Improvement of RT-PCR based detection of two closteroviruses associated with little cherry disease in sweet cherries

Abstract

The increasing availability of nucleotide sequence data from plant pathogenic fruit tree viruses led recently to the development of nucleic-acid-based detection methods as a routine tool for diagnosis. We present a simplification and modification of the previously described test procedure for the two closteroviruses Little cherry virus 1 and 2 (LChV-1, LChV-2). For LChV-1, a single tube reaction reduced unspecific amplification that led to false positive results. For the detection of LChV-2, a primer set was developed for the local isolates of the area 'Altes Land' in the northern part of Germany. To determine the optimal sampling conditions for a reliable detection of both viruses different types of tissue (bark, bud, leaves) were tested in monthly intervals.