Development of a Rapid Enzymatic Cdna Amplification Test for the Detection of Apple Scar Skin Viroid (Assvd) in Apple Trees From Eastern Anatolia, Turkey

Abstract

A validated RT-PCR method was used to investigate the presence of Apple scar skin viroid (ASSVd) in the commercial apple orchards of eastern Anatolia. Among three modified and simplified silica-capture based extraction methods, one was used for mass extraction to ascertain the presence of ASSVd. The test was initially performed from an ASSVd-infected source and then applied to total RNA preparations from fresh leaf tissues of apple trees collected from eastern Anatolia. ASSVd was found to occur in apple trees. Among 263 apple samples, 121 were positive for ASSVd. The infected trees showed no apparent disease symptoms on the leaves other than scarring on fruit skin. Overall incidence of ASSVd was 46% in eastern Anatolia. The presence of ASSVd reported for the first time in Turkey. Among three total RNA preparation methods, Method II was determined to be the best procedure for large scale routine analysis. The improved test can be used in a certification or clean stock program to contribute to the prevention of the spread of ASSVd in the eastern Anatolia.