Alkaline, Thermostable,Thermophilic and Chelator Resistant Type II Pullulanase from Native Isolate Anoxybacillus rupiensis strain NP2

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6 TH

_{INTERNATIONAL MOLECULAR}

BIOLOGY and BIOTECHNOLOGY

CONGRESS

22 - 25 December 2017

ABSTRACT BOOK

“ SCIENCE CENTER OF TURKEY ”

NOBEL

SCIENCE

www.molbiotech.gen.tr

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CONTENT

Foreword ...3

Organizing Committee ...4

Scientific Committee ...5

Oral Presentation ... 8-53

Poster Presentation ... 54-91

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Global populaton growth in the 21st century and earth’s limited natural resources

present great challenges to all of humanty. This Conference aims to highlight advances

in fundamental and applied biological research that may provide answers to a sustianable

future. Molbiotech will cover advances in Molecular Biology and Biotechnology

in Agriculture, Microbiology, Plant, Animal, Aquatic, Environment, Medicine

and Industry.

We are very happy that you are attanding this meeting.

Please let us know if there is anything further we can do.

Sincerely,

Prof. Dr. Mehmet Karatas

Chair of Congress

Necmettin Erbakan Universty

Department of Botechnology

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Organization Committee

Chairman of Congress

Prof. Dr. Mehmet KARATAŞ, Necmettin Erbakan University, Turkey

Congress Coordinator

Prof. Dr. Bashkim ZIBERI, University of Tetova, Macedonia

Chairman of Organizing Committee

Assoc. Prof. Dr. Zahit IQBAL, Isra University, Pakistan

Secretariat

Dr. Özhan ŞİMŞEK, University of Çukurova, Turkey

Shkurte LUMA, State University of Tetova, Macedonia

Mehtap AYDIN, Nobel Science and Research Center, Turkey

Organizing Commitee Members

Betul AKCESME, International University of Sarajevo, Bosnia and Herzegovina

Erna Karalija, University of Sarajevo, Bosnia and Herzegovina

Erhan Sulejmani , University of Tetova, Macedonia

Qenan Ferati, University of Tetova, Macedonia

Xhezair Abdiu, University of Tetova, Macedonia

Mohammed Mouradi GHARAKHLO, University of Zanjan, Iran

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Scientific Committee

Prof. Dr. Azra Bećiraj, University of Bihać, Bihać, Bosnia and Herzegovina

Prof. Dr. Güleray AĞAR, Atatürk University, TURKEY

Prof. Dr. Ijaz Javed HASAN, University of Agriculture Faisalabad, Pakistan

Prof. Dr. Halid Makić, University of Bihać, Bihać, Bosnia and Herzegovina

Prof. Dr. Mustafa Kamal, Karachi University, Pakistan

Prof. Dr. Nasir Bexheti , University of Tetova, Macedonia

Prof. Dr. Nexhbedin Beadini , University of Tetova, Macedonia

Prof. Dr. Sabino Aurelio BUFO, University of Basilicata, Italy

Prof. Dr. Kasim Bajrovic, University of Sarajevo, Bosnia and Herzegovina

Prof. Dr. Sezai TÜRKEL, Uludağ University, Turkey

Prof. Dr. Yıldız AKA KAÇAR, Çukurova University, Turkey

Prof. Dr. Zafar IQBAL , Karachi University, Pakistan

Prof. Dr. Izet Eminović, University of Sarajevo, Bosnia and Herzegovina

Assoc. Prof. Dr. Atilla KARSİ, Mississippi State University, USA

Assoc. Prof. Dr. Azra Skender, University of Bihać, Bosnia and Herzegovina

Assoc. Prof. Dr. Ali Rıza AWAN, University of Veterinary and Animal Sci, Pakistan

Assoc. Prof. Dr. Hesat Aliu, University of Tetova, Macedonia

Assoc. Prof. Muhammad Qasim Shahid, South China Agricultural University, China

Assoc. Prof. Dr. Sabina SEMİZ, University of Sarajevo, Bosnia and Herzegovina

Assoc. Prof. Dr. Jasmina Ibrahimpašić, University of Bihać, Bosnia and Herzegovina

Assoc. Prof. Dr. Zahit IQBAL, Isra University, Pakistan

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ABSTRACT for

ORAL

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RNA-SEQ TECHNOLOGIES IN PLANT SCIENCES

Yıldız Aka Kaçar

1

1_{Faculty of Agriculture, Department of Horticulture, Çukurova University, Adana, Turkey} ykacar@cu.edu.tr

Abstract

Plant breeding is an ancient technology with dynamic current techniques and an exciting future. There are a number of restraints to conventional plant breeding which are especially limiting in tree fruits with their long juvenile period, large plant size, and which are represented by unique, highly-selected heterozygous genotypes. Biotechnology offers to minimize disadvantages of classical breeding techniques. In this sense, plant breeding refers to the purposeful genetic improvement of plant crops through various techniques in-cluding selection, hybridization, mutation induction, and molecular techniques. Among molecular techniques, sequencing technology have been used for many years and recently a new concept titled “RNA-Seq” have been started to performed to understand molecular mechanisms in plants. RNA-Seq analysis is an effective tool to understand which genes involved and expressed in different mechanisms and organs/cells of a plant. Recently, many articles have been published using RNA-Seq in plants.

Key words: Anthers, Microspore, Uni nuclear, DAPI, B5 medium

N-GLYCAN RELEASE MANIPULATION OF A NOVEL

ENDO-Β-N-ACETYLGLUCOSAMINIDASE BY USING GENETICALLY ATTACHED TAGS

Sercan Karav

1

1_{Department of Molecular Biology and Genetics, Canakkale Onsekiz Mart University, 17100 Canakkale, Turkey.}

sercankarav@gmail.com

Abstract

Glycans have very interesting biological roles (in addition to their contribution to the functions of glycoproteins) such as prebiotic activity, antimicrobial, cell to cell and cell to microbe signaling. Glycans are also selectively consumed by probiotics such as Bifidobacterium longum subsp. Infantis ATCC 15697 (B.infantis). In addition to their mutual functions, specific glycans such as sialylated glycans play important role in brain development and learning skills of infants. In this research, recombinantly produced three form of a novel endo-β-N-acetylglucosaminidase (free, genetically attached Glutatiohine-S-transferase and Hexahistide) was used for the release of these bioactive N-glycans from bovine whey proteins. The performance of these forms of enzymes on total N-glycan release and the diversity of these N-glycans were compared. It was shown that, free and His-tagged EndoBI-1 showed similar enzymatic performance on the release of glycans (0.53 and 0.48 ml/mg x min-1_{, respectively), whereas GST-tagged enzyme showed a low enzyme activity with 0.07} ml/mg x min-1_{N-glycan release efficiency. The released glycans were characterized by an advance mass spectrometry; the nano liquid} chromatography Chip Quadrupole Time of Flight Mass Spectrometry. Based on the results, free and his-tagged enzyme were able to release 38 and 37 N-glycan structures respectively, while GST-tagged enzyme resulted in only 25 N-glycan compositions. Interestingly, this difference was the result of the sialylated glycan release performance of these forms of enzymes. GST-tagged enzyme was able to release only 2 sialylated glycans, whereas both free and His-tagged enzymes resulted in 15 silaylated glycans. These results suggested that different tags can be applied in order to produce a specific glycan type enriched pool such as sialylated or neutrals.

