Evaluation of various methods for the detection of barley yellow dwarf virus by the tissue-blot immunoassay and its use for virus detection in cereals inoculated at different growth stages

Abstract

Various modifications of the tissue-blot immunoassay (TBIA) for the detection of barley yellow dwarf virus (BYDV, luteovirus) were compared. Similar results were obtained by using three different labelled molecules; goat anti-rabbit antibodies conjugated to alkaline phosphatase, protein A conjugated with alkaline phosphatase and goat anti-rabbit antibodies conjugated with colloidal gold. Blocking the nitrocellulose membrane with polyvinyl alcohol for 1 min was effective and allowed the procedure to be shortened by one hour. TBIA was sensitive enough to detect BYDV in old dry tissue wich had been soaked in water for 1 h.

BYDV was monitored by TBIA in wheat, oat and barley after inoculation at heading, flowering and grain filling growth stages. The later the inoculation date, the greater the chance of detecting the virus in stem bases rather than in the upper part of the stem. The later the inoculation the less virus moved, from the inoculated tiller to other tillers of the same plant.