Two main apple growing provinces (Van and Malatya) of East Anatolia were surveyed for the presence of the Apple mosaic virus (ApMV). Dot-blot hybridization and RT-PCR tests were implemented to investigate the incidence of ApMV, testing a total of 481 samples collected from commercial apple orchards. | Turkish Journal of Agriculture and Forestry Research Article Turk J Agric For (2013) 37: 711-718 © TÜBİTAK doi: Survey of Apple mosaic virus in apple-growing provinces of East Anatolia (Malatya and Van) by RNA probe hybridization assay and RT-PCR 1 2, 2 Gülüstan KORKMAZ , Hikmet Murat SİPAHİOĞLU *, Mustafa USTA Ministry of Agriculture, Department of Plant Protection, Rural Affairs of Van, Van, Turkey 2 Department of Plant Protection, Faculty of Agriculture, Yüzüncü Yıl University, Van, Turkey 1 Received: Accepted: Published Online: Printed: Abstract: Two main apple growing provinces (Van and Malatya) of East Anatolia were surveyed for the presence of the Apple mosaic virus (ApMV). Dot-blot hybridization and RT-PCR tests were implemented to investigate the incidence of ApMV, testing a total of 481 samples collected from commercial apple orchards. Digoxigenin-labeled RNA probes of the ApMV were synthesized from the cloned PCR products and applied in dot-blot hybridization to detect the virus in RNA extracts isolated from fresh leaf tissues. A validated RNA probe (dot-blot) hybridization method was adopted to investigate the presence of ApMV infections in the apple orchards of East Anatolia. In order to determine the most suitable mass extraction methods and to ascertain the presence of virus, 3 RNA preparation procedures (QIAGEN RNA extraction kit, silica capture, and citric buffer) were tested. Among the tested extraction methods, silica capture was determined as the most suitable extraction method for dot-blot hybridization assays. ApMV was found in the surveyed locations with an average incidence of . The infected trees showed apparent disease symptoms on the leaves of apple trees. The coat protein (CP) genes of 2 viral isolates selected from Malatya (ApMV-G and ApMV-M) were cloned and their complete nucleotide sequences and deduced amino acids