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PLANT GROWTH PROMOTING PROPERTIES OF ENDOPHYTIC BACTERIA ISOLATED

FROM IN VITRO CULTURES OF PEACH AND PEAR ROOTSTOCKS

Fakhra Liaqat

1

_{Rengin Eltem}

2

1_{Ege University, Graduate School of Natural and Applied Sciences, Department of Biotechnology, Izmir, Turkey}

2_{Ege University, Faculty of Engineering, Department of Bioengineering, Izmir, Turkey}

fakhra243@gmail.com

Abstract

Endophytes are microorganisms which live symbiotically with almost all varieties of plant and in turn helping the plant in a number of ways. Instead of satisfactory surface sterilization approaches, repeatedly occurring bacterial growth on in vitro rootstock cultures of peach and pear was identified and isolated as endophytic bacteria in our current study. Five different isolates from peach rootstocks were molecularly identified by 16S rRNA gene sequencing as Brevundimonas diminuta, Leifsonia shinshuensis, Sphingomonas parapaucimobilis Brevundimonas vesicularis, Agrobacterium tumefaciens while two endophytic isolates of pear were identified as Pseudoxanthomonas mexicana, and Stenotrophomonas rhizophilia. Identified endophytes were also screened for their plant growth promoting potential in terms of indoleacetic acid (IAA) production, nitrogen fixation, phosphate solubilization and siderophore production. All seven endophytic isolates have shown positive results for IAA, nitrogen fixation and phosphate solubilization tests. However, two out of seven isolates showed positive results for siderophore production. On the basis of these growth promoting competences, isolated endophytes can be anticipated to have significant influence on the growth of host plants. Future studies required to determine the antimicrobial susceptibility profile and potential application of these isolates in biological control, microbial biofertilizers and degradative enzyme production.

Keywords: Endophyte, Peach rootstock, Pear rootstock, Plant growth promoting properties,

CYTOTOXIC EFFECTS OF LINARIACONFERTIFLORA EXTRACTS ON K562

MYLEOMONOCYTIC LEUKEMIA CELL LINE

Faruk Süzergöz

1

_{Ayfer Özçelik}

1

_{H. A. Zafer Sak}

2

1_{Department of Biology, Faculty of Art and Science, Harran University, Sanliurfa, Turkey} 1_{Department of Chest Disease, Faculty of Medicine, Harran University, Sanliurfa, Turkey}

suzergoz@harran.edu.tr

Abstract

The determination of the therapeutic use capacities of plant sources has recently become an important topic for drug research. Turkey is very rich in terms of endemic plants; it is a great necessity to investigate into this richness in a way that contributes tohuman health, science and country economy. In this study, the cytotoxic effects of Linariaconfertifloraplant extracts against cancer cells were investigated.Methanolic extracts of flowers, leaves, stem and roots of L. confertiflora,endemic for Turkey, were prepared. Methanolic extracts were tested on K562 myelocytic leukemia cells. The MTT method was used to determine the IC50 values of the methanolic extracts that wasprepared from each parts of plant. The plant extracts were used at 1000 μg/ml, 100 μg/ml, 10 μg/ml, 1 μg/ml doses in triplicate order. The strongest cytotoxic effects for L. confertiflora were obtained from the stem extract of the plant. The IC50 value ofL. confertiflora was1.76 μg/ml. Although our findings need to be supported by further studies, it has been observed that L. confertiflora have promising potential for cancer therapy. The screening of the therapeutic properties of the endemic plants in our country has great importance in terms of human health and our country’s economy.

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LOW-AFFINITY CAMP PHOSPHODIESTERASE ACTIVITY ESSENTIAL FOR NTH1 GENE

EXPRESSION IN SACCHAROMYCES CEREVISIAE

Tülay Turgut Genç

1

_{Mehmet Şerafeddin Solak}

2

1_{Department of Biology, Faculty of Arts and Science, Çanakkale Onsekiz Mart University, Çanakkale, Turkey} 2_{Graduate School of Natural and Applied Sciences, Çanakkale Onsekiz Mart University, Çanakkale, Turkey}

tturgutgenc@comu.edu.tr

Abstract

Saccharomyces cerevisiae includes two genes, PDE1 and PDE2. PDE1 encodes low affinity phosphodiesterase that controls glucose and intracellular acidification-induced cAMP signalling and PDE2 encodes high affinity phosphodiesterase that protects the yeast cells from extracellular cAMP, controls the basal level of cAMP. The major determinant of stress resistance of the yeast S. cerevisiae is the Ras-cAMP pathway. Also it is already known that NTH1 is a multiple stress response gene. The regulation of NTH1 gene expression involves different signaling pathways including many transcriptional activators and repressors that regulate the expression of NTH1 gene at the transcriptional level. Despite the fact that regulation of this gene is controlled by a complicated pathway, the role of these activators and repressors in this pathway are not known yet. So that in our research we analyzed the effect of low affinity phosphodiesterase, Pde1, on NTH1 gene expression. The transcription of NTH1 gene decreased 2-fold in ∆pde1 yeast strains under normal growth conditions. Also NTH1 gene expression in ∆pde1 mutant yeast strains decreased 3-fold than wild type strain under nitrogen starvation. Similarly heat stres also caused to decrease gene expressin 2-fold in ∆pde1 mutant yeast strains than wild type strains. These results shows that the low affinity cyclic AMP phosphodiesterase activity and so active PKA necessary for NTH1 gene expression both in normal and stress conditions.

This work was supported by Çanakkale Onsekiz Mart University The Scientific Research Coordination Unit, Project number: FYL-2014-301.

Keywords: Saccharomyces cerevisiae, NTH1, Pde1, Stress

COMPARISON OF BISULFITE SEQUENCING AND WHOLE GENOME BISULFITE

SEQUENCING FOR EPIGENETIC RESEARCH IN PLANTS

Mehmet Karaca

1

_{Ayse Gül İnce}

2*

_{Emine U. Göçer}

1

_{Adnan Aydın}

1

1_{Department of Field Crops, Akdeniz University, 07070 Antalya, Turkey,} 2_{Vocational School of Technical Sciences, Akdeniz University, 07070 Antalya, Turkey}

*Corresponding author: aince@akdeniz.edu.tr

DNA cytosine methylation, as an important epigenetic mechanism involves in the control of all genetic functions such as DNA replication and repair, gene transposition and transcription, cell differentiation and gene silencing, and imprinting and affects vernalization, drought and salt tolerance, heterosis, bio-defense, transgenic expression and expression of foreign DNA in cell. DNA cytosine methylation is usually studied using two methods. BS is a first NGS and WGBS is a second next generation sequencing method. We used BS in several plant species including cotton and about to conduct epigenetic research with WGBS method. In this study we prove information on advantages and disadvantages of BS and WGBS methods for researchers employing epigenetic research. Briefly, BS method stats with treatment of genomic DNA with sodium bisulfite, unmethylated cytosine residues are converted to uracil whereas 5-methylcytosine (5mC) remains unaffected. After PCR amplification, uracil residues are converted to thymine. PCR amplified products are cloned and sequenced using Sanger sequences method. WGBS starts with random fragmentation of DNA into small fragments and adapters are ligated to the fragments. DNA fragments are sodium bisulfite treated and are enriched with PCR. Size selected library fragments are sequenced using one of the NGS technologies. These two sequencing techniques although quite different in many aspects could be used in the same research. In the present study we discuss some technical aspects of these first and second generation bisulfite sequencing methods with drawings and tables.

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IN VITRO PROPAGATION OF TRUE SEEDS OF POTATO (SOLANUM TUBEROSUM L.)

Gülsüm Öztürk

1

1_{Ege University, Faculty of Agriculture, Department of Field Crops, Bornova-Izmir, TURKEY}

gulsum.ozturk@ege.edu.tr

Abstract

The purpose of the study was to select suitable medium to propagate potato true seeds to be used in the various studies in the future. The study was conducted in the Tissue Culture Laboratory and potato production seedbeds of the Field Crops Department of the Faculty of Agriculture of the Ege University in 2016 and 2017. Tubers of the potato genotype 101 (Nif), a release of the Department, were grown in seedbeds and the fruits were collected. The fruits were squeezed in water and the true seeds were obtained and stored to break their dormancy. The true seeds were cultured in the test tubes of the MS medium (control) and other 25 different nutrient medium supplemented with GA₃, IAA, BAP (0.1, 0.5, 1, 2, 3 mg/l), IAA, BAP (0.1, 0.5, 1, 2, 3 mg/l)+GA₃(0.1 mg/l). The regenerations of plantlets were evaluated. Based on the results; the highest plantlet height was obtained from the MS+0.1 mg/l IAA medium (8.4 cm); the highest regenerant number from the MS+1, 2 mg/l BAP medium and the MS+2, 3 mg/l BAP+0.1 mg/l GA₃ medium with the range of 1.83 and 2.00. The highest root length was obtained from the MS+3 mg/l IAA+0.1 mg/l GA₃ medium (3.23 cm) and the highest root number was obtained from the MS+1 mg/l IAA+ 0.1 mg/l GA₃medium (3.0).

