Development of a non-radioactive molecular hybridization probe for detecting Strawberry mottle virus in strawberry

Abstract

The vegetative propagation of strawberries favors transmission of systemic pathogens, such as viruses, which are one of the main yield-limiting factors for this crop. More than 20 viruses have been described as infecting this species; one of the most frequent is the Strawberry mottle virus (SMoV), which is responsible for significant economic losses. SMoV is usually detected by reverse transcription polymerase chain reaction (RT-PCR), given that serum is not available for serological tests. In this study, a non-radioactive molecular probe was developed for SMoV detection. The cDNA was synthesized by RT-PCR using specific primers designed from the 3'UTR region of the viral genome. The cloned cDNA segment was labeled and used as a probe. Six RNA extraction protocols were evaluated, and the modified cetyltrimethylammonium bromide (CTAB) method showed the highest sensitivity level. Leaves at different phenological stages and petioles were evaluated; the highest reaction was observed in old leaves and in petioles.