Development of a DNA macroarray for the detection and identification of fungal pathogens causing decline of young grapevines.

Urbez-Torres, J.R., Haag, P., Bowen, P.A., Lowery, T., Lowery, D.T., and O'Gorman, D.T. (2015). "Development of a DNA macroarray for the detection and identification of fungal pathogens causing decline of young grapevines.", Phytopathology, 105(10), pp. 1373-1388. doi : 10.1094/PHYTO-03-15-0069-R  Access to full text

Abstract

Young vine decline (YVD) is a complex disease caused by at least 48 different fungi and responsible for important economic losses to the grapevine industry worldwide. YVD fungi are known to occur in planting material. Hence, detection prior to planting is critical to assure longevity of newly established vineyards. A DNA-macroarray based on reverse dot blot hybridization containing 102 oligonucleotides complementary to portions of the β-tubulin region was developed for detection of YVD fungi. Specificity of the array was first evaluated against 138 pure fungal cultures representing 72 different taxa from eight genera, including 37 YVD species. A total of 61 species including 34 YVD pathogens were detected and identified by the array. The detection limit of the array was below 0.1 pg of genomic DNA. The array was validated against artificially inoculated canes and soil, and commercial planting material, showing the latter a high incidence of YVD fungi in nursery plants otherwise not detected by traditional plating and culturing. This DNA array showed to be a rapid and specific tool to simultaneously detect and identify most YVD fungi in a single test, which has the potential to be used in commercial diagnostics and/or by the grapevine nursery industry to determine the health status of the planting material.

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