Keywords: True seed potato, plant growth regulators, in vitro propagation

INVESTIGATION OF BORON-RELATED GENES IN A HIGHLY BORON TOLERANT

ISOLATE SIMILAR TO PSEUDOMONAS ZHAODONGENSIS

Bekir Çöl Esra Dibek Merve Sezer Nihan Akgüç

Muğla SK University, Science Faculty, Biology Department, Muğla, Turkey

Abstract

Excess boric acid present life-threatening effects to the living organisms and are known to destroy most of them at certain concentrations. The level of boric acid levels in the Boron mines of Turkey is at the extreme amounts reaching even thousands of ppm values in some locations. The bacteria isolates that are found in Boron mines in Turkey in that regard must have acquired over the course of their evolutionary journey the necessary genes and molecular mechanisms employed to be used as extreme measures to live and function in such harsh conditions.

This current study aimed at proposing some gene activities found in a highly boron tolerant gram negative bacterium isolate belonging to Pseudomonas genus. Phylogenetic tree constructed with the 16SrRNA gene sequence of the isolate and closely related other taxa obtained from 16SrRNA database has indicated that the isolate is within the same branch with Pseudomonas zhaodongensis.

Using this bacterium as the model system and genomic-library selection-screening approach, we have investigated the genes that may be implicated in boron tolerance, and using Escherichia coli DH10b strain as the receiving bacterium, found a number of eleven recombinant plasmids carrying the genes from the Pseudomonas zhaodongensis-similar isolate. Sanger sequencing of the insert regions and bioinformatic analyzes suggested partly or fully the following activities namely sugar ABC transporter permease, membrane-bound lytic murein transglycosylase, excinuclease ABC subunit B, endonuclease III, integrase, 2-oxoglutarate dehydrogenase E1 component, pseudouridylate synthase I, putative cation efflux system protein (Na+/ H+ ion antiporter family protein), flagellin modification protein FlmH, Ca2+/Na+ antiporter (K+dependent Na+ exchanger-like protein), spore coat polysaccharide biosynthesis protein (glycosyltransferase), thiamine-phosphate pyrophosphorylase, glutamate-1-semialdehyde 2,1-aminomutase and bifunctional hydroxy-methylpyrimidine kinase.

Acknowledgement: This study is supported by The Scientific and Technological Research Council of Turkey (TÜBİTAK, projects

114Z987 and 112T614).

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PHYLOGENETIC ANALYSIS OF NONOMURAEA SP. GG64 ISOLATED FROM THE ACIGÖL

SOIL, NEVŞEHIR

Talha Gençbay

1

_{Kamil Işik}

1

_{Ahmet Rıdvan Topkara}

1

_{Salih Saricaoğlu}

2

1_{Department of Biology, Faculty of Art and Science, Ondokuz Mayis University, Samsun, Turkey.} 2_{Ahi Evran University, School of Health, Nursing Department, Kırşehir}

mr.gencbay@hotmail.com

Abstract

Actinomycetes are widely distributed in terrestrial and aquatic ecosystems, especially in soil, where they play a crucial role in the recy-cling of refractory biomaterials by decomposition and humus formation. The species of Nonomuraea are widely distributed in soil. In addition, some of them show the potential ability to produce bioactive metabolites such as maduramycin, pyralomicin, brartemicin, daunomycin and gluvirucin B₀. The strain GG64 was isolated from a soil sample collected from Nevşehir, Turkey, using czapek’s dox agar. This onging study was aimed to determine taxonomic position of Nonomuraea sp. GG64 isolated from soil of the Acıgöl using by polyphasic approach. Strain GG64 was isolated using the standard dilution plate method. Obtained results from 16S rRNA gene sequence comparison showed that the strain GG64 was most closely related to the type strains of Nonomuraea rhizophila (16/1463;

98.91%), Nonomuraea solani (20/1461; 98.63%) and Nonomuraea monospora (22/1415; 98.45%). Based on the results of 16S rRNA gene sequence analyses, the isolate GG64 will have a novel species position in the literature after completion of polar lipids isoprenoid quinones analysis and phentypic characterization. A polyphasic approach will promptly be implemented to determine exact positions of the strain GG64 from Turkish soil.

Keywords: Actinobacteria, Nonomuraea , 16S rRNA gene sequencing

Acknowledgements: This research was supported by TÜBİTAK. (Project no: 115Z792)

CYTOTOXIC EFFECT OF EPIRUBICIN-HCL ON PARENTAL AND EPIRUBICIN-HCI

RESISTANT LIVER CANCER CELLS

Ayşe ERDOĞAN

1

_{Aysun ÖZKAN}

2

1_{Alanya Alaaddin Keykubat University, Faculty of Engineering, Genetic and Bioengineering Department, 07425, Alanya, Antalya,} Turkey

2_{Akdeniz University, Science Faculty, Biology Department, 07058, Antalya, Turkey} ayse.erdogan@alanya.edu.tr

Abstract

In this study, the cytotoxic effect of epirubicin-HCl, a derivative of doxorubicin anthracycline, was investigated in human liver cancer (P-Hep G2) and epirubicin-HCl resistant liver cancer (R-Hep G2) cells in which biotransformation of several substances and carcinogenic effect components were performed. Epirubicin-HCl (drug) resistant Hep G2 cells were obtained by exposing parental Hep G2 cells to epirubicin-HCl at increasing concentrations, step by step. The cytotoxic effect of epirubicin-HCl on parental and drug resistant Hep G2 cells for 72 h was demonstrated using the Cell Titer-BlueR_{Cell viability assay kit. Parental and drug resistant Hep G2 cells were exposed} to different concentration of epirubicin-HCl. The cytotoxic effect of epirubicin-HCl increased with concentrations on both parental and drug resistant cells. Parental Hep G2 cells were found to be more sensitive to the epirubicin-HCl cytotoxic effect than drug resistant Hep G2 cells. The results we have obtained are the first scientific study to demonstrate the differences in responses of epirubicin-HCl resistant Hep G2 cells and parental Hep G2 cells to epirubicin-HCl. The cytotoxic effect of epirubicin-HCl has changed depending on the epirubicin-HCl concentration and the drug resistance properties of the cells.

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IN VITRO PROPAGATION OF TRUE SEEDS OF POTATO (SOLANUM TUBEROSUM L.)

Gulsum Ozturk

1

Abstract

Keywords: True seed potato, plant growth regulators, in vitro propagation

GENETIC RELATIONSHIPS AMONG BUMBLE BEE (B. TERRESTRIS TERRESTRIS)

POPULATIONS IN TERMS OF MTDNA IN ANTALYA REGION

Kemal Karabağ

Department of Agricultural Biotechnology, Faculty of Agriculture, Akdeniz University, Antalya, 07058, Turkey karabag@akdeniz.edu.tr

Abstract

The purpose of this study is to determine the genetic relationships among commercial cultivated B. terrestris bee colonies and natural populations via mtDNA-CO1 using SNP haplotypes. Approximately 250 thousand commercial colonies are used extensively in greenhouses activities in the Antalya region. Also, Antalya is a region where natural bombus colonies are dense and therefore a rich genetic resource. However, hybridization with natural populations is a matter of course because the cultivated colonies from greenhouses to nature. Due to these reasons the situation needs to be clarified. For this purpose, worker bee samples were collected as project material from 3 commercial producers, from 3 greenhouse centers and from 3 natural areas which are at least 30 km away from the nearest greenhouse areas. Total of 34 SNP haplotypes were detected in a total of 237 DNA sequences that amplified in PCR using the B. Terrestris mtDNA-CO1 (Cytochrome Oxydase-1) gene-specific primers.

