Characterization of Strawberry latent ringspot virus (SLRSV) on strawberry in Egypt
Characterization of Strawberry latent ringspot virus (SLRSV) on strawberry in Egypt
Amira A. Mazyad1; A. A. Kheder1; Ahmed K. El-Attar1; W. Amer.2; Mahasen, H. Ismail.2; Amal, A.F.1
ABSTRACT
Strawberry latent ringspot virus (SLRSV) was isolated from symptomless strawberry plants and identified with a specific antiserum (Loewe Biochemica GmbH) using Double Antibody Sandwich ELISA (DAS-ELISA). Virus survey was carried out during 2013 - 2014 in different locations on commercial strawberry fields. The percentages of infection were 3.7, 4.5, 15.7 and 20% in El-Behera, El-Kalubeia, El-Ismalia and El-Menofia respectively. SLRSV was transmitted either by Xiphinema americanumas nematode vector or mechanically from infected strawberry plants onto 16 host species belonging to seven families. Typical leaf curling, chlorosis, vein clearing and stunting were appeared on indicator host (cucumber) 3 weeks post inoculation. Stability experiments of SLRSV showed that the thermal inactivation point was 48-62°C; the dilution end point was 10-4 to 10-6 and the longevity in vitro 10 - 18 days at room temperature. The diagnosis of SLRSV in the infected tissues of strawberry plants was performed using both serological (DAS-ELISA) and molecular assays. Reverse transcription polymerase chain reaction (RT-PCR) was used to amplify 497 bp fragment using PCR primers specific for the viral coat protein gene as a tool for molecular diagnosis. The PCR detection was confirmed with direct DNA sequencing and phylogenetic analysis for the coat protein gene. Further insurance of SLRSV infection was performed using light microscopy which showed presence of amorphous inclusion bodies, electron microscopy and chemical analysis.
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