Browsing by Author "Gunay, Abidin"

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First Investigation of Five Tobacco Viruses Using Pcr Based Methods in Tobacco Plants Grown in Adiyaman, Turkey
(Parlar Scientific Publications (p S P), 2020) Gunay, Abidin; Usta, Mustafa
Survey studies were carried out in in 2018 in Adiyaman, Turkey, where tobacco is traditionally produced, to detect the viruses that infect tobacco including Tobacco mosaic tobamovirus (TMV), Cucumber mosaic cucumovirus (CMV), Potato potyvirus Y (PVY), Tobacco vein banding mosaic potyvirus (TVBMV), and Tobacco etch potyvirus (TEV). Accordingly, 450 fresh tobacco leaves were simultaneously analyzed by the multiplex RT-PCR method for the presence of tobacco viruses. As a result of molecular analysis, CMV (8.8%), TMV (5.5%), and TMV + CMV mixed infection (0.2%) were detected in 40 of the tobacco samples. TEV, PVY, and TVMBV viruses were not detected in any of the samples. The overall infection incidence of tobacco viruses in Adiyaman province was found to be 14.4%. Two isolates were randomly selected from each detected virus and their coat protein (CP) genes were cloned into the prokaryotic cloning vector and transformed into E. coli. Recombinant plasmids obtained from positive clones were sequenced by Next Generation Sequencing (NGS) system to reveal the viral (CP) sequence. Nucleic acid sequence similarity analyses revealed that the TMV Adiyaman isolates showed similarity in the range of 74.17-99.38% and the CMV Adiyaman isolates in the range of 75.61-99.54%. In the phylogenetic analyis using (CP) sequences, tobacco-CMV isolates were found to be in phylogenetic affinity with the Greece (AY450854) and Japan (AB448696) isolates, while both tobacco-TMV isolates showed a phylogenetic affinity with the Serbia isolate (GQ340671). According to the current literature, this is the first molecular survey study on tobacco viruses in Adiyaman and Turkey. With the present study, the molecular char acterization of the isolates selected with the presence of CMV and TMV in the tobacco plant has been re- ported for the first time.
The Molecular Characterization of the Coat Protein Sequence and Differentiation of Cmv- Subgroup I on Tobacco From Native Flora in Turkey
(Univ Agr Sci & veterinary Med Cluj-napoca, 2020) Usta, Mustafa; Guller, Abdullah; Gunay, Abidin
Cucumber mosaic virus (CMV) has a broad plant-host range and a wide ecological zone distribution. Virus-like symptoms were observed on tobacco fields of Adiyaman province (Turkey) showing conspicuous mottling, greenish mosaic patterns and severe malformations of leaves. A total of forty tobacco samples tested positive against CMV by reverse transcription polymerase chain reaction (RT-PCR) using coat protein gene specific primers. Five randomly chosen CMV isolates were cloned into pGEM T-Easy vector and transformed into Escherichia coli JM109 strain. The recombinant bacterial clones containing insert-DNA were further purified and sequenced bidirectionally. In multiplex-RT-PCR studies carried out, it was found that all 40 CMV isolates belong to Subgroup I by resulting a 593 bp long DNA fragments. CMV subgroup IA was found to predominate in 4 out of 5 tobacco samples and CMV subgroup IB was found in 1 out of 5 CMV-positive samples by comparing the isolates with CMV reference isolates in phylogenetic tree. However, no Subgroup II sequences were found by multiplex RT-PCR using discriminating primers. The nucleic acid sequences were analyzed for the investigation of diversity of coat protein (CP) sequences of 5 CMV isolates. The sequence similarity ranged from 94.2-100% with the CMV subgroup I isolates infecting diverse plants in other regions of the world. The evolutionary tree revealed that the CMV IA Adiyaman isolates exhibited a genetic affinity with Australian and Spanish isolates. However, the CMV IB Adiyaman isolate showed a close genetic relationship with only the Australian isolates. To our knowledge, this study shows for the first time the occurrence of CMV IA and IB isolates infecting cultured tobacco plants in Adiyaman province.