Phylogenetic analyzes using these haplotypes have shown that natural populations and commercial populations are clearly separated from each other. However, it has been observed that the commercial populations and the natural bee populations in the intensive greenhouses regions were genetically close to each other. From these results, it was concluded that the probability of gene flow to natural populations from commercial colonies.

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MOLECULAR CLONING OF STREPTOMYCES MOBARAENSIS

PRO-TRANSGLUTAMINASE GENE IN YEAST

Aysun Türkanoğlu Özçelik

1

_{Fatma Ersöz}

2

_{Mehmet İnan}

1,2

1_{Food Safety and Agricultural Research Center, Akdeniz University, Antalya, Turkey} 2_{Department of Food Engineering, Faculty of Engineering, Akdeniz University, Antalya, Turkey}

aysunozcelik@akdeniz.edu.tr

Abstract

Transglutaminases (EC 2.3.2.13) are enzymes that catalyse the acyl transfer reaction between the γ- carboxyl groups of protein-bound glutamine and primary amines. Transglutaminases promote the covalent modifications of proteins becuase of this property they are widely used in food, medicine, textile and leather industries. For instance, microbial transglutaminase (MTGase) from Streptomyces mobaraensis supports the crosslinking of meat and fish proteins and gelation in yoghurt and cheese. In addition, this enzyme is used for the improvement of rheological properties and mechanical strength of foods so the use of food additives will be reduced by this way. P. pastoris is a single cell methylotrophic yeast and grows rapidly on inexpensive media containing methanol, glucose, glycerol or ethanol as a sole carbon source. It is capable of many post-translational modifications, important for eukaryotic proteins, such as proteolytic processing, disulfide bridge formation and glycosylation. These properties make Pichia expression system more adventageous than bacteria for recombinant protein production.

In this study, Streptomyces mobaraensis MTGase gene was cloned into pGAPZαA vector. The ligation products were transformed into the E.coli XL1blue compotent cells. Selected transformant cells were digested with the restriction enzymes to confirm the cloning. Verified plasmids were transformed into the Pichia pastoris X33 compotent cells with the electroporation method to express recombinant MTGase protein.

Keywords: Pichia pastoris, microbial transglutaminase, GAP promoter

EMBRYOGENIC CALLUS INDUCTION, ORGANOGENESIS AND REGENERATION OF

PANCRATIUM MARITIMUM

Sara YASEMIN

1*

_{Nezihe KOKSAL}

2

_{Saadet BUYUKALACA}

2

1_{Siirt University, Faculty of Agriculture, Department of Horticulture, Kezer, Siirt, Turkey} 2_{Cukurova University, Faculty of Agriculture, Department of Horticulture, Balcalı, Adana, Turkey}

syasemin@mailzf.cu.edu.tr

Abstract

Pancratium maritimum (sea daffodil) L., a member of Amaryllidaceae family, is growing in sandy areas on coastal regions. Sea daffodil is defined as endangered because of increasing tourism and urbanization activities. Micropropagation aims to protect and maintain in endangered plants. Three different explant types (leaf, root and mature zygotic embryos) of P. maritimum were cultured on MS medium containing different plant growth regulators [2,4 D (1, 2, 4 mg l-1_{) and BAP (1 mg l}-1_{)] for callus induction. Callus formation rate,} callus growth rate, embryogenic callus rate, callus structure and callus color intensity were determined. After determination of induced calli, shoot formation was observed on MS medium including 2 mg l-1_{BAP and 0.2 and 0.5 mg l}-1_{2,4 D. Consequently, the best callus} induction were determined as zygotic embryo explants and medium containing together 2,4 D and BAP rather than only 2,4 D. In terms of shoot formation from callus, both MS media supplied with BAP (2 mg l-1_{) and 2,4D (0.5 and 0.2 mg l}-1_{) were successful (%90). In this} study, efficient embryogenic callus induction, organogenesis and regeneration protocols were developed for P. maritimum.

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MULTI-FRAGMENT MELTING ANALYSIS SYSTEM (MFMAS); A NOVEL APPROACH FOR

IDENTIFICATION OF LACTOBACILLUS SPECIES

Zülal Kesmen

1

_{Özge Kılıç}

1

_{Fatma Özge Özkök}

2

_{Mete Çelik}

2

1_{Erciyes University, Engineering Faculty, Department of Food Engineering, Kayseri, Turkey} 2_{Erciyes University, Engineering Faculty, Department of Computer Engineering, Kayseri, Turkey}

zkesmen@erciyes.edu.tr

Lactobacillus genus is one of the most studied group within lactic acid bacteria due to their indispensable functions in the production of fermented food products as well as their potential as starter and probiotic culture. Therefore, there is an increasing need for rapid and reliable methods, to be used in the identification of Lactobacillus species. For this reason, we developed a novel approach, MFMAS-Lactobacillus based on the simultaneous analysis of the characteristic melting curves of multiple DNA fragments that have inter-specific sequence heterogeneity. In this system, amplification and then melting analysis is individually performed for each target DNA fragment and a fingerprint is produced for each strain analyzed by combining the melting curve data of all the target fragments. MFMAS-Lactobacillus is a software-assisted system that allows visualizing, analyzing and storing fingerprints obtained from the melting analysis of multi-target DNA fragments. The identification of Lactobacillus species is performed by comparing the fingerprints of unknown species with those of known species registered in the database in a short time without any additional process. The system also produces a matching score, showing the identification quality according to the similarity rate between the data of known and unknown species. In conclusion, it is considered that the MFMAS-Lactobacillus is a promising system for one-step identification of the Lactobacillus species isolated from foods and process environments.

Keywords: Multi Fragment Melting Analysis System, (MFMAS), Lactobacillus, species identification

SNF2 HAS A ROLE IN TRANSCRIPTIONAL REGULATION OF NTH1 GENE IN

SACCHAROMYCES CEREVISIAE

Tülay TURGUT GENÇ

1

_{Nihan AKINCI}

1

_{and Sezai TÜRKEL}

2

_.

1_{Çanakkale Onsekiz Mart University, Faculty of Arts and Sciences, Department of Biology, 17020 Çanakkale, Turkey.} 2_{Uludag University, Faculty of Arts and Sciences, Department of Molecular Biology and Genetics, 16059- Bursa, Turkey.}

akincinihan@gmail.com

Abstract

Trehalose is a multi-functional sugar for S. cerevisiae. Trehalose continuously recycled to keep steady state level in yeast cytoplasm. Neutral trehalase enzyme which degrades trehalose, is essential for the trehalose recycling. Neutral trehalase is encoded by NTH1 and NTH2 genes. It is known that transcription NTH1 is activated by Msn2/4, stress activated transcription factors, that involves in the regulation of large set of genes in S. cerevisiae in response to physiological stresses. To be involved in continuous recycling of trehalose, NTH1 gene is also transcribed at steady state levels even under normal growth conditions. It is known that promotor regions of transcriptionally active genes are cleared by chromatin modifying complexes to provide access to transcription initiation complex. In this study, we have analyzed if the chromatin modifying complexes involves in the transcription of NTH1 gene under normal and also under stress inducing growth conditions. We tested the effect of Snf2p, catalytic subunit of Swi/Snf complex, on the transcriptional regulation of NTH1. We have found that NTH1 gene expression in Δsnf2 mutants was 6-fold higher than wild type yeast strain. Under nitrogen starvation NTH1 gene expression increased 8-fold in wild type yeast strain but no change was observed in Δsnf2 mutants. Interestingly, NTH1 gene expression increased 4- fold in wild type yeast strain and 2- fold in Δsnf2 mutant strain after addition glutamin, rich nitrogen source. Our results indicated that Swi/Snf nucleosome mobilizing complex is essential for the transcriptional regulation of NTH1 gene.

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PHYLOGENETIC TREES AND GENETIC VARIATION OF KAPPA-CASEIN (Κ-CASEIN)

GENE IN TURKISH GREY STEPPE CATTLE BREED.

