Grapevine Leafroll Associated Virus 3 ELISA Development Kit

The vegetative propagation of grapevine facilitates multiple viral infections, with different symptoms which vary according to combinations of cultivar or host species with viral species. The aims of this research were to detect and identify the viral species infecting two grapevine species/cultivars: one symptomatic and one symptomless. DsRNA from both samples was assayed by RT-PCR using 17 pairs of specific primers for detection of the Grapevine virus A (GVA), Grapevine virus B (GVB), Grapevine virus D (GVD), Grapevine fleck virus (GFkV), Grapevine fanleaf virus (GFLV), Arabis mosaic virus (ArMV), Grapevine chrome mosaic virus (GCMV), Rupestris stem pitting-associated virus (RSPaV) and Grapevine leafrollassociated virus 1-4 (GLRaV-1 to -4), besides three degenerate primer pairs. For each primer pair at least one amplicon was cloned and sequenced. Symtomatic and symptomless plants were multiple infected by RSPaV, GLRaV-2 and/or GLRaV-3. The nucleotide sequences of seven isolates of RSPaV, three of GLRaV-2 and two of GLRaV-3 showed identities higher than 90% with the homologous viral species and allowed to identify possible viral strains in infected samples. These results highlight the necessity of viral diagnosis based on specific assays to determine grapevine sanitary status.

Grapevine leafroll-associated virus 3 (GLRaV-3) is distributed in all grapevine-growing regions worldwide, resulting in significant economic losses. In this study, ribavirin (15 and 25 mu g/mL, R15 and R25), high temperature (37 degrees C, T), and a combined treatment (R15 + T and R25 + T) were used to eliminate the virus from in vitro GLRaV-3-infected Vitis champini plantlets. The phytotoxicity of ribavirin on plantlets was found in R15 and R25 lasted 30 days, but the damage of high temperature on grapevine plantlets was found in T, R15 + T and R25 + T at the beginning of treatment. Moreover, high temperature could enhance the growth of grapevine plantlets. The survival rate was lowest for the regenerated T plants (23.3%). In contrast, the survival rates for the regenerated plants that underwent the chemotherapy (R15 and R25) and the combined treatment (R15 + T and R25 + T) were 47.0% (31/66) and 41.0% (50/122), respectively. Reverse transcription polymerase chain reaction and quantitative real-time PCR assays were used to evaluate the efficiency of the GLRaV-3 elimination in the regenerated grapevine plants. The combined treatment was most effective for eliminating GLRaV-3. Specifically, GLRaV-3 was detected in all regenerated plants after the 40-day R15 and R25 treatments, whereas the virus elimination rates for the 20-day R15 + T and R25 + T treatments exceeded 80%. Additionally, the virus elimination rate for the regenerated T plants was 42.9% for the same treatment periods.