Determination of the bdv1 gene providing tolerance to barley yellow dwarf virus mav (bydv-mav) in some wheat genotypes by molecular marker

Objective: It was aimed to investigate the presence of the Bdv1 gene, which is known to provide moderate resistance to Barley yellow dwarf virus, with a molecular marker. For this purpose, first of all, the presence of the Bdv1 gene in 156 mature wheat genotypes was investigated with the WMS130 marker, which was determined by comparing the WMS130 and Lr34-D markers in terms of applicability. Material and Methods: Within the scope of this thesis study, studies were carried out on 156 dormant stage wheat genotypes and the positive control Anza wheat genotype. A preliminary study was carried out with a total of five wheat genotypes, including the positive control Anza wheat plant, for the determination of isolation starting material (seed/leaf), determination of DNA/RNA isolation methods and molecular marker selection. Within the scope of the preliminary study, DNA isolation from seeds, whole DNA and TNA isolations from leaves were carried out. PCR/RT-PCR analyzes were performed using the isolated DNA/RNAs as templates and the molecular marker was selected according to these results. After evaluating the markers in terms of applicability, reliability and cost, as well as evaluating the performance/success and purity/quality of the isolation methods, it was found appropriate to use leaf DNA isolation and the WMS130 marker to investigate the presence of the Bdv1 gene in all genotypes. After the PCR analyzes performed with the WMS130 marker on the genotypes, the PCR products of 2 samples of the two wheat genotypes Maden and Dağdaş94 that gave the best profile images were sent to the sequence. In addition, the PCR product obtained by TNA isolation from the leaf of Anza variety and using the Lr34-D marker was included in the sequence. Results: With the preliminary studies carried out to investigate the presence of the Bdv1 gene in genotypes, the most suitable isolation starting material (seed/leaf), the most suitable DNA/RNA isolation method and marker specific to the marker were determined. According to preliminary study findings; In the PCR analyzes performed with the Lr34-D marker, when DNA isolation from the leaf and TNA isolation from the leaf were compared, it was determined that the use of TNA isolation was appropriate. According to the PCR results performed with this TNA isolation, a 398 bp band specific to the Bdv1 gene was obtained. No results were obtained from PCR analysis performed with DNA obtained from the leaf. When the WMS130 marker was used, a 130 bp molecular size band specific to the Bdv1 gene was obtained in the PCR analysis performed with DNA obtained from seed DNA isolation and leaf DNA isolation. Thus, it was determined that both isolation methods were suitable for the WMS130 marker. Considering the purity and quality of DNA in isolation, it was found that the leaf DNA isolation method was most suitable for the WMS130 marker. According to the results of the preliminary study, considering the results obtained from both Lr34-D and WMS130 markers and the applicability, reliability and cost of the markers, it was deemed appropriate to use leaf DNA isolation and the WMS130 marker to investigate the presence of the Bdv1 gene in genotypes. As a result of DNA isolation from the leaf and sequencing of the PCR product using the WMS130 marker, Maden genotype was identified as Rhizoctonia solani and Dağdaş94 genotype was identified as Ascomycete sp. It was found to be associated with. The sequence result of the PCR product performed with the Lr34-D marker of the Anza variety revealed that it is related to Triticum dicoccoides registered with number KU711570.1 and Triticum monococcum registered with number KU711538.1. Conclusion: As a result of analysis performed using leaf DNA isolation and WMS130 marker in 156 established stage wheat genotypes, Bdv1 gene was found in 104 wheat genotypes. However, resistance/tolerance to BYDV MAV may have emerged after evaluating the phenotypic responses of the genotypes. As a result of the sequence analysis, Dağdaş94 genotype is Ascomycete sp. and Maden genotype was found to be associated with R. solani. It was concluded that WMS130, which is a versatile marker, is not only specific for detecting the Bdv1 gene, but also can be used to identify genes that provide resistance to some fungal diseases. It is thought that the Lr34-D marker will be a suitable alternative in the detection of the Bdv1 gene in wheat genotypes in future studies. Key Words: Bdv1 gene, BYDV MAV, Luteovirus mavhordei, SSR marker, Virus resistance.