect variation among sample. A total of 55 polymorphic loci were found using 65 primers. Two distinct cluster groups were clearly established among populations. The minimum variation was detected between Pinarbasi and Bornova (GD = 0.192) populations and the maximum was found between Icmeler and Aydin populations (GD = 0.926). As two dominant markers, RAPD and ISSRs are effective and promising marker systems for detecting genetic variation.

SNP Discovery by Illumina-Based Transcriptome Sequencing of the Olive and the Genetic Characterization of Turkish Olive Genotypes Revealed by AFLP, SSR and SNP Markers

Background The olive tree (Olea europaea L.) is a diploid (2n = 2x = 46) outcrossing species mainly grown in the Mediterranean area, where it is the most important oil-producing crop. Because of its economic, cultural and ecological importance, various DNA markers have been used in the olive to characterize and elucidate homonyms, synonyms and unknown accessions. However, a comprehensive characterization and a full sequence of its transcriptome are unavailable, leading to the importance of an efficient large-scale single nucleotide polymorphism (SNP) discovery in olive. The objectives of this study were (1) to discover olive SNPs using next-generation sequencing and to identify SNP primers for cultivar identification and (2) to characterize 96 olive genotypes originating from different regions of Turkey. Methodology/Principal Findings Next-generation sequencing technology was used with five distinct olive genotypes and generated cDNA, producing 126,542,413 reads using an Illumina Genome Analyzer IIx. Following quality and size trimming, the high-quality reads were assembled into 22,052 contigs with an average length of 1,321 bases and 45 singletons. The SNPs were filtered and 2,987 high-quality putative SNP primers were identified. The assembled sequences and singletons were subjected to BLAST similarity searches and annotated with a Gene Ontology identifier. To identify the 96 olive genotypes, these SNP primers were applied to the genotypes in combination with amplified fragment length polymorphism (AFLP) and simple sequence repeats (SSR) markers. Conclusions/Significance This study marks the highest number of SNP markers discovered to date from olive genotypes using transcriptome sequencing. The developed SNP markers will provide a useful source for molecular genetic studies, such as genetic diversity and characterization, high density quantitative trait locus (QTL) analysis, association mapping and map-based gene cloning in the olive. High levels of genetic variation among Turkish olive genotypes revealed by SNPs, AFLPs and SSRs allowed us to characterize the Turkish olive genotype.

Identification QTLs Controlling Genes for Se Uptake in Lentil Seeds

Lentil (Lens culinaris Medik.) is an excellent source of protein and carbohydrates and is also rich in essential trace elements for the human diet. Selenium (Se) is an essential micronutrient for human health and nutrition, providing protection against several diseases and regulating important biological systems. Dietary intake of 55 μg of Se per day is recommended for adults, with inadequate Se intake causing significant health problems. The objective of this study was to identify and map quantitative trait loci (QTL) of genes controlling Se accumulation in lentil seeds using a population of 96 recombinant inbred lines (RILs) developed from the cross “PI 320937” × “Eston” grown in three different environments for two years (2012 and 2013). Se concentration in seed varied between 119 and 883 μg/kg. A linkage map consisting of 1,784 markers (4 SSRs, and 1,780 SNPs) was developed. The map spanned a total length of 4,060.6 cM, consisting of 7 linkage groups (LGs) with an average distance of 2.3 cM between adjacent markers. Four QTL regions and 36 putative QTL markers, with LOD scores ranging from 3.00 to 4.97, distributed across two linkage groups (LG2 and LG5) were associated with seed Se concentration, explaining 6.3–16.9% of the phenotypic variation.

Association mapping for five agronomic traits in the common bean (Phaseolus vulgaris L.).

BACKGROUND The common bean is the most important grain legume and a major source of protein in many developing countries. We analysed the following traits: pod fibre (PF), seeds per pod (SPP), plant type (PT), growth habit (GH), and days to flowering (DF) for a set of diverse common bean accessions and determined whether such traits were associated with amplified fragment length polymorphism (AFLP), simple sequence repeat (SSR) and single nucleotide polymorphism (SNP) markers. RESULTS In this study, 66 common bean genotypes were used and genotyped with 233 AFLP, 105 SNP and 80 SSR markers. The association analysis between markers and five traits was performed using a General Linear Model (GLM) in Trait Analysis by aSSociation, Evolution and Linkage (TASSEL). The population structure was determined using the STRUCTURE software, and seven groups (K = 7) were identified among genotypes. The associations for such traits were identified and quantified; 62 markers were associated with the five traits. CONCLUSION This study demonstrated that association mapping using a reasonable number of markers, distributed across the genome and with the appropriate number of individuals harboured to detect DNA markers linked to the traits of PF, SPP, PT, GH and DF in common bean.

Induction of Gentiana cruciata hairy roots and their secondary metabolites

Gentiana cruciata L. (cross gentian) is a medicinal and ornamental plant. The root extracts of this species are known to exhibit many curative properties. The natural Gentiana populations are exposed to great danger because of their uncontrolled usage. In this study, hairy roots from Gentiana cruciata L. stem and leaf explants belonging to three different clones were induced by inoculation with four different Agrobacterium rhizogenes wild strains namely A4, 15834, 8196 and R1000. Induction of the root transformation was significantly dependent on the explant type used. On the other hand, the genotype and bacterial strain had no significant effect on hairy root formation. Hairy root formation percentages of the explants varied between 5.6–33.3% in the stem explants, and between 0.0–6.7% in the leaf explants. Transformations of the hairy roots were confirmed by PCR using rolC specific primers, and revealed the absence of contaminating A. rhizogenes with virC primers. Total of twelve hairy root clones were obtained, and their secondary metabolite content was also analyzed by HPLC. Quantitative results exhibited that gentiopicroside was the most abundant compound in all root samples. Furthermore, metabolites such as loganic acid, swertiamarin, and sweroside were also identified and quantified in the samples.

