Development of RT-PCR method for detecting GCLV by specific primers

摘要

The method was developed for the detection and resolution of the Garlic common latent virus that causes a serious disease of economically important plants of Allium species, in the Potyvirus presence particularly. GCLV was detected in garlic from Asia, Europe, South America and also from North America. GCLV together with SLV were the most widespread in the collection, as they occurred in 82 and 83% of the accessions in the Czech Republic as was described earlier. GCLV belong to ssRNA positive-strandviruses and is encoded of a single molecule of linear ssRNA of ~8.6 kb, RT-PCR was developed because it is a suitable detection method for RNA viruses. For the development of the RT-PCR method for detecting GCLV two primers were designed which amplify part of genome which encode gene NABP (nucleic acid binding proteins family). These primers amplify a product of approximately 782 base pairs. The product can be easily visualized on agarose gel electrophoresis. RT-PCR conditions were fully optimized for the detection GCLV in infected plants. This is a report on the development of a method for quick and easy, sensitive and economically reasonable virus detection GCLV.