Development of a technique for the detection of Eutypa lata from grapevines by PCR-RFLP assay
The prevalence of Eutypa dieback in vineyards in the Western Cape has serious economic implications for the South African wine industry due to the severe damage and extensive losses it can incur. Eutypa lata is the pathogen that has been commonly isolated from infected tissue. This led to research aimed at developing a screening technique that would quickly and reliably detect E. lata from infected grapevines. The technique, reverse dot blot hybridisation, used to address this matter was not without its problems.
A screening technique, reverse dot blot hybridisation, was successfully employed for the rapid and accurate detection of plant pathogens, however, problems were encountered in making the technique amenable to the detection of pathogens directly from infected tissue. This prompted an investigation into alternative methods that could be developed for the detection of E. lata. A multiplex approach that combines PCR amplification with specific primers with a restriction fragment length polymorphism (RFLP) assay could eliminate the need for culturing for pure material and detecting closely related fungi which could lead to a false diagnosis.
Safodien, S, Crous, P W, Botha A, Smit, W A and Halleen, F. 2007. Eutypa dieback of grapevines in South Africa. Paper presented at the 45th Congress of the Southern African Society of Plant Pathology. 21-24 January, Benoni, South Africa