ALTERED LEVEL OF APURINIC/APYRIMIDINIC ENDONUCLEASE/REDOX FACTOR-1 (APE/Ref-1) mRNA IN THE HIPPOCAMPUS OF OVARIECTOMIZED RATS TREATED BY RALOXIFENE AGAINST KAINIC ACID

1. Accumulated clinical evidence suggests that selective oestrogen receptor modulators (SERM), such as raloxifene, may be neuroprotective. Oxidative stress is a likely molecular mechanism in the neurotoxicity of kainic acid (KA), an excitotoxic substance. The expression levels of the apurinic/apyrimidinic endonuclease/redox factor‐1 (APE/Ref‐1) gene seem to correlate with cellular sensitivity to reactive oxygen species (ROS) and a reduction in the expression of APE/Ref‐1 may cause oxidative DNA damage.

Genetic assessment of common bean (Phaseolus vulgaris L.) accessions by peroxidase gene-based markers.

BACKGROUND Peroxidase, a plant-specific oxidoreductase, is a heme-containing glycoprotein encoded by a large multigenic family in plants. Plant peroxidases (POXs, EC 1.11.1.7) play important roles in many self-defense interactions in plants. Here, 67 common bean (Phaseolus vulgaris L.) genotypes were studied using a POX gene-based marker method. Comparison of POX genes could resolve evolutionary relationships in common bean. RESULTS Eighty fragments were obtained with 20 primer pairs that amplified one (POX8c) to eight (ATP29) bands, with a mean of four bands per primer pair. The average (polymorphic information content) PIC value for the POX products was 0.40. The maximum variation (93%) was found between Turkey (#33) and India (#52) and between Antalya (#33) and India (#53). The minimum variation (0%) was found among four pairs: Bozdag (#2) and Karadeniz (#38), Kirklareli (#11) and Turkey (#15, 16, 43), Bandirma (#13) and Turkey (#15, 16, 43), and Kirklareli (#10) and Bandirma (#22). UPGMA was used to discriminate the common bean genotypes into five clusters, while STRUCTURE software was used to investigate the genetic population structure. CONCLUSION The results showed that POX gene family markers can be used to study genotypic diversity and provide new information for breeding programs and common bean improvement practices.

Genetic variation among pathotypes of Verticillium dahliae Kleb. from cotton in western Turkey revealed by AFLP

Abstract Cotton (Gossypium hirsutum L.) is crucial for the textile industry worldwide. Among the diseases attacking cotton, Verticillium wilt caused by Verticillium dahliae Kleb. is the most significant. Isolates of V. dahliae can be classified into defoliating and non-defoliating pathotypes. Thirty-two isolates of the non-defoliating pathotype and one isolate of a virulent, defoliating pathotype were analysed by the amplified fragment length polymorphism (AFLP) method. Three hundred and forty AFLP fragments were obtained with nine primer combinations. The number of total bands per primer pair ranged from 16 to 81, with an average of 37.7. The average polymorphism information content (PIC) value for the AFLP products was 0.50. Using the genotypic data, genetic distance analysis was performed. The maximum variation was found between isolates (Vd11) Nazilli and (Vd16) Soke, at a value of 0.79 and the minimum variation was found between isolates (Vd20) Aydin and (Vd14) Soke, at 0.24. The unweighted paired group method with arithmetic averages cluster analysis (UPGMA) was used to discriminate the V. dahliae isolates into five subgroups. Defoliating pathotypes (Vd33) from Soke province formed a single subgroup. As a result, it was found that there was significant variation among Verticillium isolates. AFLP analysis is an efficient and effective marker technology for determining genetic relationships among Verticillium isolates.

Some characteristics and isolation of novel thermostable β-galactosidase from Thermus oshimai DSM 12092

The β-galactosidases belong to the class of hydrolytic enzymes and have been used as lactose hyrolysis. The enzyme is used in reducing lactose milk production, an outstanding industrial product used by a large lactoseintolerant population. This is the first detailed report of some characteristics of β-galactosidase and the gene encoding β-galactosidase in Thermus oshimai DSM 12092. The growth rate (μ, 1/h), and the doubling time (tD, h) for T. oshimai grown both in shaking flasks and in a bioreactor were determined. The optimal temperature and pH for β-galactosidase were determined as 75°C and 7.4, respectively. This enzyme was thermostable and was retained by more than 70% at 90°C for 3 h. The β-galactosidase from T. oshimai DSM 12092 was more stable in basic pH and Zn2+ was the most effective divalent cation. Also, 2 steps of purification consisting of ammonium sulfate precipitation and gel filtration were employed and purified 32-fold.

Low genetic diversity in wild emmer (T. turgidum L. subsp. dicoccoides (Körn. ex Asch. et Graebn.) Thell.) from South-eastern Turkey revealed by Restriction Fragment Length Polymorphism

The present work was carried out to study genetic diversity among 17 populations of wild emmer wheat sampled from South-eastern Turkey, considered to be an important region for domestication of wheat. Eleven RFLP clones and 4 restriction enzymes combinations were used to probe the genomic DNA. A total of 151 polymorphic loci were obtained from the enzyme-probe combinations. The Genetic Distance (GD) values were from 0.019 (Gaziantep-3 and Sanliurfa-4) to 0.200 (Gaziantep-1 and K. Maras). Cluster analysis results showed that populations formed 2 clades within the dendrogram. Population Gaziantep-1 was unique and genetically most diverse from the remaining 16 populations. The results of average genetic distance (GD) among populations suggested that narrow genetic variability exist among 17 populations in the present study.