Adem KABASAKAL

1

1_{Susurluk Vocational School, Balikesir University, Balikesir, Turkey.} ademkabasakal@hotmail.com

Abstract

Kappa-casein is an important fruction of casein in milk proteins. It is composed of kappa-casein of 80% of total casein. It is thought that there is a link between the κ-casein alleles of milk cattle with milk quantity and milk quality the composition of milk. As a matter of fact, it is observed that cows with κ-casein B allele give more milk to A allele. In this study, genomic DNA was isolated from blood samples taken from Turkish Grey steppe breed cattle. Polymerase chain reaction (PCR) was employed to amplify the κ-casein and POMC fragment from each genomic DNA sample. These fragments obtained were sequenced. Sequence analyses proved that there are A, A1, B, G2 and H alleles of κ-casein gene in Turkish Grey cattle breeds . A1 and H alleles of bovine κ-casein are described in Bos indicus and the G2 alleles are described in Bos grunniens. When phylogenetic trees are examined, it is seen that the κ-casein A and B alleles are located in opposite regions. It is seen that the phylogenetic trees are in a multi-branched structure and the outer groups used are distributed in the phylogenetic tree. Consequently, These results indicates that there may be a consanguinity between Grey cattle breed (Bos taurus) and both Bos indicus and Bos grunniens and were analysed this phylogenetic trees exhibited that in terms of milk yield there is a selection pressure on kappa casein gene of dairy cattle breeds.

Keywords: Turkish Grey Steppe Cattle Breed, κ-casein gene, Phylogenetic tree.

ISOLATION OF N

₂

-FIXING BACTERIA FROM AFŞIN-ELBISTAN LIGNITE MINE AND

THEIR MOLECULAR IDENTIFICATION

Medine Güllüce

1

_{Selma Sezen}

2

_{Hakan Özkan}

3

_{Mehmet Karadayı}

1

_{Burak Alaylar}

4

Mine İsaoğlu

4

_{Ceyda Işık}

4

1 _{Department of Biology, Faculty of Science, Atatürk University, Erzurum, Turkey} 2_{Institute of Health Sciences, Atatürk University, Erzurum, Turkey}

3 _{Department of Molecular Biology and Genetics, Faculty of Science, Atatürk University, Erzurum, Turkey} 4_{Institute of Natural and Applied Sciences, Atatürk University, Erzurum, Turkey}

selma-sezen@hotmail.com

Abstract

Nitrogen found in the structure of organic molecules as a critical element has a very crucial role in plant growth. Therefore, importance of studies on nitrogen-fixing bacteria has been increasing day by day. In this context, lignite samples for isolation of N₂-fixing strains were collected from Afşin-Elbistan lignite mining fields. The samples were asseptically transferred to the research laboratory and dilution series were prepared between 10-1_{and 10}-7_{. These dilutions were spread on Nitrogen Free Medium (NFM) agar plates, incubated for 48} h at 28 ºC. After the incubation period, the bacterial colonies growing on NFM were evaluated as N₂-fixing strains. Then, these active strains were identified by using conventional and molecular techniques. According to the results, 5 distinguished bacterial colonies were grown on NFM agar plates as N₂-fixing strains. Data of the 16S rDNA gene sequencing showed that these strains grouped in Arthrobacter, Bacillus, Pseudomonas and Rhodococcus genera and they were identified as 18: Arthrobacter humicola (%100), SLM-30: Bacillus subtilis (%100), SLM-8: Pseudomonas mandelli (%100), SLM-24: Pseudomonas mandelli (%100) and SLM-17: Rhodococcus sp. (%99).

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BIOSYNTHESIS OF CYNARA SCOLYMUS (ARTICHOKE) MEDIATED SILVER

NANOPARTICLES AND EVALUATION OF THEIR ANTIBACTERIAL ACTIVITY

Fatih ERCİ

1

_{Rabia ÇAKIR KOÇ}

2

_{Emrah TORLAK}

3

_{İbrahim IŞILDAK}

2

1_{Department of Biotechnology, Faculty of Science, Necmettin Erbakan University, Konya, Turkey}

2 _{Department of Bioengineering, Faculty of Chemical and Metallurgical Engineering, Yildiz Technical University, Istanbul, Turkey} 3_{Department of Molecular Biology and Genetics, Faculty of Science, Necmettin Erbakan University, Konya, Turkey}

ferci@konya.edu.tr

Abstract

Nanomaterials have various characteristics which make them excellent tools in various applications; for instance, their large surface area to volume ratio provides them with new mechanical, chemical, optical, magnetic, and electro-optical and magneto-optical features. Typically, non-biological methods for fabrication of nanoparticles involve external reducing agents and toxic organic solvents, which pose potential environmental and biological risks. Therefore, the use of biotechnology in the development of fast, clean, non-toxic and eco-friendly process for nanoparticle synthesis has gained great importance. Here, obtain C. scolymus mediated silver nanoparticles was synthesized by adding 5 ml of C. scolymus leaf extract to 100 mL of 1 mM AgNO3 solution. The reaction mixture was stirred at 60 °C (350 rpm), and light brown to dark brown color change indicated that silver ions could be reduced to silver nanoparticles. The synthesized nanoparticles were characterized by UV–vis spectroscopy, scanning electron microscope (SEM), energy dispersive X-ray spectroscopy (EDX) and transmission electron microscope (TEM) analysis. Also, zeta potential, particle size, polydispersity index (PDI) of nanoparticles were determined by using nanosizer. UV–vis spectra of the leaf extract-AgNO3 mixture reveal clear peak at 458 nm, which were consistent with the literature reports for AgNPs. The spherical nature of synthesized AgNPs was revealed by Scanning Electron Microscopy (SEM). TEM results show that biosynthesized nanoparticles have particle diameter with an average of 35 nm. The in vitro susceptibilities of the selected bacteria toward synthesized nanoparticles were determined by disc diffusion and broth microdilution methods. These biologically synthesized AgNPs from leaf extracts of C. scolymus displayed significant antibacterial activity against two different gram-negative and gram-positive bacteria.

Keywords: Biosynthesis, Artichoke, Silver nanoparticles, Antibacterial activity

DETERMINATION OF PROTECTIVE EFFECT OF Β-ESTRADIOL AGAINST SALINITY

STRESS USING REMAP TECHNIQUE IN TRITICUM AESTIVUM L.

Semra Yagci

1

_{Guleray Agar}

1

_{Murat Aydın}

2

_{Mahmut Sinan Taspinar}

2

1_{Department of Biology, Faculty of Science, Ataturk University, 25240, Erzurum, Turkey}

2_{Department of Agricultural Biotechnology, Faculty of Agriculture, Ataturk University, Erzurum, 25240, Turkey} semra.yagci@icloud.com

Abstract

Salt stress is one of major abiotic stresses, reduced crop productivity. High salinity causes both hyperionic and hyperosmotic stress effects, and the consequence of these can be plant death. Several studies have reported that salt stress alters gene expression by DNA methylation and histone modification and retrotransposons activates in higher plants. Mammalian sex hormones (MSHs) such as

β-estradiol, androsterone, testosterone or progesterone, present in 60-80% of the plant species. Exogenous MSHs may be positive effects

on plant growth and development. The regulatory abilities of mammalian sex hormones in plants makes possible their use in practice. In this study, we investigated the protective effect of β-estradiol against DNA damage induced by salinity stress in (Triticum aestivum L.) wheat using REMAP (Retrotransposon-Microsatellite Amplified Polymorphism) technique. The plants were divided into 4 different treatment groups, β-estradiol solutions were sprayed onto the plants and other groups were treated with β-estradiol together NaCl solutions. These results showed that salinity stress caused decreasing of GTS (Genomic Template Stability) and increasing DNA damage. However, these effects of salinity stress seen at higher levels decreased after treatment with different concentration of β-estradiol. The results of this experiment have clearly shown that β-estradiol could be used effectively to protect wheat seedlings from the destructive effects of salinity stress.

