Detection of two strains of grapevine leafroll-associated virus 2

Two strains of grapevine leafroll-associated virus 2 (GLRaV-2) were obtained by mechanical transmission from grapevines to Nicotiana benthantiana. The strains, designated 94/970 and 93/955, consistently differed with regard to the development of symptoms. The first induced chlorotic and occasional white-necrotic local lesions while the second induced chlorotic followed by metallic-opalescent, solid necrotic local lesions. The strains were indistinguishable with regard to the molecular weight of their capsid proteins or serologically. A difference in the pattern of minor dsRNA bands was consistently observed

Grapevine Leafroll-Associated Virus 2 (GLRaV-2)- Mechanical Transmission, Purification, Production and Properties of Antisera, Detection by ELISA

Grapevine leafroll-associated virus 2 (GLRaV-2) was transmitted from Vitis vinifera cv. Muscat d'Alexandrie to Nicotiana benthamiana following inoculation with grapevine leaf petioles extract concentrated by ultracentrifugation. Virus-infected N. benthamiana showed symptoms of chlorotic local lesions, systemic vein clearing followed by yellowing, stem necrosis and death of the plant.  The plants were shown to be infected only with GLRa V-2. Antisera produced to GLRa V-2 strongly decorated particles of the virus in immunoelectron microscopy (IEM) and clearly and specifically detected GLRaV-2 in concentrated extracts of infected grapevines in Western Blots. The antisera were used with success for the specific and sensitive detection of GLRaV-2 by ELISA.  Treatment of purified GLRaV-2 preparations with glutaraldehyde before immunisation markedly improved the quality of the antisera