PJB-2017-729
COMPARISON OF REAL TIME PCR, CLASSICAL PCR and MOLECULAR HYBRIDIZATION TECHNIQUES FOR DIAGNOSIS OF GYSVD-1 (Grapevine yellow speckle viroid-1) in GRAPEVINE
ismail can paylan
Abstract
GYSVd-1 is found in the important production areas, is very easily spread mechanically by contaminated cutting tools, infected graft, propagation materials and causes severe damage to quality and quantity of grapevines. Rapid diagnosis of viroid infections may be obtained by bioassays, polyacrylamide gel electrophoresis (PAGE), molecular techniques such as polymerase chain reaction (PCR) and hybridization assays. In this study we aimed to compare three molecular methods including molecular hybridization, real-time pcr and classical pcr in order to detect GYSVd-1 in grapevine. In the diagnostic analyzes conducted with 50 potentially positive grapevine samples, GYSVd-1 was detected in 45 of 50 samples by one or more detection techniques. 18 samples were found to be positive by all methods. Other 18 samples gave positive results in Real Time PCR and conventional PCR analyzes but negative in Molecular Hybridization tests. 2 samples were positively determined by Real time PCR and Molecular Hybridization tests. 6 grapevine samples were determined only with real time PCR tests and 1 sample with only classic PCR tests. Three molecular methods for detecting GYSVd-1 compared in this work and we conclude that Real time PCR was the most sensitive amplification method followed by classical PCR and Molecular Hybridization. Also Real-time PCR method is less time consuming than classical PCR and hybridization methods.
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