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TNF-α MEDIATED UPREGULATION OF THE HUMAN ADAMTS-3 GENE EXPRESSION IN

OSTEOBLAST-LIKE CELLS

Meltem Alper

1

_{Ehed Muhammed Aymaz}

2

_{A.Tuğşen Aydemir}

3

_{Feray Köçkar}

2

1 _{Present address: University of Aksaray, Vocational School of Technical Sciences, 68100 Aksaray, Turkey} 2_{University of Balikesir, Department of Molecular Biology, Cagis, 10145 Balikesir, Turkey}

3_{Present Adress: International Biomedicine and Genome Institute, İzmir, Turkey} biologmeltem@hotmail.com

Abstract

The proinflammatory cytokine TNF-α has a critical role in cartilage destruction in arthritis. TNF-α induces cartilage specific expression of MMPs and ADAMTSs. Overexpression of these metalloproteinases results exaggerated degradation of cartilage matrix, which promotes progression of osteoarthritis. Among the ADAMTS family, procollagen N proteinases involves in cartilage remodelling. A member of this group, ADAMTS-3, is mainly expressed in cartilage and involves in the maturation of type II procollagen. ADAMTS-3 gene expression was also increases in osteoarthritis, myocardial infarction and breast cancer. Expression levels of some ADAMTS family members under proinflammatory cytokine stimulation have been investigated in some chondrocyte tissues or cell lines but, despite its importance in biosynthetic processing of fibrillar procollagens and extracellular matrix, how ADAMTS-3 gene expression change under TNF-α stimulation has still not elucidated in osteosarcoma cells.

In the present study we explored effects of TNF-α to transcriptional activity of the human ADAMTS-3 gene promoter in osteoblast like cells, Saos-2. Cells were transiently transfected with ADAMTS-3 promoter fragments and the reporter gene activity in these cells that were either left untreated or exposed to TNF-α was determined. Effect of the TNF-α on the steady-state levels of ADAMTS-3 mRNA and protein were also determined by qRT-PCR and western blotting for different time intervals.

Our data provides for the first time that TNF-α is a positive regulator of ADAMTS-3 gene expression. Performing additional studies on aggrecanase activity of ADAMTS-3 novel therapeutic interventions for degenerative diseases would be developed.

Keywords: ADAMTS-3, TNF-α, Saos-2, gene regulation.

ESTABLISHMENT OF IN VITRO MICROPROPAGATION PROTOCOL OF AXILLARY AND

LATERAL BUD EXPLANTS IN LAUREL (LAURUS NOBILIS L.)

Sheida Daneshvar Royandazagh

1

_{Elif Ceren Pehlivan}

1

_{Hasret Parmak}

2

_,

Halime Tuba Yildirim

2

1_{Namık Kemal University, Faculty of Agriculture, Department of Agricultural Biotechnology, Tekirdağ, Turkey} 2_{Namık Kemal University, Graduate School of Natural and Applied Sciences, Tekirdağ, Turkey}

eckalinkara@nku.edu.tr

Abstract

Laurel (Laurus nobilis L.) which has up to 5-10 meters length, belongs to the diecious plants’ family and especially grows in Mediterranean climate zone as an evergreen plant. In Turkey, it naturally grows as an important non-arboreal forest plant in Aegean, Mediterranean, and the Black Sea shore. It is used in pharmaceutical, food and cosmetic industries because of its oil and essences profile and also used in landscaping. Because of non-arboreal forest plant such as laurel is used different purposes, it is a necessity to develop mass production technologies and cultivate plants suitable for different usage. The aim of this study is to determine the micropropagation protocol of Laurel seeds germinated in vitro conditions and to make a new approach of cultivation of Laurel plant. Axillary and lateral bud explants were the main explant materials which were taken from in vitro shoots obtained from seeds. The initial explants were cultured on MS (Murashige and Skoog) medium containing BAP (6-benzylaminopurine), NAA (naphthalene acetic acid) and GA₃(gibberellic acid A3) combinations at different concentrations. After 42 days, shoot regeneration data were collected. And then in vitro shoots were subcultured on shoot multiplication medium with BAP and GA₃ combinations at different concentrations for elongation of shoots, multiplication of vegetative parts and for maintaining the viability of shoots. According to the results of the micropropagation experiment, the highest percentage of viability was obtained with MS media including 1.5 mg L-1_{BAP + 1 mg L}-1_{NAA + 0.2 mg L}-1_GA

3 and 2 mg L-1BAP + 0.5 mg L-1NAA +0.2 mg L-1_GA

3. This work was supported by TUBITAK-3001, Project number 215O251 and carried out in the Plant Tissue and Cell Culture Laboratory of Department of Agricultural Biotechnology, Faculty of Agricultural, Namik Kemal University with the seeds supplied from Laurel producers.

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CHARACTERIZATION OF SURFACE AND SYNTHESIS OF TITANIA- SILICA

NANOPARTICLES AND REMOVAL OF CR(III) CO(II) AND PB(II) IONS AND HEAVY

METAL DAMAGE TO PLANTS.

Aysel Çimen

Karamanoğlu Mehmetbey University, Faculty of Science, Department of Chemistry, 70200 Karaman, Turkey ayselcimen42@hotmail.com.

Abstract

TiO₂/SiO₂ was prepared by means of the impregnation method. The prepared TiO₂–SiO₂ catalyst demonstrated a remarkable photocatalytic activity toward degradation of Cr(III), Co(II) and Pb(II). The effect of reaction time, calcination temperature and time, amount of catalyst and solution concentration on the structural and chemical properties of the catalyst was investigated. The catalytic performance was found to depend essentially on the catalyst and target concentrations and the reaction time. The synthesized catalyst was characterized by a variety of techniques including surface area measurement, X-ray diffraction analysis (XRD), scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) measurements. The characterization of prepared TiO₂– SiO₂ catalyst, its photocatalytic performance and applications of wastewater containing Cr(III), Co(II) and Pb(II) are presented and thoroughly discussed. The contribution of this project to environmental cleanliness was discussed.

Keyword : Co(II), Pb(II), Cr(III), Heavy metal, Photodegradation, Surface bond conjugation, Titania/silica.

THE EFFECT OF DIFFERENT CRYOPROTECTANT COMBINATIONS ON THE VIABILITY

OF BOVINE CARTILAGE CELLS

Emel Tüten Sevim

1

_{Sezen Arat}

2*

1_{Agricultural Biotechnology Department, Graduate School of Natural and Applied Sciences, Akdeniz University, Antalya, Turkey} 2_{Agricultural Biotechnology Department, Faculty of Agriculture, Namık Kemal University, Tekirdağ, Turkey}

sarat@nku.edu.tr

Abstract

Cryopreservation is based on the ability of certain small molecules to enter cells and prevent dehydration and formation of intracellular ice crystals, which can cause cell death and destruction of cell organelles during the freezing process. One common cryoprotective agent is dimethyl sulfoxide (DMSO) and it is used for protection of most type of cells and tissues. A sugars are also used for freeze-drying methods of cryopreservation. They stabilise membranes by interacting with the polar head groups of phospholipids and increase the osmolality of the extracellular space that results in cell dehydration and lower the incidence of intracellular ice. The aim of the study is to cryopreserve bovine cartilage cells by using different cryoprotectant combinations and to investigate the effect of cryoprotective agent on cell viability by using flow cytometer. We investigated the effect of serum and sugars presence used in combination with DMSO for cryopreservation. While the ratio of necrotic and apoptotic cells was increased when the serum ratio in the freezing solution decreased. The highest cell viability was obtained from freezing solution containing 10% DMSO, 40% serum, in dextran 40 or dextrose.

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CONSUMPTION OF ADDED FRUCTOSE IN DIET AND ITS RELATION WITH OBESITY,

METABOLIC SYNDROME AND DIABETES

Gökhan SADİ

Karamanoglu Mehmetbey University, K.Ö. Science Faculty, Department of Biology, Karaman, Turkey sadi@kmu.edu.tr

Abstract

Carbohydrates constitute about 50-55% of the energy taken in a daily diet in adults. Today, the consumption of fructose, which is used as a sweetener in foods, has increased considerably with a change in eating habits. Free fructose used in sweeteners and high fructose corn syrup are the most important sources of dietary fructose. They are commonly used in all types of sweetened drinks, especially carbonated ones (acid drinks, fruit juice, cold tea, fruity soda etc), chocolate, cake, jam-marmalade and other jelly-like foods. It is widely used by food manufacturers because it is sweeter and cheaper than sucrose, enhance flavor and prevent crystallization. Studies in recent years have pointed out that fructose does not produce a feeling of satiety like other sugars and therefore causes more energy intake. Since, insulin is not secreted after its consumption, individuals develop more eating behaviors. Fructose is metabolized more in the liver compared to glucose, and easily converted to triglyceride and cholesterol by de novo synthesis. Increased lipogenesis of fructose in liver might lead obesity, insulin resistance, impaired glucose tolerance, diabetes, hyperlipidemia, cardiovascular diseases, gout and metabolic syndrome. In this study, consumption of fructose-containing foods and their effects on health will be discussed in a wide range of contexts together with recent findings in literature.

