Browsing by Author "Güller, A."

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Detection and Characterization of Two Phytoplasma Lineages on Cucumber (Cucumis Sativus L.) With Same Symptomatology Based on Virtual Rflp and Nucleotide Sequence Analysis of 16s Rdna
(Centenary University, 2017) Usta, M.; Güller, A.; Sipahioğlu, H.M.
Phytoplasma-like symptoms were observed in cucumbers (Cucumis sativus L.) in Van province of Turkey. The major symptoms observed were severe dwarfing, witches’ broom, rosetting, little leaf, and sterility of plants. Genomic DNA of 8 symptomatic and non-symptomatic plant leaves was isolated for the detection of pathogenic DNA. Of the 8 cucumber leaf samples tested by nested polymerase chain reaction (Nested-PCR), the four yielded the expected 1.25-kb DNA fragments when using universal primer pairs R16mF2/R16mR1 and R16F2n/R16R2. Randomly selected two DNA bands were further cloned into a proper plasmid vector. The recombinant plasmid DNA was sequenced bidirectionally. BLAST and virtual restriction fragment length polymorphism (RFLP) analyses of the 16S rDNA sequence revealed the presence of the “Candidatus Phytoplasma solani” (similarity coefficient 1.00) (GenBank accession no: KX977570) in one of the severely symptomatic cucumber samples and the “Candidatus Phytoplasma trifolii” (similarity coefficient 0.98) (GenBank accession no.: KR080212) in the other. The isolates were designated as Van-trifolii and Van-solani isolates, respectively. No significant differences were observed between the two different phytoplasmas’ symptomatology. To the authors’ knowledge, this is the first report of ‘Ca. P. trifolii’ and ‘Ca. P. solani’ in cucumbers in Turkey. © 2017, Centenary University. All rights reserved.
First Report of ‘candidatus Phytoplasma Australasia’ Strain Related To Witches’-Broom of Tomato in Türkiye
(Centenary University, 2023) Usta, M.; Güller, A.; Sipahioğlu, H.M.
Phytoplasmas are dangerous bacteria severely infecting agricultural production worldwide. In the present study, the identification of phytoplasmas infecting tomato plants showing symptoms such as small leaves, flower abnormalities, stunting, witches' broom, and reddening was performed. Five plants, two symptomatic and three asymptomatic, were tested to verify phytoplasma infection. Total DNA isolated from 5 leaf samples was used as a template for PCR reactions. The phytoplasma agents were confirmed in the two symptomatic samples. BLASTn search of 16S rRNA of two sequences shared identity similarity of 99.84% with ‘Candidatus Phytoplasma australasia’. Computer-simulated virtual RFLP profiles show that the 16S rRNA sequences is identical to the reference pattern of the 16SrII-D subgroup, with a similarity coefficient of 1.00. Based on BLAST, virtual RFLP, and phylogenetic dendrogram, the identified phytoplasma strains are enclosed in the 16SrII-D subgroup. This is the first report of tomato witches' broom disease related to 16SrII-D subgroup phytoplasma strains in the Antalya province of Türkiye. © 2023, Centenary University. All rights reserved.
Molecular Identification of ‘candidatus Phytoplasma Solani’ Using Secy and Vmp1 Genes in Tomato Plants From Van Province
(Centenary University, 2021) Usta, M.; Güller, A.; Demirel, S.
Tomato (Solanum lycopersicum) is one of the most financially important vegetable crops. It is a species belonging to the Solanaceae family and is cultivated in many countries, including Turkey. The natural presence of ‘Candidatus Phytoplasma solani’ (‘Ca. P. solani’) from the Stolbur group (16SrXII) in tomato plants is extensively characterized based on the 16S rRNA gene worldwide. Tomato plants displaying abnormality and sterility of flower, purpling, and bushy appearance were observed in the Van province, Turkey. DNA extraction from tomato leaves was performed, and the extracted DNA was used to amplify 16S rRNA sequences using universal primer pairs by double PCR assays. After confirming the presence of the probable agent (‘Ca. P. solani’), nested PCR testing was performed using an appropriate primer set amplifying the SecY and Vmp1 genes of the same DNAs. Amplified PCR yields were then cloned into a pGEM T-Easy vector and sequenced by New Generation System (NGS). Sequenced 1438 bp nucleotides for Vmp1 gene (MN104838) and 905 bp nucleotides for SecY gene (MN125054) revealed 97.38% and 100% maximum nucleotide similarity with formerly published Vmp1 and SecY gene sequences of ‘Ca. P. solani’ species belonged to the Stolbur group (16Sr-XII), respectively. According to the phylogenetic tree created using our gene sequences from ‘Ca. P. solani’, Vmp1, and SecY gene sequences showed a highly phylogenetic affinity with the same sequences of the same agent from Serbia and France, respectively. To the best of our knowledge, this study is the first report to reveal the phylogenetic relationships of ‘Ca. P. solani’ in stolbur group (16Sr XII-A) using SecY and Vmp1 genes in the tomato plant in Turkey based on the non-ribosomal genes. © 2021, Centenary University. All rights reserved.
Presence of 'candidatus Phytoplasma Trifolii' in Pepper Plants Showing Yellowing and Bushy Appearance From Iǧdir Province of Turkey
(Ege Universitesi, 2022) Güller, A.; Usta, M.; Korkmaz, G.; Demirel, S.; Kurt, Z.
Objective: The objective of this study was to investigate the group/subgroup of phytoplasma agent in peppers showing phytoplasma symptoms. Material and Methods: In this study, plants collected from Iǧdir province in 2020 were analyzed using direct and nested PCR tests, and BLASTn, iphyClassifier, Mega 7, and pDRAW32 programs were used. Results: In the tests performed, approximately 1.2 kb of DNA fragments specific to phytoplasma were obtained. The 16S rRNA nucleotide sequence (1254 bp in length) (OM663745) revealed that it was showed more than 99.44% nucleotide similarity to other 'Ca. P. trifolii' members. The tentative RFLP and phylogenetic analyzes performed proved the 'Ca. P. trifolii' the infection from the Clover proliferation group (16SrVI) group and subgroup A in symptomatic pepper plants. Conclusion: The presence of 'Ca. P. trifolii' in naturally infected peppers in Iǧdir province of Turkey was detected using PCR-RFLP and cladistic analysis. © 2021 Authors.