Key words: Fructose, Obesity, Diabetes, Metabolic Syndrome

MOLECULAR DETECTION OF POTENTIAL AFLATOXIGENIC ASPERGILLUS FLAVUS

Işılay Lavkor

1*

1_{Biological Control Research Institute, Adana, Turkey} isilay.lavkor@tarim.gov.tr

Abstract

Aflatoxins are produced as secondary metabolites by Aspergillus flavus. They have the ability to contaminate soil, air, plants by aflatoxi-genic Aspergillus flavus. The aim of the study was the detection of 173 potential aflatoxiaflatoxi-genic A. flavus from soil, air and infected peanut plants in the peanut planting area in Adana in 2016. In order to determination of potential aflatoxigenic A. flavus, IGS-F/R primer pairs were used to target the intergenic spacer (IGS) for the aflatoxin biosynthesis genes (aflJ and aflR). PCR products of fungal isolates were restricted with BglII enzyme within method of RFLP. As a result of PCR analysis, 121 fungal DNA were amplified fragment of 674 bp. In all examined cases, 121 (70%) out of 173 samples succesfully amplified the aflatoxin biosynthesis genes. Conclusion from this study in-dicate that, PCR-RFLP analysis of 121 isolates identified as A. flavus and aflatoxin biosynthesis genes were determined in these isolates. In this study, it was also demonstrated that identification of potential aflatoxigenic A. flavus by PCR-RFLP and determination of aflJ and

aflR genes have been found accomplished with using IGS-F/R primer pairs. Keywords: aflJ, aflR, aflatoxin, Aspergillus flavus IGS, PCR-RFLP

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DETERMINING SOME SUPPRESSOR AND ONCOGENIC MICRORNA’S IN COLORECTAL

CANCER

Faruk Suzergoz

1

_{Dilek Acar}

1

_{Sema Altuntaş}

1

_{Emine Teker}

1

_{H. A. Zafer SAK}

2

suzergoz@harran.edu.tr

Abstract

Early diagnosis is very important in colon cancer as well as in other types of cancer. Parallel to recent scientific developments, biomarkers are being used for the diagnosis and treatment of cancers. In our investigations of miRNA expressions in healthy tissues and tumor tissues in colon cancer cases and their ability to be used for early diagnosis were evaluated. Six normal and tumor tissue sections from the colon of the patients withcolorectal cancer who had undergone surgical procedure were used. Expression levels of miRNA-145, miRNA-192, miRNA-21 and miRNA-92 were determined in normal and tumor tissue specimens. To determine miRNA expression levels, cycle threshold (Ct) values were analyzed using Real Time PCR and fold changes were calculated from the obtained Ct values. As a result of miRNA analysis, miRNA-21 and miRNA-92 expressions were higher levels in tumor tissue than normal tissue whereas miRNA-145 and miRNA-192 expressions were higher levels in normal tissue compared to tumor tissue. Variable expressions of miRNA-21 and miRNA-92 known as the suppressor character, and miRNA-145 and miRNA-192 which plays a role in metastasis regulation, can give us information about the capacities that can be used in the early detection of colorectal cancers.

Keywords:Colon Cancer, MicroRNA expression, Real Time PCR.

ABA RELATED GENES EXPRESSION AND BIOCHEMICAL CHANGES RESPONSE TO

DROUGHT IN WATERMELON

M. Emre Erez

1

_{M. Zeki Karipcin}

2

_{Behcet İnal}

3

_{Serdar Altintaş}

1_{Department of Biology, Siirt University, Faculty of Arts and Science, Siirt, Turkey}

2_{Department of Horticulture, Siirt University, Faculty of Agriculture, Siirt, Turkey}

3_{Agricultural Biotechnology, Siirt University, Faculty of Agriculture, Siirt, Turkey} 4_{Agricultural Biotechnology, Ankara University, Institute of Biotechnology, Ankara, Turkey}

emreerez@hotmail.com

Abstract

Drought is one of the most important factors limiting plant production in worldwide. However, in order to be able to cope with this stress, plants have developed some mechanisms that generate physiological and biochemical responses at the cellular and organism level. In the present study, changes in three different watermelon genotypes (resistant, moderate and susceptible) exposed to drought stress were investigated. In this context, expression levels of ABA pathway related genes depending on ABA accumulation in the leaves were measured by q-RT PCR (Q-NAC, ORE1, SAG12, SAG13, CER1, LTP3, PYL9, KCS2, SWEET15, WSD1, AtNAP). In addition, the activities of some antioxidant enzymes, which are thought to be effective in the mechanism of drought, have been measured.

The above-mentioned genes associated with ABA pathway in the leaves of sensitive, moderate and tolerant watermelon genotypes were measured by q-RT PCR for the first time and significant expression differences were found between all three genotypes. Expression levels of genes associated with ABA synthase were found to be altered in response to drought stress. Also, SOD and catalase activities, which are thought to be effective in drought stress, and chlorophyll and total chlorophyll contents of genotypes have also been measured and found to show a significant change between genotypes. Therefore, in the context of this study, some important genes and physio-biochemical parameters have been revealed which will elucidate the mechanism of drought in watermelon.

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MOLECULAR DIAGNOSIS OF HOP LATENT VIROID (HLVD) IN HOP PLANTS

Pakize Gök Güler

1

_{Nuket Önelge}

2

1 _{Biological Control Research Institute Adana}

2_{Çukurova University Faculty of Agriculture Department of Plant Protection Adana} pakize.gokguler@tarim.gov.tr

Abstract

Hop plant (Humulus lupulus) of the Cannabaceae family, is a dioecious perennial climbing plant native to Asia, North America, and Europe as well as grown commercially in many countries for use in brewing and pharmaceutical industry. This study was carried out in plant samples picked up from Pazaryeri region of Bilecik province where hop plants are produced in large quantities. Samples were collected randomly from plants both symptomatic and non-symptomatic in 2015 and 2016 production periods. A total of 210 samples were collected from commercial areas of Erciyes, Brewers Gold, Aroma and Ege cultivars. 210 RNAs from collected samples were extracted with the GeneJET Plant RNA Purification Mini Kit. PCR mixture was prepared with specific primer according to Thermo one-step RT-PCR kit protocol for Hop latent viroid (HLVd), and applied PCR process under thermocycle conditions with the binding temperature reported for the agent by Eastwell and Nelson (2007). HLVd agents were identified in 20 of 210 samples collected in two years. The agent formed a band at 260 bp level on 2% agarose gel. The PCR product was purified by selection from HLV-infected samples and sent to sequence analysis. Blast analyzes and dendrograms of the obtained sequences were performed using the Mega 7 program. BLAST analyses of 3 samples resulted in 97% - 98% similarity to isolates from the NCBI database, with the highest similarity match rate observed in KT600318 (Belgium) and EF613181 (China) isolates with 98%.

Keywords: Hop plant, Humulus lupulus, Viroid, HLVd

CYTOTOXIC AND ANTIPROLIFERATIVE PROPERTIES OF FLUORINE BEARED

SALICYLALDIMINES ON A549 CELL LINE

H. A. Zafer SAK

1

_{Faruk Suzergoz}

2

_{Veli T Kasumov}

3

_{Ebrar Abacıoğlu}

2

drsak19@harran.edu.tr

Abstract

Although research continues on many new methods, chemotherapy is still one of the most important treatment options for cancer. Despite the importance of chemotherapy, an ideal chemotherapeutic drug has not yet been discovered. Studies with Schiff bases suggest that newly synthesized Schiff base compounds have promising anticancer efficacy in cancer treatment. In this study, the anticancer effects of a series of fluorinated Schiff base compounds on A549 lung carcinoma cells were investigated. Schiff bases were synthesized on the basis of anilines and salicyl aldehyde (Compound 1-10). A549 lung carcinoma cells, representing lung cancer, were used to determine anticancer effects of the compounds. Colorimetric MTT assay was used to determine the IC50 values of the compounds. In order to determine the proliferative index (PI) values of the compounds, flow cytometric CFSE assay was used, which was the dye shared by two sister cells during cell division. To determine apoptosis development in cells, active caspase-3 expression and cell morphology analyzes were performed. Among the compounds tested, it was determined that compound 8 (F_3,4,5-SAL, IC50: 1.50 µM) had the strongest cytotoxic activity. It was also determined that compound 1 and compound 2 had potent cytotoxic effects (F_2,3-SAL, IC50: 2.36 µM and F_2,4-SAL, IC50: 2.31 µM, respectively). The strongest antiproliferative effect on A549 cells was achieved with compound 9 (F₄-SAL, PI: 7.56) carrying 4 fluorine atoms. Caspase-3 expression and morphologic changes such as chromatin condensation and marginalization, forming apoptotic bodies indicate the role of apoptosis in cell cytotoxicity. It has been suggested that some fluorine bearing salicylanilimines have a high potential for the treatment of lung cancer but that further analysis is needed.

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THE EFFECT OF MAGNETIC FIELD STRENGTH ON AGROBACTERIUM

TUMEFACIENS-MEDIATED GENE TRANSFER IN POTATO (SOLANUM TUBEROSUM L.)

Mustafa Yıldız

1

_{Ramazan Beyaz}

2

_{Mehtap Gürsoy}

3

1_{Department of Field Crops, Faculty of Agriculture, Ankara University, Dışkapı, Ankara, Turkey} 2_{Department of Soil and Plant Nutrition, Faculty of Agriculture, Ahi Evran University, Bağbaşı, Kırşehir, Turkey}

3_{Güzelyurt Vocational School, Aksaray University, Güzelyurt, Aksaray, Turkey} myildiz@ankara.edu.tr

Abstract

This study was conducted to investigate the effect of magnetic field strength on Agrobacterium tumefaciens-mediated gene transfer in potato (Solanum tuberosum L.). In the study, sterile seedlings of cv. ‘Marabel’ as plant material and GV2260 line of Agrobacterium tumefaciens having ‘pBIN 19’ plasmid were used. Seedlings were treated with different magnetic field strengths (0-control, 75, 150 and 300 mT) for 24 h before transformation. Stem explants having axillary meristems were used in gene transfer. Explants were imbibed in 50 ml steril water containing 500 µl bacterial solution and they were placed in a rotary shaker (180 rpm) for 3 hours at 28ºC for inoculation. 50 mg l-1_{kanamycin and 500 mg l}-1_{duocid were added in selection medium. Inoculated stem explants were then cultured on MS} medium for co-cultivation during 2 days containing. Then explants were transferred to selection medium having 50 mg/l kanamycin and 500 mg/l duocid. The presence of the npt-II gene was verified by PCR analysis in candidate plants. The highest results with respect to number of plants growing in selection medium, number of plants transferred to soil, number of of plants growing in soil, PCR (+) plants were obtained from 150 mT magnetic field strength while the lowest values were recorded in control. This study was supported by TÜBİTAK with the project numbered “113O280”.

Keywords: Agrobacterium tumefaciens, gene transfer, magnetic field strength, potato, shoot regeneration

BIODEGRADATION OF TEREPHTHALIC ACID IN CSTR STIRRED TANK BY

LYOPHILIZED MICROORGANISMS AFTER 1-YEAR STORAGE

Didem EROĞLU

1 2

_{,Güven ÖZDEMİR}

2

1Ege University Application and Research Center for Testing and Analysis, İzmir,Turkey. 2Ege University, Faculty of Science, Department of Biology, Bornova, İzmir, Turkey.

didem.erglu@gmail.com

Abstract

Terephthalic acid is especially a raw material used in the production of plastics. The release of wastewater containing terephthalic acid has a negative effect on human health. For this reason, biodegradation of it is necessary. Freeze-drying is a commonly used method to preserve bacteria. This study, it was determined viability of bacterial cultures and biodegradation activity after lyophilization. In this study, the terephthalic acid biodegrading bacterial cultures lyophilized with protectant. Microorganisms were stored at 4°C for 1 - years were added to the activated sludge system. After terephthalic acid was added to the activated sludge system at 40 ppm, respectively. Also, control system was carried out.

Lyophilized product was determined viability after 1 - year storage. The initial cell concentration was changed from 10⁸ to 10⁷. Also, biodegradation of terephthalic acid by lyophilized microorganisms were determined by HPLC analysis. It was showed that the degradation ratios of terephthalic acids; 100%. It was shown that the biodegradation rate in the microorganism-added reactor give better result than in the control reactor.

Keywords:Terephthalic acid, Reactor( CSTR) , HPLC

Acknowledgements :The authors wish to thank Republic of Turkey, Ministry of Science, Industry of Technology under the grant

SANTEZ-00719 STZ 2014 and Ege University, Scientific Research Projects Fund, Project no: (BAP 2016 FEN 008 ) for the financial support of this study. The bacteria used in this article was produced within the project above. The support program of the Scientific and Technological Research Council of Turkey.(TÜBİTAK-BİDEB 2211-D) supported this study.

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A FUNGAL ISOLATE WITH BIOTECHNOLOGICAL IMPORTANCE FOR ETHANOL

PRODUCTION FROM LIGNOCELLULOSIC AGRICULTURAL WASTES

Hakan Özkan

1

_{Mehmet Karadayı}

2

_{Medine Güllüce}

2

_{Mine İsaoğlu}

3

_{Burak Alaylar}

3

Selma Sezen

4

1 _{Department of Molecular Biology and Genetics, Faculty of Science, Atatürk University, Erzurum, Turkey} 2 _{Department of Biology, Faculty of Science, Atatürk University, Erzurum, Turkey}

1_{Institute of Natural and Applied Sciences, Atatürk University, Erzurum, Turkey} 3_{Institute of Health Sciences, Atatürk University, Erzurum, Turkey}

mine.isaoglu@gmail.com

Abstract

Agricultural wastes, mainly composed of cellulose, hemicellulose, and lignin, are one of the most abundantly available raw material on the Earth. Biological processing of various agricultural wastes is considered as a key to unlock the future bio-energy and various biotechnological developments. In the present study, a fungal isolate (MG41) was isolated from decaying woody materials collected from Erzurum by using conventional techniques. Determination of ethanol production for the isolate was done by cultivation in modified BMC medium, sunflower wastes medium (SWM), corn wastes medium (CWM) and wheat wastes medium (WWM), respectively. Ethanol levels were determined by gas chromatography method. Molecular identification of the isolate was done by using PCR with universal ITS primers, sequencing of amplicons and the BLAST analysis of NCBI database. According to the results, MG41 showed a significant lignocelluloytic activity and produced bioethanol at 9.25 g/L a concentration in modified BMC media in the end of the fermentation process for 5 days. It also produced ethanol with 26%, 22,2% and 21% efficiency in SWM, CWM and WWM, respectively. After conventional and molecular identification studies MG41 was identified as Trichoderma atroviride. In conclusion, the experimental data and results of the present study offer that MG41 isolate shows valuable features for development of new bioethanol production technologies from lignocellulosic agricultural wastes. The amount of ethanol production can be increased by optimization studies in the future.

This study was supported by Republic of Turkey – Ministry of Food, Agriculture and Livestock: TAGEM-13/ARGE/17.

Keywords: Agricultural waste, Bioethanol, Internal transcribed spacer (ITS), Lignocellulose

IN VITRO PROPAGATION OF TRUE SEEDS OF POTATO (SOLANUM TUBEROSUM L.)

Gulsum Ozturk

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Abstract